Sample records for bacterial flagellar bundling

  1. Magnetic Propulsion of Microswimmers with DNA-Based Flagellar Bundles.

    PubMed

    Maier, Alexander M; Weig, Cornelius; Oswald, Peter; Frey, Erwin; Fischer, Peer; Liedl, Tim

    2016-02-10

    We show that DNA-based self-assembly can serve as a general and flexible tool to construct artificial flagella of several micrometers in length and only tens of nanometers in diameter. By attaching the DNA flagella to biocompatible magnetic microparticles, we provide a proof of concept demonstration of hybrid structures that, when rotated in an external magnetic field, propel by means of a flagellar bundle, similar to self-propelling peritrichous bacteria. Our theoretical analysis predicts that flagellar bundles that possess a length-dependent bending stiffness should exhibit a superior swimming speed compared to swimmers with a single appendage. The DNA self-assembly method permits the realization of these improved flagellar bundles in good agreement with our quantitative model. DNA flagella with well-controlled shape could fundamentally increase the functionality of fully biocompatible nanorobots and extend the scope and complexity of active materials.

  2. Bacterial flagellar microhydrodynamics: Laminar flow over complex flagellar filaments, analog archimedean screws and cylinders, and its perturbations.

    PubMed

    Trachtenberg, Shlomo; Fishelov, Dalia; Ben-Artzi, Matania

    2003-09-01

    The flagellar filament, the bacterial organelle of motility, is the smallest rotary propeller known. It consists of 1), a basal body (part of which is the proton driven rotary motor), 2), a hook (universal joint-allowing for off-axial transmission of rotary motion), and 3), a filament (propeller-a long, rigid, supercoiled helical assembly allowing for the conversion of rotary motion into linear thrust). Helically perturbed (so-called "complex") filaments have a coarse surface composed of deep grooves and ridges following the three-start helical lines. These surface structures, reminiscent of a turbine or Archimedean screw, originate from symmetry reduction along the six-start helical lines due to dimerization of the flagellin monomers from which the filament self assembles. Using high-resolution electron microscopy and helical image reconstruction methods, we calculated three-dimensional density maps of the complex filament of Rhizobium lupini H13-3 and determined its surface pattern and boundaries. The helical symmetry of the filament allows viewing it as a stack of identical slices spaced axially and rotated by constant increments. Here we use the closed outlines of these slices to explore, in two dimensions, the hydrodynamic effect of the turbine-like boundaries of the flagellar filament. In particular, we try to determine if, and under what conditions, transitions from laminar to turbulent flow (or perturbations of the laminar flow) may occur on or near the surface of the bacterial propeller. To address these questions, we apply the boundary element method in a manner allowing the handling of convoluted boundaries. We tested the method on several simple, well-characterized cylindrical structures before applying it to real, highly convoluted biological surfaces and to simplified mechanical analogs. Our results indicate that under extreme structural and functional conditions, and at low Reynolds numbers, a deviation from laminar flow might occur on the flagellar

  3. Exchange of rotor components in functioning bacterial flagellar motor

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

    Fukuoka, Hajime; Inoue, Yuichi; Graduate School of Life Sciences, Tohoku University, Aoba-ku, Sendai 980-8577

    2010-03-26

    The bacterial flagellar motor is a rotary motor driven by the electrochemical potential of a coupling ion. The interaction between a rotor and stator units is thought to generate torque. The overall structure of flagellar motor has been thought to be static, however, it was recently proved that stators are exchanged in a rotating motor. Understanding the dynamics of rotor components in functioning motor is important for the clarifying of working mechanism of bacterial flagellar motor. In this study, we focused on the dynamics and the turnover of rotor components in a functioning flagellar motor. Expression systems for GFP-FliN, FliM-GFP,more » and GFP-FliG were constructed, and each GFP-fusion was functionally incorporated into the flagellar motor. To investigate whether the rotor components are exchanged in a rotating motor, we performed fluorescence recovery after photobleaching experiments using total internal reflection fluorescence microscopy. After photobleaching, in a tethered cell producing GFP-FliN or FliM-GFP, the recovery of fluorescence at the rotational center was observed. However, in a cell producing GFP-FliG, no recovery of fluorescence was observed. The transition phase of fluorescence intensity after full or partially photobleaching allowed the turnover of FliN subunits to be calculated as 0.0007 s{sup -1}, meaning that FliN would be exchanged in tens of minutes. These novel findings indicate that a bacterial flagellar motor is not a static structure even in functioning state. This is the first report for the exchange of rotor components in a functioning bacterial flagellar motor.« less

  4. Structural differences in the bacterial flagellar motor among bacterial species.

    PubMed

    Terashima, Hiroyuki; Kawamoto, Akihiro; Morimoto, Yusuke V; Imada, Katsumi; Minamino, Tohru

    2017-01-01

    The bacterial flagellum is a supramolecular motility machine consisting of the basal body as a rotary motor, the hook as a universal joint, and the filament as a helical propeller. Intact structures of the bacterial flagella have been observed for different bacterial species by electron cryotomography and subtomogram averaging. The core structures of the basal body consisting of the C ring, the MS ring, the rod and the protein export apparatus, and their organization are well conserved, but novel and divergent structures have also been visualized to surround the conserved structure of the basal body. This suggests that the flagellar motors have adapted to function in various environments where bacteria live and survive. In this review, we will summarize our current findings on the divergent structures of the bacterial flagellar motor.

  5. Steps in the bacterial flagellar motor.

    PubMed

    Mora, Thierry; Yu, Howard; Sowa, Yoshiyuki; Wingreen, Ned S

    2009-10-01

    The bacterial flagellar motor is a highly efficient rotary machine used by many bacteria to propel themselves. It has recently been shown that at low speeds its rotation proceeds in steps. Here we propose a simple physical model, based on the storage of energy in protein springs, that accounts for this stepping behavior as a random walk in a tilted corrugated potential that combines torque and contact forces. We argue that the absolute angular position of the rotor is crucial for understanding step properties and show this hypothesis to be consistent with the available data, in particular the observation that backward steps are smaller on average than forward steps. We also predict a sublinear speed versus torque relationship for fixed load at low torque, and a peak in rotor diffusion as a function of torque. Our model provides a comprehensive framework for understanding and analyzing stepping behavior in the bacterial flagellar motor and proposes novel, testable predictions. More broadly, the storage of energy in protein springs by the flagellar motor may provide useful general insights into the design of highly efficient molecular machines.

  6. Na+-driven bacterial flagellar motors.

    PubMed

    Imae, Y; Atsumi, T

    1989-12-01

    Bacterial flagellar motors are the reversible rotary engine which propels the cell by rotating a helical flagellar filament as a screw propeller. The motors are embedded in the cytoplasmic membrane, and the energy for rotation is supplied by the electrochemical potential of specific ions across the membrane. Thus, the analysis of motor rotation at the molecular level is linked to an understanding of how the living system converts chemical energy into mechanical work. Based on the coupling ions, the motors are divided into two types; one is the H+-driven type found in neutrophiles such as Bacillus subtilis and Escherichia coli and the other is the Na+-driven type found in alkalophilic Bacillus and marine Vibrio. In this review, we summarize the current status of research on the rotation mechanism of the Na+-driven flagellar motors, which introduces several new aspects in the analysis.

  7. Step-wise loss of bacterial flagellar torsion confers progressive phagocytic evasion.

    PubMed

    Lovewell, Rustin R; Collins, Ryan M; Acker, Julie L; O'Toole, George A; Wargo, Matthew J; Berwin, Brent

    2011-09-01

    Phagocytosis of bacteria by innate immune cells is a primary method of bacterial clearance during infection. However, the mechanisms by which the host cell recognizes bacteria and consequentially initiates phagocytosis are largely unclear. Previous studies of the bacterium Pseudomonas aeruginosa have indicated that bacterial flagella and flagellar motility play an important role in colonization of the host and, importantly, that loss of flagellar motility enables phagocytic evasion. Here we use molecular, cellular, and genetic methods to provide the first formal evidence that phagocytic cells recognize bacterial motility rather than flagella and initiate phagocytosis in response to this motility. We demonstrate that deletion of genes coding for the flagellar stator complex, which results in non-swimming bacteria that retain an initial flagellar structure, confers resistance to phagocytic binding and ingestion in several species of the gamma proteobacterial group of Gram-negative bacteria, indicative of a shared strategy for phagocytic evasion. Furthermore, we show for the first time that susceptibility to phagocytosis in swimming bacteria is proportional to mot gene function and, consequently, flagellar rotation since complementary genetically- and biochemically-modulated incremental decreases in flagellar motility result in corresponding and proportional phagocytic evasion. These findings identify that phagocytic cells respond to flagellar movement, which represents a novel mechanism for non-opsonized phagocytic recognition of pathogenic bacteria.

  8. Load-dependent assembly of the bacterial flagellar motor.

    PubMed

    Tipping, Murray J; Delalez, Nicolas J; Lim, Ren; Berry, Richard M; Armitage, Judith P

    2013-08-20

    It is becoming clear that the bacterial flagellar motor output is important not only for bacterial locomotion but also for mediating the transition from liquid to surface living. The output of the flagellar motor changes with the mechanical load placed on it by the external environment: at a higher load, the motor runs more slowly and produces higher torque. Here we show that the number of torque-generating units bound to the flagellar motor also depends on the external mechanical load, with fewer stators at lower loads. Stalled motors contained at least as many stators as rotating motors at high load, indicating that rotation is unnecessary for stator binding. Mutant stators incapable of generating torque could not be detected around the motor. We speculate that a component of the bacterial flagellar motor senses external load and mediates the strength of stator binding to the rest of the motor. The transition between liquid living and surface living is important in the life cycles of many bacteria. In this paper, we describe how the flagellar motor, used by bacteria for locomotion through liquid media and across solid surfaces, is capable of adjusting the number of bound stator units to better suit the external load conditions. By stalling motors using external magnetic fields, we also show that rotation is not required for maintenance of stators around the motor; instead, torque production is the essential factor for motor stability. These new results, in addition to previous data, lead us to hypothesize that the motor stators function as mechanosensors as well as functioning as torque-generating units.

  9. The counterbend phenomenon in flagellar axonemes and cross-linked filament bundles

    PubMed Central

    Gadêlha, Hermes; Gaffney, Eamonn A.; Goriely, Alain

    2013-01-01

    Recent observations of flagellar counterbend in sea urchin sperm show that the mechanical induction of curvature in one part of a passive flagellum induces a compensatory countercurvature elsewhere. This apparent paradoxical effect cannot be explained using the standard elastic rod theory of Euler and Bernoulli, or even the more general Cosserat theory of rods. Here, we develop a geometrically exact mechanical model to describe the statics of microtubule bundles that is capable of predicting the curvature reversal events observed in eukaryotic flagella. This is achieved by allowing the interaction of deformations in different material directions, by accounting not only for structural bending, but also for the elastic forces originating from the internal cross-linking mechanics. Large-amplitude static configurations can be described analytically, and an excellent match between the model and the observed counterbend deformation was found. This allowed a simultaneous estimation of multiple sperm flagellum material parameters, namely the cross-linking sliding resistance, the bending stiffness, and the sperm head junction compliance ratio. We further show that small variations on the empirical conditions may induce discrepancies for the evaluation of the flagellar material quantities, so that caution is required when interpreting experiments. Finally, our analysis demonstrates that the counterbend emerges as a fundamental property of sliding resistance in cross-linked filamentous polymer bundles, which also suggests that cross-linking proteins may contribute to the regulation of the flagellar waveform in swimming sperm via counterbend mechanics. PMID:23824293

  10. Step-Wise Loss of Bacterial Flagellar Torsion Confers Progressive Phagocytic Evasion

    PubMed Central

    Lovewell, Rustin R.; Collins, Ryan M.; Acker, Julie L.; O'Toole, George A.; Wargo, Matthew J.; Berwin, Brent

    2011-01-01

    Phagocytosis of bacteria by innate immune cells is a primary method of bacterial clearance during infection. However, the mechanisms by which the host cell recognizes bacteria and consequentially initiates phagocytosis are largely unclear. Previous studies of the bacterium Pseudomonas aeruginosa have indicated that bacterial flagella and flagellar motility play an important role in colonization of the host and, importantly, that loss of flagellar motility enables phagocytic evasion. Here we use molecular, cellular, and genetic methods to provide the first formal evidence that phagocytic cells recognize bacterial motility rather than flagella and initiate phagocytosis in response to this motility. We demonstrate that deletion of genes coding for the flagellar stator complex, which results in non-swimming bacteria that retain an initial flagellar structure, confers resistance to phagocytic binding and ingestion in several species of the gamma proteobacterial group of Gram-negative bacteria, indicative of a shared strategy for phagocytic evasion. Furthermore, we show for the first time that susceptibility to phagocytosis in swimming bacteria is proportional to mot gene function and, consequently, flagellar rotation since complementary genetically- and biochemically-modulated incremental decreases in flagellar motility result in corresponding and proportional phagocytic evasion. These findings identify that phagocytic cells respond to flagellar movement, which represents a novel mechanism for non-opsonized phagocytic recognition of pathogenic bacteria. PMID:21949654

  11. A macroscopic scale model of bacterial flagellar bundling

    NASA Astrophysics Data System (ADS)

    Kim, Munju; Bird, James C.; van Parys, Annemarie J.; Breuer, Kenneth S.; Powers, Thomas R.

    2003-12-01

    Escherichia coli and other bacteria use rotating helical filaments to swim. Each cell typically has about four filaments, which bundle or disperse depending on the sense of motor rotation. To study the bundling process, we built a macroscopic scale model consisting of stepper motor-driven polymer helices in a tank filled with a high-viscosity silicone oil. The Reynolds number, the ratio of viscous to elastic stresses, and the helix geometry of our experimental model approximately match the corresponding quantities of the full-scale E. coli cells. We analyze digital video images of the rotating helices to show that the initial rate of bundling is proportional to the motor frequency and is independent of the characteristic relaxation time of the filament. We also determine which combinations of helix handedness and sense of motor rotation lead to bundling.

  12. The bacterial flagellar switch complex is getting more complex

    PubMed Central

    Cohen-Ben-Lulu, Galit N; Francis, Noreen R; Shimoni, Eyal; Noy, Dror; Davidov, Yaacov; Prasad, Krishna; Sagi, Yael; Cecchini, Gary; Johnstone, Rose M; Eisenbach, Michael

    2008-01-01

    The mechanism of function of the bacterial flagellar switch, which determines the direction of flagellar rotation and is essential for chemotaxis, has remained an enigma for many years. Here we show that the switch complex associates with the membrane-bound respiratory protein fumarate reductase (FRD). We provide evidence that FRD binds to preparations of isolated switch complexes, forms a 1:1 complex with the switch protein FliG, and that this interaction is required for both flagellar assembly and switching the direction of flagellar rotation. We further show that fumarate, known to be a clockwise/switch factor, affects the direction of flagellar rotation through FRD. These results not only uncover a new component important for switching and flagellar assembly, but they also reveal that FRD, an enzyme known to be primarily expressed and functional under anaerobic conditions in Escherichia coli, nonetheless, has important, unexpected functions under aerobic conditions. PMID:18337747

  13. Simultaneous measurement of bacterial flagellar rotation rate and swimming speed.

    PubMed Central

    Magariyama, Y; Sugiyama, S; Muramoto, K; Kawagishi, I; Imae, Y; Kudo, S

    1995-01-01

    Swimming speeds and flagellar rotation rates of individual free-swimming Vibrio alginolyticus cells were measured simultaneously by laser dark-field microscopy at 25, 30, and 35 degrees C. A roughly linear relation between swimming speed and flagellar rotation rate was observed. The ratio of swimming speed to flagellar rotation rate was 0.113 microns, which indicated that a cell progressed by 7% of pitch of flagellar helix during one flagellar rotation. At each temperature, however, swimming speed had a tendency to saturate at high flagellar rotation rate. That is, the cell with a faster-rotating flagellum did not always swim faster. To analyze the bacterial motion, we proposed a model in which the torque characteristics of the flagellar motor were considered. The model could be analytically solved, and it qualitatively explained the experimental results. The discrepancy between the experimental and the calculated ratios of swimming speed to flagellar rotation rate was about 20%. The apparent saturation in swimming speed was considered to be caused by shorter flagella that rotated faster but produced less propelling force. Images FIGURE 1 FIGURE 4 PMID:8580359

  14. [Structure and function of the bacterial flagellar type III protein export system in Salmonella
].

    PubMed

    Minamino, Tohru

    2015-01-01

    The bacterial flagellum is a filamentous organelle that propels the bacterial cell body in liquid media. For construction of the bacterial flagellum beyond the cytoplasmic membrane, flagellar component proteins are transported by its specific protein export apparatus from the cytoplasm to the distal end of the growing flagellar structure. The flagellar export apparatus consists of a transmembrane export gate complex and a cytoplasmic ATPase ring complex. Flagellar substrate-specific chaperones bind to their cognate substrates in the cytoplasm and escort the substrates to the docking platform of the export gate. The export apparatus utilizes ATP and proton motive force across the cytoplasmic membrane as the energy sources to drive protein export and coordinates protein export with assembly by ordered export of substrates to parallel with their order of assembly. In this review, we summarize our current understanding of the structure and function of the flagellar protein export system in Salmonella enterica serovar Typhimurium.

  15. Fuel of the Bacterial Flagellar Type III Protein Export Apparatus.

    PubMed

    Minamino, Tohru; Kinoshita, Miki; Namba, Keiichi

    2017-01-01

    The flagellar type III export apparatus utilizes ATP and proton motive force (PMF) across the cytoplasmic membrane as the energy sources and transports flagellar component proteins from the cytoplasm to the distal growing end of the growing structure to construct the bacterial flagellum beyond the cellular membranes. The flagellar type III export apparatus coordinates flagellar protein export with assembly by ordered export of substrates to parallel with their order of the assembly. The export apparatus is composed of a PMF-driven transmembrane export gate complex and a cytoplasmic ATPase complex. Since the ATPase complex is dispensable for flagellar protein export, PMF is the primary fuel for protein unfolding and translocation. Interestingly, the export gate complex can also use sodium motive force across the cytoplasmic membrane in addition to PMF when the ATPase complex does not work properly. Here, we describe experimental protocols, which have allowed us to identify the export substrate class and the primary fuel of the flagellar type III protein export apparatus in Salmonella enterica serovar Typhimurium.

  16. Structure and Function of the Bi-Directional Bacterial Flagellar Motor

    PubMed Central

    Morimoto, Yusuke V.; Minamino, Tohru

    2014-01-01

    The bacterial flagellum is a locomotive organelle that propels the bacterial cell body in liquid environments. The flagellum is a supramolecular complex composed of about 30 different proteins and consists of at least three parts: a rotary motor, a universal joint, and a helical filament. The flagellar motor of Escherichia coli and Salmonella enterica is powered by an inward-directed electrochemical potential difference of protons across the cytoplasmic membrane. The flagellar motor consists of a rotor made of FliF, FliG, FliM and FliN and a dozen stators consisting of MotA and MotB. FliG, FliM and FliN also act as a molecular switch, enabling the motor to spin in both counterclockwise and clockwise directions. Each stator is anchored to the peptidoglycan layer through the C-terminal periplasmic domain of MotB and acts as a proton channel to couple the proton flow through the channel with torque generation. Highly conserved charged residues at the rotor–stator interface are required not only for torque generation but also for stator assembly around the rotor. In this review, we will summarize our current understanding of the structure and function of the proton-driven bacterial flagellar motor. PMID:24970213

  17. Structure and function of the bi-directional bacterial flagellar motor.

    PubMed

    Morimoto, Yusuke V; Minamino, Tohru

    2014-02-18

    The bacterial flagellum is a locomotive organelle that propels the bacterial cell body in liquid environments. The flagellum is a supramolecular complex composed of about 30 different proteins and consists of at least three parts: a rotary motor, a universal joint, and a helical filament. The flagellar motor of Escherichia coli and Salmonella enterica is powered by an inward-directed electrochemical potential difference of protons across the cytoplasmic membrane. The flagellar motor consists of a rotor made of FliF, FliG, FliM and FliN and a dozen stators consisting of MotA and MotB. FliG, FliM and FliN also act as a molecular switch, enabling the motor to spin in both counterclockwise and clockwise directions. Each stator is anchored to the peptidoglycan layer through the C-terminal periplasmic domain of MotB and acts as a proton channel to couple the proton flow through the channel with torque generation. Highly conserved charged residues at the rotor-stator interface are required not only for torque generation but also for stator assembly around the rotor. In this review, we will summarize our current understanding of the structure and function of the proton-driven bacterial flagellar motor.

  18. Themes and Variations: Regulation of RpoN-Dependent Flagellar Genes across Diverse Bacterial Species

    PubMed Central

    Tsang, Jennifer; Hoover, Timothy R.

    2014-01-01

    Flagellar biogenesis in bacteria is a complex process in which the transcription of dozens of structural and regulatory genes is coordinated with the assembly of the flagellum. Although the overall process of flagellar biogenesis is conserved among bacteria, the mechanisms used to regulate flagellar gene expression vary greatly among different bacterial species. Many bacteria use the alternative sigma factor σ 54 (also known as RpoN) to transcribe specific sets of flagellar genes. These bacteria include members of the Epsilonproteobacteria (e.g., Helicobacter pylori and Campylobacter jejuni), Gammaproteobacteria (e.g., Vibrio and Pseudomonas species), and Alphaproteobacteria (e.g., Caulobacter crescentus). This review characterizes the flagellar transcriptional hierarchies in these bacteria and examines what is known about how flagellar gene regulation is linked with other processes including growth phase, quorum sensing, and host colonization. PMID:24672734

  19. Flagellar Hook Flexibility Is Essential for Bundle Formation in Swimming Escherichia coli Cells

    PubMed Central

    Brown, Mostyn T.; Steel, Bradley C.; Silvestrin, Claudio; Wilkinson, David A.; Delalez, Nicolas J.; Lumb, Craig N.; Obara, Boguslaw; Berry, Richard M.

    2012-01-01

    Swimming Escherichia coli cells are propelled by the rotary motion of their flagellar filaments. In the normal swimming pattern, filaments positioned randomly over the cell form a bundle at the posterior pole. It has long been assumed that the hook functions as a universal joint, transmitting rotation on the motor axis through up to ∼90° to the filament in the bundle. Structural models of the hook have revealed how its flexibility is expected to arise from dynamic changes in the distance between monomers in the helical lattice. In particular, each of the 11 protofilaments that comprise the hook is predicted to cycle between short and long forms, corresponding to the inside and outside of the curved hook, once each revolution of the motor when the hook is acting as a universal joint. To test this, we genetically modified the hook so that it could be stiffened by binding streptavidin to biotinylated monomers, impeding their motion relative to each other. We found that impeding the action of the universal joint resulted in atypical swimming behavior as a consequence of disrupted bundle formation, in agreement with the universal joint model. PMID:22522898

  20. Hierarchical protein export mechanism of the bacterial flagellar type III protein export apparatus.

    PubMed

    Minamino, Tohru

    2018-06-01

    The bacterial flagellum is supramolecular motility machinery consisting of the basal body, the hook and the filament. Flagellar proteins are translocated across the cytoplasmic membrane via a type III protein export apparatus, diffuse down the central channel of the growing structure and assemble at the distal end. Flagellar assembly begins with the basal body, followed by the hook and finally the filament. The completion of hook assembly is the most important morphological checkpoint of the sequential flagellar assembly process. When the hook reaches its mature length of about 55 nm in Salmonella enterica, the type III protein export apparatus switches export specificity from proteins required for the structure and assembly of the hook to those responsible for filament assembly, thereby terminating hook assembly and initiating filament assembly. Three flagellar proteins, namely FliK, FlhB and FlhA, are responsible for this substrate specificity switching. Upon completion of the switching event, interactions among FlhA, the cytoplasmic ATPase complex and flagellar type III export chaperones establish the assembly order of the filament at the hook tip. Here, we describe our current understanding of a hierarchical protein export mechanism used in flagellar type III protein export.

  1. Mechanics of torque generation in the bacterial flagellar motor

    PubMed Central

    Mandadapu, Kranthi K.; Nirody, Jasmine A.; Berry, Richard M.; Oster, George

    2015-01-01

    The bacterial flagellar motor (BFM) is responsible for driving bacterial locomotion and chemotaxis, fundamental processes in pathogenesis and biofilm formation. In the BFM, torque is generated at the interface between transmembrane proteins (stators) and a rotor. It is well established that the passage of ions down a transmembrane gradient through the stator complex provides the energy for torque generation. However, the physics involved in this energy conversion remain poorly understood. Here we propose a mechanically specific model for torque generation in the BFM. In particular, we identify roles for two fundamental forces involved in torque generation: electrostatic and steric. We propose that electrostatic forces serve to position the stator, whereas steric forces comprise the actual “power stroke.” Specifically, we propose that ion-induced conformational changes about a proline “hinge” residue in a stator α-helix are directly responsible for generating the power stroke. Our model predictions fit well with recent experiments on a single-stator motor. The proposed model provides a mechanical explanation for several fundamental properties of the flagellar motor, including torque–speed and speed–ion motive force relationships, backstepping, variation in step sizes, and the effects of key mutations in the stator. PMID:26216959

  2. Mechanics of torque generation in the bacterial flagellar motor.

    PubMed

    Mandadapu, Kranthi K; Nirody, Jasmine A; Berry, Richard M; Oster, George

    2015-08-11

    The bacterial flagellar motor (BFM) is responsible for driving bacterial locomotion and chemotaxis, fundamental processes in pathogenesis and biofilm formation. In the BFM, torque is generated at the interface between transmembrane proteins (stators) and a rotor. It is well established that the passage of ions down a transmembrane gradient through the stator complex provides the energy for torque generation. However, the physics involved in this energy conversion remain poorly understood. Here we propose a mechanically specific model for torque generation in the BFM. In particular, we identify roles for two fundamental forces involved in torque generation: electrostatic and steric. We propose that electrostatic forces serve to position the stator, whereas steric forces comprise the actual "power stroke." Specifically, we propose that ion-induced conformational changes about a proline "hinge" residue in a stator α-helix are directly responsible for generating the power stroke. Our model predictions fit well with recent experiments on a single-stator motor. The proposed model provides a mechanical explanation for several fundamental properties of the flagellar motor, including torque-speed and speed-ion motive force relationships, backstepping, variation in step sizes, and the effects of key mutations in the stator.

  3. Bacterial flagellar capping proteins adopt diverse oligomeric states

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

    Postel, Sandra; Deredge, Daniel; Bonsor, Daniel A.

    2016-09-24

    Flagella are crucial for bacterial motility and pathogenesis. The flagellar capping protein (FliD) regulates filament assembly by chaperoning and sorting flagellin (FliC) proteins after they traverse the hollow filament and exit the growing flagellum tip. In the absence of FliD, flagella are not formed, resulting in impaired motility and infectivity. Here, we report the 2.2 Å resolution X-ray crystal structure of FliD fromPseudomonas aeruginosa, the first high-resolution structure of any FliD protein from any bacterium. Using this evidence in combination with a multitude of biophysical and functional analyses, we find thatPseudomonasFliD exhibits unexpected structural similarity to other flagellar proteins atmore » the domain level, adopts a unique hexameric oligomeric state, and depends on flexible determinants for oligomerization. Considering that the flagellin filaments on which FliD oligomers are affixed vary in protofilament number between bacteria, our results suggest that FliD oligomer stoichiometries vary across bacteria to complement their filament assemblies.« less

  4. Microscopic Analysis of Bacterial Motility at High Pressure

    PubMed Central

    Nishiyama, Masayoshi; Sowa, Yoshiyuki

    2012-01-01

    The bacterial flagellar motor is a molecular machine that converts an ion flux to the rotation of a helical flagellar filament. Counterclockwise rotation of the filaments allows them to join in a bundle and propel the cell forward. Loss of motility can be caused by environmental factors such as temperature, pH, and solvation. Hydrostatic pressure is also a physical inhibitor of bacterial motility, but the detailed mechanism of this inhibition is still unknown. Here, we developed a high-pressure microscope that enables us to acquire high-resolution microscopic images, regardless of applied pressures. We also characterized the pressure dependence of the motility of swimming Escherichia coli cells and the rotation of single flagellar motors. The fraction and speed of swimming cells decreased with increased pressure. At 80 MPa, all cells stopped swimming and simply diffused in solution. After the release of pressure, most cells immediately recovered their initial motility. Direct observation of the motility of single flagellar motors revealed that at 80 MPa, the motors generate torque that should be sufficient to join rotating filaments in a bundle. The discrepancy in the behavior of free swimming cells and individual motors could be due to the applied pressure inhibiting the formation of rotating filament bundles that can propel the cell body in an aqueous environment. PMID:22768943

  5. Cyclic di-GMP differentially tunes a bacterial flagellar motor through a novel class of CheY-like regulators.

    PubMed

    Nesper, Jutta; Hug, Isabelle; Kato, Setsu; Hee, Chee-Seng; Habazettl, Judith Maria; Manfredi, Pablo; Grzesiek, Stephan; Schirmer, Tilman; Emonet, Thierry; Jenal, Urs

    2017-11-01

    The flagellar motor is a sophisticated rotary machine facilitating locomotion and signal transduction. Owing to its important role in bacterial behavior, its assembly and activity are tightly regulated. For example, chemotaxis relies on a sensory pathway coupling chemical information to rotational bias of the motor through phosphorylation of the motor switch protein CheY. Using a chemical proteomics approach, we identified a novel family of CheY-like (Cle) proteins in Caulobacter crescentus , which tune flagellar activity in response to binding of the second messenger c-di-GMP to a C-terminal extension. In their c-di-GMP bound conformation Cle proteins interact with the flagellar switch to control motor activity. We show that individual Cle proteins have adopted discrete cellular functions by interfering with chemotaxis and by promoting rapid surface attachment of motile cells. This study broadens the regulatory versatility of bacterial motors and unfolds mechanisms that tie motor activity to mechanical cues and bacterial surface adaptation.

  6. Gap compression/extension mechanism of bacterial flagellar hook as the molecular universal joint.

    PubMed

    Furuta, Tadaomi; Samatey, Fadel A; Matsunami, Hideyuki; Imada, Katsumi; Namba, Keiichi; Kitao, Akio

    2007-03-01

    Bacterial flagellar hook acts as a molecular universal joint, transmitting torque produced by the flagellar basal body, a rotary motor, to the flagellar filament. The hook forms polymorphic supercoil structures and can be considered as an assembly of 11 circularly arranged protofilaments. We investigated the molecular mechanism of the universal joint function of the hook by a approximately two-million-atom molecular dynamics simulation. On the inner side of the supercoil, protein subunits are highly packed along the protofilament and no gaps remain for further compression, whereas subunits are slightly separated and are hydrogen bonded through one layer of water molecules on the outer side. As for the intersubunit interactions between protofilaments, subunits are packed along the 6-start helix in a left-handed supercoil whereas they are highly packed along the 5-start helix in a right-handed supercoil. We conclude that the supercoiled structures of the hook in the left- and right-handed forms make maximal use of the gaps between subunits, which we call "gap compression/extension mechanism". Mutual sliding of subunits at the subunit interface accompanying rearrangements of intersubunit hydrogen bonds is interpreted as a mechanism to allow continuous structural change of the hook during flagellar rotation at low energy cost.

  7. Structure of the bacterial flagellar hook and implication for the molecular universal joint mechanism.

    PubMed

    Samatey, Fadel A; Matsunami, Hideyuki; Imada, Katsumi; Nagashima, Shigehiro; Shaikh, Tanvir R; Thomas, Dennis R; Chen, James Z; Derosier, David J; Kitao, Akio; Namba, Keiichi

    2004-10-28

    The bacterial flagellum is a motile organelle, and the flagellar hook is a short, highly curved tubular structure that connects the flagellar motor to the long filament acting as a helical propeller. The hook is made of about 120 copies of a single protein, FlgE, and its function as a nano-sized universal joint is essential for dynamic and efficient bacterial motility and taxis. It transmits the motor torque to the helical propeller over a wide range of its orientation for swimming and tumbling. Here we report a partial atomic model of the hook obtained by X-ray crystallography of FlgE31, a major proteolytic fragment of FlgE lacking unfolded terminal regions, and by electron cryomicroscopy and three-dimensional helical image reconstruction of the hook. The model reveals the intricate molecular interactions and a plausible switching mechanism for the hook to be flexible in bending but rigid against twisting for its universal joint function.

  8. Limiting Speed of the Bacterial Flagellar Motor

    NASA Astrophysics Data System (ADS)

    Nirody, Jasmine; Berry, Richard; Oster, George

    The bacterial flagellar motor (BFM) drives swimming in a wide variety of bacterial species, making it crucial for several fundamental biological processes including chemotaxis and community formation. Recent experiments have shown that the structure of this nanomachine is more dynamic than previously believed. Specifically, the number of active torque-generating units (stators) was shown to vary across applied loads. This finding invalidates the experimental evidence reporting that limiting (zero-torque) speed is independent of the number of active stators. Here, we put forward a model for the torque generation mechanism of this motor and propose that the maximum speed of the motor increases as additional torque-generators are recruited. This is contrary to the current widely-held belief that there is a universal upper limit to the speed of the BFM. Our result arises from the assumption that stators disengage from the motor for a significant portion of their mechanochemical cycles at low loads. We show that this assumption is consistent with current experimental evidence and consolidate our predictions with arguments that a processive motor must have a high duty ratio at high loads.

  9. Real-Time Imaging of Fluorescent Flagellar Filaments of Rhizobium lupini H13-3: Flagellar Rotation and pH-Induced Polymorphic Transitions

    PubMed Central

    Scharf, Birgit

    2002-01-01

    The soil bacterium Rhizobium lupini H13-3 has complex right-handed flagellar filaments with unusual ridged, grooved surfaces. Clockwise (CW) rotation propels the cells forward, and course changes (tumbling) result from changes in filament speed instead of the more common change in direction of rotation. In view of these novelties, fluorescence labeling was used to analyze the behavior of single flagellar filaments during swimming and tumbling, leading to a model for directional changes in R. lupini. Also, flagellar filaments were investigated for helical conformational changes, which have not been previously shown for complex filaments. During full-speed CW rotation, the flagellar filaments form a propulsive bundle that pushes the cell on a straight path. Tumbling is caused by asynchronous deceleration and stops of individual filaments, resulting in dissociation of the propulsive bundle. R. lupini tumbles were not accompanied by helical conformational changes as are tumbles in other organisms including enteric bacteria. However, when pH was experimentally changed, four different polymorphic forms were observed. At a physiological pH of 7, normal flagellar helices were characterized by a pitch angle of 30°, a pitch of 1.36 μm, and a helical diameter of 0.50 μm. As pH increased from 9 to 11, the helices transformed from normal to semicoiled to straight. As pH decreased from 5 to 3, the helices transformed from normal to curly to straight. Transient conformational changes were also noted at high viscosity, suggesting that the R. lupini flagellar filament may adapt to high loads in viscous environments (soil) by assuming hydrodynamically favorable conformations. PMID:12374832

  10. Protein export through the bacterial flagellar type III export pathway.

    PubMed

    Minamino, Tohru

    2014-08-01

    For construction of the bacterial flagellum, which is responsible for bacterial motility, the flagellar type III export apparatus utilizes both ATP and proton motive force across the cytoplasmic membrane and exports flagellar proteins from the cytoplasm to the distal end of the nascent structure. The export apparatus consists of a membrane-embedded export gate made of FlhA, FlhB, FliO, FliP, FliQ, and FliR and a water-soluble ATPase ring complex consisting of FliH, FliI, and FliJ. FlgN, FliS, and FliT act as substrate-specific chaperones that do not only protect their cognate substrates from degradation and aggregation in the cytoplasm but also efficiently transfer the substrates to the export apparatus. The ATPase ring complex facilitates the initial entry of the substrates into the narrow pore of the export gate. The export gate by itself is a proton-protein antiporter that uses the two components of proton motive force, the electric potential difference and the proton concentration difference, for different steps of the export process. A specific interaction of FlhA with FliJ located in the center of the ATPase ring complex allows the export gate to efficiently use proton motive force to drive protein export. The ATPase ring complex couples ATP binding and hydrolysis to its assembly-disassembly cycle for rapid and efficient protein export cycle. This article is part of a Special Issue entitled: Protein trafficking and secretion in bacteria. Guest Editors: Anastassios Economou and Ross Dalbey. © 2013 Elsevier B.V. All rights reserved.

  11. Hybrid-fuel bacterial flagellar motors in Escherichia coli

    PubMed Central

    Sowa, Yoshiyuki; Homma, Michio; Ishijima, Akihiko; Berry, Richard M.

    2014-01-01

    The bacterial flagellar motor rotates driven by an electrochemical ion gradient across the cytoplasmic membrane, either H+ or Na+ ions. The motor consists of a rotor ∼50 nm in diameter surrounded by multiple torque-generating ion-conducting stator units. Stator units exchange spontaneously between the motor and a pool in the cytoplasmic membrane on a timescale of minutes, and their stability in the motor is dependent upon the ion gradient. We report a genetically engineered hybrid-fuel flagellar motor in Escherichia coli that contains both H+- and Na+-driven stator components and runs on both types of ion gradient. We controlled the number of each type of stator unit in the motor by protein expression levels and Na+ concentration ([Na+]), using speed changes of single motors driving 1-μm polystyrene beads to determine stator unit numbers. De-energized motors changed from locked to freely rotating on a timescale similar to that of spontaneous stator unit exchange. Hybrid motor speed is simply the sum of speeds attributable to individual stator units of each type. With Na+ and H+ stator components expressed at high and medium levels, respectively, Na+ stator units dominate at high [Na+] and are replaced by H+ units when Na+ is removed. Thus, competition between stator units for spaces in a motor and sensitivity of each type to its own ion gradient combine to allow hybrid motors to adapt to the prevailing ion gradient. We speculate that a similar process may occur in species that naturally express both H+ and Na+ stator components sharing a common rotor. PMID:24550452

  12. Changes in the flagellar bundling time account for variations in swimming behavior of flagellated bacteria in viscous media

    NASA Astrophysics Data System (ADS)

    Qu, Zijie; Temel, Fatma; Henderikx, Rene; Breuer, Kenneth

    2017-11-01

    The motility of bacteria E.coli in viscous fluids has been widely studied, although conflicting results on the effect of viscosity on swimming speed abound. The swimming mode of wild-type E.coli is idealized as a run-and-tumble sequence in which periods of straight swimming at a constant speed are randomly interrupted by a tumble, defined as a sudden change of direction with a very low speed. Using a tracking microscope, we follow cells for extended time and find that the swimming behavior of a single cell can exhibit a variety of behaviors including run-and-tumble and ``slow-random-walk'' in which the cells move at relatively low speed without the characteristic run. Although the characteristic swimming speed varies between individuals and in different polymer solutions, we find that the skewness of the speed distribution is solely a function of viscosity, and uniquely determines the ratio of the average speed to the characteristic run speed. Using Resistive Force Theory and the cell-specific measured characteristic run speed, we show that differences in the swimming behavior observed in solutions of different viscosity are due to changes in the flagellar bundling time, which increases as the viscosity rises, due to lower rotation rate of the flagellar motor. National Science Foundation.

  13. Insight into structural remodeling of the FlhA ring responsible for bacterial flagellar type III protein export

    PubMed Central

    2018-01-01

    The bacterial flagellum is a supramolecular motility machine. Flagellar assembly begins with the basal body, followed by the hook and finally the filament. A carboxyl-terminal cytoplasmic domain of FlhA (FlhAC) forms a nonameric ring structure in the flagellar type III protein export apparatus and coordinates flagellar protein export with assembly. However, the mechanism of this process remains unknown. We report that a flexible linker of FlhAC (FlhAL) is required not only for FlhAC ring formation but also for substrate specificity switching of the protein export apparatus from the hook protein to the filament protein upon completion of the hook structure. FlhAL was required for cooperative ring formation of FlhAC. Alanine substitutions of residues involved in FlhAC ring formation interfered with the substrate specificity switching, thereby inhibiting filament assembly at the hook tip. These observations lead us to propose a mechanistic model for export switching involving structural remodeling of FlhAC. PMID:29707633

  14. Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold

    PubMed Central

    Ribardo, Deborah A.; Brennan, Caitlin A.; Ruby, Edward G.; Jensen, Grant J.; Hendrixson, David R.

    2016-01-01

    Although it is known that diverse bacterial flagellar motors produce different torques, the mechanism underlying torque variation is unknown. To understand this difference better, we combined genetic analyses with electron cryo-tomography subtomogram averaging to determine in situ structures of flagellar motors that produce different torques, from Campylobacter and Vibrio species. For the first time, to our knowledge, our results unambiguously locate the torque-generating stator complexes and show that diverse high-torque motors use variants of an ancestrally related family of structures to scaffold incorporation of additional stator complexes at wider radii from the axial driveshaft than in the model enteric motor. We identify the protein components of these additional scaffold structures and elucidate their sequential assembly, demonstrating that they are required for stator-complex incorporation. These proteins are widespread, suggesting that different bacteria have tailored torques to specific environments by scaffolding alternative stator placement and number. Our results quantitatively account for different motor torques, complete the assignment of the locations of the major flagellar components, and provide crucial constraints for understanding mechanisms of torque generation and the evolution of multiprotein complexes. PMID:26976588

  15. Biotemplated flagellar nanoswimmers

    NASA Astrophysics Data System (ADS)

    Ali, Jamel; Cheang, U. Kei; Darvish, Armin; Kim, Hoyeon; Kim, Min Jun

    2017-11-01

    In this article, a porous hollow biotemplated nanoscale helix that can serve as a low Reynolds number robotic swimmer is reported. The nanorobot utilizes repolymerized bacterial flagella from Salmonella typhimurium as a nanotemplate for biomineralization. We demonstrate the ability to generate templated nanotubes with distinct helical geometries by using specific alkaline pH values to fix the polymorphic form of flagellar templates. Using uniform rotating magnetic fields to mimic the motion of the flagellar motor, we explore the swimming characteristics of these silica templated flagella and demonstrate the ability to wirelessly control their trajectories. The results suggest that the biotemplated nanoswimmer can be a cost-effective alternative to the current top-down methods used to produce helical nanorobots.

  16. The Bacterial Flagellar Type III Export Gate Complex Is a Dual Fuel Engine That Can Use Both H+ and Na+ for Flagellar Protein Export

    PubMed Central

    Minamino, Tohru; Morimoto, Yusuke V.; Hara, Noritaka; Aldridge, Phillip D.; Namba, Keiichi

    2016-01-01

    The bacterial flagellar type III export apparatus utilizes ATP and proton motive force (PMF) to transport flagellar proteins to the distal end of the growing flagellar structure for self-assembly. The transmembrane export gate complex is a H+–protein antiporter, of which activity is greatly augmented by an associated cytoplasmic ATPase complex. Here, we report that the export gate complex can use sodium motive force (SMF) in addition to PMF across the cytoplasmic membrane to drive protein export. Protein export was considerably reduced in the absence of the ATPase complex and a pH gradient across the membrane, but Na+ increased it dramatically. Phenamil, a blocker of Na+ translocation, inhibited protein export. Overexpression of FlhA increased the intracellular Na+ concentration in the presence of 100 mM NaCl but not in its absence, suggesting that FlhA acts as a Na+ channel. In wild-type cells, however, neither Na+ nor phenamil affected protein export, indicating that the Na+ channel activity of FlhA is suppressed by the ATPase complex. We propose that the export gate by itself is a dual fuel engine that uses both PMF and SMF for protein export and that the ATPase complex switches this dual fuel engine into a PMF-driven export machinery to become much more robust against environmental changes in external pH and Na+ concentration. PMID:26943926

  17. The Bacterial Flagellar Type III Export Gate Complex Is a Dual Fuel Engine That Can Use Both H+ and Na+ for Flagellar Protein Export.

    PubMed

    Minamino, Tohru; Morimoto, Yusuke V; Hara, Noritaka; Aldridge, Phillip D; Namba, Keiichi

    2016-03-01

    The bacterial flagellar type III export apparatus utilizes ATP and proton motive force (PMF) to transport flagellar proteins to the distal end of the growing flagellar structure for self-assembly. The transmembrane export gate complex is a H+-protein antiporter, of which activity is greatly augmented by an associated cytoplasmic ATPase complex. Here, we report that the export gate complex can use sodium motive force (SMF) in addition to PMF across the cytoplasmic membrane to drive protein export. Protein export was considerably reduced in the absence of the ATPase complex and a pH gradient across the membrane, but Na+ increased it dramatically. Phenamil, a blocker of Na+ translocation, inhibited protein export. Overexpression of FlhA increased the intracellular Na+ concentration in the presence of 100 mM NaCl but not in its absence, suggesting that FlhA acts as a Na+ channel. In wild-type cells, however, neither Na+ nor phenamil affected protein export, indicating that the Na+ channel activity of FlhA is suppressed by the ATPase complex. We propose that the export gate by itself is a dual fuel engine that uses both PMF and SMF for protein export and that the ATPase complex switches this dual fuel engine into a PMF-driven export machinery to become much more robust against environmental changes in external pH and Na+ concentration.

  18. Evidence for symmetry in the elementary process of bidirectional torque generation by the bacterial flagellar motor

    PubMed Central

    Nakamura, Shuichi; Kami-ike, Nobunori; Yokota, Jun-ichi P.; Minamino, Tohru; Namba, Keiichi

    2010-01-01

    The bacterial flagellar motor can rotate in both counterclockwise (CCW) and clockwise (CW) directions. It has been shown that the sodium ion-driven chimeric flagellar motor rotates with 26 steps per revolution, which corresponds to the number of FliG subunits that form part of the rotor ring, but the size of the backward step is smaller than the forward one. Here we report that the proton-driven flagellar motor of Salmonella also rotates with 26 steps per revolution but symmetrical in both CCW and CW directions with occasional smaller backward steps in both directions. Occasional shift in the stepping positions is also observed, suggesting the frequent exchange of stators in one of the 11–12 possible anchoring positions around the rotor. These observations indicate that the elementary process of torque generation by the cyclic association/dissociation of the stator with every FliG subunit along the circumference of the rotor is symmetric in CCW and CW rotation even though the structure of FliG is highly asymmetric and suggests a 180° rotation of a FliG domain for the rotor-stator interaction to reverse the direction of rotation. PMID:20876126

  19. Flagellar flows around bacterial swarms

    NASA Astrophysics Data System (ADS)

    Dauparas, Justas; Lauga, Eric

    2016-08-01

    Flagellated bacteria on nutrient-rich substrates can differentiate into a swarming state and move in dense swarms across surfaces. A recent experiment measured the flow in the fluid around an Escherichia coli swarm [Wu, Hosu, and Berg, Proc. Natl. Acad. Sci. USA 108, 4147 (2011)], 10.1073/pnas.1016693108. A systematic chiral flow was observed in the clockwise direction (when viewed from above) ahead of the swarm with flow speeds of about 10 μ m /s , about 3 times greater than the radial velocity at the edge of the swarm. The working hypothesis is that this flow is due to the action of cells stalled at the edge of a colony that extend their flagellar filaments outward, moving fluid over the virgin agar. In this work we quantitatively test this hypothesis. We first build an analytical model of the flow induced by a single flagellum in a thin film and then use the model, and its extension to multiple flagella, to compare with experimental measurements. The results we obtain are in agreement with the flagellar hypothesis. The model provides further quantitative insight into the flagella orientations and their spatial distributions as well as the tangential speed profile. In particular, the model suggests that flagella are on average pointing radially out of the swarm and are not wrapped tangentially.

  20. Bundling of elastic filaments induced by hydrodynamic interactions

    NASA Astrophysics Data System (ADS)

    Man, Yi; Page, William; Poole, Robert J.; Lauga, Eric

    2017-12-01

    Peritrichous bacteria swim in viscous fluids by rotating multiple helical flagellar filaments. As the bacterium swims forward, all its flagella rotate in synchrony behind the cell in a tight helical bundle. When the bacterium changes its direction, the flagellar filaments unbundle and randomly reorient the cell for a short period of time before returning to their bundled state and resuming swimming. This rapid bundling and unbundling is, at its heart, a mechanical process whereby hydrodynamic interactions balance with elasticity to determine the time-varying deformation of the filaments. Inspired by this biophysical problem, we present in this paper what is perhaps the simplest model of bundling whereby two or more straight elastic filaments immersed in a viscous fluid rotate about their centerline, inducing rotational flows which tend to bend the filaments around each other. We derive an integrodifferential equation governing the shape of the filaments resulting from mechanical balance in a viscous fluid at low Reynolds number. We show that such equation may be evaluated asymptotically analytically in the long-wavelength limit, leading to a local partial differential equation governed by a single dimensionless bundling number. A numerical study of the dynamics predicted by the model reveals the presence of two configuration instabilities with increasing bundling numbers: first to a crossing state where filaments touch at one point and then to a bundled state where filaments wrap along each other in a helical fashion. We also consider the case of multiple filaments and the unbundling dynamics. We next provide an intuitive physical model for the crossing instability and show that it may be used to predict analytically its threshold and adapted to address the transition to a bundling state. We then use a macroscale experimental implementation of the two-filament configuration in order to validate our theoretical predictions and obtain excellent agreement. This long

  1. Pseudomonas aeruginosa flagellar motility activates the phagocyte PI3K/Akt pathway to induce phagocytic engulfment.

    PubMed

    Lovewell, Rustin R; Hayes, Sandra M; O'Toole, George A; Berwin, Brent

    2014-04-01

    Phagocytosis of the bacterial pathogen Pseudomonas aeruginosa is the primary means by which the host controls bacterially induced pneumonia during lung infection. Previous studies have identified flagellar swimming motility as a key pathogen-associated molecular pattern (PAMP) recognized by phagocytes to initiate engulfment. Correspondingly, loss of flagellar motility is observed during chronic pulmonary infection with P. aeruginosa, and this likely reflects a selection for bacteria resistant to phagocytic clearance. However, the mechanism underlying the preferential phagocytic response to motile bacteria is unknown. Here we have identified a cellular signaling pathway in alveolar macrophages and other phagocytes that is specifically activated by flagellar motility. Genetic and biochemical methods were employed to identify that phagocyte PI3K/Akt activation is required for bacterial uptake and, importantly, it is specifically activated in response to P. aeruginosa flagellar motility. Based on these observations, the second important finding that emerged from these studies is that titration of the bacterial flagellar motility results in a proportional activation state of Akt. Therefore, the Akt pathway is responsive to, and corresponds with, the degree of bacterial flagellar motility, is independent of the actin polymerization that facilitates phagocytosis, and determines the phagocytic fate of P. aeruginosa. These findings elucidate the mechanism behind motility-dependent phagocytosis of extracellular bacteria and support a model whereby phagocytic clearance exerts a selective pressure on P. aeruginosa populations in vivo, which contributes to changes in pathogenesis during infections.

  2. Pseudomonas aeruginosa flagellar motility activates the phagocyte PI3K/Akt pathway to induce phagocytic engulfment

    PubMed Central

    Lovewell, Rustin R.; Hayes, Sandra M.; O'Toole, George A.

    2014-01-01

    Phagocytosis of the bacterial pathogen Pseudomonas aeruginosa is the primary means by which the host controls bacterially induced pneumonia during lung infection. Previous studies have identified flagellar swimming motility as a key pathogen-associated molecular pattern (PAMP) recognized by phagocytes to initiate engulfment. Correspondingly, loss of flagellar motility is observed during chronic pulmonary infection with P. aeruginosa, and this likely reflects a selection for bacteria resistant to phagocytic clearance. However, the mechanism underlying the preferential phagocytic response to motile bacteria is unknown. Here we have identified a cellular signaling pathway in alveolar macrophages and other phagocytes that is specifically activated by flagellar motility. Genetic and biochemical methods were employed to identify that phagocyte PI3K/Akt activation is required for bacterial uptake and, importantly, it is specifically activated in response to P. aeruginosa flagellar motility. Based on these observations, the second important finding that emerged from these studies is that titration of the bacterial flagellar motility results in a proportional activation state of Akt. Therefore, the Akt pathway is responsive to, and corresponds with, the degree of bacterial flagellar motility, is independent of the actin polymerization that facilitates phagocytosis, and determines the phagocytic fate of P. aeruginosa. These findings elucidate the mechanism behind motility-dependent phagocytosis of extracellular bacteria and support a model whereby phagocytic clearance exerts a selective pressure on P. aeruginosa populations in vivo, which contributes to changes in pathogenesis during infections. PMID:24487390

  3. Transcriptional Control of the Lateral-Flagellar Genes of Bradyrhizobium diazoefficiens.

    PubMed

    Mongiardini, Elías J; Quelas, J Ignacio; Dardis, Carolina; Althabegoiti, M Julia; Lodeiro, Aníbal R

    2017-08-01

    Bradyrhizobium diazoefficiens , a soybean N 2 -fixing symbiont, possesses a dual flagellar system comprising a constitutive subpolar flagellum and inducible lateral flagella. Here, we analyzed the genomic organization and biosynthetic regulation of the lateral-flagellar genes. We found that these genes are located in a single genomic cluster, organized in two monocistronic transcriptional units and three operons, one possibly containing an internal transcription start site. Among the monocistronic units is blr6846, homologous to the class IB master regulators of flagellum synthesis in Brucella melitensis and Ensifer meliloti and required for the expression of all the lateral-flagellar genes except lafA2 , whose locus encodes a single lateral flagellin. We therefore named blr6846 lafR ( la teral- f lagellar r egulator). Despite its similarity to two-component response regulators and its possession of a phosphorylatable Asp residue, lafR behaved as an orphan response regulator by not requiring phosphorylation at this site. Among the genes induced by lafR is flbT L , a class III regulator. We observed different requirements for FlbT L in the synthesis of each flagellin subunit. Although the accumulation of lafA1 , but not lafA2 , transcripts required FlbT L , the production of both flagellin polypeptides required FlbT L Moreover, the regulation cascade of this lateral-flagellar regulon appeared to be not as strictly ordered as those found in other bacterial species. IMPORTANCE Bacterial motility seems essential for the free-living style in the environment, and therefore these microorganisms allocate a great deal of their energetic resources to the biosynthesis and functioning of flagella. Despite energetic costs, some bacterial species possess dual flagellar systems, one of which is a primary system normally polar or subpolar, and the other is a secondary, lateral system that is produced only under special circumstances. Bradyrhizobium diazoefficiens , an N 2 -fixing

  4. Instability of hooks during bacterial flagellar swimming

    NASA Astrophysics Data System (ADS)

    Jabbarzadeh, Mehdi; Fu, Henry C.; Henry Fu Team

    2016-11-01

    In bacteria, a flexible hook transmits torque from the rotary motor at the cell body to the flagellum. Previously, the hook has been modeled as a Kirchhoff rod between the cell body and rotating flagellum. To study effects of the hook's flexibility on the bacteria's swimming speed and trajectory for wide range hook stiffnesses and flagellum configurations, we develop an efficient simplified spring model for the hook by linearizing the Kirchhoff rod. We treat the hydrodynamics of the cell body and helical flagellum using resistance matrices calculated by the method of regularized Stokeslets. We investigate flagellar and swimming dynamics for a range of hook flexibilities and flagellar orientations relative to the cell body and compare the results to models without hook flexibility. We investigate in detail parameters corresponding to E. coli and Vibrio alginolyticus. Generally, the flagellum changes orientation relative to the cell body, undergoing an orbit with the period of the motor rotation. We find that as the hook stiffness decreases, steady-state orbits of the flagellum first become unstable before the hook buckles, which may suggest a new mechanism of flick initiation in run-reverse-flick motility. We also find that for some parameter ranges, there are multiple stable steady state orbits, which may have implications for the tumbling and turning of bacteria.

  5. The counterbend dynamics of cross-linked filament bundles and flagella

    PubMed Central

    Coy, Rachel

    2017-01-01

    Cross-linked filament bundles, such as in cilia and flagella, are ubiquitous in biology. They are considered in textbooks as simple filaments with larger stiffness. Recent observations of flagellar counterbend, however, show that induction of curvature in one section of a passive flagellum instigates a compensatory counter-curvature elsewhere, exposing the intricate role of the diminutive cross-linking proteins at large scales. We show that this effect, a material property of the cross-linking mechanics, modifies the bundle dynamics and induces a bimodal L2 − L3 length-dependent material response that departs from the Euler–Bernoulli theory. Hence, the use of simpler theories to analyse experiments can result in paradoxical interpretations. Remarkably, the counterbend dynamics instigates counter-waves in opposition to driven oscillations in distant parts of the bundle, with potential impact on the regulation of flagellar bending waves. These results have a range of physical and biological applications, including the empirical disentanglement of material quantities via counterbend dynamics. PMID:28566516

  6. Reactions of chicken sera to recombinant Campylobacter jejuni flagellar proteins.

    PubMed

    Yeh, Hung-Yueh; Hiett, Kelli L; Line, John E

    2015-03-01

    Campylobacter jejuni is a Gram-negative spiral rod bacterium and is the leading but underreported bacterial food-borne pathogen that causes human campylobacteriosis worldwide. Raw or undercooked poultry products are regarded as a major source for human infection. C. jejuni flagella have been implicated in colonization and adhesion to the mucosal surface of chicken gastrointestinal tracts. Therefore, flagellar proteins would be the excellent targets for further investigation. In this report, we used the recombinant technology to generate a battery of C. jejuni flagellar proteins, which were purified by His tag affinity chromatography and determined antigenic profiles of these recombinant flagellar proteins using sera from chickens older than 6 weeks of age. The immunoblot results demonstrate that each chicken serum reacted to various numbers of recombinant flagellar proteins. Among these recombinant proteins, chicken sera reacted predominantly to the FlgE1, FlgK, FlhF, FliG and FliY proteins. These antibody screening results provide a rationale for further evaluation of these recombinant flagellar proteins as potential vaccines for chickens to improve food safety as well as investigation of host immune response to C. jejuni.

  7. Measurements of the Rotation of the Flagellar Motor by Bead Assay.

    PubMed

    Kasai, Taishi; Sowa, Yoshiyuki

    2017-01-01

    The bacterial flagellar motor is a reversible rotary nano-machine powered by the ion flux across the cytoplasmic membrane. Each motor rotates a long helical filament that extends from the cell body at several hundreds revolutions per second. The output of the motor is characterized by its generated torque and rotational speed. The torque can be calculated as the rotational frictional drag coefficient multiplied by the angular velocity. Varieties of methods, including a bead assay, have been developed to measure the flagellar rotation rate under various load conditions on the motor. In this chapter, we describe a method to monitor the motor rotation through a position of a 1 μm bead attached to a truncated flagellar filament.

  8. Assembly and stoichiometry of the core structure of the bacterial flagellar type III export gate complex.

    PubMed

    Fukumura, Takuma; Makino, Fumiaki; Dietsche, Tobias; Kinoshita, Miki; Kato, Takayuki; Wagner, Samuel; Namba, Keiichi; Imada, Katsumi; Minamino, Tohru

    2017-08-01

    The bacterial flagellar type III export apparatus, which is required for flagellar assembly beyond the cell membranes, consists of a transmembrane export gate complex and a cytoplasmic ATPase complex. FlhA, FlhB, FliP, FliQ, and FliR form the gate complex inside the basal body MS ring, although FliO is required for efficient export gate formation in Salmonella enterica. However, it remains unknown how they form the gate complex. Here we report that FliP forms a homohexameric ring with a diameter of 10 nm. Alanine substitutions of conserved Phe-137, Phe-150, and Glu-178 residues in the periplasmic domain of FliP (FliPP) inhibited FliP6 ring formation, suppressing flagellar protein export. FliO formed a 5-nm ring structure with 3 clamp-like structures that bind to the FliP6 ring. The crystal structure of FliPP derived from Thermotoga maritia, and structure-based photo-crosslinking experiments revealed that Phe-150 and Ser-156 of FliPP are involved in the FliP-FliP interactions and that Phe-150, Arg-152, Ser-156, and Pro-158 are responsible for the FliP-FliO interactions. Overexpression of FliP restored motility of a ∆fliO mutant to the wild-type level, suggesting that the FliP6 ring is a functional unit in the export gate complex and that FliO is not part of the final gate structure. Copurification assays revealed that FlhA, FlhB, FliQ, and FliR are associated with the FliO/FliP complex. We propose that the assembly of the export gate complex begins with FliP6 ring formation with the help of the FliO scaffold, followed by FliQ, FliR, and FlhB and finally FlhA during MS ring formation.

  9. Assembly and stoichiometry of the core structure of the bacterial flagellar type III export gate complex

    PubMed Central

    Fukumura, Takuma; Makino, Fumiaki; Dietsche, Tobias; Kinoshita, Miki; Kato, Takayuki; Wagner, Samuel; Namba, Keiichi; Imada, Katsumi

    2017-01-01

    The bacterial flagellar type III export apparatus, which is required for flagellar assembly beyond the cell membranes, consists of a transmembrane export gate complex and a cytoplasmic ATPase complex. FlhA, FlhB, FliP, FliQ, and FliR form the gate complex inside the basal body MS ring, although FliO is required for efficient export gate formation in Salmonella enterica. However, it remains unknown how they form the gate complex. Here we report that FliP forms a homohexameric ring with a diameter of 10 nm. Alanine substitutions of conserved Phe-137, Phe-150, and Glu-178 residues in the periplasmic domain of FliP (FliPP) inhibited FliP6 ring formation, suppressing flagellar protein export. FliO formed a 5-nm ring structure with 3 clamp-like structures that bind to the FliP6 ring. The crystal structure of FliPP derived from Thermotoga maritia, and structure-based photo-crosslinking experiments revealed that Phe-150 and Ser-156 of FliPP are involved in the FliP–FliP interactions and that Phe-150, Arg-152, Ser-156, and Pro-158 are responsible for the FliP–FliO interactions. Overexpression of FliP restored motility of a ∆fliO mutant to the wild-type level, suggesting that the FliP6 ring is a functional unit in the export gate complex and that FliO is not part of the final gate structure. Copurification assays revealed that FlhA, FlhB, FliQ, and FliR are associated with the FliO/FliP complex. We propose that the assembly of the export gate complex begins with FliP6 ring formation with the help of the FliO scaffold, followed by FliQ, FliR, and FlhB and finally FlhA during MS ring formation. PMID:28771466

  10. Impact of fluorescent protein fusions on the bacterial flagellar motor.

    PubMed

    Heo, M; Nord, A L; Chamousset, D; van Rijn, E; Beaumont, H J E; Pedaci, F

    2017-10-03

    Fluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side effects, but such insight is still lacking for many applications. This is particularly relevant in the study of the internal dynamics of motor proteins, where both the chemical and mechanical reaction coordinates can be affected. Fluorescent proteins fused to the stator of the Bacterial Flagellar Motor (BFM) have previously been used to unveil the motor subunit dynamics. Here we report the effects on single motors of three fluorescent proteins fused to the stators, all of which altered BFM behavior. The torque generated by individual stators was reduced while their stoichiometry remained unaffected. MotB fusions decreased the switching frequency and induced a novel bias-dependent asymmetry in the speed in the two directions. These effects could be mitigated by inserting a linker at the fusion point. These findings provide a quantitative account of the effects of fluorescent fusions to the stator on BFM dynamics and their alleviation- new insights that advance the use of fluorescent fusions to probe the dynamics of protein complexes.

  11. Inactivation of ferric uptake regulator (Fur) attenuates Helicobacter pylori J99 motility by disturbing the flagellar motor switch and autoinducer-2 production.

    PubMed

    Lee, Ai-Yun; Kao, Cheng-Yen; Wang, Yao-Kuan; Lin, Ssu-Yuan; Lai, Tze-Ying; Sheu, Bor-Shyang; Lo, Chien-Jung; Wu, Jiunn-Jong

    2017-08-01

    Flagellar motility of Helicobacter pylori has been shown to be important for the bacteria to establish initial colonization. The ferric uptake regulator (Fur) is a global regulator that has been identified in H. pylori which is involved in the processes of iron uptake and establishing colonization. However, the role of Fur in H. pylori motility is still unclear. Motility of the wild-type, fur mutant, and fur revertant J99 were determined by a soft-agar motility assay and direct video observation. The bacterial shape and flagellar structure were evaluated by transmission electron microscopy. Single bacterial motility and flagellar switching were observed by phase-contrast microscopy. Autoinducer-2 (AI-2) production in bacterial culture supernatant was analyzed by a bioluminescence assay. The fur mutant showed impaired motility in the soft-agar assay compared with the wild-type J99 and fur revertant. The numbers and lengths of flagellar filaments on the fur mutant cells were similar to those of the wild-type and revertant cells. Phenotypic characterization showed similar swimming speed but reduction in switching rate in the fur mutant. The AI-2 production of the fur mutant was dramatically reduced compared with wild-type J99 in log-phase culture medium. These results indicate that Fur positively modulates H. pylori J99 motility through interfering with bacterial flagellar switching. © 2017 John Wiley & Sons Ltd.

  12. Glucose induces delocalization of a flagellar biosynthesis protein from the flagellated pole.

    PubMed

    Park, Soyoung; Park, Young-Ha; Lee, Chang-Ro; Kim, Yeon-Ran; Seok, Yeong-Jae

    2016-09-01

    To survive in a continuously changing environment, bacteria sense concentration gradients of attractants or repellents, and purposefully migrate until a more favourable habitat is encountered. While glucose is known as the most effective attractant, the flagellar biosynthesis and hence chemotactic motility has been known to be repressed by glucose in some bacteria. To date, the only known regulatory mechanism of the repression of flagellar synthesis by glucose is via downregulation of the cAMP level, as shown in a few members of the family Enterobacteriaceae. Here we show that, in Vibrio vulnificus, the glucose-mediated inhibition of flagellar motility operates by a completely different mechanism. In the presence of glucose, EIIA(Glc) is dephosphorylated and inhibits the polar localization of FapA (flagellar assembly protein A) by sequestering it from the flagellated pole. A loss or delocalization of FapA results in a complete failure of the flagellar biosynthesis and motility. However, when glucose is depleted, EIIA(Glc) is phosphorylated and releases FapA such that free FapA can be localized back to the pole and trigger flagellation. Together, these data provide new insight into a bacterial strategy to reach and stay in the glucose-rich area. © 2016 John Wiley & Sons Ltd.

  13. Flagellar Synchronization Is a Simple Alternative to Cell Cycle Synchronization for Ciliary and Flagellar Studies

    PubMed Central

    Dutta, Soumita

    2017-01-01

    ABSTRACT The unicellular green alga Chlamydomonas reinhardtii is an ideal model organism for studies of ciliary function and assembly. In assays for biological and biochemical effects of various factors on flagellar structure and function, synchronous culture is advantageous for minimizing variability. Here, we have characterized a method in which 100% synchronization is achieved with respect to flagellar length but not with respect to the cell cycle. The method requires inducing flagellar regeneration by amputation of the entire cell population and limiting regeneration time. This results in a maximally homogeneous distribution of flagellar lengths at 3 h postamputation. We found that time-limiting new protein synthesis during flagellar synchronization limits variability in the unassembled pool of limiting flagellar protein and variability in flagellar length without affecting the range of cell volumes. We also found that long- and short-flagella mutants that regenerate normally require longer and shorter synchronization times, respectively. By minimizing flagellar length variability using a simple method requiring only hours and no changes in media, flagellar synchronization facilitates the detection of small changes in flagellar length resulting from both chemical and genetic perturbations in Chlamydomonas. This method increases our ability to probe the basic biology of ciliary size regulation and related disease etiologies. IMPORTANCE Cilia and flagella are highly conserved antenna-like organelles that found in nearly all mammalian cell types. They perform sensory and motile functions contributing to numerous physiological and developmental processes. Defects in their assembly and function are implicated in a wide range of human diseases ranging from retinal degeneration to cancer. Chlamydomonas reinhardtii is an algal model system for studying mammalian cilium formation and function. Here, we report a simple synchronization method that allows detection of

  14. Flagellar Synchronization Is a Simple Alternative to Cell Cycle Synchronization for Ciliary and Flagellar Studies.

    PubMed

    Dutta, Soumita; Avasthi, Prachee

    2017-01-01

    The unicellular green alga Chlamydomonas reinhardtii is an ideal model organism for studies of ciliary function and assembly. In assays for biological and biochemical effects of various factors on flagellar structure and function, synchronous culture is advantageous for minimizing variability. Here, we have characterized a method in which 100% synchronization is achieved with respect to flagellar length but not with respect to the cell cycle. The method requires inducing flagellar regeneration by amputation of the entire cell population and limiting regeneration time. This results in a maximally homogeneous distribution of flagellar lengths at 3 h postamputation. We found that time-limiting new protein synthesis during flagellar synchronization limits variability in the unassembled pool of limiting flagellar protein and variability in flagellar length without affecting the range of cell volumes. We also found that long- and short-flagella mutants that regenerate normally require longer and shorter synchronization times, respectively. By minimizing flagellar length variability using a simple method requiring only hours and no changes in media, flagellar synchronization facilitates the detection of small changes in flagellar length resulting from both chemical and genetic perturbations in Chlamydomonas . This method increases our ability to probe the basic biology of ciliary size regulation and related disease etiologies. IMPORTANCE Cilia and flagella are highly conserved antenna-like organelles that found in nearly all mammalian cell types. They perform sensory and motile functions contributing to numerous physiological and developmental processes. Defects in their assembly and function are implicated in a wide range of human diseases ranging from retinal degeneration to cancer. Chlamydomonas reinhardtii is an algal model system for studying mammalian cilium formation and function. Here, we report a simple synchronization method that allows detection of small

  15. H{sup +} and Na{sup +} are involved in flagellar rotation of the spirochete Leptospira

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

    Islam, Md. Shafiqul; Morimoto, Yusuke V.; Graduate School of Frontier BioSciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871

    2015-10-16

    Leptospira is a spirochete possessing intracellular flagella. Each Leptospira flagellar filament is linked with a flagellar motor composed of a rotor and a dozen stators. For many bacterial species, it is known that the stator functions as an ion channel and that the ion flux through the stator is coupled with flagellar rotation. The coupling ion varies depending on the species; for example, H{sup +} is used in Escherichia coli, and Na{sup +} is used in Vibrio spp. to drive a polar flagellum. Although genetic and structural studies illustrated that the Leptospira flagellar motor also contains a stator, the couplingmore » ion for flagellar rotation remains unknown. In the present study, we analyzed the motility of Leptospira under various pH values and salt concentrations. Leptospira cells displayed motility in acidic to alkaline pH. In the presence of a protonophore, the cells completely lost motility in acidic to neutral pH but displayed extremely slow movement under alkaline conditions. This result suggests that H{sup +} is a major coupling ion for flagellar rotation over a wide pH range; however, we also observed that the motility of Leptospira was significantly enhanced by the addition of Na{sup +}, though it vigorously moved even under Na{sup +}-free conditions. These results suggest that H{sup +} is preferentially used and that Na{sup +} is secondarily involved in flagellar rotation in Leptospira. The flexible ion selectivity in the flagellar system could be advantageous for Leptospira to survive in a wide range of environment. - Highlights: • This is a study on input energy for motility in the spirochete Leptospira. • Leptospira biflexa exhibited active motility in acidic to alkaline pH. • Both H{sup +} and Na{sup +} are involved in flagellar rotation in Leptospira. • H{sup +} is a primary energy source, but Na{sup +} can secondarily enhance motility.« less

  16. In Vitro Reconstitution of Functional Type III Protein Export and Insights into Flagellar Assembly.

    PubMed

    Terashima, Hiroyuki; Kawamoto, Akihiro; Tatsumi, Chinatsu; Namba, Keiichi; Minamino, Tohru; Imada, Katsumi

    2018-06-26

    The type III secretion system (T3SS) forms the functional core of injectisomes, protein transporters that allow bacteria to deliver virulence factors into their hosts for infection, and flagella, which are critical for many pathogens to reach the site of infection. In spite of intensive genetic and biochemical studies, the T3SS protein export mechanism remains unclear due to the difficulty of accurate measurement of protein export in vivo Here, we developed an in vitro flagellar T3S protein transport assay system using an inverted cytoplasmic membrane vesicle (IMV) for accurate and controlled measurements of flagellar protein export. We show that the flagellar T3SS in the IMV fully retains export activity. The flagellar hook was constructed inside the lumen of the IMV by adding purified component proteins externally to the IMV solution. We reproduced the hook length control and export specificity switch in the IMV consistent with that seen in the native cell. Previous in vivo analyses showed that flagellar protein export is driven by proton motive force (PMF) and facilitated by ATP hydrolysis by FliI, a T3SS-specific ATPase. Our in vitro assay recapitulated these previous in vivo observations but furthermore clearly demonstrated that even ATP hydrolysis by FliI alone can drive flagellar protein export. Moreover, this assay showed that addition of the FliH 2 /FliI complex to the assay solution at a concentration similar to that in the cell dramatically enhanced protein export, confirming that the FliH 2 /FliI complex in the cytoplasm is important for effective protein transport. IMPORTANCE The type III secretion system (T3SS) is the functional core of the injectisome, a bacterial protein transporter used to deliver virulence proteins into host cells, and bacterial flagella, critical for many pathogens. The molecular mechanism of protein transport is still unclear due to difficulties in accurate measurements of protein transport under well-controlled conditions in

  17. Kinematics of flagellar swimming in Euglena gracilis: Helical trajectories and flagellar shapes.

    PubMed

    Rossi, Massimiliano; Cicconofri, Giancarlo; Beran, Alfred; Noselli, Giovanni; DeSimone, Antonio

    2017-12-12

    The flagellar swimming of euglenids, which are propelled by a single anterior flagellum, is characterized by a generalized helical motion. The 3D nature of this swimming motion, which lacks some of the symmetries enjoyed by more common model systems, and the complex flagellar beating shapes that power it make its quantitative description challenging. In this work, we provide a quantitative, 3D, highly resolved reconstruction of the swimming trajectories and flagellar shapes of specimens of Euglena gracilis We achieved this task by using high-speed 2D image recordings taken with a conventional inverted microscope combined with a precise characterization of the helical motion of the cell body to lift the 2D data to 3D trajectories. The propulsion mechanism is discussed. Our results constitute a basis for future biophysical research on a relatively unexplored type of eukaryotic flagellar movement. Copyright © 2017 the Author(s). Published by PNAS.

  18. Flagellar motility is a key determinant of the magnitude of the inflammasome response to Pseudomonas aeruginosa.

    PubMed

    Patankar, Yash R; Lovewell, Rustin R; Poynter, Matthew E; Jyot, Jeevan; Kazmierczak, Barbara I; Berwin, Brent

    2013-06-01

    We previously demonstrated that bacterial flagellar motility is a fundamental mechanism by which host phagocytes bind and ingest bacteria. Correspondingly, loss of bacterial motility, consistently observed in clinical isolates from chronic Pseudomonas aeruginosa infections, enables bacteria to evade association and ingestion of P. aeruginosa by phagocytes both in vitro and in vivo. Since bacterial interactions with the phagocyte cell surface are required for type three secretion system-dependent NLRC4 inflammasome activation by P. aeruginosa, we hypothesized that reduced bacterial association with phagocytes due to loss of bacterial motility, independent of flagellar expression, will lead to reduced inflammasome activation. Here we report that inflammasome activation is reduced in response to nonmotile P. aeruginosa. Nonmotile P. aeruginosa elicits reduced IL-1β production as well as caspase-1 activation by peritoneal macrophages and bone marrow-derived dendritic cells in vitro. Importantly, nonmotile P. aeruginosa also elicits reduced IL-1β levels in vivo in comparison to those elicited by wild-type P. aeruginosa. This is the first demonstration that loss of bacterial motility results in reduced inflammasome activation and antibacterial IL-1β host response. These results provide a critical insight into how the innate immune system responds to bacterial motility and, correspondingly, how pathogens have evolved mechanisms to evade the innate immune system.

  19. Flagellar Motility Is a Key Determinant of the Magnitude of the Inflammasome Response to Pseudomonas aeruginosa

    PubMed Central

    Patankar, Yash R.; Lovewell, Rustin R.; Poynter, Matthew E.; Jyot, Jeevan; Kazmierczak, Barbara I.

    2013-01-01

    We previously demonstrated that bacterial flagellar motility is a fundamental mechanism by which host phagocytes bind and ingest bacteria. Correspondingly, loss of bacterial motility, consistently observed in clinical isolates from chronic Pseudomonas aeruginosa infections, enables bacteria to evade association and ingestion of P. aeruginosa by phagocytes both in vitro and in vivo. Since bacterial interactions with the phagocyte cell surface are required for type three secretion system-dependent NLRC4 inflammasome activation by P. aeruginosa, we hypothesized that reduced bacterial association with phagocytes due to loss of bacterial motility, independent of flagellar expression, will lead to reduced inflammasome activation. Here we report that inflammasome activation is reduced in response to nonmotile P. aeruginosa. Nonmotile P. aeruginosa elicits reduced IL-1β production as well as caspase-1 activation by peritoneal macrophages and bone marrow-derived dendritic cells in vitro. Importantly, nonmotile P. aeruginosa also elicits reduced IL-1β levels in vivo in comparison to those elicited by wild-type P. aeruginosa. This is the first demonstration that loss of bacterial motility results in reduced inflammasome activation and antibacterial IL-1β host response. These results provide a critical insight into how the innate immune system responds to bacterial motility and, correspondingly, how pathogens have evolved mechanisms to evade the innate immune system. PMID:23529619

  20. Structure of Salmonella FlhE, conserved member of a flagellar Type III secretion operon

    DOE PAGES

    Lee, Jaemin; Monzingo, Arthur F.; Keatinge-Clay, Adrian T.; ...

    2014-12-26

    In this paper, the bacterial flagellum is assembled by a multicomponent transport apparatus categorized as a type III secretion system. The secretion of proteins that assemble into the flagellum is driven by the proton motive force. The periplasmic protein FlhE is a member of the flhBAE operon in the majority of bacteria where FlhE is found. FlhA and FlhB are established components of the flagellar type III secretion system. The absence of FlhE results in a proton leak through the flagellar system, inappropriate secretion patterns, and cell death, indicating that FlhE regulates an important aspect of proper flagellar biosynthesis. Wemore » isolated FlhE from the periplasm of Salmonella and solved its structure to 1.5 Å resolution. The structure reveals a β-sandwich fold, with no close structural homologs. Finally, possible roles of FlhE, including that of a chaperone, are discussed.« less

  1. Structure of Salmonella FlhE, conserved member of a flagellar Type III secretion operon

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

    Lee, Jaemin; Monzingo, Arthur F.; Keatinge-Clay, Adrian T.

    In this paper, the bacterial flagellum is assembled by a multicomponent transport apparatus categorized as a type III secretion system. The secretion of proteins that assemble into the flagellum is driven by the proton motive force. The periplasmic protein FlhE is a member of the flhBAE operon in the majority of bacteria where FlhE is found. FlhA and FlhB are established components of the flagellar type III secretion system. The absence of FlhE results in a proton leak through the flagellar system, inappropriate secretion patterns, and cell death, indicating that FlhE regulates an important aspect of proper flagellar biosynthesis. Wemore » isolated FlhE from the periplasm of Salmonella and solved its structure to 1.5 Å resolution. The structure reveals a β-sandwich fold, with no close structural homologs. Finally, possible roles of FlhE, including that of a chaperone, are discussed.« less

  2. Bacteria exploit a polymorphic instability of the flagellar filament to escape from traps.

    PubMed

    Kühn, Marco J; Schmidt, Felix K; Eckhardt, Bruno; Thormann, Kai M

    2017-06-13

    Many bacterial species swim by rotating single polar helical flagella. Depending on the direction of rotation, they can swim forward or backward and change directions to move along chemical gradients but also to navigate their obstructed natural environment in soils, sediments, or mucus. When they get stuck, they naturally try to back out, but they can also resort to a radically different flagellar mode, which we discovered here. Using high-speed microscopy, we monitored the swimming behavior of the monopolarly flagellated species Shewanella putrefaciens with fluorescently labeled flagellar filaments at an agarose-glass interface. We show that, when a cell gets stuck, the polar flagellar filament executes a polymorphic change into a spiral-like form that wraps around the cell body in a spiral-like fashion and enables the cell to escape by a screw-like backward motion. Microscopy and modeling suggest that this propagation mode is triggered by an instability of the flagellum under reversal of the rotation and the applied torque. The switch is reversible and bacteria that have escaped the trap can return to their normal swimming mode by another reversal of motor direction. The screw-type flagellar arrangement enables a unique mode of propagation and, given the large number of polarly flagellated bacteria, we expect it to be a common and widespread escape or motility mode in complex and structured environments.

  3. Control of Flagellar Gene Regulation in Legionella pneumophila and Its Relation to Growth Phase▿ †

    PubMed Central

    Albert-Weissenberger, Christiane; Sahr, Tobias; Sismeiro, Odile; Hacker, Jörg; Heuner, Klaus; Buchrieser, Carmen

    2010-01-01

    The bacterial pathogen Legionella pneumophila responds to environmental changes by differentiation. At least two forms are well described: replicative bacteria are avirulent; in contrast, transmissive bacteria express virulence traits and flagella. Phenotypic analysis, Western blotting, and electron microscopy of mutants of the regulatory genes encoding RpoN, FleQ, FleR, and FliA demonstrated that flagellin expression is strongly repressed and that the mutants are nonflagellated in the transmissive phase. Transcriptome analyses elucidated that RpoN, together with FleQ, enhances transcription of 14 out of 31 flagellar class II genes, which code for the basal body, hook, and regulatory proteins. Unexpectedly, FleQ independent of RpoN enhances the transcription of fliA encoding sigma 28. Expression analysis of a fliA mutant showed that FliA activates three out of the five remaining flagellar class III genes and the flagellar class IV genes. Surprisingly, FleR does not induce but inhibits expression of at least 14 flagellar class III genes on the transcriptional level. Thus, we propose that flagellar class II genes are controlled by FleQ and RpoN, whereas the transcription of the class III gene fliA is controlled in a FleQ-dependent but RpoN-independent manner. However, RpoN and FleR might influence flagellin synthesis on a posttranscriptional level. In contrast to the commonly accepted view that enhancer-binding proteins such as FleQ always interact with RpoN to fullfill their regulatory functions, our results strongly indicate that FleQ regulates gene expression that is RpoN dependent and RpoN independent. Finally, FliA induces expression of flagellar class III and IV genes leading to the complete synthesis of the flagellum. PMID:19915024

  4. The Flagellar Regulon of Legionella—A Review

    PubMed Central

    Appelt, Sandra; Heuner, Klaus

    2017-01-01

    The Legionella genus comprises more than 60 species. In particular, Legionella pneumophila is known to cause severe illnesses in humans. Legionellaceae are ubiquitous inhabitants of aquatic environments. Some Legionellaceae are motile and their motility is important to move around in habitats. Motility can be considered as a potential virulence factor as already shown for various human pathogens. The genes of the flagellar system, regulator and structural genes, are structured in hierarchical levels described as the flagellar regulon. Their expression is modulated by various environmental factors. For L. pneumophila it was shown that the expression of genes of the flagellar regulon is modulated by the actual growth phase and temperature. Especially, flagellated Legionella are known to express genes during the transmissive phase of growth that are involved in the expression of virulence traits. It has been demonstrated that the alternative sigma-28 factor is part of the link between virulence expression and motility. In the following review, the structure of the flagellar regulon of L. pneumophila is discussed and compared to other flagellar systems of different Legionella species. Recently, it has been described that Legionella micdadei and Legionella fallonii contain a second putative partial flagellar system. Hence, the report will focus on flagellated and non-flagellated Legionella strains, phylogenetic relationships, the role and function of the alternative sigma factor (FliA) and its anti-sigma-28 factor (FlgM). PMID:29104863

  5. The Flagellar Regulon of Legionella-A Review.

    PubMed

    Appelt, Sandra; Heuner, Klaus

    2017-01-01

    The Legionella genus comprises more than 60 species. In particular, Legionella pneumophila is known to cause severe illnesses in humans. Legionellaceae are ubiquitous inhabitants of aquatic environments. Some Legionellaceae are motile and their motility is important to move around in habitats. Motility can be considered as a potential virulence factor as already shown for various human pathogens. The genes of the flagellar system, regulator and structural genes, are structured in hierarchical levels described as the flagellar regulon. Their expression is modulated by various environmental factors. For L. pneumophila it was shown that the expression of genes of the flagellar regulon is modulated by the actual growth phase and temperature. Especially, flagellated Legionella are known to express genes during the transmissive phase of growth that are involved in the expression of virulence traits. It has been demonstrated that the alternative sigma-28 factor is part of the link between virulence expression and motility. In the following review, the structure of the flagellar regulon of L. pneumophila is discussed and compared to other flagellar systems of different Legionella species. Recently, it has been described that Legionella micdadei and Legionella fallonii contain a second putative partial flagellar system. Hence, the report will focus on flagellated and non-flagellated Legionella strains, phylogenetic relationships, the role and function of the alternative sigma factor (FliA) and its anti-sigma-28 factor (FlgM).

  6. Applying torque to the Escherichia coli flagellar motor using magnetic tweezers.

    PubMed

    van Oene, Maarten M; Dickinson, Laura E; Cross, Bronwen; Pedaci, Francesco; Lipfert, Jan; Dekker, Nynke H

    2017-03-07

    The bacterial flagellar motor of Escherichia coli is a nanoscale rotary engine essential for bacterial propulsion. Studies on the power output of single motors rely on the measurement of motor torque and rotation under external load. Here, we investigate the use of magnetic tweezers, which in principle allow the application and active control of a calibrated load torque, to study single flagellar motors in Escherichia coli. We manipulate the external load on the motor by adjusting the magnetic field experienced by a magnetic bead linked to the motor, and we probe the motor's response. A simple model describes the average motor speed over the entire range of applied fields. We extract the motor torque at stall and find it to be similar to the motor torque at drag-limited speed. In addition, use of the magnetic tweezers allows us to force motor rotation in both forward and backward directions. We monitor the motor's performance before and after periods of forced rotation and observe no destructive effects on the motor. Our experiments show how magnetic tweezers can provide active and fast control of the external load while also exposing remaining challenges in calibration. Through their non-invasive character and straightforward parallelization, magnetic tweezers provide an attractive platform to study nanoscale rotary motors at the single-motor level.

  7. Applying torque to the Escherichia coli flagellar motor using magnetic tweezers

    PubMed Central

    van Oene, Maarten M.; Dickinson, Laura E.; Cross, Bronwen; Pedaci, Francesco; Lipfert, Jan; Dekker, Nynke H.

    2017-01-01

    The bacterial flagellar motor of Escherichia coli is a nanoscale rotary engine essential for bacterial propulsion. Studies on the power output of single motors rely on the measurement of motor torque and rotation under external load. Here, we investigate the use of magnetic tweezers, which in principle allow the application and active control of a calibrated load torque, to study single flagellar motors in Escherichia coli. We manipulate the external load on the motor by adjusting the magnetic field experienced by a magnetic bead linked to the motor, and we probe the motor’s response. A simple model describes the average motor speed over the entire range of applied fields. We extract the motor torque at stall and find it to be similar to the motor torque at drag-limited speed. In addition, use of the magnetic tweezers allows us to force motor rotation in both forward and backward directions. We monitor the motor’s performance before and after periods of forced rotation and observe no destructive effects on the motor. Our experiments show how magnetic tweezers can provide active and fast control of the external load while also exposing remaining challenges in calibration. Through their non-invasive character and straightforward parallelization, magnetic tweezers provide an attractive platform to study nanoscale rotary motors at the single-motor level. PMID:28266562

  8. Noise effects in bacterial motor switch

    NASA Astrophysics Data System (ADS)

    Tu, Yuhai

    2006-03-01

    The clockwise (CW) or counter clockwise (CCW) spinning of bacterial flagellar motors is controlled by the concentration of a phosphorylated protein CheY-P. In this talk, we represent the stochastic switching behavior of a bacterial flagellar motor by a dynamical two-state (CW and CCW) model, with the energy levels of the two states fluctuating in time according to the variation of the CheY-P concentration in the cell. We show that with a generic normal distribution and a modest amplitude for CheY-P concentration fluctuations, the dynamical two-state model is capable of generating a power-law distribution (as opposed to an exponential Poisson-like distribution) for the durations of the CCW states, in agreement with recent experimental observations of Korobkova et al (Nature, 428, 574(2004)). In addition, we show that the power spectrum for the flagellar motor switching time series is not determined solely by the power-law duration distribution, but also by the temporal correlation between the duration times of different CCW intervals. We point out the intrinsic connection between anomalously large fluctuations of the motor output and the overall high gain of the bacterial chemotaxis system. Suggestions for experimental verification of the dynamical two-state model will also be discussed.

  9. FliH and FliI ensure efficient energy coupling of flagellar type III protein export in Salmonella.

    PubMed

    Minamino, Tohru; Kinoshita, Miki; Inoue, Yumi; Morimoto, Yusuke V; Ihara, Kunio; Koya, Satomi; Hara, Noritaka; Nishioka, Noriko; Kojima, Seiji; Homma, Michio; Namba, Keiichi

    2016-06-01

    For construction of the bacterial flagellum, flagellar proteins are exported via its specific export apparatus from the cytoplasm to the distal end of the growing flagellar structure. The flagellar export apparatus consists of a transmembrane (TM) export gate complex and a cytoplasmic ATPase complex consisting of FliH, FliI, and FliJ. FlhA is a TM export gate protein and plays important roles in energy coupling of protein translocation. However, the energy coupling mechanism remains unknown. Here, we performed a cross-complementation assay to measure robustness of the energy transduction system of the export apparatus against genetic perturbations. Vibrio FlhA restored motility of a Salmonella ΔflhA mutant but not that of a ΔfliH-fliI flhB(P28T) ΔflhA mutant. The flgM mutations significantly increased flagellar gene expression levels, allowing Vibrio FlhA to exert its export activity in the ΔfliH-fliI flhB(P28T) ΔflhA mutant. Pull-down assays revealed that the binding affinities of Vibrio FlhA for FliJ and the FlgN-FlgK chaperone-substrate complex were much lower than those of Salmonella FlhA. These suggest that Vibrio FlhA requires the support of FliH and FliI to efficiently and properly interact with FliJ and the FlgN-FlgK complex. We propose that FliH and FliI ensure robust and efficient energy coupling of protein export during flagellar assembly. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  10. Speed of the bacterial flagellar motor near zero load depends on the number of stator units.

    PubMed

    Nord, Ashley L; Sowa, Yoshiyuki; Steel, Bradley C; Lo, Chien-Jung; Berry, Richard M

    2017-10-31

    The bacterial flagellar motor (BFM) rotates hundreds of times per second to propel bacteria driven by an electrochemical ion gradient. The motor consists of a rotor 50 nm in diameter surrounded by up to 11 ion-conducting stator units, which exchange between motors and a membrane-bound pool. Measurements of the torque-speed relationship guide the development of models of the motor mechanism. In contrast to previous reports that speed near zero torque is independent of the number of stator units, we observe multiple speeds that we attribute to different numbers of units near zero torque in both Na + - and H + -driven motors. We measure the full torque-speed relationship of one and two H + units in Escherichia coli by selecting the number of H + units and controlling the number of Na + units in hybrid motors. These experiments confirm that speed near zero torque in H + -driven motors increases with the stator number. We also measured 75 torque-speed curves for Na + -driven chimeric motors at different ion-motive force and stator number. Torque and speed were proportional to ion-motive force and number of stator units at all loads, allowing all 77 measured torque-speed curves to be collapsed onto a single curve by simple rescaling. Published under the PNAS license.

  11. Speed of the bacterial flagellar motor near zero load depends on the number of stator units

    PubMed Central

    Nord, Ashley L.; Sowa, Yoshiyuki; Steel, Bradley C.; Lo, Chien-Jung; Berry, Richard M.

    2017-01-01

    The bacterial flagellar motor (BFM) rotates hundreds of times per second to propel bacteria driven by an electrochemical ion gradient. The motor consists of a rotor 50 nm in diameter surrounded by up to 11 ion-conducting stator units, which exchange between motors and a membrane-bound pool. Measurements of the torque–speed relationship guide the development of models of the motor mechanism. In contrast to previous reports that speed near zero torque is independent of the number of stator units, we observe multiple speeds that we attribute to different numbers of units near zero torque in both Na+- and H+-driven motors. We measure the full torque–speed relationship of one and two H+ units in Escherichia coli by selecting the number of H+ units and controlling the number of Na+ units in hybrid motors. These experiments confirm that speed near zero torque in H+-driven motors increases with the stator number. We also measured 75 torque–speed curves for Na+-driven chimeric motors at different ion-motive force and stator number. Torque and speed were proportional to ion-motive force and number of stator units at all loads, allowing all 77 measured torque–speed curves to be collapsed onto a single curve by simple rescaling. PMID:29078322

  12. Characterization of C-ring component assembly in flagellar motors from amino acid coevolution

    PubMed Central

    dos Santos, Ricardo Nascimento; Khan, Shahid

    2018-01-01

    Bacterial flagellar motility, an important virulence factor, is energized by a rotary motor localized within the flagellar basal body. The rotor module consists of a large framework (the C-ring), composed of the FliG, FliM and FliN proteins. FliN and FliM contacts the FliG torque ring to control the direction of flagellar rotation. We report that structure-based models constrained only by residue coevolution can recover the binding interface of atomic X-ray dimer complexes with remarkable accuracy (approx. 1 Å RMSD). We propose a model for FliM–FliN heterodimerization, which agrees accurately with homologous interfaces as well as in situ cross-linking experiments, and hence supports a proposed architecture for the lower portion of the C-ring. Furthermore, this approach allowed the identification of two discrete and interchangeable homodimerization interfaces between FliM middle domains that agree with experimental measurements and might be associated with C-ring directional switching dynamics triggered upon binding of CheY signal protein. Our findings provide structural details of complex formation at the C-ring that have been difficult to obtain with previous methodologies and clarify the architectural principle that underpins the ultra-sensitive allostery exhibited by this ring assembly that controls the clockwise or counterclockwise rotation of flagella. PMID:29892378

  13. Ruthenium red-induced bundling of bacterial cell division protein, FtsZ.

    PubMed

    Santra, Manas Kumar; Beuria, Tushar K; Banerjee, Abhijit; Panda, Dulal

    2004-06-18

    The assembly of FtsZ plays a major role in bacterial cell division, and it is thought that the assembly dynamics of FtsZ is a finely regulated process. Here, we show that ruthenium red is able to modulate FtsZ assembly in vitro. In contrast to the inhibitory effects of ruthenium red on microtubule polymerization, we found that a substoichiometric concentration of ruthenium red strongly increased the light-scattering signal of FtsZ assembly. Further, sedimentable polymer mass was increased by 1.5- and 2-fold in the presence of 2 and 10 microm ruthenium red, respectively. In addition, ruthenium red strongly reduced the GTPase activity and prevented dilution-induced disassembly of FtsZ polymers. Electron microscopic analysis showed that 4-10 microm of ruthenium red produced thick bundles of FtsZ polymers. The significant increase in the light-scattering signal and pelletable polymer mass in the presence of ruthenium red seemed to be due to the bundling of FtsZ protofilaments into larger polymers rather than the actual increase in the level of polymeric FtsZ. Furthermore, ruthenium red was found to copolymerize with FtsZ, and the copolymerization of substoichiometric amounts of ruthenium red with FtsZ polymers promoted cooperative assembly of FtsZ that produced large bundles. Calcium inhibited the binding of ruthenium red to FtsZ. However, a concentration of calcium 1000-fold higher than that of ruthenium red was required to produce similar effects on FtsZ assembly. Ruthenium red strongly modulated FtsZ polymerization, suggesting the presence of an important regulatory site on FtsZ and suggesting that a natural ligand, which mimics the action of ruthenium red, may regulate the assembly of FtsZ in bacteria.

  14. Loose coupling in the bacterial flagellar motor

    PubMed Central

    Boschert, Ryan; Adler, Frederick R.; Blair, David F.

    2015-01-01

    Physiological properties of the flagellar rotary motor have been taken to indicate a tightly coupled mechanism in which each revolution is driven by a fixed number of energizing ions. Measurements that would directly test the tight-coupling hypothesis have not been made. Energizing ions flow through membrane-bound complexes formed from the proteins MotA and MotB, which are anchored to the cell wall and constitute the stator. Genetic and biochemical evidence points to a “power stroke” mechanism in which the ions interact with an aspartate residue of MotB to drive conformational changes in MotA that are transmitted to the rotor protein FliG. Each stator complex contains two separate ion-binding sites, raising the question of whether the power stroke is driven by one, two, or either number of ions. Here, we describe simulations of a model in which the conformational change can be driven by either one or two ions. This loosely coupled model can account for the observed physiological properties of the motor, including those that have been taken to indicate tight coupling; it also accords with recent measurements of motor torque at high load that are harder to explain in tight-coupling models. Under loads relevant to a swimming cell, the loosely coupled motor would perform about as well as a two-proton motor and significantly better than a one-proton motor. The loosely coupled motor is predicted to be especially advantageous under conditions of diminished energy supply, or of reduced temperature, turning faster than an obligatorily two-proton motor while using fewer ions. PMID:25825730

  15. A continuum theoretical model and finite elements simulation of bacterial flagellar filament phase transition.

    PubMed

    Wang, Xiaoling; Meng, Shuo; Han, Jingshi

    2017-10-03

    The Bacterial flagellar filament can undergo a polymorphic phase transition in response to both mechanical and chemical variations in vitro and in vivo environments. Under mechanical stimuli, such as viscous flow or forces induced by motor rotation, the filament changes its phase from left-handed normal (N) to right-handed semi-coiled (SC) via phase nucleation and growth. Our detailed mechanical analysis of existing experiments shows that both torque and bending moment contribute to the filament phase transition. In this paper, we establish a non-convex and non-local continuum model based on the Ginzburg-Landau theory to describe main characteristics of the filament phase transition such as new-phase nucleation, growth, propagation and the merging of neighboring interfaces. The finite element method (FEM) is adopted to simulate the phase transition under a displacement-controlled loading condition (rotation angle and bending deflection). We show that new-phase nucleation corresponds to the maximum torque and bending moment at the stuck end of the filament. The hysteresis loop in the loading and unloading curves indicates energy dissipation. When the new phase grows and propagates, torque and bending moment remain static. We also find that there is a drop in load when the two interfaces merge, indicating a concomitant reduction in the interfacial energy. Finally, the interface thickness is governed by the coefficients of the gradient of order parameters in the non-local interface energy. Our continuum theory and the finite element method provide a method to study the mechanical behavior of such biomaterials. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Polar Flagellar Motility of the Vibrionaceae

    PubMed Central

    McCarter, Linda L.

    2001-01-01

    Polar flagella of Vibrio species can rotate at speeds as high as 100,000 rpm and effectively propel the bacteria in liquid as fast as 60 μm/s. The sodium motive force powers rotation of the filament, which acts as a propeller. The filament is complex, composed of multiple subunits, and sheathed by an extension of the cell outer membrane. The regulatory circuitry controlling expression of the polar flagellar genes of members of the Vibrionaceae is different from the peritrichous system of enteric bacteria or the polar system of Caulobacter crescentus. The scheme of gene control is also pertinent to other members of the gamma purple bacteria, in particular to Pseudomonas species. This review uses the framework of the polar flagellar system of Vibrio parahaemolyticus to provide a synthesis of what is known about polar motility systems of the Vibrionaceae. In addition to its propulsive role, the single polar flagellum of V. parahaemolyticus is believed to act as a tactile sensor controlling surface-induced gene expression. Under conditions that impede rotation of the polar flagellum, an alternate, lateral flagellar motility system is induced that enables movement through viscous environments and over surfaces. Although the dual flagellar systems possess no shared structural components and although distinct type III secretion systems direct the simultaneous placement and assembly of polar and lateral organelles, movement is coordinated by shared chemotaxis machinery. PMID:11528005

  17. Specific arrangement of alpha-helical coiled coils in the core domain of the bacterial flagellar hook for the universal joint function.

    PubMed

    Fujii, Takashi; Kato, Takayuki; Namba, Keiichi

    2009-11-11

    The bacterial flagellar hook is a short, highly curved tubular structure connecting the rotary motor to the filament acting as a helical propeller. The bending flexibility of the hook allows it to work as a universal joint. A partial atomic model of the hook revealed a sliding intersubunit domain interaction along the protofilament to produce bending flexibility. However, it remained unclear how the tightly packed inner core domains can still permit axial extension and compression. We report advances in cryoEM image analysis for high-resolution, high-throughput structural analysis and a density map of the hook that reveals most of the secondary structures, including the terminal alpha helices forming a coiled coil. The orientations and axial packing interactions of these two alpha helices are distinctly different from those of the filament, allowing them to have a room for axial compression and extension for bending flexibility without impairing the mechanical stability of the hook.

  18. Flagellar Cap Protein FliD Mediates Adherence of Atypical Enteropathogenic Escherichia coli to Enterocyte Microvilli

    PubMed Central

    Sampaio, Suely C. F.; Luiz, Wilson B.; Vieira, Mônica A. M.; Ferreira, Rita C. C.; Garcia, Bruna G.; Sinigaglia-Coimbra, Rita; Sampaio, Jorge L. M.; Ferreira, Luís C. S.

    2016-01-01

    The expression of flagella correlates with different aspects of bacterial pathogenicity, ranging from adherence to host cells to activation of inflammatory responses by the innate immune system. In the present study, we investigated the role of flagella in the adherence of an atypical enteropathogenic Escherichia coli (aEPEC) strain (serotype O51:H40) to human enterocytes. Accordingly, isogenic mutants deficient in flagellin (FliC), the flagellar structural subunit; the flagellar cap protein (FliD); or the MotAB proteins, involved in the control of flagellar motion, were generated and tested for binding to differentiated Caco-2 cells. Binding of the aEPEC strain to enterocytes was significantly impaired in strains with the fliC and fliD genes deleted, both of which could not form flagella on the bacterial surface. A nonmotile but flagellated MotAB mutant also showed impaired adhesion to Caco-2 cells. In accordance with these observations, adhesion of aEPEC strain 1711-4 to Caco-2 cells was drastically reduced after the treatment of Caco-2 cells with purified FliD. In addition, incubation of aEPEC bacteria with specific anti-FliD serum impaired binding to Caco-2 cells. Finally, incubation of Caco-2 cells with purified FliD, followed by immunolabeling, showed that the protein was specifically bound to the microvillus tips of differentiated Caco-2 cells. The aEPEC FliD or anti-FliD serum also reduced the adherence of prototype typical enteropathogenic, enterohemorrhagic, and enterotoxigenic E. coli strains to Caco-2 cells. In conclusion, our findings further strengthened the role of flagella in the adherence of aEPEC to human enterocytes and disclosed the relevant structural and functional involvement of FliD in the adhesion process. PMID:26831466

  19. Load Response of the Flagellar Beat

    NASA Astrophysics Data System (ADS)

    Klindt, Gary S.; Ruloff, Christian; Wagner, Christian; Friedrich, Benjamin M.

    2016-12-01

    Cilia and flagella exhibit regular bending waves that perform mechanical work on the surrounding fluid, to propel cellular swimmers and pump fluids inside organisms. Here, we quantify a force-velocity relationship of the beating flagellum, by exposing flagellated Chlamydomonas cells to controlled microfluidic flows. A simple theory of flagellar limit-cycle oscillations, calibrated by measurements in the absence of flow, reproduces this relationship quantitatively. We derive a link between the energy efficiency of the flagellar beat and its ability to synchronize to oscillatory flows.

  20. A coordinated sequence of distinct flagellar waveforms enables a sharp flagellar turn mediated by squid sperm pH-taxis.

    PubMed

    Iida, Tomohiro; Iwata, Yoko; Mohri, Tatsuma; Baba, Shoji A; Hirohashi, Noritaka

    2017-10-11

    Animal spermatozoa navigate by sensing ambient chemicals to reach the site of fertilization. Generally, such chemicals derive from the female reproductive organs or cells. Exceptionally, squid spermatozoa mutually release and perceive carbon dioxide to form clusters after ejaculation. We previously identified the pH-taxis by which each spermatozoon can execute a sharp turn, but how flagellar dynamics enable this movement remains unknown. Here, we show that initiation of the turn motion requires a swim down a steep proton gradient (a theoretical estimation of ≥0.025 pH/s), crossing a threshold pH value of ~5.5. Time-resolved kinematic analysis revealed that the turn sequence results from the rhythmic exercise of two flagellar motions: a stereotypical flagellar 'bent-cane' shape followed by asymmetric wave propagation, which enables a sharp turn in the realm of low Reynolds numbers. This turning episode is terminated by an 'overshoot' trajectory that differs from either straight-line motility or turning. As with bidirectional pH-taxes in some bacteria, squid spermatozoa also showed repulsion from strong acid conditions with similar flagellar kinematics as in positive pH-taxis. These findings indicate that squid spermatozoa might have a unique reorientation mechanism, which could be dissimilar to that of classical egg-guided sperm chemotaxis in other marine invertebrates.

  1. Novel Single-Tube Agar-Based Test System for Motility Enhancement and Immunocapture of Escherichia coli O157:H7 by H7 Flagellar Antigen-Specific Antibodies

    PubMed Central

    Murinda, Shelton E.; Nguyen, Lien T.; Ivey, Susan J.; Almeida, Raul A.; Oliver, Stephen P.

    2002-01-01

    This paper describes a novel single-tube agar-based technique for motility enhancement and immunoimmobilization of Escherichia coli O157:H7. Motility indole ornithine medium and agar (0.4%, wt/vol) media containing either nutrient broth, tryptone broth, or tryptic soy broth (TSBA) were evaluated for their abilities to enhance bacterial motility. Twenty-six E. coli strains, including 19 O157:H7 strains, 1 O157:H− strain, and 6 generic E. coli strains, were evaluated. Test bacteria were stab inoculated in the center of the agar column, and tubes were incubated at 37°C for 18 to 96 h. Nineteen to 24 of the 26 test strains (73.1 to 92.3%) were motile in the different media. TSBA medium performed best and was employed in subsequent studies of motility enhancement and H7 flagellar immunocapture. H7 flagellar antiserum (30 and 60 μl) mixed with TSBA was placed as a band (1 ml) in the middle of an agar column separating the top (3-ml) and bottom (3-ml) agar layers. The top agar layer was inoculated with the test bacterial strains. The tubes were incubated at 37°C for 12 to 18 h and for 18 to 96 h. The specificity and sensitivity of the H7 flagellar immunocapture tests were 75 and 100%, respectively. The procedure described is simple and sensitive and could be adapted easily for routine use in laboratories that do not have sophisticated equipment and resources for confirming the presence of H7 flagellar antigens. Accurate and rapid identification of H7 flagellar antigen is critical for the complete characterization of E. coli O157:H7, owing to the immense clinical, public health, and economic significance of this food-borne pathogen. PMID:12454173

  2. Bacterial flagella grow through an injection-diffusion mechanism

    PubMed Central

    Renault, Thibaud T; Abraham, Anthony O; Bergmiller, Tobias; Paradis, Guillaume; Rainville, Simon; Charpentier, Emmanuelle; Guet, Călin C; Tu, Yuhai; Namba, Keiichi; Keener, James P; Minamino, Tohru; Erhardt, Marc

    2017-01-01

    The bacterial flagellum is a self-assembling nanomachine. The external flagellar filament, several times longer than a bacterial cell body, is made of a few tens of thousands subunits of a single protein: flagellin. A fundamental problem concerns the molecular mechanism of how the flagellum grows outside the cell, where no discernible energy source is available. Here, we monitored the dynamic assembly of individual flagella using in situ labelling and real-time immunostaining of elongating flagellar filaments. We report that the rate of flagellum growth, initially ∼1,700 amino acids per second, decreases with length and that the previously proposed chain mechanism does not contribute to the filament elongation dynamics. Inhibition of the proton motive force-dependent export apparatus revealed a major contribution of substrate injection in driving filament elongation. The combination of experimental and mathematical evidence demonstrates that a simple, injection-diffusion mechanism controls bacterial flagella growth outside the cell. DOI: http://dx.doi.org/10.7554/eLife.23136.001 PMID:28262091

  3. Bacterial flagella grow through an injection-diffusion mechanism.

    PubMed

    Renault, Thibaud T; Abraham, Anthony O; Bergmiller, Tobias; Paradis, Guillaume; Rainville, Simon; Charpentier, Emmanuelle; Guet, Călin C; Tu, Yuhai; Namba, Keiichi; Keener, James P; Minamino, Tohru; Erhardt, Marc

    2017-03-06

    The bacterial flagellum is a self-assembling nanomachine. The external flagellar filament, several times longer than a bacterial cell body, is made of a few tens of thousands subunits of a single protein: flagellin. A fundamental problem concerns the molecular mechanism of how the flagellum grows outside the cell, where no discernible energy source is available. Here, we monitored the dynamic assembly of individual flagella using in situ labelling and real-time immunostaining of elongating flagellar filaments. We report that the rate of flagellum growth, initially ∼1,700 amino acids per second, decreases with length and that the previously proposed chain mechanism does not contribute to the filament elongation dynamics. Inhibition of the proton motive force-dependent export apparatus revealed a major contribution of substrate injection in driving filament elongation. The combination of experimental and mathematical evidence demonstrates that a simple, injection-diffusion mechanism controls bacterial flagella growth outside the cell.

  4. Polymorphic transformation of helical flagella of bacteria

    NASA Astrophysics Data System (ADS)

    Lim, Sookkyung; Howard Berg Collaboration; William Ko Collaboration; Yongsam Kim Collaboration; Wanho Lee Collaboration; Charles Peskin Collaboration

    2016-11-01

    Bacteria such as E. coli swim in an aqueous environment by utilizing the rotation of flagellar motors and alternate two modes of motility, runs and tumbles. Runs are steady forward swimming driven by bundles of flagellar filaments whose motors are turning CCW; tumbles involve a reorientation of the direction of swimming triggered by motor reversals. During tumbling, the helical flagellum undergoes polymorphic transformations, which is a local change in helical pitch, helical radius, and handedness. In this work, we investigate the underlying mechanism of structural conformation and how this polymorphic transition plays a role in bacterial swimming. National Science Foundation.

  5. Design of a Comprehensive Biochemistry and Molecular Biology Experiment: Phase Variation Caused by Recombinational Regulation of Bacterial Gene Expression

    ERIC Educational Resources Information Center

    Sheng, Xiumei; Xu, Shungao; Lu, Renyun; Isaac, Dadzie; Zhang, Xueyi; Zhang, Haifang; Wang, Huifang; Qiao, Zheng; Huang, Xinxiang

    2014-01-01

    Scientific experiments are indispensable parts of Biochemistry and Molecular Biology. In this study, a comprehensive Biochemistry and Molecular Biology experiment about "Salmonella enterica" serovar Typhi Flagellar phase variation has been designed. It consisted of three parts, namely, inducement of bacterial Flagellar phase variation,…

  6. Modeling torque versus speed, shot noise, and rotational diffusion of the bacterial flagellar motor.

    PubMed

    Mora, Thierry; Yu, Howard; Wingreen, Ned S

    2009-12-11

    We present a minimal physical model for the flagellar motor that enables bacteria to swim. Our model explains the experimentally measured torque-speed relationship of the proton-driven E. coli motor at various pH and temperature conditions. In particular, the dramatic drop of torque at high rotation speeds (the "knee") is shown to arise from saturation of the proton flux. Moreover, we show that shot noise in the proton current dominates the diffusion of motor rotation at low loads. This suggests a new way to probe the discreteness of the energy source, analogous to measurements of charge quantization in superconducting tunnel junctions.

  7. The Flagellar Hook Protein, FlgE, of Salmonella enterica Serovar Typhimurium Is Posttranscriptionally Regulated in Response to the Stage of Flagellar Assembly

    PubMed Central

    Bonifield, Heather R.; Yamaguchi, Shigeru; Hughes, Kelly T.

    2000-01-01

    We investigated the posttranscriptional regulation of flgE, a class 2 gene that encodes the hook subunit protein of the flagella. RNase protection assays demonstrated that the flgE gene was transcribed at comparable levels in numerous strains defective in known steps of flagellar assembly. However, Western analyses of these strains demonstrated substantial differences in FlgE protein levels. Although wild-type FlgE levels were observed in strains with deletions of genes encoding components of the switch complex and the flagellum-specific secretion apparatus, no protein was detected in a strain with deletions of the rod, ring, and hook-associated proteins. To determine whether FlgE levels were affected by the stage of hook–basal-body assembly, Western analysis was performed on strains with mutations at individual loci encompassed by the deletion. FlgE protein was undetectable in rod mutants, intermediate in ring mutants, and wild type in hook-associated protein mutants. The lack of negative regulation in switch complex and flagellum-specific secretion apparatus deletion mutants blocked for flagellar construction prior to rod assembly suggests that these structures play a role in the negative regulation of FlgE. Quantitative Western analyses of numerous flagellar mutants indicate that FlgE levels reflect the stage at which flagellar assembly is blocked. These data provide evidence for negative posttranscriptional regulation of FlgE in response to the stage of flagellar assembly. PMID:10869084

  8. Divalent Cation Control of Flagellar Motility in African Trypanosomes

    NASA Astrophysics Data System (ADS)

    Westergard, Anna M.; Hutchings, Nathan R.

    2005-03-01

    Changes in calcium concentration have been shown to dynamically affect flagellar motility in several eukaryotic systems. The African trypanosome is a monoflagellated protozoan parasite and the etiological agent of sleeping sickness. Although cell motility has been implicated in disease progression, very little is currently known about biochemical control of the trypanosome flagellum. In this study, we assess the effects of extracellular changes in calcium and nickel concentration on trypanosome flagellar movement. Using a flow through chamber, we determine the relative changes in motility in individual trypanosomes in response to various concentrations of calcium and nickel, respectively. Extracellular concentrations of calcium and nickel (as low as 100 micromolar) significantly inhibit trypanosome cell motility. The effects are reversible, as indicated by the recovery of motion after removal of the calcium or nickel from the chamber. We are currently investigating the specific changes in flagellar oscillation and coordination that result from calcium and nickel, respectively. These results verify the presence of a calcium-responsive signaling mechanism(s) that regulates flagellar beat in trypanosomes.

  9. Ocean acidification reduces sperm flagellar motility in broadcast spawning reef invertebrates.

    PubMed

    Morita, Masaya; Suwa, Ryota; Iguchi, Akira; Nakamura, Masako; Shimada, Kazuaki; Sakai, Kazuhiko; Suzuki, Atsushi

    2010-05-01

    Ocean acidification is now recognized as a threat to marine ecosystems; however, the effect of ocean acidification on fertilization in marine organisms is still largely unknown. In this study, we focused on sperm flagellar motility in broadcast spawning reef invertebrates (a coral and a sea cucumber). Below pH 7.7, the pH predicted to occur within the next 100 years, sperm flagellar motility was seriously impaired in these organisms. Considering that sperm flagellar motility is indispensable for transporting the paternal haploid genome for fertilization, fertilization taking place in seawater may decline in the not too distant future. Urgent surveys are necessary for a better understanding of the physiological consequences of ocean acidification on sperm flagellar motility in a wide range of marine invertebrates.

  10. Interactions of the chemotaxis signal protein CheY with bacterial flagellar motors visualized by evanescent wave microscopy.

    PubMed

    Khan, S; Pierce, D; Vale, R D

    The chemotaxis signal protein CheY of enteric bacteria shuttles between transmembrane methyl-accepting chemotaxis protein (MCP) receptor complexes and flagellar basal bodies [1]. The basal body C-rings, composed of the FliM, FliG and FliN proteins, form the rotor of the flagellar motor [2]. Phosphorylated CheY binds to isolated FliM [3] and may also interact with FliG [4], but its binding to basal bodies has not been measured. Using the chemorepellent acetate to phosphorylate and acetylate CheY [5], we have measured the covalent-modification-dependent binding of a green fluorescent protein-CheY fusion (GFP-CheY) to motor assemblies in bacteria lacking MCP complexes by evanescent wave microscopy [6]. At acetate concentrations that cause solely clockwise rotation, GFP-CheY molecules bound to native basal bodies or to overproduced rotor complexes with a stoichiometry comparable to the number of C-ring subunits. GFP-CheY did not bind to rotors lacking FIiM/FliN, showing that these subunits are essential for the association. This assay provides a new means of monitoring protein-protein interactions in signal transduction pathways in living cells.

  11. Regulation of flagellar assembly by glycogen synthase kinase 3 in Chlamydomonas reinhardtii.

    PubMed

    Wilson, Nedra F; Lefebvre, Paul A

    2004-10-01

    Chlamydomonas reinhardtii controls flagellar assembly such that flagella are of an equal and predetermined length. Previous studies demonstrated that lithium, an inhibitor of glycogen synthase kinase 3 (GSK3), induced flagellar elongation, suggesting that a lithium-sensitive signal transduction pathway regulated flagellar length (S. Nakamura, H. Takino, and M. K. Kojima, Cell Struct. Funct. 12:369-374, 1987). Here, we demonstrate that lithium treatment depletes the pool of flagellar proteins from the cell body and that the heterotrimeric kinesin Fla10p accumulates in flagella. We identify GSK3 in Chlamydomonas and demonstrate that its kinase activity is inhibited by lithium in vitro. The tyrosine-phosphorylated, active form of GSK3 was enriched in flagella and GSK3 associated with the axoneme in a phosphorylation-dependent manner. The level of active GSK3 correlated with flagellar length; early during flagellar regeneration, active GSK3 increased over basal levels. This increase in active GSK3 was rapidly lost within 30 min of regeneration as the level of active GSK3 decreased relative to the predeflagellation level. Taken together, these results suggest a possible role for GSK3 in regulating the assembly and length of flagella.

  12. Modeling Torque Versus Speed, Shot Noise, and Rotational Diffusion of the Bacterial Flagellar Motor

    PubMed Central

    Mora, Thierry; Yu, Howard; Wingreen, Ned S.

    2010-01-01

    We present a minimal physical model for the flagellar motor that enables bacteria to swim. Our model explains the experimentally measured torque-speed relationship of the proton-driven E. coli motor at various pH and temperature conditions. In particular, the dramatic drop of torque at high rotation speeds (the “knee”) is shown to arise from saturation of the proton flux. Moreover, we show that shot noise in the proton current dominates the diffusion of motor rotation at low loads. This suggests a new way to probe the discreteness of the energy source, analogous to measurements of charge quantization in superconducting tunnel junctions. PMID:20366231

  13. Direction of flagellar rotation in bacterial cell envelopes.

    PubMed Central

    Ravid, S; Eisenbach, M

    1984-01-01

    Cell envelopes with functional flagella, isolated from wild-type strains of Escherichia coli and Salmonella typhimurium by formation of spheroplasts with penicillin and subsequent osmotic lysis, demonstrate counterclockwise (CCW)-biased rotation when energized with an electron donor for respiration, DL-lactate. Since the direction of flagellar rotation in bacteria is central to the expression of chemotaxis, we studied the cause of this bias. Our main observations were: (i) spheroplasts acquired a clockwise (CW) bias if instead of being lysed they were further incubated with penicillin; (ii) repellents temporarily caused CW rotation of tethered bacteria and spheroplasts but not of their derived cell envelopes; (iii) deenergizing CW-rotating cheV bacteria by KCN or arsenate treatment caused CCW bias; (iv) cell envelopes isolated from CW-rotating cheC and cheV mutants retained the CW bias, unlike envelopes isolated from cheB and cheZ mutants, which upon cytoplasmic release lost this bias and acquired CCW bias; and (v) an inwardly directed, artificially induced proton current rotated tethered envelopes in CCW direction, but an outwardly directed current was unable to rotate the envelopes. It is concluded that (i) a cytoplasmic constituent is required for the expression of CW rotation (or repression of CCW rotation) in strains which are not defective in the switch; (ii) in the absence of this cytoplasmic constituent, the motor is not reversible in such strains, and it probably is mechanically constricted so as to permit CCW sense of rotation only; (iii) the requirement of CW rotation for ATP is not at the level of the motor or the switch but at one of the preceding functional steps of the chemotaxis machinery; (iv) the cheC and cheV gene products are associated with the cytoplasmic membrane; and (v) direct interaction between the switch-motor system and the repellent sensors is improbable. Images PMID:6370958

  14. Flagellar region 3b supports strong expression of integrated DNA and the highest chromosomal integration efficiency of the Escherichia coli flagellar regions.

    PubMed

    Juhas, Mario; Ajioka, James W

    2015-07-01

    The Gram-negative bacterium Escherichia coli is routinely used as the chassis for a variety of biotechnology and synthetic biology applications. Identification and analysis of reliable chromosomal integration and expression target loci is crucial for E. coli engineering. Chromosomal loci differ significantly in their ability to support integration and expression of the integrated genetic circuits. In this study, we investigate E. coli K12 MG1655 flagellar regions 2 and 3b. Integration of the genetic circuit into seven and nine highly conserved genes of the flagellar regions 2 (motA, motB, flhD, flhE, cheW, cheY and cheZ) and 3b (fliE, F, G, J, K, L, M, P, R), respectively, showed significant variation in their ability to support chromosomal integration and expression of the integrated genetic circuit. While not reducing the growth of the engineered strains, the integrations into all 16 target sites led to the loss of motility. In addition to high expression, the flagellar region 3b supports the highest efficiency of integration of all E. coli K12 MG1655 flagellar regions and is therefore potentially the most suitable for the integration of synthetic genetic circuits. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  15. Motility and Flagellar Glycosylation in Clostridium difficile▿ †

    PubMed Central

    Twine, Susan M.; Reid, Christopher W.; Aubry, Annie; McMullin, David R.; Fulton, Kelly M.; Austin, John; Logan, Susan M.

    2009-01-01

    In this study, intact flagellin proteins were purified from strains of Clostridium difficile and analyzed using quadrupole time of flight and linear ion trap mass spectrometers. Top-down studies showed the flagellin proteins to have a mass greater than that predicted from the corresponding gene sequence. These top-down studies revealed marker ions characteristic of glycan modifications. Additionally, diversity in the observed masses of glycan modifications was seen between strains. Electron transfer dissociation mass spectrometry was used to demonstrate that the glycan was attached to the flagellin protein backbone in O linkage via a HexNAc residue in all strains examined. Bioinformatic analysis of C. difficile genomes revealed diversity with respect to glycan biosynthesis gene content within the flagellar biosynthesis locus, likely reflected by the observed flagellar glycan diversity. In C. difficile strain 630, insertional inactivation of a glycosyltransferase gene (CD0240) present in all sequenced genomes resulted in an inability to produce flagellar filaments at the cell surface and only minor amounts of unmodified flagellin protein. PMID:19749038

  16. A quantitative description of flagellar movement in golden hamster spermatozoa.

    PubMed

    Ishijima, S; Mohri, H

    1985-01-01

    Flagellar movement of golden hamster spermatozoa obtained from the testis and the caput and cauda epididymides was observed by a light microscope while holding them at their heads with a micropipette. Flagellar movement of capacitated spermatozoa and of reactivated spermatozoa demembranated with Triton X-100 was also observed. Testicular and caput epididymal spermatozoa showed weak movement in Tyrode's solution, whereas cauda epididymal spermatozoa showed vigorous movement. The flagellar bends of the cauda epididymal spermatozoa were almost planar. Capacitated spermatozoa moved with waves of a large amplitude. Demembranated spermatozoa reactivated with ATP only had a latent period before the initiation of flagellar movement, and beat at low frequency, whereas demembranated spermatozoa reactivated with both ATP and cAMP began to move immediately at high frequency. Thrust and hydrodynamic power output were calculated using the parameters for the typical waveforms of cauda epididymal spermatozoa before and after capacitation. The possible role of the large amplitude beat in capacitated spermatozoa is discussed. A comparison of the 'principal' and 'reverse' bends in golden hamster sperm flagella as defined by Woolley (1977) with those in sea urchin sperm flagella suggests that the so-called 'principal' bend in golden hamster sperm flagella corresponds to the reverse bend in sea urchin sperm flagella and vice versa.

  17. Lipopolysaccharide-Specific but Not Anti-Flagellar Immunoglobulin A Monoclonal Antibodies Prevent Salmonella enterica Serotype Enteritidis Invasion and Replication within HEp-2 Cell Monolayers

    PubMed Central

    Iankov, Ianko D.; Petrov, Dragomir P.; Mladenov, Ivan V.; Haralambieva, Iana H.; Mitov, Ivan G.

    2002-01-01

    The protective potential of immunoglobulin A (IgA) monoclonal antibodies (MAbs) directed against O and H antigens of Salmonella enterica serotype Enteritidis to prevent bacterial adhesion to and invasion of HEp-2 cells was evaluated. Although anti-flagellar IgA MAbs showed strong agglutinating capacities, they did not protect cell monolayers. In contrast, IgA MAbs specific for the O:9 epitope of Salmonella lipopolysaccharide antigen alone prevented S. enterica serotype Enteritidis entry and replication within HEp-2 cells, and the protection was not mediated by direct binding of antibodies to bacterial adhesins or by agglutination of microorganisms. PMID:11854252

  18. Isolation and characterization of flagellar filament from zoospores of Dermatophilus congolensis.

    PubMed

    Hiraizumi, Mieko; Tagawa, Yuichi

    2014-09-17

    Highly motile zoospores from Dermatophilus congolensis bovine isolates from clinical dermatophilosis in Japan were obtained by culturing at 27°C in an ambient atmosphere on heart infusion agar supplemented with 5% defibrinated sheep blood for 72h or in heart infusion broth for 48h with gentle shaking. After vigorous mechanical agitation of the zoospore suspension, the flagellar filaments detached from motile zoospores and were isolated in the clear gelatinous part of the final pellet by differential centrifugation. Typical morphology of a flagellar filament, with a width of approximately 15nm, was observed in the isolated flagellar filament by electron microscopy. A single major protein (flagellin) band with an apparent molecular mass of 35kDa was detected in the flagellar filament of D. congolensis strain AM-1 and that of 33kDa was detected in strain IT-2 by SDS-PAGE. In immunoblot analysis of whole-cell proteins from seven isolates of D. congolensis, antiserum to strain AM-1 zoospores reacted with the 35-kDa antigen band of strain AM-1, but not with any antigen band of other strains in a similar molecular mass range. In contrast, antiserum to strain IT-2 zoospores reacted with antigen bands at 33kDa from six strains, except strain AM-1. Similar strain-specific reactions of these anti-zoospore sera with isolated flagellar filaments from strains AM-1 and IT-2 were confirmed by immunoblot, indicating the presence of antigenic variations of flagellins of D. congolensis zoospores. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Intracellular Ca2+ threshold reversibly switches flagellar beat off and on.

    PubMed

    Sánchez-Cárdenas, C; Montoya, F; Navarrete, F A; Hernández-Cruz, A; Corkidi, G; Visconti, P E; Darszon, A

    2018-06-08

    Sperm motility is essential for fertilization. The asymmetry of flagellar beat in spermatozoa is finely regulated by intracellular calcium concentration ([Ca2+]i). Recently, we demonstrated that the application of high concentrations (10-20 μM) of the Ca2+ ionophore A23187 promotes sperm immobilization after 10 minutes, and its removal thereafter allows motility recovery, hyperactivation and fertilization. In addition, the same ionophore treatment overcomes infertility observed in sperm from Catsper1-/-, Slo3-/- and Adcy10-/-, but not PMCA4-/-, which strongly suggest that regulation of [Ca2+]i is mandatory for sperm motility and hyperactivation. In this study we found that prior to inducing sperm immobilization, high A23187 concentrations (10 μM) increase flagellar beat. While 5-10 μM A23187 substantially elevates [Ca2+]i and rapidly immobilizes sperm in a few minutes, smaller concentrations (0.5 and 1 μM) provoke smaller [Ca2+]i increases and sperm hyperactivation, confirming that [Ca2+]i increases act as a motility switch. Until now the [Ca2+]i thresholds that switch motility on and off were not fully understood. To study the relationship between [Ca2+]i and flagellar beating, we developed an automatic tool that allows the simultaneous measurement of these two parameters. Individual spermatozoa were treated with A23187 which is then washed to evaluate [Ca2+]i and flagellar beat recovery using the implemented method. We observe that [Ca2+]i must decrease below a threshold concentration range to facilitate subsequent flagellar beat recovery and sperm motility.

  20. The Flagellar Protein FliL Is Essential for Swimming in Rhodobacter sphaeroides▿ †

    PubMed Central

    Suaste-Olmos, Fernando; Domenzain, Clelia; Mireles-Rodríguez, José Cruz; Poggio, Sebastian; Osorio, Aurora; Dreyfus, Georges; Camarena, Laura

    2010-01-01

    In this work we characterize the function of the flagellar protein FliL in Rhodobacter sphaeroides. Our results show that FliL is essential for motility in this bacterium and that in its absence flagellar rotation is highly impaired. A green fluorescent protein (GFP)-FliL fusion forms polar and lateral fluorescent foci that show different spatial dynamics. The presence of these foci is dependent on the expression of the flagellar genes controlled by the master regulator FleQ, suggesting that additional components of the flagellar regulon are required for the proper localization of GFP-FliL. Eight independent pseudorevertants were isolated from the fliL mutant strain. In each of these strains a single nucleotide change in motB was identified. The eight mutations affected only three residues located on the periplasmic side of MotB. Swimming of the suppressor mutants was not affected by the presence of the wild-type fliL allele. Pulldown and yeast two-hybrid assays showed that that the periplasmic domain of FliL is able to interact with itself but not with the periplasmic domain of MotB. From these results we propose that FliL could participate in the coupling of MotB with the flagellar rotor in an indirect fashion. PMID:20889747

  1. Coevolved Mutations Reveal Distinct Architectures for Two Core Proteins in the Bacterial Flagellar Motor

    PubMed Central

    Pandini, Alessandro; Kleinjung, Jens; Rasool, Shafqat; Khan, Shahid

    2015-01-01

    Switching of bacterial flagellar rotation is caused by large domain movements of the FliG protein triggered by binding of the signal protein CheY to FliM. FliG and FliM form adjacent multi-subunit arrays within the basal body C-ring. The movements alter the interaction of the FliG C-terminal (FliGC) “torque” helix with the stator complexes. Atomic models based on the Salmonella entrovar C-ring electron microscopy reconstruction have implications for switching, but lack consensus on the relative locations of the FliG armadillo (ARM) domains (amino-terminal (FliGN), middle (FliGM) and FliGC) as well as changes during chemotaxis. The generality of the Salmonella model is challenged by the variation in motor morphology and response between species. We studied coevolved residue mutations to determine the unifying elements of switch architecture. Residue interactions, measured by their coevolution, were formalized as a network, guided by structural data. Our measurements reveal a common design with dedicated switch and motor modules. The FliM middle domain (FliMM) has extensive connectivity most simply explained by conserved intra and inter-subunit contacts. In contrast, FliG has patchy, complex architecture. Conserved structural motifs form interacting nodes in the coevolution network that wire FliMM to the FliGC C-terminal, four-helix motor module (C3-6). FliG C3-6 coevolution is organized around the torque helix, differently from other ARM domains. The nodes form separated, surface-proximal patches that are targeted by deleterious mutations as in other allosteric systems. The dominant node is formed by the EHPQ motif at the FliMMFliGM contact interface and adjacent helix residues at a central location within FliGM. The node interacts with nodes in the N-terminal FliGc α-helix triad (ARM-C) and FliGN. ARM-C, separated from C3-6 by the MFVF motif, has poor intra-network connectivity consistent with its variable orientation revealed by structural data. ARM-C could

  2. Intraflagellar transport particle size scales inversely with flagellar length: revisiting the balance-point length control model.

    PubMed

    Engel, Benjamin D; Ludington, William B; Marshall, Wallace F

    2009-10-05

    The assembly and maintenance of eukaryotic flagella are regulated by intraflagellar transport (IFT), the bidirectional traffic of IFT particles (recently renamed IFT trains) within the flagellum. We previously proposed the balance-point length control model, which predicted that the frequency of train transport should decrease as a function of flagellar length, thus modulating the length-dependent flagellar assembly rate. However, this model was challenged by the differential interference contrast microscopy observation that IFT frequency is length independent. Using total internal reflection fluorescence microscopy to quantify protein traffic during the regeneration of Chlamydomonas reinhardtii flagella, we determined that anterograde IFT trains in short flagella are composed of more kinesin-associated protein and IFT27 proteins than trains in long flagella. This length-dependent remodeling of train size is consistent with the kinetics of flagellar regeneration and supports a revised balance-point model of flagellar length control in which the size of anterograde IFT trains tunes the rate of flagellar assembly.

  3. Visualization of bacterial flagella dynamics in a viscous shear flow

    NASA Astrophysics Data System (ADS)

    Ali, Jamel; Kim, Minjun

    2016-11-01

    We report on the dynamics of tethered bacterial flagella in an applied viscous shear flow and analyze their behavior using image processing. Flagellin proteins were repolymerized into flagellar filaments functionalized with biotin at their proximal end, and allowed to self-assemble within a micro channel coated with streptavidin. It was observed that all attached flagellar filaments aligned with the steady shear flow of various polymeric solutions. Furthermore it was observed that many of the filaments were stretched, and at elevated flow rates began to undergo polymorphic transformations, which were initiated at one end of the flagellum. When undergoing a change to a different helical form the flagellum was observed to transform to an oppositely handed helix, as to counteract the viscous torque imparted by the shear flow. It was also observed that some flagellar filaments did not undergo polymorphic transformations, but rotated about their helical axis. The rate of this rotation appears to be a function of the applied flow rate. These results expand on previous experimental work and aid in the development of a novel platform that harnesses the autonomic response of a 'forest' of bacterial flagella for engineering applications. This work was funded by NSF Grant CMMI-1000255, KEIT MOTIE Grant No. 10052980, and with Government support under and awarded by DoD, Air Force Office of Scientific Research, National Defense Science and Engineering Graduate (NDSEG) Fellowship, 32 CFR 168a.

  4. The dynamic instability in the hook/flagellum system that triggers bacterial flicks

    NASA Astrophysics Data System (ADS)

    Jabbarzadeh, Mehdi; Fu, Henry

    2017-11-01

    Dynamical bending, buckling, and polymorphic transformations of the flagellum are known to affect bacterial motility, but run-reverse-flick motility of monotrichous bacteria also involves the even more flexible hook, which connects the flagellum to the cell body. Here, we identify the dynamic buckling mechanism that produces flicks in Vibrio alginolyticus. Estimates of forces and torques on the hook from experimental observations suggest that flicks are triggered at stresses below the hook's static Euler buckling criterion. Using an accurate linearization of the Kirchoff rod model for the hook in a model of a swimming bacterium with rigid flagellum, we show that as hook stiffness decreases there is a transition from on-axis flagellar rotation with small hook deflections to flagellar precession with large deflections. When flagellum flexibility is incorporated, the precession is disrupted by significant flagellar bending - i.e., incipient flicks. The predicted onset of dynamic instabilities corresponds well with experimentally observed flick events.

  5. Transcriptional regulation of coordinate changes in flagellar mRNAs during differentiation of Naegleria gruberi amoebae into flagellates

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

    Lee, J.H.; Walsh, C.J.

    1988-06-01

    The nuclear run-on technique was used to measure the rate of transcription of flagellar genes during the differentiation of Naegleria gruberi amebae into flagellates. Synthesis of mRNAs for the axonemal proteins ..cap alpha..- and BETA-tubulin and flagellar calmodulin, as well as a coordinately regulated poly(A)/sup +/ RNA that codes for an unidentified protein, showed transient increases averaging 22-fold. The rate of synthesis of two poly(A)/sup +/ RNAs common to ameobae and flagellates was low until the transcription of the flagellar genes began to decline, at which time synthesis of the RNAs found in ameobae increased 3- to 10-fold. The observedmore » changes in the rate of transcription can account quantitatively for the 20-fold increase in flagellar mRNA concentration during the differentiation. The data for the flagellar calmodulin gene demonstrate transcriptional regulation for a nontubulin axonemal protein. The data also demonstrate at least two programs of transcriptional regulation during the differentiation and raise the intriguing possibility that some significant fraction of the nearly 200 different proteins of the flagellar axoneme is transcriptionally regulated during the 1 h it takes N. gruberi amebae to form visible flagella.« less

  6. Common Evolutionary Origin for the Rotor Domain of Rotary Atpases and Flagellar Protein Export Apparatus

    PubMed Central

    Kishikawa, Jun-ichi; Ibuki, Tatsuya; Nakamura, Shuichi; Nakanishi, Astuko; Minamino, Tohru; Miyata, Tomoko; Namba, Keiichi; Konno, Hiroki; Ueno, Hiroshi; Imada, Katsumi; Yokoyama, Ken

    2013-01-01

    The V1- and F1- rotary ATPases contain a rotor that rotates against a catalytic A3B3 or α3β3 stator. The rotor F1-γ or V1-DF is composed of both anti-parallel coiled coil and globular-loop parts. The bacterial flagellar type III export apparatus contains a V1/F1-like ATPase ring structure composed of FliI6 homo-hexamer and FliJ which adopts an anti-parallel coiled coil structure without the globular-loop part. Here we report that FliJ of Salmonella enterica serovar Typhimurium shows a rotor like function in Thermus thermophilus A3B3 based on both biochemical and structural analysis. Single molecular analysis indicates that an anti-parallel coiled-coil structure protein (FliJ structure protein) functions as a rotor in A3B3. A rotary ATPase possessing an F1-γ-like protein generated by fusion of the D and F subunits of V1 rotates, suggesting F1-γ could be the result of a fusion of the genes encoding two separate rotor subunits. Together with sequence comparison among the globular part proteins, the data strongly suggest that the rotor domains of the rotary ATPases and the flagellar export apparatus share a common evolutionary origin. PMID:23724081

  7. Bacterial flagella and Type III secretion: case studies in the evolution of complexity.

    PubMed

    Pallen, M J; Gophna, U

    2007-01-01

    Bacterial flagella at first sight appear uniquely sophisticated in structure, so much so that they have even been considered 'irreducibly complex' by the intelligent design movement. However, a more detailed analysis reveals that these remarkable pieces of molecular machinery are the product of processes that are fully compatible with Darwinian evolution. In this chapter we present evidence for such processes, based on a review of experimental studies, molecular phylogeny and microbial genomics. Several processes have played important roles in flagellar evolution: self-assembly of simple repeating subunits, gene duplication with subsequent divergence, recruitment of elements from other systems ('molecular bricolage'), and recombination. We also discuss additional tentative new assignments of homology (FliG with MgtE, FliO with YscJ). In conclusion, rather than providing evidence of intelligent design, flagellar and non-flagellar Type III secretion systems instead provide excellent case studies in the evolution of complex systems from simpler components.

  8. Role of calmodulin and calcineurin in regulating flagellar motility and wave polarity in Leishmania.

    PubMed

    Mukhopadhyay, Aakash Gautam; Dey, Chinmoy Sankar

    2017-11-01

    We have previously reported the involvement of cyclic AMP in regulating flagellar waveforms in Leishmania. Here, we investigated the roles of calcium, calmodulin, and calcineurin in flagellar motility regulation in L. donovani. Using high-speed videomicroscopy, we show that calcium-independent calmodulin and calcineurin activity is necessary for motility in Leishmania. Inhibition of calmodulin and calcineurin induced ciliary beats interrupting flagellar beating in both live (in vivo) and ATP-reactivated (in vitro) parasites. Our results indicate that signaling mediated by calmodulin and calcineurin operates antagonistically to cAMP signaling in regulating the waveforms of Leishmania flagellum. These two pathways are possibly involved in maintaining the balance between the two waveforms, essential for responding to environmental cues, survival, and infectivity.

  9. Tuning bacterial hydrodynamics with magnetic fields

    NASA Astrophysics Data System (ADS)

    Pierce, C. J.; Mumper, E.; Brown, E. E.; Brangham, J. T.; Lower, B. H.; Lower, S. K.; Yang, F. Y.; Sooryakumar, R.

    2017-06-01

    Magnetotactic bacteria are a group of motile prokaryotes that synthesize chains of lipid-bound, magnetic nanoparticles called magnetosomes. This study exploits their innate magnetism to investigate previously unexplored facets of bacterial hydrodynamics at surfaces. Through use of weak, uniform, external magnetic fields and local, micromagnetic surface patterns, the relative strength of hydrodynamic, magnetic, and flagellar force components is tuned through magnetic control of the bacteria's orientation. The resulting swimming behaviors provide a means to experimentally determine hydrodynamic parameters and offer a high degree of control over large numbers of living microscopic entities. The implications of this controlled motion for studies of bacterial motility near surfaces and for micro- and nanotechnology are discussed.

  10. Numerical studies of bacterial-carpet microflows

    NASA Astrophysics Data System (ADS)

    Huber, Greg; Tillberg, Dan; Powers, Thomas R.

    2004-03-01

    Bacterial carpets are arrays of motile bacteria attached to two-dimensional surfaces. Improved understanding of carpet flows is important in the design of microfluidic devices and transport systems powered by bacterial flagellar motion. In recent experiments by the group of Howard Berg, cells of swarming S. marcescens are stuck to the surface, with most of their flagella free to rotate in the fluid. These studies show modified transport and greatly enhanced diffusion near the active carpet surface. We present theoretical models of the flagella-driven flow, bridging the nano- to the macro-scale, simulate the diffusion and advection of passive tracers, and compare the numerical results with the tracking data of Berg et al.

  11. Sodium-driven energy conversion for flagellar rotation of the earliest divergent hyperthermophilic bacterium.

    PubMed

    Takekawa, Norihiro; Nishiyama, Masayoshi; Kaneseki, Tsuyoshi; Kanai, Tamotsu; Atomi, Haruyuki; Kojima, Seiji; Homma, Michio

    2015-08-05

    Aquifex aeolicus is a hyperthermophilic, hydrogen-oxidizing and carbon-fixing bacterium that can grow at temperatures up to 95 °C. A. aeolicus has an almost complete set of flagellar genes that are conserved in bacteria. Here we observed that A. aeolicus has polar flagellum and can swim with a speed of 90 μm s(-1) at 85 °C. We expressed the A. aeolicus mot genes (motA and motB), which encode the torque generating stator proteins of the flagellar motor, in a corresponding mot nonmotile mutant of Escherichia coli. Its motility was slightly recovered by expression of A. aeolicus MotA and chimeric MotB whose periplasmic region was replaced with that of E. coli. A point mutation in the A. aeolicus MotA cytoplasmic region remarkably enhanced the motility. Using this system in E. coli, we demonstrate that the A. aeolicus motor is driven by Na(+). As motor proteins from hyperthermophilic bacteria represent the earliest motor proteins in evolution, this study strongly suggests that ancient bacteria used Na(+) for energy coupling of the flagellar motor. The Na(+)-driven flagellar genes might have been laterally transferred from early-branched bacteria into late-branched bacteria and the interaction surfaces of the stator and rotor seem not to change in evolution.

  12. Arsenate arrests flagellar rotation in cytoplasm-free envelopes of bacteria.

    PubMed Central

    Margolin, Y; Barak, R; Eisenbach, M

    1994-01-01

    The effect of arsenate on flagellar rotation in cytoplasm-free flagellated envelopes of Escherichia coli and Salmonella typhimurium was investigated. Flagellar rotation ceased as soon as the envelopes were exposed to arsenate. Inclusion of phosphate intracellularly (but not extracellular) prevented the inhibition by arsenate. In a parallel experiment, the rotation was not affected by inclusion of an ATP trap (hexokinase and glucose) within the envelopes. It is concluded that arsenate affects the motor in a way other than reversible deenergization. This may be an irreversible damage to the cell or direct inhibition of the motor by arsenate. The latter possibility suggests that a process of phosphorylation or phosphate binding is involved in the motor function. PMID:8071237

  13. Asymmetry in the clockwise and counterclockwise rotation of the bacterial flagellar motor

    PubMed Central

    Yuan, Junhua; Fahrner, Karen A.; Turner, Linda; Berg, Howard C.

    2010-01-01

    Cells of Escherichia coli are able to swim up gradients of chemical attractants by modulating the direction of rotation of their flagellar motors, which spin alternately clockwise (CW) and counterclockwise (CCW). Rotation in either direction has been thought to be symmetric and exhibit the same torques and speeds. The relationship between torque and speed is one of the most important measurable characteristics of the motor, used to distinguish specific mechanisms of motor rotation. Previous measurements of the torque–speed relationship have been made with cells lacking the response regulator CheY that spin their motors exclusively CCW. In this case, the torque declines slightly up to an intermediate speed called the “knee speed” after which it falls rapidly to zero. This result is consistent with a “power-stroke” mechanism for torque generation. Here, we measure the torque–speed relationship for cells that express large amounts of CheY and only spin their motors CW. We find that the torque decreases linearly with speed, a result remarkably different from that for CCW rotation. We obtain similar results for wild-type cells by reexamining data collected in previous work. We speculate that CCW rotation might be optimized for runs, with higher speeds increasing the ability of cells to sense spatial gradients, whereas CW rotation might be optimized for tumbles, where the object is to change cell trajectories. But why a linear torque–speed relationship might be optimum for the latter purpose we do not know. PMID:20615986

  14. Genome-wide transcriptional analysis of flagellar regeneration in Chlamydomonas reinhardtii identifies orthologs of ciliary disease genes

    NASA Technical Reports Server (NTRS)

    Stolc, Viktor; Samanta, Manoj Pratim; Tongprasit, Waraporn; Marshall, Wallace F.

    2005-01-01

    The important role that cilia and flagella play in human disease creates an urgent need to identify genes involved in ciliary assembly and function. The strong and specific induction of flagellar-coding genes during flagellar regeneration in Chlamydomonas reinhardtii suggests that transcriptional profiling of such cells would reveal new flagella-related genes. We have conducted a genome-wide analysis of RNA transcript levels during flagellar regeneration in Chlamydomonas by using maskless photolithography method-produced DNA oligonucleotide microarrays with unique probe sequences for all exons of the 19,803 predicted genes. This analysis represents previously uncharacterized whole-genome transcriptional activity profiling study in this important model organism. Analysis of strongly induced genes reveals a large set of known flagellar components and also identifies a number of important disease-related proteins as being involved with cilia and flagella, including the zebrafish polycystic kidney genes Qilin, Reptin, and Pontin, as well as the testis-expressed tubby-like protein TULP2.

  15. Flagellar glycosylation in Clostridium botulinum.

    PubMed

    Twine, Susan M; Paul, Catherine J; Vinogradov, Evgeny; McNally, David J; Brisson, Jean-Robert; Mullen, James A; McMullin, David R; Jarrell, Harold C; Austin, John W; Kelly, John F; Logan, Susan M

    2008-09-01

    Flagellins from Clostridium botulinum were shown to be post-translationally modified with novel glycan moieties by top-down MS analysis of purified flagellin protein from strains of various toxin serotypes. Detailed analyses of flagellin from two strains of C. botulinum demonstrated that the protein is modified by a novel glycan moiety of mass 417 Da in O-linkage. Bioinformatic analysis of available C. botulinum genomes identified a flagellar glycosylation island containing homologs of genes recently identified in Campylobacter coli that have been shown to be responsible for the biosynthesis of legionaminic acid derivatives. Structural characterization of the carbohydrate moiety was completed utilizing both MS and NMR spectroscopy, and it was shown to be a novel legionaminic acid derivative, 7-acetamido-5-(N-methyl-glutam-4-yl)-amino-3,5,7,9-tetradeoxy-D-glycero-alpha-D-galacto-nonulosonic acid, (alphaLeg5GluNMe7Ac). Electron transfer dissociation MS with and without collision-activated dissociation was utilized to map seven sites of O-linked glycosylation, eliminating the need for chemical derivatization of tryptic peptides prior to analysis. Marker ions for novel glycans, as well as a unique C-terminal flagellin peptide marker ion, were identified in a top-down analysis of the intact protein. These ions have the potential for use in for rapid detection and discrimination of C. botulinum cells, indicating botulinum neurotoxin contamination. This is the first report of glycosylation of Gram-positive flagellar proteins by the 'sialic acid-like' nonulosonate sugar, legionaminic acid.

  16. Temperature Dependences of Torque Generation and Membrane Voltage in the Bacterial Flagellar Motor

    PubMed Central

    Inoue, Yuichi; Baker, Matthew A.B.; Fukuoka, Hajime; Takahashi, Hiroto; Berry, Richard M.; Ishijima, Akihiko

    2013-01-01

    In their natural habitats bacteria are frequently exposed to sudden changes in temperature that have been shown to affect their swimming. With our believed to be new methods of rapid temperature control for single-molecule microscopy, we measured here the thermal response of the Na+-driven chimeric motor expressed in Escherichia coli cells. Motor torque at low load (0.35 μm bead) increased linearly with temperature, twofold between 15°C and 40°C, and torque at high load (1.0 μm bead) was independent of temperature, as reported for the H+-driven motor. Single cell membrane voltages were measured by fluorescence imaging and these were almost constant (∼120 mV) over the same temperature range. When the motor was heated above 40°C for 1–2 min the torque at high load dropped reversibly, recovering upon cooling below 40°C. This response was repeatable over as many as 10 heating cycles. Both increases and decreases in torque showed stepwise torque changes with unitary size ∼150 pN nm, close to the torque of a single stator at room temperature (∼180 pN nm), indicating that dynamic stator dissociation occurs at high temperature, with rebinding upon cooling. Our results suggest that the temperature-dependent assembly of stators is a general feature of flagellar motors. PMID:24359752

  17. Design of a comprehensive biochemistry and molecular biology experiment: phase variation caused by recombinational regulation of bacterial gene expression.

    PubMed

    Sheng, Xiumei; Xu, Shungao; Lu, Renyun; Isaac, Dadzie; Zhang, Xueyi; Zhang, Haifang; Wang, Huifang; Qiao, Zheng; Huang, Xinxiang

    2014-01-01

    Scientific experiments are indispensable parts of Biochemistry and Molecular Biology. In this study, a comprehensive Biochemistry and Molecular Biology experiment about Salmonella enterica serovar Typhi Flagellar phase variation has been designed. It consisted of three parts, namely, inducement of bacterial Flagellar phase variation, antibody agglutination test, and PCR analysis. Phase variation was observed by baterial motility assay and identified by antibody agglutination test and PCR analysis. This comprehensive experiment can be performed to help students improve their ability to use the knowledge acquired in Biochemistry and Molecular Biology. Copyright © 2014 by The International Union of Biochemistry and Molecular Biology.

  18. Evidence-based care bundles for preventing surgical site infections in spinal instrumentation surgery.

    PubMed

    Yamada, Koji; Abe, Hiroaki; Higashikawa, Akiro; Tonosu, Juichi; Kuniya, Takashi; Nakajima, Koji; Fujii, Haruko; Niwa, Kazuki; Shinozaki, Tomohiro; Watanabe, Kenichi; Sakae, Tanaka; Okazaki, Hiroshi

    2018-05-22

    Retrospective study, using prospectively collected data. To evaluate the impact of evidence-based care bundles for preventing surgical site infections (SSI) in spinal instrumentation surgery. About half of all SSIs are preventable via evidence-based methods. For successful SSI prevention, the bacterial load must be minimized, and methicillin-resistant Staphylococcus aureus (MRSA) protection must be maximized. However, it is difficult to cover all of these requirements by single preventative method. We screened consecutive patients scheduled for spinal instrumentation surgeries at a single tertiary referral hospital for high surgical, SSI, and MRSA colonization risks. Evidence-based care bundles were implemented for high risk patients and included: 1) additional vancomycin prophylaxis, 2) diluted povidone-iodine irrigation, and 3) nasal and body decontamination. Patient demographics, comorbidities, operative features, and SSIs reported to the Japanese Nosocomial Infections Surveillance system were prospectively obtained in the same method by the same assessor and were used for the analyses. The results were compared before and after the application of the bundle. There were 1,042 spinal instrumentation surgeries (741 before and 301 after care bundles) performed from November 2010 to December 2015. Of 301 surgeries, 57 cases (18.9%) received care bundles. There were no significant differences in patient backgrounds before and after the intervention. The SSI rate decreased significantly from 3.8% to 0.7% (P < 0.01) after the intervention, with an overall 82% relative risk reduction. A significant protective effect was observed in the multivariate analysis (adjusted odds ratio 0.18, 95% confidence interval: 0.04-0.77, P = 0.02). There were no MRSA-related SSIs among those that received care bundles, even though MRSA was the predominant pathogen in the study population. Evidence-based care bundles, applied in selected high-risk spinal instrumentation cases

  19. KHARON Is an Essential Cytoskeletal Protein Involved in the Trafficking of Flagellar Membrane Proteins and Cell Division in African Trypanosomes*

    PubMed Central

    Sanchez, Marco A.; Tran, Khoa D.; Valli, Jessica; Hobbs, Sam; Johnson, Errin; Gluenz, Eva; Landfear, Scott M.

    2016-01-01

    African trypanosomes and related kinetoplastid parasites selectively traffic specific membrane proteins to the flagellar membrane, but the mechanisms for this trafficking are poorly understood. We show here that KHARON, a protein originally identified in Leishmania parasites, interacts with a putative trypanosome calcium channel and is required for its targeting to the flagellar membrane. KHARON is located at the base of the flagellar axoneme, where it likely mediates targeting of flagellar membrane proteins, but is also on the subpellicular microtubules and the mitotic spindle. Hence, KHARON is probably a multifunctional protein that associates with several components of the trypanosome cytoskeleton. RNA interference-mediated knockdown of KHARON mRNA results in failure of the calcium channel to enter the flagellar membrane, detachment of the flagellum from the cell body, and disruption of mitotic spindles. Furthermore, knockdown of KHARON mRNA induces a lethal failure of cytokinesis in both bloodstream (mammalian host) and procyclic (insect vector) life cycle stages, and KHARON is thus critical for parasite viability. PMID:27489106

  20. Second-chance signal transduction explains cooperative flagellar switching.

    PubMed

    Zot, Henry G; Hasbun, Javier E; Minh, Nguyen Van

    2012-01-01

    The reversal of flagellar motion (switching) results from the interaction between a switch complex of the flagellar rotor and a torque-generating stationary unit, or stator (motor unit). To explain the steeply cooperative ligand-induced switching, present models propose allosteric interactions between subunits of the rotor, but do not address the possibility of a reaction that stimulates a bidirectional motor unit to reverse direction of torque. During flagellar motion, the binding of a ligand-bound switch complex at the dwell site could excite a motor unit. The probability that another switch complex of the rotor, moving according to steady-state rotation, will reach the same dwell site before that motor unit returns to ground state will be determined by the independent decay rate of the excited-state motor unit. Here, we derive an analytical expression for the energy coupling between a switch complex and a motor unit of the stator complex of a flagellum, and demonstrate that this model accounts for the cooperative switching response without the need for allosteric interactions. The analytical result can be reproduced by simulation when (1) the motion of the rotor delivers a subsequent ligand-bound switch to the excited motor unit, thereby providing the excited motor unit with a second chance to remain excited, and (2) the outputs from multiple independent motor units are constrained to a single all-or-none event. In this proposed model, a motor unit and switch complex represent the components of a mathematically defined signal transduction mechanism in which energy coupling is driven by steady-state and is regulated by stochastic ligand binding. Mathematical derivation of the model shows the analytical function to be a general form of the Hill equation (Hill AV (1910) The possible effects of the aggregation of the molecules of haemoglobin on its dissociation curves. J Physiol 40: iv-vii).

  1. Constraints on models for the flagellar rotary motor.

    PubMed Central

    Berg, H C

    2000-01-01

    Most bacteria that swim are propelled by flagellar filaments, each driven at its base by a rotary motor embedded in the cell wall and cytoplasmic membrane. A motor is about 45 nm in diameter and made up of about 20 different kinds of parts. It is assembled from the inside out. It is powered by a proton (or in some species, a sodium-ion) flux. It steps at least 400 times per revolution. At low speeds and high torques, about 1000 protons are required per revolution, speed is proportional to protonmotive force, and torque varies little with temperature or hydrogen isotope. At high speeds and low torques, torque increases with temperature and is sensitive to hydrogen isotope. At room temperature, torque varies remarkably little with speed from about -100 Hz (the present limit of measurement) to about 200 Hz, and then it declines rapidly reaching zero at about 300 Hz. These are facts that motor models should explain. None of the existing models for the flagellar rotary motor completely do so. PMID:10836502

  2. MotD of Sinorhizobium meliloti and Related α-Proteobacteria Is the Flagellar-Hook-Length Regulator and Therefore Reassigned as FliK

    PubMed Central

    Eggenhofer, Elke; Rachel, Reinhard; Haslbeck, Martin; Scharf, Birgit

    2006-01-01

    The flagella of the soil bacterium Sinorhizobium meliloti differ from the enterobacterial paradigm in the complex filament structure and modulation of the flagellar rotary speed. The mode of motility control in S. meliloti has a molecular corollary in two novel periplasmic motility proteins, MotC and MotE, that are present in addition to the ubiquitous MotA/MotB energizing proton channel. A fifth motility gene is located in the mot operon downstream of the motB and motC genes. Its gene product was originally designated MotD, a cytoplasmic motility protein having an unknown function. We report here reassignment of MotD as FliK, the regulator of flagellar hook length. The FliK gene is one of the few flagellar genes not annotated in the contiguous flagellar regulon of S. meliloti. Characteristic for its class, the 475-residue FliK protein contains a conserved, compactly folded Flg hook domain in its carboxy-terminal region. Deletion of fliK leads to formation of prolonged flagellar hooks (polyhooks) with missing filament structures. Extragenic suppressor mutations all mapped in the cytoplasmic region of the transmembrane export protein FlhB and restored assembly of a flagellar filament, and thus motility, in the presence of polyhooks. The structural properties of FliK are consistent with its function as a substrate specificity switch of the flagellar export apparatus for switching from rod/hook-type substrates to filament-type substrates. PMID:16513744

  3. Human sperm steer with second harmonics of the flagellar beat.

    PubMed

    Saggiorato, Guglielmo; Alvarez, Luis; Jikeli, Jan F; Kaupp, U Benjamin; Gompper, Gerhard; Elgeti, Jens

    2017-11-10

    Sperm are propelled by bending waves traveling along their flagellum. For steering in gradients of sensory cues, sperm adjust the flagellar waveform. Symmetric and asymmetric waveforms result in straight and curved swimming paths, respectively. Two mechanisms causing spatially asymmetric waveforms have been proposed: an average flagellar curvature and buckling. We image flagella of human sperm tethered with the head to a surface. The waveform is characterized by a fundamental beat frequency and its second harmonic. The superposition of harmonics breaks the beat symmetry temporally rather than spatially. As a result, sperm rotate around the tethering point. The rotation velocity is determined by the second-harmonic amplitude and phase. Stimulation with the female sex hormone progesterone enhances the second-harmonic contribution and, thereby, modulates sperm rotation. Higher beat frequency components exist in other flagellated cells; therefore, this steering mechanism might be widespread and could inspire the design of synthetic microswimmers.

  4. Flagellar central pair assembly in Chlamydomonas reinhardtii

    PubMed Central

    2013-01-01

    Background Most motile cilia and flagella have nine outer doublet and two central pair (CP) microtubules. Outer doublet microtubules are continuous with the triplet microtubules of the basal body, are templated by the basal body microtubules, and grow by addition of new subunits to their distal (“plus”) ends. In contrast, CP microtubules are not continuous with basal body microtubules, raising the question of how these microtubules are assembled and how their polarity is established. Methods CP assembly in Chlamydomonas reinhardtii was analyzed by electron microscopy and wide-field and super-resolution immunofluorescence microscopy. To analyze CP assembly independently from flagellar assembly, the CP-deficient katanin mutants pf15 or pf19 were mated to wild-type cells. HA-tagged tubulin and the CP-specific protein hydin were used as markers to analyze de novo CP assembly inside the formerly mutant flagella. Results In regenerating flagella, the CP and its projections assemble near the transition zone soon after the onset of outer doublet elongation. During de novo CP assembly in full-length flagella, the nascent CP was first apparent in a subdistal region of the flagellum. The developing CP replaces a fibrous core that fills the axonemal lumen of CP-deficient flagella. The fibrous core contains proteins normally associated with the C1 CP microtubule and proteins involved in intraflagellar transport (IFT). In flagella of the radial spoke-deficient mutant pf14, two pairs of CPs are frequently present with identical correct polarities. Conclusions The temporal separation of flagellar and CP assembly in dikaryons formed by mating CP-deficient gametes to wild-type gametes revealed that the formation of the CP does not require proximity to the basal body or transition zone, or to the flagellar tip. The observations on pf14 provide further support that the CP self-assembles without a template and eliminate the possibility that CP polarity is established by interaction

  5. Two flagellar BAR domain proteins in Trypanosoma brucei with stage-specific regulation

    PubMed Central

    Cicova, Zdenka; Dejung, Mario; Skalicky, Tomas; Eisenhuth, Nicole; Hanselmann, Steffen; Morriswood, Brooke; Figueiredo, Luisa M.; Butter, Falk; Janzen, Christian J.

    2016-01-01

    Trypanosomes are masters of adaptation to different host environments during their complex life cycle. Large-scale proteomic approaches provide information on changes at the cellular level, and in a systematic way. However, detailed work on single components is necessary to understand the adaptation mechanisms on a molecular level. Here, we have performed a detailed characterization of a bloodstream form (BSF) stage-specific putative flagellar host adaptation factor Tb927.11.2400, identified previously in a SILAC-based comparative proteome study. Tb927.11.2400 shares 38% amino acid identity with TbFlabarin (Tb927.11.2410), a procyclic form (PCF) stage-specific flagellar BAR domain protein. We named Tb927.11.2400 TbFlabarin-like (TbFlabarinL), and demonstrate that it originates from a gene duplication event, which occurred in the African trypanosomes. TbFlabarinL is not essential for the growth of the parasites under cell culture conditions and it is dispensable for developmental differentiation from BSF to the PCF in vitro. We generated TbFlabarinL-specific antibodies, and showed that it localizes in the flagellum. Co-immunoprecipitation experiments together with a biochemical cell fractionation suggest a dual association of TbFlabarinL with the flagellar membrane and the components of the paraflagellar rod. PMID:27779220

  6. Genetic dissection of the consensus sequence for the class 2 and class 3 flagellar promoters

    PubMed Central

    Wozniak, Christopher E.; Hughes, Kelly T.

    2008-01-01

    Summary Computational searches for DNA binding sites often utilize consensus sequences. These search models make assumptions that the frequency of a base pair in an alignment relates to the base pair’s importance in binding and presume that base pairs contribute independently to the overall interaction with the DNA binding protein. These two assumptions have generally been found to be accurate for DNA binding sites. However, these assumptions are often not satisfied for promoters, which are involved in additional steps in transcription initiation after RNA polymerase has bound to the DNA. To test these assumptions for the flagellar regulatory hierarchy, class 2 and class 3 flagellar promoters were randomly mutagenized in Salmonella. Important positions were then saturated for mutagenesis and compared to scores calculated from the consensus sequence. Double mutants were constructed to determine how mutations combined for each promoter type. Mutations in the binding site for FlhD4C2, the activator of class 2 promoters, better satisfied the assumptions for the binding model than did mutations in the class 3 promoter, which is recognized by the σ28 transcription factor. These in vivo results indicate that the activator sites within flagellar promoters can be modeled using simple assumptions but that the DNA sequences recognized by the flagellar sigma factor require more complex models. PMID:18486950

  7. Modulation of Chlamydomonas reinhardtii flagellar motility by redox poise

    PubMed Central

    Wakabayashi, Ken-ichi; King, Stephen M.

    2006-01-01

    Redox-based regulatory systems are essential for many cellular activities. Chlamydomonas reinhardtii exhibits alterations in motile behavior in response to different light conditions (photokinesis). We hypothesized that photokinesis is signaled by variations in cytoplasmic redox poise resulting from changes in chloroplast activity. We found that this effect requires photosystem I, which generates reduced NADPH. We also observed that photokinetic changes in beat frequency and duration of the photophobic response could be obtained by altering oxidative/reductive stress. Analysis of reactivated cell models revealed that this redox poise effect is mediated through the outer dynein arms (ODAs). Although the global redox state of the thioredoxin-related ODA light chains LC3 and LC5 and the redox-sensitive Ca2+-binding subunit of the docking complex DC3 did not change upon light/dark transitions, we did observe significant alterations in their interactions with other flagellar components via mixed disulfides. These data indicate that redox poise directly affects ODAs and suggest that it may act in the control of flagellar motility. PMID:16754958

  8. The flagellar apparatus of heteroloboseans.

    PubMed

    Brugerolle, Guy; Simpson, Alastair G B

    2004-01-01

    The flagellar apparatus of four heterolobosean species Percolomonas descissus, Percolomonas sulcatus, Tetramitus rostratus, and Naegleria gruberi were examined. P. descissus lives in oxygen-poor water. It is a quadriflagellated cell with a ventral groove. The two pairs of basal bodies are connected to an apical structure from which the peripheral dorso-lateral microtubules and a short striated rhizoplast originate. There is one major microtubular root, R1, which originates from the posterior basal body pair and splits into left and right portions that support the sides of the ventral groove. The anterior pair of basal bodies is associated with a root of four to five microtubules that runs to the left of the groove. This organisation is similar to that previously reported for Psalteriomonas, Lyromonas, and Percolomonas cosmopolitus. Percolomonas sulcatus has two parallel pairs of basal bodies, each of which is associated with a well-developed R1 root. These roots divide to give two distinct left portions and one merged right portion that support the margins of the slit-like ventral groove. Tetramitus rostratus has two pairs of basal bodies, several rhizoplast fibres, and two R1 roots. Each R1 root supports one wall of the ventral groove. Naegleria gruberi may have two pairs of basal bodies, each associated with a microtubular root and one long rhizoplast fibre. From available data, a 'double bikont'-like organisation of the heterolobosean flagellar apparatus is inferred, where both of the eldest basal bodies have largely 'mature' complements of microtubular roots. The cytoskeletal organisation of heteroloboseans is compared to those of (other) excavates. Our structural data and existing molecular phylogenies weaken the case that Percolomonas, Psalteriomonas, and Lyromonas are phylogenetically separable from other heteroloboseans, undermining many of the highest-level taxa proposed for these organisms, including Percolozoa, Striatorhiza, Percolomonada, Percolomonadea

  9. A chemotactic signaling surface on CheY defined by suppressors of flagellar switch mutations.

    PubMed Central

    Roman, S J; Meyers, M; Volz, K; Matsumura, P

    1992-01-01

    CheY is the response regulator protein that interacts with the flagellar switch apparatus to modulate flagellar rotation during chemotactic signaling. CheY can be phosphorylated and dephosphorylated in vitro, and evidence indicates that CheY-P is the activated form that induces clockwise flagellar rotation, resulting in a tumble in the cell's swimming pattern. The flagellar switch apparatus is a complex macromolecular structure composed of at least three gene products, FliG, FliM, and FliN. Genetic analysis of Escherichia coli has identified fliG and fliM as genes in which mutations occur that allele specifically suppress cheY mutations, indicating interactions among these gene products. We have generated a class of cheY mutations selected for dominant suppression of fliG mutations. Interestingly, these cheY mutations dominantly suppressed both fliG and fliM mutations; this is consistent with the idea that the CheY protein interacts with both switch gene products during signaling. Biochemical characterization of wild-type and suppressor CheY proteins did not reveal altered phosphorylation properties or evidence for phosphorylation-dependent CheY multimerization. These data indicate that suppressor CheY proteins are specifically altered in the ability to transduce chemotactic signals to the switch at some point subsequent to phosphorylation. Physical mapping of suppressor amino acid substitutions on the crystal structure of CheY revealed a high degree of spatial clustering, suggesting that this region of CheY is a signaling surface that transduces chemotactic signals to the switch. Images PMID:1400175

  10. Effective viscosity of a suspension of flagellar-beating microswimmers: Three-dimensional modeling

    NASA Astrophysics Data System (ADS)

    Jibuti, Levan; Zimmermann, Walter; Rafaï, Salima; Peyla, Philippe

    2017-11-01

    Micro-organisms usually can swim in their liquid environment by flagellar or ciliary beating. In this numerical work, we analyze the influence of flagellar beating on the orbits of a swimming cell in a shear flow. We also calculate the effect of the flagellar beating on the rheology of a dilute suspension of microswimmers. A three-dimensional model is proposed for Chlamydomonas Reinhardtii swimming with a breaststroke-like beating of two anterior flagella modeled by two counter-rotating fore beads. The active swimmer model reveals unusual angular orbits in a linear shear flow. Namely, the swimmer sustains orientations transiently across the flow. Such behavior is a result of the interplay between shear flow and the swimmer's periodic beating motion of flagella, which exert internal torques on the cell body. This peculiar behavior has some significant consequences on the rheological properties of the suspension. We calculate Einstein's viscosity of the suspension composed of such isolated modeled microswimmers (dilute case) in a shear flow. We use numerical simulations based on a Rotne-Prager-like approximation for hydrodynamic interaction between simplified flagella and the cell body. The results show an increased intrinsic viscosity for active swimmer suspensions in comparison to nonactive ones as well as a shear thinning behavior in accordance with previous experimental measurements [Phys. Rev. Lett. 104, 098102 (2010), 10.1103/PhysRevLett.104.098102].

  11. The C-terminal periplasmic domain of MotB is responsible for load-dependent control of the number of stators of the bacterial flagellar motor.

    PubMed

    Castillo, David J; Nakamura, Shuichi; Morimoto, Yusuke V; Che, Yong-Suk; Kami-Ike, Nobunori; Kudo, Seishi; Minamino, Tohru; Namba, Keiichi

    2013-01-01

    The bacterial flagellar motor is made of a rotor and stators. In Salmonella it is thought that about a dozen MotA/B complexes are anchored to the peptidoglycan layer around the motor through the C-terminal peptidoglycan-binding domain of MotB to become active stators as well as proton channels. MotB consists of 309 residues, forming a single transmembrane helix (30-50), a stalk (51-100) and a C-terminal peptidoglycan-binding domain (101-309). Although the stalk is dispensable for torque generation by the motor, it is required for efficient motor performance. Residues 51 to 72 prevent premature proton leakage through the proton channel prior to stator assembly into the motor. However, the role of residues 72-100 remains unknown. Here, we analyzed the torque-speed relationship of the MotB(Δ72-100) motor. At a low speed near stall, this mutant motor produced torque at the wild-type level. Unlike the wild-type motor, however, torque dropped off drastically by slight decrease in external load and then showed a slow exponential decay over a wide range of load by its further reduction. Since it is known that the stator is a mechano-sensor and that the number of active stators changes in a load-dependent manner, we interpreted this unusual torque-speed relationship as anomaly in load-dependent control of the number of active stators. The results suggest that residues 72-100 of MotB is required for proper load-dependent control of the number of active stators around the rotor.

  12. The role of the dynein light intermediate chain in retrograde IFT and flagellar function in Chlamydomonas

    PubMed Central

    Reck, Jaimee; Schauer, Alexandria M.; VanderWaal Mills, Kristyn; Bower, Raqual; Tritschler, Douglas; Perrone, Catherine A.; Porter, Mary E.

    2016-01-01

    The assembly of cilia and flagella depends on the activity of two microtubule motor complexes, kinesin-2 and dynein-2/1b, but the specific functions of the different subunits are poorly defined. Here we analyze Chlamydomonas strains expressing different amounts of the dynein 1b light intermediate chain (D1bLIC). Disruption of D1bLIC alters the stability of the dynein 1b complex and reduces both the frequency and velocity of retrograde intraflagellar transport (IFT), but it does not eliminate retrograde IFT. Flagellar assembly, motility, gliding, and mating are altered in a dose-dependent manner. iTRAQ-based proteomics identifies a small subset of proteins that are significantly reduced or elevated in d1blic flagella. Transformation with D1bLIC-GFP rescues the mutant phenotypes, and D1bLIC-GFP assembles into the dynein 1b complex at wild-type levels. D1bLIC-GFP is transported with anterograde IFT particles to the flagellar tip, dissociates into smaller particles, and begins processive retrograde IFT in <2 s. These studies demonstrate the role of D1bLIC in facilitating the recycling of IFT subunits and other proteins, identify new components potentially involved in the regulation of IFT, flagellar assembly, and flagellar signaling, and provide insight into the role of D1bLIC and retrograde IFT in other organisms. PMID:27251063

  13. Anatomical approach to permanent His bundle pacing: Optimizing His bundle capture.

    PubMed

    Vijayaraman, Pugazhendhi; Dandamudi, Gopi

    2016-01-01

    Permanent His bundle pacing is a physiological alternative to right ventricular pacing. In this article we describe our approach to His bundle pacing in patients with AV nodal and intra-Hisian conduction disease. It is essential for the implanters to understand the anatomic variations of the His bundle course and its effect on the type of His bundle pacing achieved. We describe several case examples to illustrate our anatomical approach to permanent His bundle pacing in this article. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Spirochete motility and morpholgy

    NASA Astrophysics Data System (ADS)

    Charon, Nyles

    2004-03-01

    . burgdorferi during chemotaxis. In translational motility, the bundles of periplasmic flagella rotate in opposite directions. When not translating, they rotate in the same direction, and the cells flex. We present evidence that asymmetrical rotation of the bundles during translation does not depend upon the chemotaxis signal transduction system. The histidine kinase CheA is known to be an essential component in the signaling pathway for bacterial chemotaxis. Mutants of cheA in flagellated bacteria continually rotate their flagella in one direction. B. burgdorferi has two copies of cheA. We reasoned that if chemotaxis were essential for asymmetrical rotation of the flagellar bundles, and if the flagellar motors at both cell ends were identical, inactivation of the two cheA genes should result in cells that constant flex. To test this hypothesis, the signaling pathway was completely blocked by construction of a double cheA mutant. This mutant was completely deficient in chemotaxis. Rather than flexing, it failed to reverse, and it continually translated only in one direction. The results indicate that asymmetrical rotation does not depend upon the chemotaxis system but rather upon differences between the two flagellar bundles. We propose that certain factors within the spirochete localize at flagellar motors at one end of the cell to effect this asymmetry (3). References: 1. Charon, N.W. and S.F. Goldstein. 2002. The genetics of motility and chemotaxis of a fascinating group of bacteria: the spirochetes. Ann. Rev. Genetics. 36: 47-73. 2. Motaleb M.A., L. Corum, J.L Bono, A.F. Elias, P. Rosa, D.S. Samuels, N.W. Charon. 2000. Borrelia burgdorferi periplasmic flagella have both skeletal and motility functions. Proc Natl Acad Sci. 2000 97:10899-10904. 3. Li, C. R. Bakker, M. Motaleb, F. Cabello, M.L. Sartakova, and N.W. Charon. 2002. Asymmetrical flagellar rotation in Borrelia burgdorferi non-chemotaxis mutants. Proc. Natl. Acad. Sci. 99:6169-6174.

  15. Magnetic Actuation of Self-assembled Bacteria Inspired Nanoswimmers

    NASA Astrophysics Data System (ADS)

    Ali, Jamel; Cheang, U. Kei; Martindale, James D.; Jabbarzadeh, Mehdi; Fu, Henry C.; Kim, Min Jun

    2017-11-01

    Currently, there is growing interest in developing nanoscale swimmers for biological and biomedical tasks. Of particular interest is the development of soft stimuli-responsive nanorobots to probe cellular and sub-cellular environments. While there have been a few reports of nanoscale robotic swimmers, which have shown potential to be used for these tasks, they often lack multifuctionality. In particular, no man-made soft nanoscale material has been able to match the ability of natural bacterial flagella to undergo rapid and reversible morphological changes in response to multiple forms of environmental stimuli. Towards this end, we report self-assembled stimuli-responsive nanoscale robotic swimmers composed of single or multiple bacterial flagella and attached to magnetic nanoparticles. We visualize the movement of flagella using high resolution fluorescence microscopy while controlling these swimmers via a magnetic control system. Differences in in propulsion before and after the change in flagellar form are observed. Furthermore, we demonstrate the ability to induce flagellar bundling in multiflagellated nanoswimmers. This work was funded by the National Science Foundation (DMR 1712061 and CMMI 1737682 to M.J.K. and DMR 1650970 and CBET 1651031 to H.C.F.), and the Korea Evaluation Institute of Industrial Technology (MOTIE) (NO. 10052980) award to M.J.K.

  16. Polyelectrolyte bundles

    NASA Astrophysics Data System (ADS)

    Limbach, H. J.; Sayar, M.; Holm, C.

    2004-06-01

    Using extensive Molecular Dynamics simulations we study the behavior of polyelectrolytes with hydrophobic side chains, which are known to form cylindrical micelles in aqueous solution. We investigate the stability of such bundles with respect to hydrophobicity, the strength of the electrostatic interaction, and the bundle size. We show that for the parameter range relevant for sulfonated poly-para-phenylenes (PPP) one finds a stable finite bundle size. In a more generic model we also show the influence of the length of the precursor oligomer on the stability of the bundles. We also point out that our model has close similarities to DNA solutions with added condensing agents, hinting to the possibility that the size of DNA aggregates is under certain circumstances thermodynamically limited.

  17. An Element of Determinism in a Stochastic Flagellar Motor Switch

    PubMed Central

    Xie, Li; Altindal, Tuba; Wu, Xiao-Lun

    2015-01-01

    Marine bacterium Vibrio alginolyticus uses a single polar flagellum to navigate in an aqueous environment. Similar to Escherichia coli cells, the polar flagellar motor has two states; when the motor is counter-clockwise, the cell swims forward and when the motor is clockwise, the cell swims backward. V. alginolyticus also incorporates a direction randomization step at the start of the forward swimming interval by flicking its flagellum. To gain an understanding on how the polar flagellar motor switch is regulated, distributions of the forward Δf and backward Δb intervals are investigated herein. We found that the steady-state probability density functions, P(Δf) and P(Δb), of freely swimming bacteria are strongly peaked at a finite time, suggesting that the motor switch is not Poissonian. The short-time inhibition is sufficiently strong and long lasting, i.e., several hundred milliseconds for both intervals, which is readily observed and characterized. Treating motor reversal dynamics as a first-passage problem, which results from conformation fluctuations of the motor switch, we calculated P(Δf) and P(Δb) and found good agreement with the measurements. PMID:26554590

  18. An Element of Determinism in a Stochastic Flagellar Motor Switch.

    PubMed

    Xie, Li; Altindal, Tuba; Wu, Xiao-Lun

    2015-01-01

    Marine bacterium Vibrio alginolyticus uses a single polar flagellum to navigate in an aqueous environment. Similar to Escherichia coli cells, the polar flagellar motor has two states; when the motor is counter-clockwise, the cell swims forward and when the motor is clockwise, the cell swims backward. V. alginolyticus also incorporates a direction randomization step at the start of the forward swimming interval by flicking its flagellum. To gain an understanding on how the polar flagellar motor switch is regulated, distributions of the forward Δf and backward Δb intervals are investigated herein. We found that the steady-state probability density functions, P(Δf) and P(Δb), of freely swimming bacteria are strongly peaked at a finite time, suggesting that the motor switch is not Poissonian. The short-time inhibition is sufficiently strong and long lasting, i.e., several hundred milliseconds for both intervals, which is readily observed and characterized. Treating motor reversal dynamics as a first-passage problem, which results from conformation fluctuations of the motor switch, we calculated P(Δf) and P(Δb) and found good agreement with the measurements.

  19. Emergence of flagellar beating from the collective behavior of individual ATP-powered dyneins

    NASA Astrophysics Data System (ADS)

    Namdeo, S.; Onck, P. R.

    2016-10-01

    Flagella are hair-like projections from the surface of eukaryotic cells, and they play an important role in many cellular functions, such as cell-motility. The beating of flagella is enabled by their internal architecture, the axoneme, and is powered by a dense distribution of motor proteins, dyneins. The dyneins deliver the required mechanical work through the hydrolysis of ATP. Although the dynein-ATP cycle, the axoneme microstructure, and the flagellar-beating kinematics are well studied, their integration into a coherent picture of ATP-powered flagellar beating is still lacking. Here we show that a time-delayed negative-work-based switching mechanism is able to convert the individual sliding action of hundreds of dyneins into a regular overall beating pattern leading to propulsion. We developed a computational model based on a minimal representation of the axoneme consisting of two representative doublet microtubules connected by nexin links. The relative sliding of the microtubules is incorporated by modeling two groups of ATP-powered dyneins, each responsible for sliding in opposite directions. A time-delayed switching mechanism is postulated, which is key in converting the local individual sliding action of multiple dyneins into global beating. Our results demonstrate that an overall nonreciprocal beating pattern can emerge with time due to the spatial and temporal coordination of the individual dyneins. These findings provide insights in the fundamental working mechanism of axonemal dyneins and could possibly open new research directions in the field of flagellar motility.

  20. Emergence of flagellar beating from the collective behavior of individual ATP-powered dyneins.

    PubMed

    Namdeo, S; Onck, P R

    2016-10-01

    Flagella are hair-like projections from the surface of eukaryotic cells, and they play an important role in many cellular functions, such as cell-motility. The beating of flagella is enabled by their internal architecture, the axoneme, and is powered by a dense distribution of motor proteins, dyneins. The dyneins deliver the required mechanical work through the hydrolysis of ATP. Although the dynein-ATP cycle, the axoneme microstructure, and the flagellar-beating kinematics are well studied, their integration into a coherent picture of ATP-powered flagellar beating is still lacking. Here we show that a time-delayed negative-work-based switching mechanism is able to convert the individual sliding action of hundreds of dyneins into a regular overall beating pattern leading to propulsion. We developed a computational model based on a minimal representation of the axoneme consisting of two representative doublet microtubules connected by nexin links. The relative sliding of the microtubules is incorporated by modeling two groups of ATP-powered dyneins, each responsible for sliding in opposite directions. A time-delayed switching mechanism is postulated, which is key in converting the local individual sliding action of multiple dyneins into global beating. Our results demonstrate that an overall nonreciprocal beating pattern can emerge with time due to the spatial and temporal coordination of the individual dyneins. These findings provide insights in the fundamental working mechanism of axonemal dyneins and could possibly open new research directions in the field of flagellar motility.

  1. Polycation induced actin bundles.

    PubMed

    Muhlrad, Andras; Grintsevich, Elena E; Reisler, Emil

    2011-04-01

    Three polycations, polylysine, the polyamine spermine and the polycationic protein lysozyme were used to study the formation, structure, ionic strength sensitivity and dissociation of polycation-induced actin bundles. Bundles form fast, simultaneously with the polymerization of MgATP-G-actins, upon the addition of polycations to solutions of actins at low ionic strength conditions. This indicates that nuclei and/or nascent filaments bundle due to attractive, electrostatic effect of polycations and the neutralization of repulsive interactions of negative charges on actin. The attractive forces between the filaments are strong, as shown by the low (in nanomolar range) critical concentration of their bundling at low ionic strength. These bundles are sensitive to ionic strength and disassemble partially in 100 mM NaCl, but both the dissociation and ionic strength sensitivity can be countered by higher polycation concentrations. Cys374 residues of actin monomers residing on neighboring filaments in the bundles can be cross-linked by the short span (5.4Å) MTS-1 (1,1-methanedyl bismethanethiosulfonate) cross-linker, which indicates a tight packing of filaments in the bundles. The interfilament cross-links, which connect monomers located on oppositely oriented filaments, prevent disassembly of bundles at high ionic strength. Cofilin and the polysaccharide polyanion heparin disassemble lysozyme induced actin bundles more effectively than the polylysine-induced bundles. The actin-lysozyme bundles are pathologically significant as both proteins are found in the pulmonary airways of cystic fibrosis patients. Their bundles contribute to the formation of viscous mucus, which is the main cause of breathing difficulties and eventual death in this disorder. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Flagellar filament bio-templated inorganic oxide materials - towards an efficient lithium battery anode

    NASA Astrophysics Data System (ADS)

    Beznosov, Sergei N.; Veluri, Pavan S.; Pyatibratov, Mikhail G.; Chatterjee, Abhijit; Macfarlane, Douglas R.; Fedorov, Oleg V.; Mitra, Sagar

    2015-01-01

    Designing a new generation of energy-intensive and sustainable electrode materials for batteries to power a variety of applications is an imperative task. The use of biomaterials as a nanosized structural template for these materials has the potential to produce hitherto unachievable structures. In this report, we have used genetically modified flagellar filaments of the extremely halophilic archaea species Halobacterium salinarum to synthesize nanostructured iron oxide composites for use as a lithium-ion battery anode. The electrode demonstrated a superior electrochemical performance compared to existing literature results, with good capacity retention of 1032 mAh g-1 after 50 cycles and with high rate capability, delivering 770 mAh g-1 at 5 A g-1 (~5 C) discharge rate. This unique flagellar filament based template has the potential to provide access to other highly structured advanced energy materials in the future.

  3. Polymorphism in Bacterial Flagella Suspensions

    NASA Astrophysics Data System (ADS)

    Schwenger, Walter J.

    Bacterial flagella are a type of biological polymer studied for its role in bacterial motility and the polymorphic transitions undertaken to facilitate the run and tumble behavior. The naturally rigid, helical shape of flagella gives rise to novel colloidal dynamics and material properties. This thesis studies methods in which the shape of bacterial flagella can be controlled using in vitro methods and the changes the shape of the flagella have on both single particle dynamics and bulk material properties. We observe individual flagellum in both the dilute and semidilute regimes to observe the effects of solvent condition on the shape of the filament as well as the effect the filament morphology has on reptation through a network of flagella. In addition, we present rheological measurements showing how the shape of filaments effects the bulk material properties of flagellar suspensions. We find that the individual particle dynamics in suspensions of flagella can vary with geometry from needing to reptate linearly via rotation for helical filaments to the prevention of long range diffusion for block copolymer filaments. Similarly, for bulk material properties of flagella suspensions, helical geometries show a dramatic enhancement in elasticity over straight filaments while block copolymers form an elastic gel without the aid of crosslinking agents.

  4. Characterization of a Chlamydomonas Insertional Mutant that Disrupts Flagellar Central Pair Microtubule-associated Structures

    PubMed Central

    Mitchell, David R.; Sale, Winfield S.

    1999-01-01

    Two alleles at a new locus, central pair–associated complex 1 (CPC1), were selected in a screen for Chlamydomonas flagellar motility mutations. These mutations disrupt structures associated with central pair microtubules and reduce flagellar beat frequency, but do not prevent changes in flagellar activity associated with either photophobic responses or phototactic accumulation of live cells. Comparison of cpc1 and pf6 axonemes shows that cpc1 affects a row of projections along C1 microtubules distinct from those missing in pf6, and a row of thin fibers that form an arc between the two central pair microtubules. Electron microscopic images of the central pair in axonemes from radial spoke–defective strains reveal previously undescribed central pair structures, including projections extending laterally toward radial spoke heads, and a diagonal link between the C2 microtubule and the cpc1 projection. By SDS-PAGE, cpc1 axonemes show reductions of 350-, 265-, and 79-kD proteins. When extracted from wild-type axonemes, these three proteins cosediment on sucrose gradients with three other central pair proteins (135, 125, and 56 kD) in a 16S complex. Characterization of cpc1 provides new insights into the structure and biochemistry of the central pair apparatus, and into its function as a regulator of dynein-based motility. PMID:9922455

  5. Structure of the torque ring of the flagellar motor and the molecular basis for rotational switching.

    PubMed

    Lee, Lawrence K; Ginsburg, Michael A; Crovace, Claudia; Donohoe, Mhairi; Stock, Daniela

    2010-08-19

    The flagellar motor drives the rotation of flagellar filaments at hundreds of revolutions per second, efficiently propelling bacteria through viscous media. The motor uses the potential energy from an electrochemical gradient of cations across the cytoplasmic membrane to generate torque. A rapid switch from anticlockwise to clockwise rotation determines whether a bacterium runs smoothly forward or tumbles to change its trajectory. A protein called FliG forms a ring in the rotor of the flagellar motor that is involved in the generation of torque through an interaction with the cation-channel-forming stator subunit MotA. FliG has been suggested to adopt distinct conformations that induce switching but these structural changes and the molecular mechanism of switching are unknown. Here we report the molecular structure of the full-length FliG protein, identify conformational changes that are involved in rotational switching and uncover the structural basis for the formation of the FliG torque ring. This allows us to propose a model of the complete ring and switching mechanism in which conformational changes in FliG reverse the electrostatic charges involved in torque generation.

  6. The Armadillo Repeat Protein PF16 Is Essential for Flagellar Structure and Function in Plasmodium Male Gametes

    PubMed Central

    Ferguson, David J. P.; Bunting, Karen A.; Xu, Zhengyao; Bailes, Elizabeth; Sinden, Robert E.; Holder, Anthony A.; Smith, Elizabeth F.; Coates, Juliet C.; Rita Tewari

    2010-01-01

    Malaria, caused by the apicomplexan parasite Plasmodium, threatens 40% of the world's population. Transmission between vertebrate and insect hosts depends on the sexual stages of the life-cycle. The male gamete of Plasmodium parasite is the only developmental stage that possesses a flagellum. Very little is known about the identity or function of proteins in the parasite's flagellar biology. Here, we characterise a Plasmodium PF16 homologue using reverse genetics in the mouse malaria parasite Plasmodium berghei. PF16 is a conserved Armadillo-repeat protein that regulates flagellar structure and motility in organisms as diverse as green algae and mice. We show that P. berghei PF16 is expressed in the male gamete flagellum, where it plays a crucial role maintaining the correct microtubule structure in the central apparatus of the axoneme as studied by electron microscopy. Disruption of the PF16 gene results in abnormal flagellar movement and reduced fertility, but does not lead to complete sterility, unlike pf16 mutations in other organisms. Using homology modelling, bioinformatics analysis and complementation studies in Chlamydomonas, we show that some regions of the PF16 protein are highly conserved across all eukaryotes, whereas other regions may have species-specific functions. PF16 is the first ARM-repeat protein characterised in the malaria parasite genus Plasmodium and this study opens up a novel model for analysis of Plasmodium flagellar biology that may provide unique insights into an ancient organelle and suggest novel intervention strategies to control the malaria parasite. PMID:20886115

  7. Flagellar filament bio-templated inorganic oxide materials - towards an efficient lithium battery anode.

    PubMed

    Beznosov, Sergei N; Veluri, Pavan S; Pyatibratov, Mikhail G; Chatterjee, Abhijit; MacFarlane, Douglas R; Fedorov, Oleg V; Mitra, Sagar

    2015-01-13

    Designing a new generation of energy-intensive and sustainable electrode materials for batteries to power a variety of applications is an imperative task. The use of biomaterials as a nanosized structural template for these materials has the potential to produce hitherto unachievable structures. In this report, we have used genetically modified flagellar filaments of the extremely halophilic archaea species Halobacterium salinarum to synthesize nanostructured iron oxide composites for use as a lithium-ion battery anode. The electrode demonstrated a superior electrochemical performance compared to existing literature results, with good capacity retention of 1032 mAh g(-1) after 50 cycles and with high rate capability, delivering 770 mAh g(-1) at 5 A g(-1) (~5 C) discharge rate. This unique flagellar filament based template has the potential to provide access to other highly structured advanced energy materials in the future.

  8. Flagellar oscillation: a commentary on proposed mechanisms.

    PubMed

    Woolley, David M

    2010-08-01

    Eukaryotic flagella and cilia have a remarkably uniform internal 'engine' known as the '9+2' axoneme. With few exceptions, the function of cilia and flagella is to beat rhythmically and set up relative motion between themselves and the liquid that surrounds them. The molecular basis of axonemal movement is understood in considerable detail, with the exception of the mechanism that provides its rhythmical or oscillatory quality. Some kind of repetitive 'switching' event is assumed to occur; there are several proposals regarding the nature of the 'switch' and how it might operate. Herein I first summarise all the factors known to influence the rate of the oscillation (the beating frequency). Many of these factors exert their effect through modulating the mean sliding velocity between the nine doublet microtubules of the axoneme, this velocity being the determinant of bend growth rate and bend propagation rate. Then I explain six proposed mechanisms for flagellar oscillation and review the evidence on which they are based. Finally, I attempt to derive an economical synthesis, drawing for preference on experimental research that has been minimally disruptive of the intricate structure of the axoneme. The 'provisional synthesis' is that flagellar oscillation emerges from an effect of passive sliding direction on the dynein arms. Sliding in one direction facilitates force-generating cycles and dynein-to-dynein synchronisation along a doublet; sliding in the other direction is inhibitory. The direction of the initial passive sliding normally oscillates because it is controlled hydrodynamically through the alternating direction of the propulsive thrust. However, in the absence of such regulation, there can be a perpetual, mechanical self-triggering through a reversal of sliding direction due to the recoil of elastic structures that deform as a response to the prior active sliding. This provisional synthesis may be a useful basis for further examination of the problem.

  9. Purification and Characterization of the Bacterial Flagellar Basal Body from Salmonella enterica.

    PubMed

    Aizawa, Shin-Ichi

    2017-01-01

    The bacterial flagellum is a motility organelle. The flagellum is composed of three main structures: the basal body as a rotary engine embedded in the cellular membranes and cell wall, the long external filament that acts as a propeller, and the hook acting as a universal joint that connects them. I describe protocols for the purification of the filament and hook-basal body from Salmonella enterica serovar Typhimurium.

  10. Bundled payments in orthopedic surgery.

    PubMed

    Bushnell, Brandon D

    2015-02-01

    As a result of reading this article, physicians should be able to: 1. Describe the concept of bundled payments and the potential applications of bundled payments in orthopedic surgery. 2. For specific situations, outline a clinical episode of care, determine the participants in a bundling situation, and define care protocols and pathways. 3. Recognize the importance of resource utilization management, quality outcome measurement, and combined economic-clinical value in determining the value of bundled payment arrangements. 4. Identify the implications of bundled payments for practicing orthopedists, as well as the legal issues and potential future directions of this increasingly popular alternative payment method. Bundled payments, the idea of paying a single price for a bundle of goods and services, is a financial concept familiar to most American consumers because examples appear in many industries. The idea of bundled payments has recently gained significant momentum as a financial model with the potential to decrease the significant current costs of health care. Orthopedic surgery as a field of medicine is uniquely positioned for success in an environment of bundled payments. This article reviews the history, logistics, and implications of the bundled payment model relative to orthopedic surgery. Copyright 2015, SLACK Incorporated.

  11. Flagellar dynamics reveal the distribution of chemotactic signaling molecule CheY-P in E. coli

    NASA Astrophysics Data System (ADS)

    Bano, Roshni; Mears, Patrick; Chemla, Yann; Golding, Ido

    E. colicells swim in a random walk consisting of ''runs'' - during which the flagella that propel the cell rotate counter-clockwise (CCW) - and ''tumbles''- during which one or more flagella rotate clockwise (CW). The tumbling frequency is modulated by the phosphorylation state of the signaling molecule CheY, which depends on the cell's environment. Phosphorylated CheY (CheY-P) binds to a flagellar motor and engenders a change in rotation state from CCW to CW. Despite advances in methods used to observe chemotactic signaling, it remains a challenge to measure the CheY-P level in cells directly. Here, we used an optical trap assay coupled with fluorescence microscopy to observe the dynamics of fluorescently labelled flagella in individual cells. By measuring the distribution of flagellar states in multi-flagellated cells and using our recent finding that each flagellar motor independently measures the cellular CheY-P concentration, we are able to extract the probability distribution of the CheY-P level in the cell. This analysis reveals the magnitude of fluctuations in chemotactic signaling in the live cell. We further investigate how this CheY-P distribution changes when cells encounter chemical gradients and perform chemotaxis. This work was supported by the National Science Foundation (NSF) through the Centre for Physics of Living Cells (CPLC).

  12. The phylogeny of swimming kinematics: The environment controls flagellar waveforms in sperm motility

    NASA Astrophysics Data System (ADS)

    Guasto, Jeffrey; Burton, Lisa; Zimmer, Richard; Hosoi, Anette; Stocker, Roman

    2013-11-01

    In recent years, phylogenetic and molecular analyses have dominated the study of ecology and evolution. However, physical interactions between organisms and their environment, a fundamental determinant of organism ecology and evolution, are mediated by organism form and function, highlighting the need to understand the mechanics of basic survival strategies, including locomotion. Focusing on spermatozoa, we combined high-speed video microscopy and singular value decomposition analysis to quantitatively compare the flagellar waveforms of eight species, ranging from marine invertebrates to humans. We found striking similarities in sperm swimming kinematics between genetically dissimilar organisms, which could not be uncovered by phylogenetic analysis. The emergence of dominant waveform patterns across species are suggestive of biological optimization for flagellar locomotion and point toward environmental cues as drivers of this convergence. These results reinforce the power of quantitative kinematic analysis to understand the physical drivers of evolution and as an approach to uncover new solutions for engineering applications, such as micro-robotics.

  13. Bacterial copper storage proteins.

    PubMed

    Dennison, Christopher; David, Sholto; Lee, Jaeick

    2018-03-30

    Copper is essential for most organisms as a cofactor for key enzymes involved in fundamental processes such as respiration and photosynthesis. However, copper also has toxic effects in cells, which is why eukaryotes and prokaryotes have evolved mechanisms for safe copper handling. A new family of bacterial proteins uses a Cys-rich four-helix bundle to safely store large quantities of Cu(I). The work leading to the discovery of these proteins, their properties and physiological functions, and how their presence potentially impacts the current views of bacterial copper handling and use are discussed in this review. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Flagellar filament bio-templated inorganic oxide materials – towards an efficient lithium battery anode

    PubMed Central

    Beznosov, Sergei N.; Veluri, Pavan S.; Pyatibratov, Mikhail G.; Chatterjee, Abhijit; MacFarlane, Douglas R.; Fedorov, Oleg V.; Mitra, Sagar

    2015-01-01

    Designing a new generation of energy-intensive and sustainable electrode materials for batteries to power a variety of applications is an imperative task. The use of biomaterials as a nanosized structural template for these materials has the potential to produce hitherto unachievable structures. In this report, we have used genetically modified flagellar filaments of the extremely halophilic archaea species Halobacterium salinarum to synthesize nanostructured iron oxide composites for use as a lithium-ion battery anode. The electrode demonstrated a superior electrochemical performance compared to existing literature results, with good capacity retention of 1032 mAh g−1 after 50 cycles and with high rate capability, delivering 770 mAh g−1 at 5 A g−1 (~5 C) discharge rate. This unique flagellar filament based template has the potential to provide access to other highly structured advanced energy materials in the future. PMID:25583370

  15. Bundle Branch Block

    MedlinePlus

    ... known cause. Causes can include: Left bundle branch block Heart attacks (myocardial infarction) Thickened, stiffened or weakened ... myocarditis) High blood pressure (hypertension) Right bundle branch block A heart abnormality that's present at birth (congenital) — ...

  16. Fluid mechanics of swimming bacteria with multiple flagella.

    PubMed

    Kanehl, Philipp; Ishikawa, Takuji

    2014-04-01

    It is known that some kinds of bacteria swim by forming a bundle of their multiple flagella. However, the details of flagella synchronization as well as the swimming efficiency of such bacteria have not been fully understood. In this study, swimming of multiflagellated bacteria is investigated numerically by the boundary element method. We assume that the cell body is a rigid ellipsoid and the flagella are rigid helices suspended on flexible hooks. Motors apply constant torque to the hooks, rotating the flagella either clockwise or counterclockwise. Rotating all flagella clockwise, bundling of all flagella is observed in every simulated case. It is demonstrated that the counter rotation of the body speeds up the bundling process. During this procedure the flagella synchronize due to hydrodynamic interactions. Moreover, the results illustrated that during running the multiflagellated bacterium shows higher propulsive efficiency (distance traveled per one flagellar rotation) over a bacterium with a single thick helix. With an increasing number of flagella the propulsive efficiency increases, whereas the energetic efficiency decreases, which indicates that efficiency is something multiflagellated bacteria are assigning less priority to than to motility. These findings form a fundamental basis in understanding bacterial physiology and metabolism.

  17. Association of Lis1 with outer arm dynein is modulated in response to alterations in flagellar motility

    PubMed Central

    Rompolas, Panteleimon; Patel-King, Ramila S.; King, Stephen M.

    2012-01-01

    The cytoplasmic dynein regulatory factor Lis1, which induces a persistent tight binding to microtubules and allows for transport of cargoes under high-load conditions, is also present in motile cilia/flagella. We observed that Lis1 levels in flagella of Chlamydomonas strains that exhibit defective motility due to mutation of various axonemal substructures were greatly enhanced compared with wild type; this increase was absolutely dependent on the presence within the flagellum of the outer arm dynein α heavy chain/light chain 5 thioredoxin unit. To assess whether cells might interpret defective motility as a “high-load environment,” we reduced the flagellar beat frequency of wild-type cells through enhanced viscous load and by reductive stress; both treatments resulted in increased levels of flagellar Lis1, which altered the intrinsic beat frequency of the trans flagellum. Differential extraction of Lis1 from wild-type and mutant axonemes suggests that the affinity of outer arm dynein for Lis1 is directly modulated. In cytoplasm, Lis1 localized to two punctate structures, one of which was located near the base of the flagella. These data reveal that the cell actively monitors motility and dynamically modulates flagellar levels of the dynein regulatory factor Lis1 in response to imposed alterations in beat parameters. PMID:22855525

  18. An analytical fiber bundle model for pullout mechanics of root bundles

    NASA Astrophysics Data System (ADS)

    Cohen, D.; Schwarz, M.; Or, D.

    2011-09-01

    Roots in soil contribute to the mechanical stability of slopes. Estimation of root reinforcement is challenging because roots form complex biological networks whose geometrical and mechanical characteristics are difficult to characterize. Here we describe an analytical model that builds on simple root descriptors to estimate root reinforcement. Root bundles are modeled as bundles of heterogeneous fibers pulled along their long axes neglecting root-soil friction. Analytical expressions for the pullout force as a function of displacement are derived. The maximum pullout force and corresponding critical displacement are either derived analytically or computed numerically. Key model inputs are a root diameter distribution (uniform, Weibull, or lognormal) and three empirical power law relations describing tensile strength, elastic modulus, and length of roots as functions of root diameter. When a root bundle with root tips anchored in the soil matrix is pulled by a rigid plate, a unique parameter, ?, that depends only on the exponents of the power law relations, dictates the order in which roots of different diameters break. If ? < 1, small roots break first; if ? > 1, large roots break first. When ? = 1, all fibers break simultaneously, and the maximum tensile force is simply the roots' mean force times the number of roots in the bundle. Based on measurements of root geometry and mechanical properties, the value of ? is less than 1, usually ranging between 0 and 0.7. Thus, small roots always fail first. The model shows how geometrical and mechanical characteristics of roots and root diameter distribution affect the pullout force, its maximum and corresponding displacement. Comparing bundles of roots that have similar mean diameters, a bundle with a narrow variance in root diameter will result in a larger maximum force and a smaller displacement at maximum force than a bundle with a wide diameter distribution. Increasing the mean root diameter of a bundle without

  19. Disruption of the A-Kinase Anchoring Domain in Flagellar Radial Spoke Protein 3 Results in Unregulated Axonemal cAMP-dependent Protein Kinase Activity and Abnormal Flagellar Motility

    PubMed Central

    Gaillard, Anne R.; Fox, Laura A.; Rhea, Jeanne M.; Craige, Branch

    2006-01-01

    Biochemical studies of Chlamydomonas flagellar axonemes revealed that radial spoke protein (RSP) 3 is an A-kinase anchoring protein (AKAP). To determine the physiological role of PKA anchoring in the axoneme, an RSP3 mutant, pf14, was transformed with an RSP3 gene containing a mutation in the PKA-binding domain. Analysis of several independent transformants revealed that the transformed cells exhibit an unusual phenotype: a fraction of the cells swim normally; the remainder of the cells twitch feebly or are paralyzed. The abnormal/paralyzed motility is not due to an obvious deficiency of radial spoke assembly, and the phenotype cosegregates with the mutant RSP3. We postulated that paralysis was due to failure in targeting and regulation of axonemal cAMP-dependent protein kinase (PKA). To test this, reactivation experiments of demembranated cells were performed in the absence or presence of PKA inhibitors. Importantly, motility in reactivated cell models mimicked the live cell phenotype with nearly equal fractions of motile and paralyzed cells. PKA inhibitors resulted in a twofold increase in the number of motile cells, rescuing paralysis. These results confirm that flagellar RSP3 is an AKAP and reveal that a mutation in the PKA binding domain results in unregulated axonemal PKA activity and inhibition of normal motility. PMID:16571668

  20. Salmonella Enteritidis flagellar mutants have a colonization benefit in the chicken oviduct.

    PubMed

    Kilroy, Sofie; Raspoet, Ruth; Martel, An; Bosseler, Leslie; Appia-Ayme, Corinne; Thompson, Arthur; Haesebrouck, Freddy; Ducatelle, Richard; Van Immerseel, Filip

    2017-02-01

    Egg borne Salmonella Enteritidis is still a major cause of human food poisoning. Eggs can become internally contaminated following colonization of the hen's oviduct. In this paper we aimed to analyze the role of flagella of Salmonella Enteritidis in colonization of the hen's oviduct. Using a transposon library screen we showed that mutants lacking functional flagella are significantly more efficient in colonizing the hen's oviduct in vivo. A micro-array analysis proved that transcription of a number of flagellar genes is down-regulated inside chicken oviduct cells. Flagella contain flagellin, a pathogen associated molecular pattern known to bind to Toll-like receptor 5, activating a pro-inflammatory cascade. In vitro tests using primary oviduct cells showed that flagellin is not involved in invasion. Using a ligated loop model, a diminished inflammatory reaction was seen in the oviduct resulting from injection of an aflagellated mutant compared to the wild-type. It is hypothesized that Salmonella Enteritidis downregulates flagellar gene expression in the oviduct and consequently prevents a flagellin-induced inflammatory response, thereby increasing its oviduct colonization efficiency. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Bacterially induced precipitation of CaCO sub 3 : An example from studies of cyanobacterial mats

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

    Chafetz, H.S.

    1990-04-30

    Bacteria induce the precipitation of calcium carbonate in the laboratory and in nature by altering their chemical environment. Geologists are recognizing the possibility that bacterially induced precipitates may form significant mineral deposits, unfortunately, there are currently no sound criteria by which they can be recognized in recent sediments, or in the rock record. Cultures of aerobic and facultative bacteria from cyanobacterial mats on Andros Island, Bahamas, and Baffin Bay, Texas, induced the precipitation of calcium carbonate under controlled conditions. Crusts, the largest features formed, are composed of 5--200{mu}m diameter bundles which are, in turn, composed of numerous individual crystals. Themore » smallest observed features are 0.1--0.4{mu}m spheres and rods which comprise some individual crystals and crystal bundles. Crystal bundles resembling rhombohedra, tetragonal disphenoids, tetragonal dipyramids, and calcite dumbbells appear to be uniquely bacterial in origin, and they have all been observed in recent sediments. Swollen rods, discs, curved dumbbells, and 50--200{mu}m optically continuous crystals resembling brushes may be uniquely bacterial in origin, however, they have not been reported by other laboratories nor observed in natural settings. Presence of any of these forms in recent sediments should be taken as strong evidence for bacterial influence. Spheres and aragonite dumbbells have also been observed in natural environments, however, they are not always bacterial in origin. Precipitation of calcium carbonate occurs preferentially on dead cyanobacteria in the presence of bacteria. Lithification of algal mats to form stromatolites may take place in the zone of decaying organic matter due to bacterial activity.« less

  2. Function of Proline Residues of MotA in Torque Generation by the Flagellar Motor of Escherichia coli

    PubMed Central

    Braun, Timothy F.; Poulson, Susan; Gully, Jonathan B.; Empey, J. Courtney; Van Way, Susan; Putnam, Angélica; Blair, David F.

    1999-01-01

    Bacterial flagellar motors obtain energy for rotation from the membrane gradient of protons or, in some species, sodium ions. The molecular mechanism of flagellar rotation is not understood. MotA and MotB are integral membrane proteins that function in proton conduction and are believed to form the stator of the motor. Previous mutational studies identified two conserved proline residues in MotA (Pro 173 and Pro 222 in the protein from Escherichia coli) and a conserved aspartic acid residue in MotB (Asp 32) that are important for function. Asp 32 of MotB probably forms part of the proton path through the motor. To learn more about the roles of the conserved proline residues of MotA, we examined motor function in Pro 173 and Pro 222 mutants, making measurements of torque at high load, speed at low and intermediate loads, and solvent-isotope effects (D2O versus H2O). Proton conduction by wild-type and mutant MotA-MotB channels was also assayed, by a growth defect that occurs upon overexpression. Several different mutations of Pro 173 reduced the torque of the motor under high load, and a few prevented motor rotation but still allowed proton flow through the MotA-MotB channels. These and other properties of the mutants suggest that Pro 173 has a pivotal role in coupling proton flow to motor rotation and is positioned in the channel near Asp 32 of MotB. Replacements of Pro 222 abolished function in all assays and were strongly dominant. Certain Pro 222 mutant proteins prevented swimming almost completely when expressed at moderate levels in wild-type cells. This dominance might be caused by rotor-stator jamming, because it was weaker when FliG carried a mutation believed to increase rotor-stator clearance. We propose a mechanism for torque generation, in which specific functions are suggested for the proline residues of MotA and Asp32 of MotB. PMID:10348868

  3. Structural Insights into Membrane Targeting by the Flagellar Calcium-binding Protein (FCaBP) a Myristoylated and Palmitoylated Calcium Sensor in Trypanosoma cruzi

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

    J Wingard; J Ladner; M Vanarotti

    2011-12-31

    The flagellar calcium-binding protein (FCaBP) of the protozoan Trypanosoma cruzi is targeted to the flagellar membrane where it regulates flagellar function and assembly. As a first step toward understanding the Ca{sup 2+}-induced conformational changes important for membrane-targeting, we report here the x-ray crystal structure of FCaBP in the Ca{sup 2+}-free state determined at 2.2{angstrom} resolution. The first 17 residues from the N terminus appear unstructured and solvent-exposed. Residues implicated in membrane targeting (Lys-19, Lys-22, and Lys-25) are flanked by an exposed N-terminal helix (residues 26-37), forming a patch of positive charge on the protein surface that may interact electrostatically withmore » flagellar membrane targets. The four EF-hands in FCaBP each adopt a 'closed conformation' similar to that seen in Ca{sup 2+}-free calmodulin. The overall fold of FCaBP is closest to that of grancalcin and other members of the penta EF-hand superfamily. Unlike the dimeric penta EF-hand proteins, FCaBP lacks a fifth EF-hand and is monomeric. The unstructured N-terminal region of FCaBP suggests that its covalently attached myristoyl group at the N terminus may be solvent-exposed, in contrast to the highly sequestered myristoyl group seen in recoverin and GCAP1. NMR analysis demonstrates that the myristoyl group attached to FCaBP is indeed solvent-exposed in both the Ca{sup 2+}-free and Ca{sup 2+}-bound states, and myristoylation has no effect on protein structure and folding stability. We propose that exposed acyl groups at the N terminus may anchor FCaBP to the flagellar membrane and that Ca{sup 2+}-induced conformational changes may control its binding to membrane-bound protein targets..« less

  4. Flagellar generated flow mediates attachment of Giardia lamblia

    NASA Astrophysics Data System (ADS)

    Urbach, Jeffrey; Luo, Haibei; Picou, Theodore; McAllister, Ryan; Elmendorf, Heidi

    2011-03-01

    Giardia lamblia is a protozoan parasite responsible for widespread diarrheal disease in humans and animals worldwide. Attachment to the host intestinal mucosa and resistance to peristalsis is necessary for establishing infection, but the physical basis for this attachment is poorly understood. We report results from TIRF and confocal fluorescence microscopy that demonstrate that the regular beating of the posterior flagella generate a flow through the ventral disk, a suction-cup shaped structure that is against the substrate during attachment. Finite element simulations are used to compare the negative pressure generated by the flow to the measured attachment force and the expected performance of the flagellar pump. NIH grant 1R21AI062934-0.

  5. Testing the time-of-flight model for flagellar length sensing.

    PubMed

    Ishikawa, Hiroaki; Marshall, Wallace F

    2017-11-07

    Cilia and flagella are microtubule-based organelles that protrude from the surface of most cells, are important to the sensing of extracellular signals, and make a driving force for fluid flow. Maintenance of flagellar length requires an active transport process known as intraflagellar transport (IFT). Recent studies reveal that the amount of IFT injection negatively correlates with the length of flagella. These observations suggest that a length-dependent feedback regulates IFT. However, it is unknown how cells recognize the length of flagella and control IFT. Several theoretical models try to explain this feedback system. We focused on one of the models, the "time-of-flight" model, which measures the length of flagella on the basis of the travel time of IFT protein in the flagellar compartment. We tested the time-of-flight model using Chlamydomonas dynein mutant cells, which show slower retrograde transport speed. The amount of IFT injection in dynein mutant cells was higher than that in control cells. This observation does not support the prediction of the time-of-flight model and suggests that Chlamydomonas uses another length-control feedback system rather than that described by the time-of-flight model. © 2017 Ishikawa and Marshall. 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).

  6. A solid-state control system for dynein-based ciliary/flagellar motility

    PubMed Central

    2013-01-01

    Ciliary and flagellar beating requires the coordinated action of multiple dyneins with different enzymatic and motor properties. In this issue, Yamamoto et al. (2013. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201211048) identify the MIA (modifier of inner arms) complex within the Chlamydomonas reinhardtii axoneme that physically links to a known regulatory structure and provides a signaling conduit from the radial spokes to an inner arm dynein essential for waveform determination. PMID:23569213

  7. MotI (DgrA) acts as a molecular clutch on the flagellar stator protein MotA in Bacillus subtilis

    PubMed Central

    Subramanian, Sundharraman; Gao, Xiaohui; Dann, Charles E.; Kearns, Daniel B.

    2017-01-01

    Stator elements consisting of MotA4MotB2 complexes are anchored to the cell wall, extend through the cell membrane, and interact with FliG in the cytoplasmic C ring rotor of the flagellum. The cytoplasmic loop of MotA undergoes proton-driven conformational changes that drive flagellar rotation. Functional regulators inhibit motility by either disengaging or jamming the stator–rotor interaction. Here we show that the YcgR homolog MotI (formerly DgrA) of Bacillus subtilis inhibits motility like a molecular clutch that disengages MotA. MotI-inhibited flagella rotated freely by Brownian motion, and suppressor mutations in MotA that were immune to MotI inhibition were located two residues downstream of the critical force generation site. The 3D structure of MotI bound to c-di-GMP was solved, and MotI-fluorescent fusions localized as transient MotA-dependent puncta at the membrane when induced at subinhibitory levels. Finally, subinhibitory levels of MotI expression resulted in incomplete inhibition and proportional decreases in swimming speed. We propose a model in which flagellar stators are disengaged and sequestered from the flagellar rotor when bound by MotI. PMID:29196522

  8. Listeria monocytogenes DNA glycosylase AdiP affects flagellar motility, biofilm formation, virulence, and stress responses

    USDA-ARS?s Scientific Manuscript database

    The temperature-dependent alteration of flagellar motility gene expression is critical for the foodborne pathogen Listeria monocytogenes to respond to a changing environment. In this study, a genetic determinant, L. monocytogenes f2365_0220 (lmof2365_0220), encoding a putative protein that is struct...

  9. On the concept of a filtered bundle

    NASA Astrophysics Data System (ADS)

    Bruce, Andrew James; Grabowska, Katarzyna; Grabowski, Janusz

    We present the notion of a filtered bundle as a generalization of a graded bundle. In particular, we weaken the necessity of the transformation laws for local coordinates to exactly respect the weight of the coordinates by allowing more general polynomial transformation laws. The key examples of such bundles include affine bundles and various jet bundles, both of which play fundamental roles in geometric mechanics and classical field theory. We also present the notion of double filtered bundles which provide natural generalizations of double vector bundles and double affine bundles. Furthermore, we show that the linearization of a filtered bundle — which can be seen as a partial polarization of the admissible changes of local coordinates — is well defined.

  10. Bacterially induced precipitation of CaCO 3: An example from studies of cyanobacterial mats. Final report

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

    Chafetz, Henry S.

    1990-04-30

    Bacteria induce the precipitation of calcium carbonate in the laboratory and in nature by altering their chemical environment. Geologists are recognizing the possibility that bacterially induced precipitates may form significant mineral deposits, unfortunately, there are currently no sound criteria by which they can be recognized in recent sediments, or in the rock record. Cultures of aerobic and facultative bacteria from cyanobacterial mats on Andros Island, Bahamas, and Baffin Bay, Texas, induced the precipitation of calcium carbonate under controlled conditions. Crusts, the largest features formed, are composed of 5--200μm diameter bundles which are, in turn, composed of numerous individual crystals. Themore » smallest observed features are 0.1--0.4μm spheres and rods which comprise some individual crystals and crystal bundles. Crystal bundles resembling rhombohedra, tetragonal disphenoids, tetragonal dipyramids, and calcite dumbbells appear to be uniquely bacterial in origin, and they have all been observed in recent sediments. Swollen rods, discs, curved dumbbells, and 50--200μm optically continuous crystals resembling brushes may be uniquely bacterial in origin, however, they have not been reported by other laboratories nor observed in natural settings. Presence of any of these forms in recent sediments should be taken as strong evidence for bacterial influence. Spheres and aragonite dumbbells have also been observed in natural environments, however, they are not always bacterial in origin. Precipitation of calcium carbonate occurs preferentially on dead cyanobacteria in the presence of bacteria. Lithification of algal mats to form stromatolites may take place in the zone of decaying organic matter due to bacterial activity.« less

  11. "Bundling" in Learning.

    ERIC Educational Resources Information Center

    Spiegel, U.; Templeman, J.

    1996-01-01

    Applies the literature of bundling, tie-in sales, and vertical integration to higher education. Students are often required to purchase a package of courses, some of which are unrelated to their major. This kind of bundling policy can be utilized as a profit-maximizing strategy for universities exercising a degree of monopolistic power. (12…

  12. The Glycosylphosphatidylinositol-PLC in Trypanosoma brucei Forms a Linear Array on the Exterior of the Flagellar Membrane Before and After Activation

    PubMed Central

    Hanrahan, Orla; Webb, Helena; O'Byrne, Robert; Brabazon, Elaine; Treumann, Achim; Sunter, Jack D.; Carrington, Mark; Voorheis, H. Paul

    2009-01-01

    Bloodstream forms of Trypanosoma brucei contain a glycosylphosphatidylinositol-specific phospholipase C (GPI-PLC) that cleaves the GPI-anchor of the variable surface glycoprotein (VSG). Its location in trypanosomes has been controversial. Here, using confocal microscopy and surface labelling techniques, we show that the GPI-PLC is located exclusively in a linear array on the outside of the flagellar membrane, close to the flagellar attachment zone, but does not co-localize with the flagellar attachment zone protein, FAZ1. Consequently, the GPI-PLC and the VSG occupy the same plasma membrane leaflet, which resolves the topological problem associated with the cleavage reaction if the VSG and the GPI-PLC were on opposite sides of the membrane. The exterior location requires the enzyme to be tightly regulated to prevent VSG release under basal conditions. During stimulated VSG release in intact cells, the GPI-PLC did not change location, suggesting that the release mechanism involves lateral diffusion of the VSG in the plane of the membrane to the fixed position of the GPI-PLC. PMID:19503825

  13. Tuning Bacterial Hydrodynamics with Magnetic Fields: A Path to Bacterial Robotics

    NASA Astrophysics Data System (ADS)

    Pierce, Christopher; Mumper, Eric; Brangham, Jack; Wijesinghe, Hiran; Lower, Stephen; Lower, Brian; Yang, Fengyuan; Sooryakumar, Ratnasingham

    Magnetotactic Bacteria (MTB) are a group of motile prokaryotes that synthesize chains of lipid-bound, magnetic nano-particles. In this study, the innate magnetism of these flagellated swimmers is exploited to explore their hydrodynamics near confining surfaces, using the magnetic field as a tuning parameter. With weak (Gauss), uniform, external, magnetic ?elds and the field gradients arising from micro-magnetic surface patterns, the relative strength of hydrodynamic, magnetic and ?agellar force components is tuned through magnetic control of the bacteria's orientation and position. In addition to direct measurement of several hydrodynamic quantities related to the motility of individual cells, their tunable dynamics reveal a number of novel, highly controllable swimming behaviors with potential value in micro-robotics applications. Specifically, the experiments permit the MTB cells to be directed along parallel or divergent trajectories, suppress their flagellar forces through magnetic means, and induce transitions between planar, circulating trajectories and drifting, vertically oriented ``top-like'' motion. The implications of the work for fundamental hydrodynamics research as well as bacterially driven robotics applications will be discussed.

  14. Signal detection by active, noisy hair bundles

    NASA Astrophysics Data System (ADS)

    O'Maoiléidigh, Dáibhid; Salvi, Joshua D.; Hudspeth, A. J.

    2018-05-01

    Vertebrate ears employ hair bundles to transduce mechanical movements into electrical signals, but their performance is limited by noise. Hair bundles are substantially more sensitive to periodic stimulation when they are mechanically active, however, than when they are passive. We developed a model of active hair-bundle mechanics that predicts the conditions under which a bundle is most sensitive to periodic stimulation. The model relies only on the existence of mechanotransduction channels and an active adaptation mechanism that recloses the channels. For a frequency-detuned stimulus, a noisy hair bundle's phase-locked response and degree of entrainment as well as its detection bandwidth are maximized when the bundle exhibits low-amplitude spontaneous oscillations. The phase-locked response and entrainment of a bundle are predicted to peak as functions of the noise level. We confirmed several of these predictions experimentally by periodically forcing hair bundles held near the onset of self-oscillation. A hair bundle's active process amplifies the stimulus preferentially over the noise, allowing the bundle to detect periodic forces less than 1 pN in amplitude. Moreover, the addition of noise can improve a bundle's ability to detect the stimulus. Although, mechanical activity has not yet been observed in mammalian hair bundles, a related model predicts that active but quiescent bundles can oscillate spontaneously when they are loaded by a sufficiently massive object such as the tectorial membrane. Overall, this work indicates that auditory systems rely on active elements, composed of hair cells and their mechanical environment, that operate on the brink of self-oscillation.

  15. Computer simulation of flagellar movement. VI. Simple curvature-controlled models are incompletely specified.

    PubMed

    Brokaw, C J

    1985-10-01

    Computer simulation is used to examine a simple flagellar model that will initiate and propagate bending waves in the absence of viscous resistances. The model contains only an elastic bending resistance and an active sliding mechanism that generates reduced active shear moment with increasing sliding velocity. Oscillation results from a distributed control mechanism that reverses the direction of operation of the active sliding mechanism when the curvature reaches critical magnitudes in either direction. Bend propagation by curvature-controlled flagellar models therefore does not require interaction with the viscous resistance of an external fluid. An analytical examination of moment balance during bend propagation by this model yields a solution curve giving values of frequency and wavelength that satisfy the moment balance equation and give uniform bend propagation, suggesting that the model is underdetermined. At 0 viscosity, the boundary condition of 0 shear rate at the basal end of the flagellum during the development of new bends selects the particular solution that is obtained by computer simulations. Therefore, the details of the pattern of bend initiation at the basal end of a flagellum can be of major significance in determining the properties of propagated bending waves in the distal portion of a flagellum. At high values of external viscosity, the model oscillates at frequencies and wavelengths that give approximately integral numbers of waves on the flagellum. These operating points are selected because they facilitate the balance of bending moments at the ends of the model, where the external viscous moment approaches 0. These mode preferences can be overridden by forcing the model to operate at a predetermined frequency. The strong mode preferences shown by curvature-controlled flagellar models, in contrast to the weak or absent mode preferences shown by real flagella, therefore do not demonstrate the inapplicability of the moment-balance approach

  16. Bundled payment fails to gain a foothold In California: the experience of the IHA bundled payment demonstration.

    PubMed

    Ridgely, M Susan; de Vries, David; Bozic, Kevin J; Hussey, Peter S

    2014-08-01

    To determine whether bundled payment could be an effective payment model for California, the Integrated Healthcare Association convened a group of stakeholders (health plans, hospitals, ambulatory surgery centers, physician organizations, and vendors) to develop, through a consensus process, the methods and means of implementing bundled payment. In spite of a high level of enthusiasm and effort, the pilot did not succeed in its goal to implement bundled payment for orthopedic procedures across multiple payers and hospital-physician partners. An evaluation of the pilot documented a number of barriers, such as administrative burden, state regulatory uncertainty, and disagreements about bundle definition and assumption of risk. Ultimately, few contracts were signed, which resulted in insufficient volume to test hypotheses about the impact of bundled payment on quality and costs. Although bundled payment failed to gain a foothold in California, the evaluation provides lessons for future bundled payment initiatives. Project HOPE—The People-to-People Health Foundation, Inc.

  17. The sensory transduction pathways in bacterial chemotaxis

    NASA Technical Reports Server (NTRS)

    Taylor, Barry L.

    1989-01-01

    Bacterial chemotaxis is a useful model for investigating in molecular detail the behavioral response of cells to changes in their environment. Peritrichously flagellated bacteria such as coli and typhimurium swim by rotating helical flagella in a counterclockwise direction. If flagellar rotation is briefly reversed, the bacteria tumble and change the direction of swimming. The bacteria continuously sample the environment and use a temporal sensing mechanism to compare the present and immediate past environments. Bacteria respond to a broad range of stimuli including changes in temperature, oxygen concentration, pH and osmotic strength. Bacteria are attracted to potential sources of nutrition such as sugars and amino acids and are repelled by other chemicals. In the methylation-dependent pathways for sensory transduction and adaptation in E. coli and S. typhimurium, chemoeffectors bind to transducing proteins that span the plasma membrane. The transducing proteins are postulated to control the rate of autophosphorylation of the CheA protein, which in turn phosphorylates the CheY protein. The phospho-CheY protein binds to the switch on the flagellar motor and is the signal for clockwise rotation of the motor. Adaptation to an attractant is achieved by increasing methylation of the transducing protein until the attractant stimulus is cancelled. Responses to oxygen and certain sugars involve methylation-independent pathways in which adaption occurs without methylation of a transducing protein. Taxis toward oxygen is mediated by the electron transport system and changes in the proton motive force. Recent studies have shown that the methylation-independent pathway converges with the methylation-dependent pathway at or before the CheA protein.

  18. A Conservative Amino Acid Mutation in the Master Regulator FleQ Renders Pseudomonas aeruginosa Aflagellate

    PubMed Central

    Jain, Ruchi; Kazmierczak, Barbara I.

    2014-01-01

    Flagellar-based motility plays a critical role in Pseudomonas aeruginosa pathogenesis, influencing both the establishment of bacterial infection and the host's response to the pathogen. Nonetheless, aflagellate clinical strains are often isolated from acutely and chronically infected patients and include the virulent laboratory strain PA103. We determined that PA103's aflagellate phenotype is the result of a single amino acid change (G240V) in the master flagellar regulator, FleQ. This mutation, which lies just outside the Walker B box of FleQ, abrogates the ability of FleQ to positively regulate flagellar gene expression. Reversal of this seemingly conservative amino acid substitution is sufficient to restore swimming motility to PA103, despite the presence of mutations in other flagellar genes of PA103. We also investigated the consequences of restoring flagellar assembly on PA103 virulence. Although a negative correlation between flagellar assembly and Type 3 secretion system (T3SS) expression has been reported previously, we did not observe downregulation of T3SS expression or function in Fla+ PA103. Restoration of flagellar assembly did, however, amplify IL-1 signals measured during murine pulmonary infection and was associated with increased bacterial clearance. These experiments suggest that loss of flagellar motility may primarily benefit PA103 by attenuating pathogen recognition and clearance during acute infection. PMID:24827992

  19. Robust peptide bundles designed computationally

    NASA Astrophysics Data System (ADS)

    Haider, Michael; Zhang, Huixi Violet; Kiick, Kristi; Saven, Jeffery; Pochan, Darrin

    Peptides are ideal candidates for the design and controlled assembly of nanoscale materials due to their potential to assemble with atomistic precision as in biological systems. Unlike other work utilizing natural proteins and structural motifs, this effort is completely de novo in order to build arbitrary structures with desired size for the specific placement and separation of functional groups. We have successfully computationally designed soluble, coiled coil, peptide, tetramer bundles which are robust and stable. Using circular dichroism we demonstrated the thermal stability of these bundles as well as confirmed their alpha helical and coiled coil nature. The stability of these bundles arises from the computational design of the coiled coil interior core residues. The coiled coil tetramer was confirmed to be the dominant species by analytical ultra-centrifugation sedimentation studies. We also established how these bundles behave in solution using small angle neutron scattering. The form factor of the bundles is well represented by a cylinder model and their behavior at high concentrations is modeled using a structure factor for aggregates of the cylinders. All of these experiments support our claim that the designed coiled coil bundles were achieved in solution. NSF DMREF 1234161.

  20. Proposed model for the flagellar rotary motor with shear stress transmission

    PubMed Central

    Mitsui, Toshio; Ohshima, Hiroyuki

    2012-01-01

    Most bacteria that swim are propelled by flagellar filaments, which are driven by a rotary motor powered by proton flux. The motor consists of the rotor and the stator. The stator consists of about 8 MotA-Mot B complex. There seems to be no definite information about the structure between the rotor and the stator, and it is examined whether the experimental data can be explained based upon the following assumptions. (a) There is viscoelastic medium between the rotor and the stator. (b) MotA-MotB complex has an electric dipole moment and produces shear stress in the electric field by a proton in the channel. Calculation results based upon these assumptions are in good agreement with the following experimental observations. (1) One revolution of the flagellar rotation consists of a constant number of steps. (2) The rotation velocity of the rotor is proportional to the trans-membrane potential difference. (3) When the rotational velocity of a flagellum is changed by adjusting the viscosity of the outer fluid, the torque for the cell to rotate a flagellum is practically constant but sharply decreases when the rotational velocity increases over a critical value. (4) The rotation direction remains the same when the sign of the electrochemical potential gradient is reversed. (5) The cell produces constant torque to rotate the flagellum even when the cell is rotated by externally applied torque. (6) A simple switch mechanism is proposed for chemotaxis. PMID:27493532

  1. Moisture separator reheater with round tube bundle

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

    Byerley, W. M.

    1984-11-27

    A moisture separator reheater having a central chamber with cylindrical wall protions and a generally round tube bundle, the tube bundle having arcuate plates disposed on each side of the bundle which form a wrapper on each side of the bundle and having a tongue and groove juncture between the wrapper and cylindrical wall portions to provide a seal therebetween and a track for installing and removing the tube bundle from the central chamber.

  2. Fiber bundle phase conjugate mirror

    DOEpatents

    Ward, Benjamin G.

    2012-05-01

    An improved method and apparatus for passively conjugating the phases of a distorted wavefronts resulting from optical phase mismatch between elements of a fiber laser array are disclosed. A method for passively conjugating a distorted wavefront comprises the steps of: multiplexing a plurality of probe fibers and a bundle pump fiber in a fiber bundle array; passing the multiplexed output from the fiber bundle array through a collimating lens and into one portion of a non-linear medium; passing the output from a pump collection fiber through a focusing lens and into another portion of the non-linear medium so that the output from the pump collection fiber mixes with the multiplexed output from the fiber bundle; adjusting one or more degrees of freedom of one or more of the fiber bundle array, the collimating lens, the focusing lens, the non-linear medium, or the pump collection fiber to produce a standing wave in the non-linear medium.

  3. Curcumin Reduces the Motility of Salmonella enterica Serovar Typhimurium by Binding to the Flagella, Thereby Leading to Flagellar Fragility and Shedding

    PubMed Central

    Balakrishnan, Arjun; Negi, Vidya Devi; Sakorey, Deepika; Chandra, Nagasuma

    2016-01-01

    ABSTRACT One of the important virulence properties of the pathogen is its ability to travel to a favorable environment, cross the viscous mucus barrier (intestinal barrier for enteric pathogens), and reach the epithelia to initiate pathogenesis with the help of an appendage, like flagella. Nonetheless, flagella can act as an “Achilles heel,” revealing the pathogen's presence to the host through the stimulation of innate and adaptive immune responses. We assessed whether curcumin, a dietary polyphenol, could alter the motility of Salmonella, a foodborne pathogen. It reduced the motility of Salmonella enterica serovar Typhimurium by shortening the length of the flagellar filament (from ∼8 μm to ∼5 μm) and decreasing its density (4 or 5 flagella/bacterium instead of 8 or 9 flagella/bacterium). Upon curcumin treatment, the percentage of flagellated bacteria declined from ∼84% to 59%. However, no change was detected in the expression of the flagellin gene and protein. A fluorescence binding assay demonstrated binding of curcumin to the flagellar filament. This might make the filament fragile, breaking it into smaller fragments. Computational analysis predicted the binding of curcumin, its analogues, and its degraded products to a flagellin molecule at an interface between domains D1 and D2. Site-directed mutagenesis and a fluorescence binding assay confirmed the binding of curcumin to flagellin at residues ASN120, ASP123, ASN163, SER164, ASN173, and GLN175. IMPORTANCE This work, to our knowledge the first report of its kind, examines how curcumin targets flagellar density and affects the pathogenesis of bacteria. We found that curcumin does not affect any of the flagellar synthesis genes. Instead, it binds to the flagellum and makes it fragile. It increases the torsional stress on the flagellar filament that then breaks, leaving fewer flagella around the bacteria. Flagella, which are crucial ligands for Toll-like receptor 5, are some of the most important

  4. Dispersions of attractive semiflexible fiberlike colloidal particles from bacterial cellulose microfibrils.

    PubMed

    Kuijk, Anke; Koppert, Remco; Versluis, Peter; van Dalen, Gerard; Remijn, Caroline; Hazekamp, Johan; Nijsse, Jaap; Velikov, Krassimir P

    2013-11-26

    We prepared dispersions from bacterial cellulose microfibrils (CMF) of a commercial Nata de Coco source. We used an ultra-high-energy mechanical deagglomeration process that is able to disperse the CMFs from the pellicle in which they are organized in an irregular network. Because of the strong attractions between the CMFs, the dispersion remained highly heterogeneous, consisting of fiber bundles, flocs, and voids spanning tens to hundreds of micrometers depending on concentration. The size of these flocs increased with CMF concentration, the size of the bundles stayed constant, and the size of the voids decreased. The observed percolation threshold in MFC dispersions is lower than the theoretical prediction, which is accounted for by the attractive interactions in the system. Because bacterial cellulose is chemically very pure, it can be used to study the interaction of attractive and highly shape-anisotropic, semiflexible fiberlike colloidal particles.

  5. The flaA locus of Bacillus subtilis is part of a large operon coding for flagellar structures, motility functions, and an ATPase-like polypeptide.

    PubMed Central

    Albertini, A M; Caramori, T; Crabb, W D; Scoffone, F; Galizzi, A

    1991-01-01

    We cloned and sequenced 8.3 kb of Bacillus subtilis DNA corresponding to the flaA locus involved in flagellar biosynthesis, motility, and chemotaxis. The DNA sequence revealed the presence of 10 complete and 2 incomplete open reading frames. Comparison of the deduced amino acid sequences to data banks showed similarities of nine of the deduced products to a number of proteins of Escherichia coli and Salmonella typhimurium for which a role in flagellar functioning has been directly demonstrated. In particular, the sequence data suggest that the flaA operon codes for the M-ring protein, components of the motor switch, and the distal part of the basal-body rod. The gene order is remarkably similar to that described for region III of the enterobacterial flagellar regulon. One of the open reading frames was translated into a protein with 48% amino acid identity to S. typhimurium FliI and 29% identity to the beta subunit of E. coli ATP synthase. PMID:1828465

  6. Interplay between the Localization and Kinetics of Phosphorylation in Flagellar Pole Development of the Bacterium Caulobacter crescentus

    PubMed Central

    Tropini, Carolina; Huang, Kerwyn Casey

    2012-01-01

    Bacterial cells maintain sophisticated levels of intracellular organization that allow for signal amplification, response to stimuli, cell division, and many other critical processes. The mechanisms underlying localization and their contribution to fitness have been difficult to uncover, due to the often challenging task of creating mutants with systematically perturbed localization but normal enzymatic activity, and the lack of quantitative models through which to interpret subtle phenotypic changes. Focusing on the model bacterium Caulobacter crescentus, which generates two different types of daughter cells from an underlying asymmetric distribution of protein phosphorylation, we use mathematical modeling to investigate the contribution of the localization of histidine kinases to the establishment of cellular asymmetry and subsequent developmental outcomes. We use existing mutant phenotypes and fluorescence data to parameterize a reaction-diffusion model of the kinases PleC and DivJ and their cognate response regulator DivK. We then present a systematic computational analysis of the effects of changes in protein localization and abundance to determine whether PleC localization is required for correct developmental timing in Caulobacter. Our model predicts the developmental phenotypes of several localization mutants, and suggests that a novel strain with co-localization of PleC and DivJ could provide quantitative insight into the signaling threshold required for flagellar pole development. Our analysis indicates that normal development can be maintained through a wide range of localization phenotypes, and that developmental defects due to changes in PleC localization can be rescued by increased PleC expression. We also show that the system is remarkably robust to perturbation of the kinetic parameters, and while the localization of either PleC or DivJ is required for asymmetric development, the delocalization of one of these two components does not prevent

  7. Structure and Activity of the Flagellar Rotor Protein FliY

    PubMed Central

    Sircar, Ria; Greenswag, Anna R.; Bilwes, Alexandrine M.; Gonzalez-Bonet, Gabriela; Crane, Brian R.

    2013-01-01

    Rotating flagella propel bacteria toward favorable environments. Sense of rotation is determined by the intracellular response regulator CheY, which when phosphorylated (CheY-P) interacts directly with the flagellar motor. In many different types of bacteria, the CheC/CheX/FliY (CXY) family of phosphatases terminates the CheY-P signal. Unlike CheC and CheX, FliY is localized in the flagellar switch complex, which also contains the stator-coupling protein FliG and the target of CheY-P, FliM. The 2.5 Å resolution crystal structure of the FliY catalytic domain from Thermotoga maritima bears strong resemblance to the middle domain of FliM. Regions of FliM that mediate contacts within the rotor compose the phosphatase active sites in FliY. Despite the similarity between FliY and FliM, FliY does not bind FliG and thus is unlikely to be a substitute for FliM in the center of the switch complex. Solution studies indicate that FliY dimerizes through its C-terminal domains, which resemble the Escherichia coli switch complex component FliN. FliY differs topologically from the E. coli chemotaxis phosphatase CheZ but appears to utilize similar structural motifs for CheY dephosphorylation in close analogy to CheX. Recognition properties and phosphatase activities of site-directed mutants identify two pseudosymmetric active sites in FliY (Glu35/Asn38 and Glu132/Asn135), with the second site (Glu132/Asn135) being more active. A putative N-terminal CheY binding domain conserved with FliM is not required for binding CheY-P or phosphatase activity. PMID:23532838

  8. A method for the purification of bacterial flagellin that allows simple upscaling.

    PubMed

    Hiriart, Yanina; Errea, Agustina; González Maciel, Dolores; Lopez, Juan Carlos; Rumbo, Martin

    2012-01-01

    There is a growing interest in enterobacterial flagellins that may result in a demand to produce flagellin on an industrial scale for possible applications as an adjuvant, immunomodulatory agent or vaccine antigen. Traditionally, small-scale production of flagellin has occurred in the laboratory by flagellar shearing of bacterial surfaces and subsequent ultracentrifugation. The main drawback of this method is the need to use low-agitation cultures to avoid the loss of flagella due to shearing during culture. In the present work, we describe a scalable protocol for the production of flagellin with higher yields than traditional laboratory-scale protocols. The use of cross-flow filtration to concentrate bacterial cultures combines extensive shearing of flagella with a reduction in volume, greatly simplifying downstream processing. This technique also allows the use of highly-agitated culture conditions because any sheared flagella are retained in the bacterial concentrate. Flagella obtained with this procedure showed in vivo and in vitro innate activating capacities similar to those of flagella produced at laboratory scale. This procedure is flexible, allowing an increase in production scale, an enhancement of flagellin yield and no requirement for expensive equipment.

  9. Enzymatic hydrolysis of biomimetic bacterial cellulose-hemicellulose composites.

    PubMed

    Penttilä, Paavo A; Imai, Tomoya; Hemming, Jarl; Willför, Stefan; Sugiyama, Junji

    2018-06-15

    The production of biofuels and other chemicals from lignocellulosic biomass is limited by the inefficiency of enzymatic hydrolysis. Here a biomimetic composite material consisting of bacterial cellulose and wood-based hemicelluloses was used to study the effects of hemicelluloses on the enzymatic hydrolysis with a commercial cellulase mixture. Bacterial cellulose synthesized in the presence of hemicelluloses, especially xylan, was found to be more susceptible to enzymatic hydrolysis than hemicellulose-free bacterial cellulose. The reason for the easier hydrolysis could be related to the nanoscale structure of the substrate, particularly the packing of cellulose microfibrils into ribbons or bundles. In addition, small-angle X-ray scattering was used to show that the average nanoscale morphology of bacterial cellulose remained unchanged during the enzymatic hydrolysis. The reported easier enzymatic hydrolysis of bacterial cellulose produced in the presence of wood-based xylan offers new insights to overcome biomass recalcitrance through genetic engineering. Copyright © 2018 Elsevier Ltd. All rights reserved.

  10. An AlgU-regulated antisense transcript encoded within the Pseudomonas syringae fleQ gene has a positive effect on motility

    USDA-ARS?s Scientific Manuscript database

    Bacterial flagella production is controlled by a multi-tiered regulatory system that coordinates expression of 40-50 subunits and correct assembly of these complicated structures. Flagellar expression is environmentally controlled, presumably to optimize the benefits and liabilities of flagellar ex...

  11. From molecular evolution to biobricks and synthetic modules: a lesson by the bacterial flagellum.

    PubMed

    Altegoer, Florian; Schuhmacher, Jan; Pausch, Patrick; Bange, Gert

    2014-10-01

    The bacterial flagellum is a motility structure and represents one of the most sophisticated nanomachines in the biosphere. Here, we review the current knowledge on the flagellum, its architecture with respect to differences between Gram-negative and Gram-positive bacteria and other species-specific variations (e.g. the flagellar filament protein, Flagellin). We further focus on the mechanism by which the two nucleotide-binding proteins FlhF and FlhG ensure the correct reproduction of flagella place and number (the flagellation pattern). We will finish the review with an overview of current biotechnological applications, and a perspective of how understanding flagella can contribute to developing modules for synthetic approaches.

  12. Expression, purification and biochemical characterization of the cytoplasmic loop of PomA, a stator component of the Na+ driven flagellar motor

    PubMed Central

    Abe-Yoshizumi, Rei; Kobayashi, Shiori; Gohara, Mizuki; Hayashi, Kokoro; Kojima, Chojiro; Kojima, Seiji; Sudo, Yuki; Asami, Yasuo; Homma, Michio

    2013-01-01

    Flagellar motors embedded in bacterial membranes are molecular machines powered by specific ion flows. Each motor is composed of a stator and a rotor and the interactions of those components are believed to generate the torque. Na+ influx through the PomA/PomB stator complex of Vibrio alginolyticus is coupled to torque generation and is speculated to trigger structural changes in the cytoplasmic domain of PomA that interacts with a rotor protein in the C-ring, FliG, to drive the rotation. In this study, we tried to overproduce the cytoplasmic loop of PomA (PomA-Loop), but it was insoluble. Thus, we made a fusion protein with a small soluble tag (GB1) which allowed us to express and characterize the recombinant protein. The structure of the PomA-Loop seems to be very elongated or has a loose tertiary structure. When the PomA-Loop protein was produced in E. coli, a slight dominant effect was observed on motility. We conclude that the cytoplasmic loop alone retains a certain function. PMID:27493537

  13. Robust Mapping of Incoherent Fiber-Optic Bundles

    NASA Technical Reports Server (NTRS)

    Roberts, Harry E.; Deason, Brent E.; DePlachett, Charles P.; Pilgrim, Robert A.; Sanford, Harold S.

    2007-01-01

    A method and apparatus for mapping between the positions of fibers at opposite ends of incoherent fiber-optic bundles have been invented to enable the use of such bundles to transmit images in visible or infrared light. The method is robust in the sense that it provides useful mapping even for a bundle that contains thousands of narrow, irregularly packed fibers, some of which may be defective. In a coherent fiber-optic bundle, the input and output ends of each fiber lie at identical positions in the input and output planes; therefore, the bundle can be used to transmit images without further modification. Unfortunately, the fabrication of coherent fiber-optic bundles is too labor-intensive and expensive for many applications. An incoherent fiber-optic bundle can be fabricated more easily and at lower cost, but it produces a scrambled image because the position of the end of each fiber in the input plane is generally different from the end of the same fiber in the output plane. However, the image transmitted by an incoherent fiber-optic bundle can be unscrambled (or, from a different perspective, decoded) by digital processing of the output image if the mapping between the input and output fiber-end positions is known. Thus, the present invention enables the use of relatively inexpensive fiber-optic bundles to transmit images.

  14. Rumen Bacterial Degradation of Forage Cell Walls Investigated by Electron Microscopy

    PubMed Central

    Akin, Danny E.; Amos, Henry E.

    1975-01-01

    The association of rumen bacteria with specific leaf tissues of the forage grass Kentucky-31 tall fescue (Festuca arundinacea Schreb.) during in vitro degradation was investigated by transmission and scanning electron microscopy. Examination of degraded leaf cross-sections revealed differential rates of tissue degradation in that the cell walls of the mesophyll and pholem were degraded prior to those of the outer bundle sheath and epidermis. Rumen bacteria appeared to degrade the mesophyll, in some cases, and phloem without prior attachment to the plant cell walls. The degradation of bundle sheath and epidermal cell walls appeared to be preceded by attachment of bacteria to the plant cell wall. Ultrastructural features apparently involved in the adhesion of large cocci to plant cells were observed by transmission and scanning electron microscopy. The physical association between plant and rumen bacterial cells during degradation apparently varies with tissue types. Bacterial attachment, by extracellular features in some microorganisms, is required prior to degradation of the more resistant tissues. Images PMID:16350017

  15. Strategies for prevention of ventilator-associated pneumonia: bundles, devices, and medications for improved patient outcomes.

    PubMed

    Alroumi, Fahad; Sarwar, Akmal; Grgurich, Philip E; Lei, Yuxiu; Hudcova, Jana; Craven, Donald E

    2012-02-01

    Ventilator-associated pneumonia is associated with significant patient morbidity, mortality, and increased health care costs. In the current economic climate, it is crucial to implement cost-effective prevention strategies that have proven efficacy. Multiple prevention measures have been proposed by various expert panels. Global strategies have focused on infection control, and reduction of lower airway colonization with bacterial pathogens, intubation, duration of mechanical ventilation, and length of stay in the intensive care unit. Routine use of the Institute for Healthcare Improvement ventilator care bundle is widespread, and has been clearly demonstrated to be an effective method for reducing the incidence of ventilator-associated pneumonia. In this article, we examine specific aspects of the Institute for Healthcare Improvement bundle, better-designed endotracheal tubes, use of antibiotics and probiotics, and treatment of ventilator-associated tracheobronchitis to prevent ventilator-associated pneumonia.

  16. Warps, grids and curvature in triple vector bundles

    NASA Astrophysics Data System (ADS)

    Flari, Magdalini K.; Mackenzie, Kirill

    2018-06-01

    A triple vector bundle is a cube of vector bundle structures which commute in the (strict) categorical sense. A grid in a triple vector bundle is a collection of sections of each bundle structure with certain linearity properties. A grid provides two routes around each face of the triple vector bundle, and six routes from the base manifold to the total manifold; the warps measure the lack of commutativity of these routes. In this paper we first prove that the sum of the warps in a triple vector bundle is zero. The proof we give is intrinsic and, we believe, clearer than the proof using decompositions given earlier by one of us. We apply this result to the triple tangent bundle T^3M of a manifold and deduce (as earlier) the Jacobi identity. We further apply the result to the triple vector bundle T^2A for a vector bundle A using a connection in A to define a grid in T^2A . In this case the curvature emerges from the warp theorem.

  17. Bundle Security Protocol for ION

    NASA Technical Reports Server (NTRS)

    Burleigh, Scott C.; Birrane, Edward J.; Krupiarz, Christopher

    2011-01-01

    This software implements bundle authentication, conforming to the Delay-Tolerant Networking (DTN) Internet Draft on Bundle Security Protocol (BSP), for the Interplanetary Overlay Network (ION) implementation of DTN. This is the only implementation of BSP that is integrated with ION.

  18. Double-bundle ACL reconstruction can improve rotational stability.

    PubMed

    Yagi, Masayoshi; Kuroda, Ryosuke; Nagamune, Kouki; Yoshiya, Shinichi; Kurosaka, Masahiro

    2007-01-01

    Double-bundle anterior cruciate ligament (ACL) reconstruction reproduces anteromedial and posterolateral bundles, and thus has theoretical advantages over conventional single-bundle reconstruction in controlling rotational torque in vitro. However, its superiority in clinical practice has not been proven. We analyzed rotational stability with three reconstruction techniques in 60 consecutive patients who were randomly divided into three groups (double-bundle, anteromedial single-bundle, posterolateral single-bundle). In the reconstructive procedure, the hamstring tendon was harvested and used as a free tendon graft. Followup examinations were performed 1 year after surgery. Anteroposterior laxity of the knee was examined with a KT-1000 arthrometer, whereas rotatory instability, as elicited by the pivot shift test, was assessed using a new measurement system incorporating three-dimensional electromagnetic sensors. Routine clinical evaluations, including KT examination, demonstrated no differences among the three groups. However, using the new measurement system, patients with double-bundle ACL reconstruction showed better pivot shift control of complex instability than patients with anteromedial and posterolateral single-bundle reconstruction.

  19. Unique ATPase site architecture triggers cis-mediated synchronized ATP binding in heptameric AAA+-ATPase domain of flagellar regulatory protein FlrC.

    PubMed

    Dey, Sanjay; Biswas, Maitree; Sen, Udayaditya; Dasgupta, Jhimli

    2015-04-03

    Bacterial enhancer-binding proteins (bEBPs) oligomerize through AAA(+) domains and use ATP hydrolysis-driven energy to isomerize the RNA polymerase-σ(54) complex during transcriptional initiation. Here, we describe the first structure of the central AAA(+) domain of the flagellar regulatory protein FlrC (FlrC(C)), a bEBP that controls flagellar synthesis in Vibrio cholerae. Our results showed that FlrC(C) forms heptamer both in nucleotide (Nt)-free and -bound states without ATP-dependent subunit remodeling. Unlike the bEBPs such as NtrC1 or PspF, a novel cis-mediated "all or none" ATP binding occurs in the heptameric FlrC(C), because constriction at the ATPase site, caused by loop L3 and helix α7, restricts the proximity of the trans-protomer required for Nt binding. A unique "closed to open" movement of Walker A, assisted by trans-acting "Glu switch" Glu-286, facilitates ATP binding and hydrolysis. Fluorescence quenching and ATPase assays on FlrC(C) and mutants revealed that although Arg-349 of sensor II, positioned by trans-acting Glu-286 and Tyr-290, acts as a key residue to bind and hydrolyze ATP, Arg-319 of α7 anchors ribose and controls the rate of ATP hydrolysis by retarding the expulsion of ADP. Heptameric state of FlrC(C) is restored in solution even with the transition state mimicking ADP·AlF3. Structural results and pulldown assays indicated that L3 renders an in-built geometry to L1 and L2 causing σ(54)-FlrC(C) interaction independent of Nt binding. Collectively, our results underscore a novel mechanism of ATP binding and σ(54) interaction that strives to understand the transcriptional mechanism of the bEBPs, which probably interact directly with the RNA polymerase-σ(54) complex without DNA looping. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. [Biofilm: set-up and organization of a bacterial community].

    PubMed

    Filloux, Alain; Vallet, Isabelle

    2003-01-01

    Bacterial attachment on various surfaces mostly takes place in the form of specialised bacterial communities, referred to as biofilm. The biofilm is formed through series of interactions between cells and adherence to surface, resulting in an organised structure. In this review we have been using Pseudomonas aeruginosa as a model microorganism to describe the series of events that occurred during this developmental process. P. aeruginosa is an opportunistic pathogen that has a wide variety of hosts and infectious sites. In addition to biofilm formation in certain tissues, inert surfaces, such as catheters, are also target for bacterial biofilm development. The use of convenient genetic screens has made possible the identification of numerous biofilm-defective mutants, which have been characterised further. These studies have allowed the proposal for a global model, in which key events are described for the different stages of biofilm formation. Briefly, flagellar mobility is crucial for approaching the surface, whereas type IV pili motility is preponderant for surface colonisation and microcolonies formation. These microcolonies are finally packed together and buried in an exopolysaccharide matrix to form the differentiated bio-film. It is obvious that the different stages of biofilm formation also involved perception of environmental stimuli. These stimuli, and their associated complex regulatory networks, have still to be fully characterised to understand the bacterial strategy, which initiates biofilm formation. One such regulatory system, called Quorum sensing, is one of the key player in the initial differentiation of biofilm. Finally, a better understanding, at the molecular level, of biofilm establishment and persistence should help for the design of antimicrobials that prevent bacterial infections.

  1. Comparison of computational results of the SABRE LMFBR pin bundle blockage code with data from well-instrumented out-of-pile test bundles (THORS bundles 3A and 5A)

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

    Dearing, J.F.

    The Subchannel Analysis of Blockages in Reactor Elements (SABRE) computer code, developed by the United Kingdom Atomic Energy Authority, is currently the only practical tool available for performing detailed analyses of velocity and temperature fields in the recirculating flow regions downstream of blockages in liquid-metal fast breeder reactor (LMFBR) pin bundles. SABRE is a subchannel analysis code; that is, it accurately represents the complex geometry of nuclear fuel pins arranged on a triangular lattice. The results of SABRE computational models are compared here with temperature data from two out-of-pile 19-pin test bundles from the Thermal-Hydraulic Out-of-Reactor Safety (THORS) Facility atmore » Oak Ridge National Laboratory. One of these bundles has a small central flow blockage (bundle 3A), while the other has a large edge blockage (bundle 5A). Values that give best agreement with experiment for the empirical thermal mixing correlation factor, FMIX, in SABRE are suggested. These values of FMIX are Reynolds-number dependent, however, indicating that the coded turbulent mixing correlation is not appropriate for wire-wrap pin bundles.« less

  2. Verification of the FBR fuel bundle-duct interaction analysis code BAMBOO by the out-of-pile bundle compression test with large diameter pins

    NASA Astrophysics Data System (ADS)

    Uwaba, Tomoyuki; Ito, Masahiro; Nemoto, Junichi; Ichikawa, Shoichi; Katsuyama, Kozo

    2014-09-01

    The BAMBOO computer code was verified by results for the out-of-pile bundle compression test with large diameter pin bundle deformation under the bundle-duct interaction (BDI) condition. The pin diameters of the examined test bundles were 8.5 mm and 10.4 mm, which are targeted as preliminary fuel pin diameters for the upgraded core of the prototype fast breeder reactor (FBR) and for demonstration and commercial FBRs studied in the FaCT project. In the bundle compression test, bundle cross-sectional views were obtained from X-ray computer tomography (CT) images and local parameters of bundle deformation such as pin-to-duct and pin-to-pin clearances were measured by CT image analyses. In the verification, calculation results of bundle deformation obtained by the BAMBOO code analyses were compared with the experimental results from the CT image analyses. The comparison showed that the BAMBOO code reasonably predicts deformation of large diameter pin bundles under the BDI condition by assuming that pin bowing and cladding oval distortion are the major deformation mechanisms, the same as in the case of small diameter pin bundles. In addition, the BAMBOO analysis results confirmed that cladding oval distortion effectively suppresses BDI in large diameter pin bundles as well as in small diameter pin bundles.

  3. Are there intracellular Ca2+ oscillations correlated with flagellar beating in human sperm? A three vs. two-dimensional analysis.

    PubMed

    Corkidi, G; Montoya, F; Hernández-Herrera, P; Ríos-Herrera, W A; Müller, M F; Treviño, C L; Darszon, A

    2017-09-01

    Are there intracellular Ca2+ ([Ca2+]i) oscillations correlated with flagellar beating in human sperm? The results reveal statistically significant [Ca2+]i oscillations that are correlated with the human sperm flagellar beating frequency, when measured in three-dimensions (3D). Fast [Ca2+]i oscillations that are correlated to the beating flagellar frequency of cells swimming in a restricted volume have been detected in hamster sperm. To date, such findings have not been confirmed in any other mammalian sperm species. An important question that has remained regarding these observations is whether the fast [Ca2+]i oscillations are real or might they be due to remaining defocusing effects of the Z component arising from the 3D beating of the flagella. Healthy donors whose semen samples fulfill the WHO criteria between the age of 18-28 were selected. Cells from at least six different donors were utilized for analysis. Approximately the same number of experimental and control cells were analyzed. Motile cells were obtained by the swim-up technique and were loaded with Fluo-4 (Ca2+ sensitive dye) or with Calcein (Ca2+ insensitive dye). Ni2+ was used as a non-specific plasma membrane Ca2+ channel blocker. Fluorescence data and flagella position were acquired in 3D. Each cell was recorded for up to 5.6 s within a depth of 16 microns with a high speed camera (coupled to an image intensifier) acquiring at a rate of 3000 frames per second, while an oscillating objective vibrated at 90 Hz via a piezoelectric device. From these samples, eight experimental and nine control sperm cells were analyzed in both 2D and 3D. We have implemented a new system that allows [Ca2+]i measurements of the human sperm flagellum beating in 3D. These measurements reveal statistically significant [Ca2+]i oscillations that correlate with the flagellar beating frequency. These oscillations may arise from intracellular sources and/or Ca2+ transporters, as they were insensitive to external Ni2+, a non

  4. Computational imaging through a fiber-optic bundle

    NASA Astrophysics Data System (ADS)

    Lodhi, Muhammad A.; Dumas, John Paul; Pierce, Mark C.; Bajwa, Waheed U.

    2017-05-01

    Compressive sensing (CS) has proven to be a viable method for reconstructing high-resolution signals using low-resolution measurements. Integrating CS principles into an optical system allows for higher-resolution imaging using lower-resolution sensor arrays. In contrast to prior works on CS-based imaging, our focus in this paper is on imaging through fiber-optic bundles, in which manufacturing constraints limit individual fiber spacing to around 2 μm. This limitation essentially renders fiber-optic bundles as low-resolution sensors with relatively few resolvable points per unit area. These fiber bundles are often used in minimally invasive medical instruments for viewing tissue at macro and microscopic levels. While the compact nature and flexibility of fiber bundles allow for excellent tissue access in-vivo, imaging through fiber bundles does not provide the fine details of tissue features that is demanded in some medical situations. Our hypothesis is that adapting existing CS principles to fiber bundle-based optical systems will overcome the resolution limitation inherent in fiber-bundle imaging. In a previous paper we examined the practical challenges involved in implementing a highly parallel version of the single-pixel camera while focusing on synthetic objects. This paper extends the same architecture for fiber-bundle imaging under incoherent illumination and addresses some practical issues associated with imaging physical objects. Additionally, we model the optical non-idealities in the system to get lower modelling errors.

  5. Altering the speract-induced ion permeability changes that generate flagellar Ca2+ spikes regulates their kinetics and sea urchin sperm motility.

    PubMed

    Wood, Christopher D; Nishigaki, Takuya; Tatsu, Yoshiro; Yumoto, Noboru; Baba, Shoji A; Whitaker, Michael; Darszon, Alberto

    2007-06-15

    Speract, an egg-derived sperm-activating peptide, induces changes in intracellular Ca2+, Na+, pH, cAMP, cGMP, and membrane potential in sperm of the sea urchin Strongylocentrotus purpuratus. Ca2+ is a key regulator of motility in all sperm and, in many marine species, is required for generating turns interspersed with straighter swimming paths that are essential for chemotaxis towards the egg. We show that speract triggers a train of increases in flagellar Ca2+, and that each individual Ca2+ fluctuation induces a transient increase in flagellar asymmetry that leads to a turn. We also find that modifying the amplitude, duration and interval between individual Ca2+ fluctuations by treating sperm with niflumic acid, an inhibitor of Ca2+-activated Cl(-) channels, correspondingly alters the properties of the sperm turns. We conclude that Ca2+ entry through a fast flagellar pathway not only induces sperm turns, but the kinetics of Ca2+ entry may shape the nature of these turns, and that these kinetics are tuned by other channels, possibly including Cl(-) channels. In addition, the speract-induced changes in sperm motility closely resemble those seen during chemotaxis in other marine organisms, yet speract is not a chemoattractant. This implies the Ca2+-induced motility changes are necessary but not sufficient for chemotaxis.

  6. Evaluating big deal journal bundles.

    PubMed

    Bergstrom, Theodore C; Courant, Paul N; McAfee, R Preston; Williams, Michael A

    2014-07-01

    Large commercial publishers sell bundled online subscriptions to their entire list of academic journals at prices significantly lower than the sum of their á la carte prices. Bundle prices differ drastically between institutions, but they are not publicly posted. The data that we have collected enable us to compare the bundle prices charged by commercial publishers with those of nonprofit societies and to examine the types of price discrimination practiced by commercial and nonprofit journal publishers. This information is of interest to economists who study monopolist pricing, librarians interested in making efficient use of library budgets, and scholars who are interested in the availability of the work that they publish.

  7. Flagellar Kinematics and Swimming of Algal Cells in Viscoelastic Fluids

    PubMed Central

    Qin, B.; Gopinath, A.; Yang, J.; Gollub, J. P.; Arratia, P. E.

    2015-01-01

    The motility of microorganisms is influenced greatly by their hydrodynamic interactions with the fluidic environment they inhabit. We show by direct experimental observation of the bi-flagellated alga Chlamydomonas reinhardtii that fluid elasticity and viscosity strongly influence the beating pattern - the gait - and thereby control the propulsion speed. The beating frequency and the wave speed characterizing the cyclical bending are both enhanced by fluid elasticity. Despite these enhancements, the net swimming speed of the alga is hindered for fluids that are sufficiently elastic. The origin of this complex response lies in the interplay between the elasticity-induced changes in the spatial and temporal aspects of the flagellar cycle and the buildup and subsequent relaxation of elastic stresses during the power and recovery strokes. PMID:25778677

  8. Pulsating flow past a tube bundle

    NASA Astrophysics Data System (ADS)

    Molochnikov, V. M.; Mikheev, N. I.; Vazeev, T. A.; Paereliy, A. A.

    2017-11-01

    Visualization of the pulsating cross-flow past the in-line and staggered tube bundles has been performed. The frequency and amplitude of forced flow pulsations and the tube pitch in the bundle varied in the experiments. The main attention was focused on the flow pattern in the near wake of the third-row tube. The most indicative regimes of flow past a tube in a bundle have been revealed depending on forced flow unsteadiness parameters. The obtained data have been generalized in the flow maps in the space of dimensionless frequency (Strouhal number, St) and relative pulsation amplitude, β, individually for the in-line and staggered tube arrangement. Three most indicative regimes of pulsating flow past the tubes in a bundle have been singled out in each flow map.

  9. Learning and evolution in bacterial taxis: an operational amplifier circuit modeling the computational dynamics of the prokaryotic 'two component system' protein network.

    PubMed

    Di Paola, Vieri; Marijuán, Pedro C; Lahoz-Beltra, Rafael

    2004-01-01

    Adaptive behavior in unicellular organisms (i.e., bacteria) depends on highly organized networks of proteins governing purposefully the myriad of molecular processes occurring within the cellular system. For instance, bacteria are able to explore the environment within which they develop by utilizing the motility of their flagellar system as well as a sophisticated biochemical navigation system that samples the environmental conditions surrounding the cell, searching for nutrients or moving away from toxic substances or dangerous physical conditions. In this paper we discuss how proteins of the intervening signal transduction network could be modeled as artificial neurons, simulating the dynamical aspects of the bacterial taxis. The model is based on the assumption that, in some important aspects, proteins can be considered as processing elements or McCulloch-Pitts artificial neurons that transfer and process information from the bacterium's membrane surface to the flagellar motor. This simulation of bacterial taxis has been carried out on a hardware realization of a McCulloch-Pitts artificial neuron using an operational amplifier. Based on the behavior of the operational amplifier we produce a model of the interaction between CheY and FliM, elements of the prokaryotic two component system controlling chemotaxis, as well as a simulation of learning and evolution processes in bacterial taxis. On the one side, our simulation results indicate that, computationally, these protein 'switches' are similar to McCulloch-Pitts artificial neurons, suggesting a bridge between evolution and learning in dynamical systems at cellular and molecular levels and the evolutive hardware approach. On the other side, important protein 'tactilizing' properties are not tapped by the model, and this suggests further complexity steps to explore in the approach to biological molecular computing.

  10. Nonlinear amplitude dynamics in flagellar beating

    NASA Astrophysics Data System (ADS)

    Oriola, David; Gadêlha, Hermes; Casademunt, Jaume

    2017-03-01

    The physical basis of flagellar and ciliary beating is a major problem in biology which is still far from completely understood. The fundamental cytoskeleton structure of cilia and flagella is the axoneme, a cylindrical array of microtubule doublets connected by passive cross-linkers and dynein motor proteins. The complex interplay of these elements leads to the generation of self-organized bending waves. Although many mathematical models have been proposed to understand this process, few attempts have been made to assess the role of dyneins on the nonlinear nature of the axoneme. Here, we investigate the nonlinear dynamics of flagella by considering an axonemal sliding control mechanism for dynein activity. This approach unveils the nonlinear selection of the oscillation amplitudes, which are typically either missed or prescribed in mathematical models. The explicit set of nonlinear equations are derived and solved numerically. Our analysis reveals the spatio-temporal dynamics of dynein populations and flagellum shape for different regimes of motor activity, medium viscosity and flagellum elasticity. Unstable modes saturate via the coupling of dynein kinetics and flagellum shape without the need of invoking a nonlinear axonemal response. Hence, our work reveals a novel mechanism for the saturation of unstable modes in axonemal beating.

  11. Nonlinear amplitude dynamics in flagellar beating.

    PubMed

    Oriola, David; Gadêlha, Hermes; Casademunt, Jaume

    2017-03-01

    The physical basis of flagellar and ciliary beating is a major problem in biology which is still far from completely understood. The fundamental cytoskeleton structure of cilia and flagella is the axoneme, a cylindrical array of microtubule doublets connected by passive cross-linkers and dynein motor proteins. The complex interplay of these elements leads to the generation of self-organized bending waves. Although many mathematical models have been proposed to understand this process, few attempts have been made to assess the role of dyneins on the nonlinear nature of the axoneme. Here, we investigate the nonlinear dynamics of flagella by considering an axonemal sliding control mechanism for dynein activity. This approach unveils the nonlinear selection of the oscillation amplitudes, which are typically either missed or prescribed in mathematical models. The explicit set of nonlinear equations are derived and solved numerically. Our analysis reveals the spatio-temporal dynamics of dynein populations and flagellum shape for different regimes of motor activity, medium viscosity and flagellum elasticity. Unstable modes saturate via the coupling of dynein kinetics and flagellum shape without the need of invoking a nonlinear axonemal response. Hence, our work reveals a novel mechanism for the saturation of unstable modes in axonemal beating.

  12. Nonlinear amplitude dynamics in flagellar beating

    PubMed Central

    Casademunt, Jaume

    2017-01-01

    The physical basis of flagellar and ciliary beating is a major problem in biology which is still far from completely understood. The fundamental cytoskeleton structure of cilia and flagella is the axoneme, a cylindrical array of microtubule doublets connected by passive cross-linkers and dynein motor proteins. The complex interplay of these elements leads to the generation of self-organized bending waves. Although many mathematical models have been proposed to understand this process, few attempts have been made to assess the role of dyneins on the nonlinear nature of the axoneme. Here, we investigate the nonlinear dynamics of flagella by considering an axonemal sliding control mechanism for dynein activity. This approach unveils the nonlinear selection of the oscillation amplitudes, which are typically either missed or prescribed in mathematical models. The explicit set of nonlinear equations are derived and solved numerically. Our analysis reveals the spatio-temporal dynamics of dynein populations and flagellum shape for different regimes of motor activity, medium viscosity and flagellum elasticity. Unstable modes saturate via the coupling of dynein kinetics and flagellum shape without the need of invoking a nonlinear axonemal response. Hence, our work reveals a novel mechanism for the saturation of unstable modes in axonemal beating. PMID:28405357

  13. Episodic payments (bundling): PART I.

    PubMed

    Jacofsky, D J

    2017-10-01

    Episodic, or bundled payments, is a concept now familiar to most in the healthcare arena, but the models are often misunderstood. Under a traditional fee-for-service model, each provider bills separately for their services which creates financial incentives to maximise volumes. Under a bundled payment, a single entity, often referred to as a convener (maybe the hospital, the physician group, or a third party) assumes the risk through a payer contract for all services provided within a defined episode of care, and receives a single (bundled) payment for all services provided for that episode. The time frame around the intervention is variable, but defined in advance, as are included and excluded costs. Timing of the actual payment in a bundle may either be before the episode occurs (prospective payment model), or after the end of the episode through a reconciliation (retrospective payment model). In either case, the defined costs over the defined time frame are borne by the convener. Cite this article: Bone Joint J 2017;99-B:1280-5. ©2017 The British Editorial Society of Bone & Joint Surgery.

  14. Equilibrium polyelectrolyte bundles with different multivalent counterion concentrations

    NASA Astrophysics Data System (ADS)

    Sayar, Mehmet; Holm, Christian

    2010-09-01

    We present the results of molecular-dynamics simulations on the salt concentration dependence of the formation of polyelectrolyte bundles in thermodynamic equilibrium. Extending our results on salt-free systems we investigate here deficiency or excess of trivalent counterions in solution. Our results reveal that the trivalent counterion concentration significantly alters the bundle size and size distribution. The onset of bundle formation takes place at earlier Bjerrum length values with increasing trivalent counterion concentration. For the cases of 80%, 95%, and 100% charge compensation via trivalent counterions, the net charge of the bundles decreases with increasing size. We suggest that competition among two different mechanisms, counterion condensation and merger of bundles, leads to a nonmonotonic change in line-charge density with increasing Bjerrum length. The investigated case of having an abundance of trivalent counterions by 200% prohibits such a behavior. In this case, we find that the difference in effective line-charge density of different size bundles diminishes. In fact, the system displays an isoelectric point, where all bundles become charge neutral.

  15. Evaluating big deal journal bundles

    PubMed Central

    Bergstrom, Theodore C.; Courant, Paul N.; McAfee, R. Preston; Williams, Michael A.

    2014-01-01

    Large commercial publishers sell bundled online subscriptions to their entire list of academic journals at prices significantly lower than the sum of their á la carte prices. Bundle prices differ drastically between institutions, but they are not publicly posted. The data that we have collected enable us to compare the bundle prices charged by commercial publishers with those of nonprofit societies and to examine the types of price discrimination practiced by commercial and nonprofit journal publishers. This information is of interest to economists who study monopolist pricing, librarians interested in making efficient use of library budgets, and scholars who are interested in the availability of the work that they publish. PMID:24979785

  16. Cations Modulate Actin Bundle Mechanics, Assembly Dynamics, and Structure.

    PubMed

    Castaneda, Nicholas; Zheng, Tianyu; Rivera-Jacquez, Hector J; Lee, Hyun-Ju; Hyun, Jaekyung; Balaeff, Alexander; Huo, Qun; Kang, Hyeran

    2018-04-12

    Actin bundles are key factors in the mechanical support and dynamic reorganization of the cytoskeleton. High concentrations of multivalent counterions promote bundle formation through electrostatic attraction between actin filaments that are negatively charged polyelectrolytes. In this study, we evaluate how physiologically relevant divalent cations affect the mechanical, dynamic, and structural properties of actin bundles. Using a combination of total internal reflection fluorescence microscopy, transmission electron microscopy, and dynamic light scattering, we demonstrate that divalent cations modulate bundle stiffness, length distribution, and lateral growth. Molecular dynamics simulations of an all-atom model of the actin bundle reveal specific actin residues coordinate cation-binding sites that promote the bundle formation. Our work suggests that specific cation interactions may play a fundamental role in the assembly, structure, and mechanical properties of actin bundles.

  17. Mutations in Genes Involved in the Flagellar Export Apparatus of the Solvent-Tolerant Pseudomonas putida DOT-T1E Strain Impair Motility and Lead to Hypersensitivity to Toluene Shocks

    PubMed Central

    Segura, Ana; Duque, Estrella; Hurtado, Ana; Ramos, Juan L.

    2001-01-01

    Pseudomonas putida DOT-T1E is a solvent-tolerant strain able to grow in the presence of 1% (vol/vol) toluene in the culture medium. Random mutagenesis with mini-Tn5-′phoA-Km allowed us to isolate a mutant strain (DOT-T1E-42) that formed blue colonies on Luria-Bertani medium supplemented with 5-bromo-4-chloro-3-indolylphosphate and that, in contrast to the wild-type strain, was unable to tolerate toluene shocks (0.3%, vol/vol). The mutant strain exhibited patterns of tolerance or sensitivity to a number of antibiotics, detergents, and chelating agents similar to those of the wild-type strain. The mutation in this strain therefore seemed to specifically affect toluene tolerance. Cloning and sequencing of the mutation revealed that the mini-Tn5-′phoA-Km was inserted within the fliP gene, which is part of the fliLMNOPQRflhBA cluster, a set of genes that encode flagellar structure components. FliP is involved in the export of flagellar proteins, and in fact, the P. putida fliP mutant was nonmotile. The finding that, after replacing the mutant allele with the wild-type one, the strain recovered the wild-type pattern of toluene tolerance and motility unequivocally assigned FliP a function in solvent resistance. An flhB knockout mutant, another gene component of the flagellar export apparatus, was also nonmotile and hypersensitive to toluene. In contrast, a nonpolar mutation at the fliL gene, which encodes a cytoplasmic membrane protein associated with the flagellar basal body, yielded a nonmotile yet toluene-resistant strain. The results are discussed regarding a possible role of the flagellar export apparatus in the transport of one or more proteins necessary for toluene tolerance in P. putida DOT-T1E to the periplasm. PMID:11418551

  18. Square-core bundles for astronomical imaging

    NASA Astrophysics Data System (ADS)

    Bryant, Julia J.; Bland-Hawthorn, Joss

    2012-09-01

    Optical fibre imaging bundles (hexabundles) are proving to be the next logical step for large galaxy surveys as they offer spatially-resolved spectroscopy of galaxies and can be used with conventional fibre positioners. Hexabundles have been effectively demonstrated in the Sydney-AAO Multi-object IFS (SAMI) instrument at the Anglo- Australian Telescope[5]. Based on the success of hexabundles that have circular cores, we have characterised a bundle made instead from square-core fibres. Square cores naturally pack more evenly, which reduces the interstitial holes and can increase the covering, or filling fraction. Furthermore the regular packing simplifies the process of combining and dithering the final images. We discuss the relative issues of filling fraction, focal ratio degradation (FRD), and cross-talk, and find that square-core bundles perform well enough to warrant further development as a format for imaging fibre bundles.

  19. The Lipid Raft Proteome of African Trypanosomes Contains Many Flagellar Proteins.

    PubMed

    Sharma, Aabha I; Olson, Cheryl L; Engman, David M

    2017-08-24

    Lipid rafts are liquid-ordered membrane microdomains that form by preferential association of 3-β-hydroxysterols, sphingolipids and raft-associated proteins often having acyl modifications. We isolated lipid rafts of the protozoan parasite Trypanosoma brucei and determined the protein composition of lipid rafts in the cell. This analysis revealed a striking enrichment of flagellar proteins and several putative signaling proteins in the lipid raft proteome. Calpains and intraflagellar transport proteins, in particular, were found to be abundant in the lipid raft proteome. These findings provide additional evidence supporting the notion that the eukaryotic cilium/flagellum is a lipid raft-enriched specialized structure with high concentrations of sterols, sphingolipids and palmitoylated proteins involved in environmental sensing and cell signaling.

  20. Localized Statistics for DW-MRI Fiber Bundle Segmentation

    PubMed Central

    Lankton, Shawn; Melonakos, John; Malcolm, James; Dambreville, Samuel; Tannenbaum, Allen

    2013-01-01

    We describe a method for segmenting neural fiber bundles in diffusion-weighted magnetic resonance images (DWMRI). As these bundles traverse the brain to connect regions, their local orientation of diffusion changes drastically, hence a constant global model is inaccurate. We propose a method to compute localized statistics on orientation information and use it to drive a variational active contour segmentation that accurately models the non-homogeneous orientation information present along the bundle. Initialized from a single fiber path, the proposed method proceeds to capture the entire bundle. We demonstrate results using the technique to segment the cingulum bundle and describe several extensions making the technique applicable to a wide range of tissues. PMID:23652079

  1. Morphology, Growth, and Size Limit of Bacterial Cells

    NASA Astrophysics Data System (ADS)

    Jiang, Hongyuan; Sun, Sean X.

    2010-07-01

    Bacterial cells utilize a living peptidoglycan network (PG) to separate the cell interior from the surroundings. The shape of the cell is controlled by PG synthesis and cytoskeletal proteins that form bundles and filaments underneath the cell wall. The PG layer also resists turgor pressure and protects the cell from osmotic shock. We argue that mechanical influences alter the chemical equilibrium of the reversible PG assembly and determine the cell shape and cell size. Using a mechanochemical approach, we show that the cell shape can be regarded as a steady state of a growing network under the influence of turgor pressure and mechanical stress. Using simple elastic models, we predict the size of common spherical and rodlike bacteria. The influence of cytoskeletal bundles such as crescentin and MreB are discussed within the context of our model.

  2. Monoubiquitination Inhibits the Actin Bundling Activity of Fascin*

    PubMed Central

    Lin, Shengchen; Lu, Shuang; Mulaj, Mentor; Fang, Bin; Keeley, Tyler; Wan, Lixin; Hao, Jihui; Muschol, Martin; Sun, Jianwei; Yang, Shengyu

    2016-01-01

    Fascin is an actin bundling protein that cross-links individual actin filaments into straight, compact, and stiff bundles, which are crucial for the formation of filopodia, stereocillia, and other finger-like membrane protrusions. The dysregulation of fascin has been implicated in cancer metastasis, hearing loss, and blindness. Here we identified monoubiquitination as a novel mechanism that regulates fascin bundling activity and dynamics. The monoubiquitination sites were identified to be Lys247 and Lys250, two residues located in a positive charge patch at the actin binding site 2 of fascin. Using a chemical ubiquitination method, we synthesized chemically monoubiquitinated fascin and determined the effects of monoubiquitination on fascin bundling activity and dynamics. Our data demonstrated that monoubiquitination decreased the fascin bundling EC50, delayed the initiation of bundle assembly, and accelerated the disassembly of existing bundles. By analyzing the electrostatic properties on the solvent-accessible surface of fascin, we proposed that monoubiquitination introduced steric hindrance to interfere with the interaction between actin filaments and the positively charged patch at actin binding site 2. We also identified Smurf1 as a E3 ligase regulating the monoubiquitination of fascin. Our findings revealed a previously unidentified regulatory mechanism for fascin, which will have important implications for the understanding of actin bundle regulation under physiological and pathological conditions. PMID:27879315

  3. Monoubiquitination Inhibits the Actin Bundling Activity of Fascin.

    PubMed

    Lin, Shengchen; Lu, Shuang; Mulaj, Mentor; Fang, Bin; Keeley, Tyler; Wan, Lixin; Hao, Jihui; Muschol, Martin; Sun, Jianwei; Yang, Shengyu

    2016-12-30

    Fascin is an actin bundling protein that cross-links individual actin filaments into straight, compact, and stiff bundles, which are crucial for the formation of filopodia, stereocillia, and other finger-like membrane protrusions. The dysregulation of fascin has been implicated in cancer metastasis, hearing loss, and blindness. Here we identified monoubiquitination as a novel mechanism that regulates fascin bundling activity and dynamics. The monoubiquitination sites were identified to be Lys 247 and Lys 250 , two residues located in a positive charge patch at the actin binding site 2 of fascin. Using a chemical ubiquitination method, we synthesized chemically monoubiquitinated fascin and determined the effects of monoubiquitination on fascin bundling activity and dynamics. Our data demonstrated that monoubiquitination decreased the fascin bundling EC 50 , delayed the initiation of bundle assembly, and accelerated the disassembly of existing bundles. By analyzing the electrostatic properties on the solvent-accessible surface of fascin, we proposed that monoubiquitination introduced steric hindrance to interfere with the interaction between actin filaments and the positively charged patch at actin binding site 2. We also identified Smurf1 as a E3 ligase regulating the monoubiquitination of fascin. Our findings revealed a previously unidentified regulatory mechanism for fascin, which will have important implications for the understanding of actin bundle regulation under physiological and pathological conditions. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Heat Transfer Analysis in Wire Bundles for Aerospace Vehicles

    NASA Technical Reports Server (NTRS)

    Rickman, S. L.; Iamello, C. J.

    2016-01-01

    Design of wiring for aerospace vehicles relies on an understanding of "ampacity" which refers to the current carrying capacity of wires, either, individually or in wire bundles. Designers rely on standards to derate allowable current flow to prevent exceedance of wire temperature limits due to resistive heat dissipation within the wires or wire bundles. These standards often add considerable margin and are based on empirical data. Commercial providers are taking an aggressive approach to wire sizing which challenges the conventional wisdom of the established standards. Thermal modelling of wire bundles may offer significant mass reduction in a system if the technique can be generalized to produce reliable temperature predictions for arbitrary bundle configurations. Thermal analysis has been applied to the problem of wire bundles wherein any or all of the wires within the bundle may carry current. Wire bundles present analytical challenges because the heat transfer path from conductors internal to the bundle is tortuous, relying on internal radiation and thermal interface conductance to move the heat from within the bundle to the external jacket where it can be carried away by convective and radiative heat transfer. The problem is further complicated by the dependence of wire electrical resistivity on temperature. Reduced heat transfer out of the bundle leads to higher conductor temperatures and, hence, increased resistive heat dissipation. Development of a generalized wire bundle thermal model is presented and compared with test data. The steady state heat balance for a single wire is derived and extended to the bundle configuration. The generalized model includes the effects of temperature varying resistance, internal radiation and thermal interface conductance, external radiation and temperature varying convective relief from the free surface. The sensitivity of the response to uncertainties in key model parameters is explored using Monte Carlo analysis.

  5. Crossed Module Bundle Gerbes; Classification, String Group and Differential Geometry

    NASA Astrophysics Data System (ADS)

    Jurčo, Branislav

    We discuss nonabelian bundle gerbes and their differential geometry using simplicial methods. Associated to any crossed module there is a simplicial group NC, the nerve of the 1-category defined by the crossed module and its geometric realization |NC|. Equivalence classes of principal bundles with structure group |NC| are shown to be one-to-one with stable equivalence classes of what we call crossed module gerbes bundle gerbes. We can also associate to a crossed module a 2-category C'. Then there are two equivalent ways how to view classifying spaces of NC-bundles and hence of |NC|-bundles and crossed module bundle gerbes. We can either apply the W-construction to NC or take the nerve of the 2-category C'. We discuss the string group and string structures from this point of view. Also a simplicial principal bundle can be equipped with a simplicial connection and a B-field. It is shown how in the case of a simplicial principal NC-bundle these simplicial objects give the bundle gerbe connection and the bundle gerbe B-field.

  6. Cost-Effectiveness of a Central Venous Catheter Care Bundle

    PubMed Central

    Halton, Kate A.; Cook, David; Paterson, David L.; Safdar, Nasia; Graves, Nicholas

    2010-01-01

    Background A bundled approach to central venous catheter care is currently being promoted as an effective way of preventing catheter-related bloodstream infection (CR-BSI). Consumables used in the bundled approach are relatively inexpensive which may lead to the conclusion that the bundle is cost-effective. However, this fails to consider the nontrivial costs of the monitoring and education activities required to implement the bundle, or that alternative strategies are available to prevent CR-BSI. We evaluated the cost-effectiveness of a bundle to prevent CR-BSI in Australian intensive care patients. Methods and Findings A Markov decision model was used to evaluate the cost-effectiveness of the bundle relative to remaining with current practice (a non-bundled approach to catheter care and uncoated catheters), or use of antimicrobial catheters. We assumed the bundle reduced relative risk of CR-BSI to 0.34. Given uncertainty about the cost of the bundle, threshold analyses were used to determine the maximum cost at which the bundle remained cost-effective relative to the other approaches to infection control. Sensitivity analyses explored how this threshold alters under different assumptions about the economic value placed on bed-days and health benefits gained by preventing infection. If clinicians are prepared to use antimicrobial catheters, the bundle is cost-effective if national 18-month implementation costs are below $1.1 million. If antimicrobial catheters are not an option the bundle must cost less than $4.3 million. If decision makers are only interested in obtaining cash-savings for the unit, and place no economic value on either the bed-days or the health benefits gained through preventing infection, these cost thresholds are reduced by two-thirds. Conclusions A catheter care bundle has the potential to be cost-effective in the Australian intensive care setting. Rather than anticipating cash-savings from this intervention, decision makers must be prepared

  7. Cost-effectiveness of a central venous catheter care bundle.

    PubMed

    Halton, Kate A; Cook, David; Paterson, David L; Safdar, Nasia; Graves, Nicholas

    2010-09-17

    A bundled approach to central venous catheter care is currently being promoted as an effective way of preventing catheter-related bloodstream infection (CR-BSI). Consumables used in the bundled approach are relatively inexpensive which may lead to the conclusion that the bundle is cost-effective. However, this fails to consider the nontrivial costs of the monitoring and education activities required to implement the bundle, or that alternative strategies are available to prevent CR-BSI. We evaluated the cost-effectiveness of a bundle to prevent CR-BSI in Australian intensive care patients. A Markov decision model was used to evaluate the cost-effectiveness of the bundle relative to remaining with current practice (a non-bundled approach to catheter care and uncoated catheters), or use of antimicrobial catheters. We assumed the bundle reduced relative risk of CR-BSI to 0.34. Given uncertainty about the cost of the bundle, threshold analyses were used to determine the maximum cost at which the bundle remained cost-effective relative to the other approaches to infection control. Sensitivity analyses explored how this threshold alters under different assumptions about the economic value placed on bed-days and health benefits gained by preventing infection. If clinicians are prepared to use antimicrobial catheters, the bundle is cost-effective if national 18-month implementation costs are below $1.1 million. If antimicrobial catheters are not an option the bundle must cost less than $4.3 million. If decision makers are only interested in obtaining cash-savings for the unit, and place no economic value on either the bed-days or the health benefits gained through preventing infection, these cost thresholds are reduced by two-thirds. A catheter care bundle has the potential to be cost-effective in the Australian intensive care setting. Rather than anticipating cash-savings from this intervention, decision makers must be prepared to invest resources in infection control to

  8. Evidence for Loss of a Partial Flagellar Glycolytic Pathway during Trypanosomatid Evolution

    PubMed Central

    Brown, Robert W. B.; Collingridge, Peter W.; Gull, Keith; Rigden, Daniel J.; Ginger, Michael L.

    2014-01-01

    Classically viewed as a cytosolic pathway, glycolysis is increasingly recognized as a metabolic pathway exhibiting surprisingly wide-ranging variations in compartmentalization within eukaryotic cells. Trypanosomatid parasites provide an extreme view of glycolytic enzyme compartmentalization as several glycolytic enzymes are found exclusively in peroxisomes. Here, we characterize Trypanosoma brucei flagellar proteins resembling glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoglycerate kinase (PGK): we show the latter associates with the axoneme and the former is a novel paraflagellar rod component. The paraflagellar rod is an essential extra-axonemal structure in trypanosomes and related protists, providing a platform into which metabolic activities can be built. Yet, bioinformatics interrogation and structural modelling indicate neither the trypanosome PGK-like nor the GAPDH-like protein is catalytically active. Orthologs are present in a free-living ancestor of the trypanosomatids, Bodo saltans: the PGK-like protein from B. saltans also lacks key catalytic residues, but its GAPDH-like protein is predicted to be catalytically competent. We discuss the likelihood that the trypanosome GAPDH-like and PGK-like proteins constitute molecular evidence for evolutionary loss of a flagellar glycolytic pathway, either as a consequence of niche adaptation or the re-localization of glycolytic enzymes to peroxisomes and the extensive changes to glycolytic flux regulation that accompanied this re-localization. Evidence indicating loss of localized ATP provision via glycolytic enzymes therefore provides a novel contribution to an emerging theme of hidden diversity with respect to compartmentalization of the ubiquitous glycolytic pathway in eukaryotes. A possibility that trypanosome GAPDH-like protein additionally represents a degenerate example of a moonlighting protein is also discussed. PMID:25050549

  9. Topological distribution of four-alpha-helix bundles.

    PubMed Central

    Presnell, S R; Cohen, F E

    1989-01-01

    The four-alpha-helix bundle, a common structural motif in globular proteins, provides an excellent forum for the examination of predictive constraints for protein backbone topology. An exhaustive examination of the Brookhaven Crystallographic Protein Data Bank and other literature sources has lead to the discovery of 20 putative four-alpha-helix bundles. Application of an analytical method that examines the difference between solvent-accessible surface areas in packed and partially unpacked bundles reduced the number of structures to 16. Angular requirements further reduced the list of bundles to 13. In 12 of these bundles, all pairs of neighboring helices were oriented in an anti-parallel fashion. This distribution is in accordance with structure types expected if the helix macro dipole effect makes a substantial contribution to the stability of the native structure. The characterizations and classifications made in this study prompt a reevaluation of constraints used in structure prediction efforts. Images PMID:2771946

  10. Nonabelian Bundle Gerbes, Their Differential Geometry and Gauge Theory

    NASA Astrophysics Data System (ADS)

    Aschieri, Paolo; Cantini, Luigi; Jurčo, Branislav

    2005-03-01

    Bundle gerbes are a higher version of line bundles, we present nonabelian bundle gerbes as a higher version of principal bundles. Connection, curving, curvature and gauge transformations are studied both in a global coordinate independent formalism and in local coordinates. These are the gauge fields needed for the construction of Yang-Mills theories with 2-form gauge potential.

  11. Nanomechanics of Pectin-Linked β-Lactoglobulin Nanofibril Bundles.

    PubMed

    Loveday, Simon M; Gunning, A Patrick

    2018-06-14

    Nanofibrils of β-lactoglobulin can be assembled into bundles by site-specific noncovalent cross-linking with high-methoxyl pectin (Hettiarachchi et al. Soft Matter 2016, 12, 756). Here we characterized the nanomechanical properties of bundles using atomic force microscopy and force spectroscopy. Bundles had Gaussian cross sections and a mean height of 17.4 ± 1.4 nm. Persistence lengths were calculated using image analysis with the mean-squared end-to-end model. The relationship between the persistence length and the thickness had exponents of 1.69-2.30, which is consistent with previous reports for other fibril types. In force spectroscopy experiments, the bundles stretched in a qualitatively different manner to fibrils, and some of the force curves were consistent with peeling fibrils away from bundles. The flexibility of pectin-linked nanofibril bundles is likely to be tunable by modulating the stiffness and length of fibrils and the ratio of pectin to fibrils, giving rise to a wide range of structures and functionalities.

  12. Cohomology of line bundles: Applications

    NASA Astrophysics Data System (ADS)

    Blumenhagen, Ralph; Jurke, Benjamin; Rahn, Thorsten; Roschy, Helmut

    2012-01-01

    Massless modes of both heterotic and Type II string compactifications on compact manifolds are determined by vector bundle valued cohomology classes. Various applications of our recent algorithm for the computation of line bundle valued cohomology classes over toric varieties are presented. For the heterotic string, the prime examples are so-called monad constructions on Calabi-Yau manifolds. In the context of Type II orientifolds, one often needs to compute cohomology for line bundles on finite group action coset spaces, necessitating us to generalize our algorithm to this case. Moreover, we exemplify that the different terms in Batyrev's formula and its generalizations can be given a one-to-one cohomological interpretation. Furthermore, we derive a combinatorial closed form expression for two Hodge numbers of a codimension two Calabi-Yau fourfold.

  13. PDS4 Bundle Creation Governance Using BPMN

    NASA Astrophysics Data System (ADS)

    Radulescu, C.; Levoe, S. R.; Algermissen, S. S.; Rye, E. D.; Hardman, S. H.

    2015-06-01

    The AMMOS-PDS Pipeline Service (APPS) provides a Bundle Builder tool, which governs the process of creating, and ultimately generates, PDS4 bundles incrementally, as science products are being generated.

  14. New system speeds bundling of split firewood

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

    Not Available

    1979-01-01

    A firewood compacting and strapping machine is manufactured by Carolson Stapler and Shippers Supply, Omaha, and FMC Industrial Packaging Division, Philadelphia. A hydraulic compactor applies 20,000 lbs of compressive force to each bundle of split logs, reducing each package to a diameter of about 12 inches. A polypropylene band is applied and heat sealed around each bundle. Bundles are stacked on end, twenty-four to a pallet, and the entire load is banded with one horizontal strap.

  15. Crystal structure of bacterial cell-surface alginate-binding protein with an M75 peptidase motif

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

    Maruyama, Yukie; Ochiai, Akihito; Mikami, Bunzo

    Research highlights: {yields} Bacterial alginate-binding Algp7 is similar to component EfeO of Fe{sup 2+} transporter. {yields} We determined the crystal structure of Algp7 with a metal-binding motif. {yields} Algp7 consists of two helical bundles formed through duplication of a single bundle. {yields} A deep cleft involved in alginate binding locates around the metal-binding site. {yields} Algp7 may function as a Fe{sup 2+}-chelated alginate-binding protein. -- Abstract: A gram-negative Sphingomonas sp. A1 directly incorporates alginate polysaccharide into the cytoplasm via the cell-surface pit and ABC transporter. A cell-surface alginate-binding protein, Algp7, functions as a concentrator of the polysaccharide in the pit.more » Based on the primary structure and genetic organization in the bacterial genome, Algp7 was found to be homologous to an M75 peptidase motif-containing EfeO, a component of a ferrous ion transporter. Despite the presence of an M75 peptidase motif with high similarity, the Algp7 protein purified from recombinant Escherichia coli cells was inert on insulin B chain and N-benzoyl-Phe-Val-Arg-p-nitroanilide, both of which are substrates for a typical M75 peptidase, imelysin, from Pseudomonas aeruginosa. The X-ray crystallographic structure of Algp7 was determined at 2.10 A resolution by single-wavelength anomalous diffraction. Although a metal-binding motif, HxxE, conserved in zinc ion-dependent M75 peptidases is also found in Algp7, the crystal structure of Algp7 contains no metal even at the motif. The protein consists of two structurally similar up-and-down helical bundles as the basic scaffold. A deep cleft between the bundles is sufficiently large to accommodate macromolecules such as alginate polysaccharide. This is the first structural report on a bacterial cell-surface alginate-binding protein with an M75 peptidase motif.« less

  16. The Lipid Raft Proteome of African Trypanosomes Contains Many Flagellar Proteins

    PubMed Central

    Sharma, Aabha I.; Olson, Cheryl L.; Engman, David M.

    2017-01-01

    Lipid rafts are liquid-ordered membrane microdomains that form by preferential association of 3-β-hydroxysterols, sphingolipids and raft-associated proteins often having acyl modifications. We isolated lipid rafts of the protozoan parasite Trypanosoma brucei and determined the protein composition of lipid rafts in the cell. This analysis revealed a striking enrichment of flagellar proteins and several putative signaling proteins in the lipid raft proteome. Calpains and intraflagellar transport proteins, in particular, were found to be abundant in the lipid raft proteome. These findings provide additional evidence supporting the notion that the eukaryotic cilium/flagellum is a lipid raft-enriched specialized structure with high concentrations of sterols, sphingolipids and palmitoylated proteins involved in environmental sensing and cell signaling. PMID:28837104

  17. Framework for shape analysis of white matter fiber bundles.

    PubMed

    Glozman, Tanya; Bruckert, Lisa; Pestilli, Franco; Yecies, Derek W; Guibas, Leonidas J; Yeom, Kristen W

    2018-02-15

    Diffusion imaging coupled with tractography algorithms allows researchers to image human white matter fiber bundles in-vivo. These bundles are three-dimensional structures with shapes that change over time during the course of development as well as in pathologic states. While most studies on white matter variability focus on analysis of tissue properties estimated from the diffusion data, e.g. fractional anisotropy, the shape variability of white matter fiber bundle is much less explored. In this paper, we present a set of tools for shape analysis of white matter fiber bundles, namely: (1) a concise geometric model of bundle shapes; (2) a method for bundle registration between subjects; (3) a method for deformation estimation. Our framework is useful for analysis of shape variability in white matter fiber bundles. We demonstrate our framework by applying our methods on two datasets: one consisting of data for 6 normal adults and another consisting of data for 38 normal children of age 11 days to 8.5 years. We suggest a robust and reproducible method to measure changes in the shape of white matter fiber bundles. We demonstrate how this method can be used to create a model to assess age-dependent changes in the shape of specific fiber bundles. We derive such models for an ensemble of white matter fiber bundles on our pediatric dataset and show that our results agree with normative human head and brain growth data. Creating these models for a large pediatric longitudinal dataset may improve understanding of both normal development and pathologic states and propose novel parameters for the examination of the pediatric brain. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Actin-binding proteins sensitively mediate F-actin bundle stiffness

    NASA Astrophysics Data System (ADS)

    Claessens, Mireille M. A. E.; Bathe, Mark; Frey, Erwin; Bausch, Andreas R.

    2006-09-01

    Bundles of filamentous actin (F-actin) form primary structural components of a broad range of cytoskeletal processes including filopodia, sensory hair cell bristles and microvilli. Actin-binding proteins (ABPs) allow the cell to tailor the dimensions and mechanical properties of the bundles to suit specific biological functions. Therefore, it is important to obtain quantitative knowledge on the effect of ABPs on the mechanical properties of F-actin bundles. Here we measure the bending stiffness of F-actin bundles crosslinked by three ABPs that are ubiquitous in eukaryotes. We observe distinct regimes of bundle bending stiffness that differ by orders of magnitude depending on ABP type, concentration and bundle size. The behaviour observed experimentally is reproduced quantitatively by a molecular-based mechanical model in which ABP shearing competes with F-actin extension/compression. Our results shed new light on the biomechanical function of ABPs and demonstrate how single-molecule properties determine mesoscopic behaviour. The bending mechanics of F-actin fibre bundles are general and have implications for cytoskeletal mechanics and for the rational design of functional materials.

  19. Hierarchical streamline bundles.

    PubMed

    Yu, Hongfeng; Wang, Chaoli; Shene, Ching-Kuang; Chen, Jacqueline H

    2012-08-01

    Effective 3D streamline placement and visualization play an essential role in many science and engineering disciplines. The main challenge for effective streamline visualization lies in seed placement, i.e., where to drop seeds and how many seeds should be placed. Seeding too many or too few streamlines may not reveal flow features and patterns either because it easily leads to visual clutter in rendering or it conveys little information about the flow field. Not only does the number of streamlines placed matter, their spatial relationships also play a key role in understanding the flow field. Therefore, effective flow visualization requires the streamlines to be placed in the right place and in the right amount. This paper introduces hierarchical streamline bundles, a novel approach to simplifying and visualizing 3D flow fields defined on regular grids. By placing seeds and generating streamlines according to flow saliency, we produce a set of streamlines that captures important flow features near critical points without enforcing the dense seeding condition. We group spatially neighboring and geometrically similar streamlines to construct a hierarchy from which we extract streamline bundles at different levels of detail. Streamline bundles highlight multiscale flow features and patterns through clustered yet not cluttered display. This selective visualization strategy effectively reduces visual clutter while accentuating visual foci, and therefore is able to convey the desired insight into the flow data.

  20. Anatomy of the anterior cruciate ligament with regard to its two bundles.

    PubMed

    Petersen, Wolf; Zantop, Thore

    2007-01-01

    The anterior cruciate ligament (ACL) consists of two major fiber bundles, namely the anteromedial and posterolateral bundle. When the knee is extended, the posterolateral bundle (PL) is tight and the anteromedial (AM) bundle is moderately lax. As the knee is flexed, the femoral attachment of the ACL becomes a more horizontal orientation; causing the AM bundle to tighten and the PL bundle to relax. There is some degree of variability for the femoral origin of the anterome-dial and posterolateral bundle. The anteromedial bundle is located proximal and anterior in the femoral ACL origin (high and deep in the notch when the knee is flexed at 90 degrees ); the posterolateral bundle starts in the distal and posterior aspect of the femoral ACL origin (shallow and low when the knee is flexed at 90 degrees ). In the frontal plane the anteromedial bundle origin is in the 10:30 clock position and the postero-lateral bundle origin in the 9:30 clock position. At the tibial insertion the ACL fans out to form the foot region. The anteromedial bundle insertion is in the anterior part of the tibial ACL footprint, the posterolateral bundle in the posterior part. While the anteromedial bundle is the primary restraint against anterior tibial translation, the posterolateral bundle tends to stabilize the knee near full extension, particularly against rotatory loads.

  1. Bundled Payments for Care Improvement

    PubMed Central

    Hardin, Lauran; Kilian, Adam; Murphy, Elizabeth

    2017-01-01

    BACKGROUND The Centers for Medicare and Medicaid Services Innovation Center introduced the Bundled Payments for Care Improvement (BPCI) initiative in 2011 as 1 strategy to encourage healthcare organizations and clinicians to improve healthcare delivery for patients, both when they are in the hospital and after they are discharged. Mercy Health Saint Mary’s, a large urban academic medical center, engaged in BPCI primarily with a group of medical diagnosis-related groups (DRGs). OBJECTIVES In this article, we describe our experience creating a system of response for the diverse people and diagnoses that fall into the medical DRG bundles and specifically identify organizational factors for enabling successful implementation of bundled payments. RESULTS Our experience suggests that interprofessional collaboration enabled program success. CONCLUSIONS Although still in its early phases, observations from our program’s strategies and tactics may provide potential insights for organizations considering engagement in the BPCI initiative. PMID:28509721

  2. How molecular motors shape the flagellar beat

    PubMed Central

    Riedel-Kruse, Ingmar H.; Hilfinger, Andreas; Howard, Jonathon; Jülicher, Frank

    2007-01-01

    Cilia and eukaryotic flagella are slender cellular appendages whose regular beating propels cells and microorganisms through aqueous media. The beat is an oscillating pattern of propagating bends generated by dynein motor proteins. A key open question is how the activity of the motors is coordinated in space and time. To elucidate the nature of this coordination we inferred the mechanical properties of the motors by analyzing the shape of beating sperm: Steadily beating bull sperm were imaged and their shapes were measured with high precision using a Fourier averaging technique. Comparing our experimental data with wave forms calculated for different scenarios of motor coordination we found that only the scenario of interdoublet sliding regulating motor activity gives rise to satisfactory fits. We propose that the microscopic origin of such “sliding control” is the load dependent detachment rate of motors. Agreement between observed and calculated wave forms was obtained only if significant sliding between microtubules occurred at the base. This suggests a novel mechanism by which changes in basal compliance could reverse the direction of beat propagation. We conclude that the flagellar beat patterns are determined by an interplay of the basal properties of the axoneme and the mechanical feedback of dynein motors. PMID:19404446

  3. Following the Viterbi Path to Deduce Flagellar Actin-Interacting Proteins of Leishmania spp.: Report on Cofilins and Twinfilins

    NASA Astrophysics Data System (ADS)

    Pacheco, Ana Carolina L.; Araújo, Fabiana F.; Kamimura, Michel T.; Medeiros, Sarah R.; Viana, Daniel A.; Oliveira, Fátima de Cássia E.; Filho, Raimundo Araújo; Costa, Marcília P.; Oliveira, Diana M.

    2007-11-01

    For performing vital cellular processes, such as motility, eukaryotic cells rely on the actin cytoskeleton, whose structure and dynamics are tightly controlled by a large number of actin-interacting (AIP) or actin-related/regulating (ARP) proteins. Trypanosomatid protozoa, such as Leishmania, rely on their flagellum for motility and sensory reception, which are believed to allow parasite migration, adhesion, invasion and even persistence on mammalian host tissues to cause disease. Actin can determine cell stiffness and transmit force during mechanotransduction, cytokinesis, cell motility and other cellular shape changes, while the identification and analyses of AIPs can help to improve understanding of their mechanical properties on physiological architectures, such as the present case regarding Leishmania flagellar apparatus. This work conveniently apply bioinformatics tools in some refined pattern recognition techniques (such as hidden Markov models (HMMs) through the Viterbi algorithm/path) in order to improve the recognition of actin-binding/interacting activity through identification of AIPs in genomes, transcriptomes and proteomes of Leishmania species. We here report cofilin and twinfilin as putative components of the flagellar apparatus, a direct bioinformatics contribution in the secondary annotation of Leishmania and trypanosomatid genomes.

  4. Synchronization of Spontaneous Active Motility of Hair Cell Bundles

    PubMed Central

    Zhang, Tracy-Ying; Ji, Seung; Bozovic, Dolores

    2015-01-01

    Hair cells of the inner ear exhibit an active process, believed to be crucial for achieving the sensitivity of auditory and vestibular detection. One of the manifestations of the active process is the occurrence of spontaneous hair bundle oscillations in vitro. Hair bundles are coupled by overlying membranes in vivo; hence, explaining the potential role of innate bundle motility in the generation of otoacoustic emissions requires an understanding of the effects of coupling on the active bundle dynamics. We used microbeads to connect small groups of hair cell bundles, using in vitro preparations that maintain their innate oscillations. Our experiments demonstrate robust synchronization of spontaneous oscillations, with either 1:1 or multi-mode phase-locking. The frequency of synchronized oscillation was found to be near the mean of the innate frequencies of individual bundles. Coupling also led to an improved regularity of entrained oscillations, demonstrated by an increase in the quality factor. PMID:26540409

  5. Sealed fiber-optic bundle feedthrough

    DOEpatents

    Tanner, Carol E.

    2002-01-01

    A sealed fiber-optic bundle feedthrough by which a multitude of fiber-optic elements may be passed through an opening or port in a wall or structure separating two environments at different pressures or temperatures while maintaining the desired pressure or temperature in each environment. The feedthrough comprises a rigid sleeve of suitable material, a bundle of individual optical fibers, and a resin-based sealing material that bonds the individual optical fibers to each other and to the rigid sleeve.

  6. Topological T-duality for torus bundles with monodromy

    NASA Astrophysics Data System (ADS)

    Baraglia, David

    2015-05-01

    We give a simplified definition of topological T-duality that applies to arbitrary torus bundles. The new definition does not involve Chern classes or spectral sequences, only gerbes and morphisms between them. All the familiar topological conditions for T-duals are shown to follow. We determine necessary and sufficient conditions for existence of a T-dual in the case of affine torus bundles. This is general enough to include all principal torus bundles as well as torus bundles with arbitrary monodromy representations. We show that isomorphisms in twisted cohomology, twisted K-theory and of Courant algebroids persist in this general setting. We also give an example where twisted K-theory groups can be computed by iterating T-duality.

  7. Delay Tolerant Networking - Bundle Protocol Simulation

    NASA Technical Reports Server (NTRS)

    SeGui, John; Jenning, Esther

    2006-01-01

    In this paper, we report on the addition of MACHETE models needed to support DTN, namely: the Bundle Protocol (BP) model. To illustrate the useof MACHETE with the additional DTN model, we provide an example simulation to benchmark its performance. We demonstrate the use of the DTN protocol and discuss statistics gathered concerning the total time needed to simulate numerous bundle transmissions.

  8. Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR.

    PubMed Central

    Shin, S; Park, C

    1995-01-01

    During the search for unknown factors involved in motility, we have found that expression of the flagellar master operon flhDC is affected by mutations of the pta and ackA genes, encoding phosphotransacetylase and acetate kinase, respectively (S. Shin, J. Sheen, and C. Park, Korean J. Microbiol. 31:504-511, 1993). Here we describe results showing that this effect is modulated by externally added acetate, except when both pta and ackA are mutated, suggesting the role of acetyl phosphate, an intermediate of acetate metabolism, as a regulatory effector. Furthermore, the following evidence indicates that the phosphorylation of OmpR, a trans factor for osmoregulation, regulates flagellar expression. First, in a strain lacking ompR, the expression of flhDC is no longer responsive to a change in the level of acetyl phosphate. Second, an increase in medium osmolarity does not decrease flhDC expression in an ompR mutant. It is known that such an increase normally enhances OmpR phosphorylation. Third, OmpR protein binds to the DNA fragment containing the flhDC promoter, and its affinity is increased with phosphorylation by acetyl phosphate. DNase I footprinting revealed the regions of the flhDC promoter protected by OmpR in the presence or absence of phosphorylation. Therefore, we propose that the phosphorylated OmpR, generated by either osmolarity change or the internal level of acetyl phosphate, negatively regulates the expression of flagella. PMID:7642497

  9. Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR.

    PubMed

    Shin, S; Park, C

    1995-08-01

    During the search for unknown factors involved in motility, we have found that expression of the flagellar master operon flhDC is affected by mutations of the pta and ackA genes, encoding phosphotransacetylase and acetate kinase, respectively (S. Shin, J. Sheen, and C. Park, Korean J. Microbiol. 31:504-511, 1993). Here we describe results showing that this effect is modulated by externally added acetate, except when both pta and ackA are mutated, suggesting the role of acetyl phosphate, an intermediate of acetate metabolism, as a regulatory effector. Furthermore, the following evidence indicates that the phosphorylation of OmpR, a trans factor for osmoregulation, regulates flagellar expression. First, in a strain lacking ompR, the expression of flhDC is no longer responsive to a change in the level of acetyl phosphate. Second, an increase in medium osmolarity does not decrease flhDC expression in an ompR mutant. It is known that such an increase normally enhances OmpR phosphorylation. Third, OmpR protein binds to the DNA fragment containing the flhDC promoter, and its affinity is increased with phosphorylation by acetyl phosphate. DNase I footprinting revealed the regions of the flhDC promoter protected by OmpR in the presence or absence of phosphorylation. Therefore, we propose that the phosphorylated OmpR, generated by either osmolarity change or the internal level of acetyl phosphate, negatively regulates the expression of flagella.

  10. The N Terminus of Phosphodiesterase TbrPDEB1 of Trypanosoma brucei Contains the Signal for Integration into the Flagellar Skeleton ▿

    PubMed Central

    Luginbuehl, Edith; Ryter, Damaris; Schranz-Zumkehr, Judith; Oberholzer, Michael; Kunz, Stefan; Seebeck, Thomas

    2010-01-01

    The precise subcellular localization of the components of the cyclic AMP (cAMP) signaling pathways is a crucial aspect of eukaryotic intracellular signaling. In the human pathogen Trypanosoma brucei, the strict control of cAMP levels by cAMP-specific phosphodiesterases is essential for parasite survival, both in cell culture and in the infected host. Among the five cyclic nucleotide phosphodiesterases identified in this organism, two closely related isoenzymes, T. brucei PDEB1 (TbrPDEB1) (PDEB1) and TbrPDEB2 (PDEB2) are predominantly responsible for the maintenance of cAMP levels. Despite their close sequence similarity, they are distinctly localized in the cell. PDEB1 is mostly located in the flagellum, where it forms an integral part of the flagellar skeleton. PDEB2 is mainly located in the cell body, and only a minor part of the protein localizes to the flagellum. The current study, using transfection of procyclic trypanosomes with green fluorescent protein (GFP) reporters, demonstrates that the N termini of the two enzymes are essential for determining their final subcellular localization. The first 70 amino acids of PDEB1 are sufficient to specifically direct a GFP reporter to the flagellum and to lead to its detergent-resistant integration into the flagellar skeleton. In contrast, the analogous region of PDEB2 causes the GFP reporter to reside predominantly in the cell body. Mutagenesis of selected residues in the N-terminal region of PDEB2 demonstrated that single amino acid changes are sufficient to redirect the reporter from a cell body location to stable integration into the flagellar skeleton. PMID:20693305

  11. Proteomic Analysis of Intact Flagella of Procyclic Trypanosoma brucei Cells Identifies Novel Flagellar Proteins with Unique Sub-localization and Dynamics*

    PubMed Central

    Subota, Ines; Julkowska, Daria; Vincensini, Laetitia; Reeg, Nele; Buisson, Johanna; Blisnick, Thierry; Huet, Diego; Perrot, Sylvie; Santi-Rocca, Julien; Duchateau, Magalie; Hourdel, Véronique; Rousselle, Jean-Claude; Cayet, Nadège; Namane, Abdelkader; Chamot-Rooke, Julia; Bastin, Philippe

    2014-01-01

    Cilia and flagella are complex organelles made of hundreds of proteins of highly variable structures and functions. Here we report the purification of intact flagella from the procyclic stage of Trypanosoma brucei using mechanical shearing. Structural preservation was confirmed by transmission electron microscopy that showed that flagella still contained typical elements such as the membrane, the axoneme, the paraflagellar rod, and the intraflagellar transport particles. It also revealed that flagella severed below the basal body, and were not contaminated by other cytoskeletal structures such as the flagellar pocket collar or the adhesion zone filament. Mass spectrometry analysis identified a total of 751 proteins with high confidence, including 88% of known flagellar components. Comparison with the cell debris fraction revealed that more than half of the flagellum markers were enriched in flagella and this enrichment criterion was taken into account to identify 212 proteins not previously reported to be associated to flagella. Nine of these were experimentally validated including a 14-3-3 protein not yet reported to be associated to flagella and eight novel proteins termed FLAM (FLAgellar Member). Remarkably, they localized to five different subdomains of the flagellum. For example, FLAM6 is restricted to the proximal half of the axoneme, no matter its length. In contrast, FLAM8 is progressively accumulating at the distal tip of growing flagella and half of it still needs to be added after cell division. A combination of RNA interference and Fluorescence Recovery After Photobleaching approaches demonstrated very different dynamics from one protein to the other, but also according to the stage of construction and the age of the flagellum. Structural proteins are added to the distal tip of the elongating flagellum and exhibit slow turnover whereas membrane proteins such as the arginine kinase show rapid turnover without a detectible polarity. PMID:24741115

  12. Morphometrics of the Southern Green Stink Bug [Nezara viridula (L.) (Hemiptera: Pentatomidae)] Stylet Bundle.

    PubMed

    Esquivel, J F; Droleskey, R E; Ward, L A; Harvey, R B

    2018-03-29

    The southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), is a cosmopolitan pest of high-value cash crops, including cotton (Gossypium hirsutum L.; Malvales: Malvaceae). The pest can ingest and transmit disease-causing bacterial and fungal pathogens of cotton. We hypothesized that the size of the food canal may contribute to selective transmission, as observed in previous reports. The objective of this study was to examine food canal size and other morphometric parameters of the southern green stink bug stylet bundle at two locations (labium and head) to improve our understanding of factors that may contribute to pathogen transmission. For the food canal, females possessed significantly larger canals than males, major axes were significantly longer than minor axes, and canal sizes were numerically higher at the labium compared with the head. For salivary canal, mean axes lengths were similar between sexes and the head and labium. For both food and salivary canals, axes lengths were longer and area was larger at the labium compared with the head. These findings indicate the presence of a "funnel effect" with canals becoming narrower proximally. Sex and location significantly affected the size of the intact stylet bundle. Results indicate the food canal size was not a factor affecting previously observed selective passive transmission. Major and minor axes measurements, coupled with morphological observations of canal shapes and observed "funnel effect" in the food and salivary canals, improve our understanding of the hemipteran stylet bundle and its relationship with the insect's internal morphology.

  13. Time-dependent fiber bundles with local load sharing.

    PubMed

    Newman, W I; Phoenix, S L

    2001-02-01

    Fiber bundle models, where fibers have random lifetimes depending on their load histories, are useful tools in explaining time-dependent failure in heterogeneous materials. Such models shed light on diverse phenomena such as fatigue in structural materials and earthquakes in geophysical settings. Various asymptotic and approximate theories have been developed for bundles with various geometries and fiber load-sharing mechanisms, but numerical verification has been hampered by severe computational demands in larger bundles. To gain insight at large size scales, interest has returned to idealized fiber bundle models in 1D. Such simplified models typically assume either equal load sharing (ELS) among survivors, or local load sharing (LLS) where a failed fiber redistributes its load onto its two nearest flanking survivors. Such models can often be solved exactly or asymptotically in increasing bundle size, N, yet still capture the essence of failure in real materials. The present work focuses on 1D bundles under LLS. As in previous works, a fiber has failure rate following a power law in its load level with breakdown exponent rho. Surviving fibers under fixed loads have remaining lifetimes that are independent and exponentially distributed. We develop both new asymptotic theories and new computational algorithms that greatly increase the bundle sizes that can be treated in large replications (e.g., one million fibers in thousands of realizations). In particular we develop an algorithm that adapts several concepts and methods that are well-known among computer scientists, but relatively unknown among physicists, to dramatically increase the computational speed with no attendant loss of accuracy. We consider various regimes of rho that yield drastically different behavior as N increases. For 1/2< or =rho< or =1, ELS and LLS have remarkably similar behavior (they have identical lifetime distributions at rho=1) with approximate Gaussian bundle lifetime statistics and a

  14. Characterization of active hair-bundle motility by a mechanical-load clamp

    NASA Astrophysics Data System (ADS)

    Salvi, Joshua D.; Maoiléidigh, Dáibhid Ó.; Fabella, Brian A.; Tobin, Mélanie; Hudspeth, A. J.

    2015-12-01

    Active hair-bundle motility endows hair cells with several traits that augment auditory stimuli. The activity of a hair bundle might be controlled by adjusting its mechanical properties. Indeed, the mechanical properties of bundles vary between different organisms and along the tonotopic axis of a single auditory organ. Motivated by these biological differences and a dynamical model of hair-bundle motility, we explore how adjusting the mass, drag, stiffness, and offset force applied to a bundle control its dynamics and response to external perturbations. Utilizing a mechanical-load clamp, we systematically mapped the two-dimensional state diagram of a hair bundle. The clamp system used a real-time processor to tightly control each of the virtual mechanical elements. Increasing the stiffness of a hair bundle advances its operating point from a spontaneously oscillating regime into a quiescent regime. As predicted by a dynamical model of hair-bundle mechanics, this boundary constitutes a Hopf bifurcation.

  15. Quantum Bundle Description of Quantum Projective Spaces

    NASA Astrophysics Data System (ADS)

    Ó Buachalla, Réamonn

    2012-12-01

    We realise Heckenberger and Kolb's canonical calculus on quantum projective ( N - 1)-space C q [ C p N-1] as the restriction of a distinguished quotient of the standard bicovariant calculus for the quantum special unitary group C q [ SU N ]. We introduce a calculus on the quantum sphere C q [ S 2 N-1] in the same way. With respect to these choices of calculi, we present C q [ C p N-1] as the base space of two different quantum principal bundles, one with total space C q [ SU N ], and the other with total space C q [ S 2 N-1]. We go on to give C q [ C p N-1] the structure of a quantum framed manifold. More specifically, we describe the module of one-forms of Heckenberger and Kolb's calculus as an associated vector bundle to the principal bundle with total space C q [ SU N ]. Finally, we construct strong connections for both bundles.

  16. Structures of bacterial homologues of SWEET transporters in two distinct conformations.

    PubMed

    Xu, Yan; Tao, Yuyong; Cheung, Lily S; Fan, Chao; Chen, Li-Qing; Xu, Sophia; Perry, Kay; Frommer, Wolf B; Feng, Liang

    2014-11-20

    SWEETs and their prokaryotic homologues are monosaccharide and disaccharide transporters that are present from Archaea to plants and humans. SWEETs play crucial roles in cellular sugar efflux processes: that is, in phloem loading, pollen nutrition and nectar secretion. Their bacterial homologues, which are called SemiSWEETs, are among the smallest known transporters. Here we show that SemiSWEET molecules, which consist of a triple-helix bundle, form symmetrical, parallel dimers, thereby generating the translocation pathway. Two SemiSWEET isoforms were crystallized, one in an apparently open state and one in an occluded state, indicating that SemiSWEETs and SWEETs are transporters that undergo rocking-type movements during the transport cycle. The topology of the triple-helix bundle is similar yet distinct to that of the basic building block of animal and plant major facilitator superfamily (MFS) transporters (for example, GLUTs and SUTs). This finding indicates two possibilities: that SWEETs and MFS transporters evolved from an ancestral triple-helix bundle or that the triple-helix bundle represents convergent evolution. In SemiSWEETs and SWEETs, two triple-helix bundles are arranged in a parallel configuration to produce the 6- and 6 + 1-transmembrane-helix pores, respectively. In the 12-transmembrane-helix MFS transporters, four triple-helix bundles are arranged into an alternating antiparallel configuration, resulting in a much larger 2 × 2 triple-helix bundle forming the pore. Given the similarity of SemiSWEETs and SWEETs to PQ-loop amino acid transporters and to mitochondrial pyruvate carriers (MPCs), the structures characterized here may also be relevant to other transporters in the MtN3 clan. The insight gained from the structures of these transporters and from the analysis of mutations of conserved residues will improve the understanding of the transport mechanism, as well as allow comparative studies of the different superfamilies involved in sugar

  17. Bundled monocapillary optics

    DOEpatents

    Hirsch, Gregory

    2002-01-01

    A plurality of glass or metal wires are precisely etched to form the desired shape of the individual channels of the final polycapillary optic. This shape is created by carefully controlling the withdrawal speed of a group of wires from an etchant bath. The etched wires undergo a subsequent operation to create an extremely smooth surface. This surface is coated with a layer of material which is selected to maximize the reflectivity of the radiation being used. This reflective surface may be a single layer of material, or a multilayer coating for optimizing the reflectivity in a narrower wavelength interval. The collection of individual wires is assembled into a close-packed multi-wire bundle, and the wires are bonded together in a manner which preserves the close-pack configuration, irrespective of the local wire diameter. The initial wires are then removed by either a chemical etching procedure or mechanical force. In the case of chemical etching, the bundle is generally segmented by cutting a series of etching slots. Prior to removing the wire, the capillary array is typically bonded to a support substrate. The result of the process is a bundle of precisely oriented radiation-reflecting hollow channels. The capillary optic is used for efficiently collecting and redirecting the radiation from a source of radiation which could be the anode of an x-ray tube, a plasma source, the fluorescent radiation from an electron microprobe, a synchrotron radiation source, a reactor or spallation source of neutrons, or some other source.

  18. Interplanetary Overlay Network Bundle Protocol Implementation

    NASA Technical Reports Server (NTRS)

    Burleigh, Scott C.

    2011-01-01

    The Interplanetary Overlay Network (ION) system's BP package, an implementation of the Delay-Tolerant Networking (DTN) Bundle Protocol (BP) and supporting services, has been specifically designed to be suitable for use on deep-space robotic vehicles. Although the ION BP implementation is unique in its use of zero-copy objects for high performance, and in its use of resource-sensitive rate control, it is fully interoperable with other implementations of the BP specification (Internet RFC 5050). The ION BP implementation is built using the same software infrastructure that underlies the implementation of the CCSDS (Consultative Committee for Space Data Systems) File Delivery Protocol (CFDP) built into the flight software of Deep Impact. It is designed to minimize resource consumption, while maximizing operational robustness. For example, no dynamic allocation of system memory is required. Like all the other ION packages, ION's BP implementation is designed to port readily between Linux and Solaris (for easy development and for ground system operations) and VxWorks (for flight systems operations). The exact same source code is exercised in both environments. Initially included in the ION BP implementations are the following: libraries of functions used in constructing bundle forwarders and convergence-layer (CL) input and output adapters; a simple prototype bundle forwarder and associated CL adapters designed to run over an IPbased local area network; administrative tools for managing a simple DTN infrastructure built from these components; a background daemon process that silently destroys bundles whose time-to-live intervals have expired; a library of functions exposed to applications, enabling them to issue and receive data encapsulated in DTN bundles; and some simple applications that can be used for system checkout and benchmarking.

  19. Stability of the parabolic Poincaré bundle

    NASA Astrophysics Data System (ADS)

    Basu, Suratno; Biswas, Indranil; Dan, Krishanu

    Given a compact Riemann surface X and a moduli space Mα(Λ) of parabolic stable bundles on it of fixed determinant of complete parabolic flags, we prove that the Poincaré parabolic bundle on X × Mα(Λ) is parabolic stable with respect to a natural polarization on X × Mα(Λ).

  20. BiSet: Semantic Edge Bundling with Biclusters for Sensemaking.

    PubMed

    Sun, Maoyuan; Mi, Peng; North, Chris; Ramakrishnan, Naren

    2016-01-01

    Identifying coordinated relationships is an important task in data analytics. For example, an intelligence analyst might want to discover three suspicious people who all visited the same four cities. Existing techniques that display individual relationships, such as between lists of entities, require repetitious manual selection and significant mental aggregation in cluttered visualizations to find coordinated relationships. In this paper, we present BiSet, a visual analytics technique to support interactive exploration of coordinated relationships. In BiSet, we model coordinated relationships as biclusters and algorithmically mine them from a dataset. Then, we visualize the biclusters in context as bundled edges between sets of related entities. Thus, bundles enable analysts to infer task-oriented semantic insights about potentially coordinated activities. We make bundles as first class objects and add a new layer, "in-between", to contain these bundle objects. Based on this, bundles serve to organize entities represented in lists and visually reveal their membership. Users can interact with edge bundles to organize related entities, and vice versa, for sensemaking purposes. With a usage scenario, we demonstrate how BiSet supports the exploration of coordinated relationships in text analytics.

  1. Bacterial Tethering Analysis Reveals a “Run-Reverse-Turn” Mechanism for Pseudomonas Species Motility

    PubMed Central

    Qian, Chen; Wong, Chui Ching; Swarup, Sanjay

    2013-01-01

    We have developed a program that can accurately analyze the dynamic properties of tethered bacterial cells. The program works especially well with cells that tend to give rise to unstable rotations, such as polar-flagellated bacteria. The program has two novel components. The first dynamically adjusts the center of the cell's rotational trajectories. The second applies piecewise linear approximation to the accumulated rotation curve to reduce noise and separate the motion of bacteria into phases. Thus, it can separate counterclockwise (CCW) and clockwise (CW) rotations distinctly and measure rotational speed accurately. Using this program, we analyzed the properties of tethered Pseudomonas aeruginosa and Pseudomonas putida cells for the first time. We found that the Pseudomonas flagellar motor spends equal time in both CCW and CW phases and that it rotates with the same speed in both phases. In addition, we discovered that the cell body can remain stationary for short periods of time, leading to the existence of a third phase of the flagellar motor which we call “pause.” In addition, P. aeruginosa cells adopt longer run lengths, fewer pause frequencies, and shorter pause durations as part of their chemotactic response. We propose that one purpose of the pause phase is to allow the cells to turn at a large angle, where we show that pause durations in free-swimming cells positively correlate with turn angle sizes. Taken together, our results suggest a new “run-reverse-turn” paradigm for polar-flagellated Pseudomonas motility that is different from the “run-and-tumble” paradigm established for peritrichous Escherichia coli. PMID:23728820

  2. A comprehensive insight into bacterial virulence in drinking water using 454 pyrosequencing and Illumina high-throughput sequencing.

    PubMed

    Huang, Kailong; Zhang, Xu-Xiang; Shi, Peng; Wu, Bing; Ren, Hongqiang

    2014-11-01

    In order to comprehensively investigate bacterial virulence in drinking water, 454 pyrosequencing and Illumina high-throughput sequencing were used to detect potential pathogenic bacteria and virulence factors (VFs) in a full-scale drinking water treatment and distribution system. 16S rRNA gene pyrosequencing revealed high bacterial diversity in the drinking water (441-586 operational taxonomic units). Bacterial diversity decreased after chlorine disinfection, but increased after pipeline distribution. α-Proteobacteria was the most dominant taxonomic class. Alignment against the established pathogen database showed that several types of putative pathogens were present in the drinking water and Pseudomonas aeruginosa had the highest abundance (over 11‰ of total sequencing reads). Many pathogens disappeared after chlorine disinfection, but P. aeruginosa and Leptospira interrogans were still detected in the tap water. High-throughput sequencing revealed prevalence of various pathogenicity islands and virulence proteins in the drinking water, and translocases, transposons, Clp proteases and flagellar motor switch proteins were the predominant VFs. Both diversity and abundance of the detectable VFs increased after the chlorination, and decreased after the pipeline distribution. This study indicates that joint use of 454 pyrosequencing and Illumina sequencing can comprehensively characterize environmental pathogenesis, and several types of putative pathogens and various VFs are prevalent in drinking water. Copyright © 2014 Elsevier Inc. All rights reserved.

  3. Matrix remodeling between cells and cellular interactions with collagen bundle

    NASA Astrophysics Data System (ADS)

    Kim, Jihan; Sun, Bo

    When cells are surrounded by complex environment, they continuously probe and interact with it by applying cellular traction forces. As cells apply traction forces, they can sense rigidity of their local environment and remodel the matrix microstructure simultaneously. Previous study shows that single human carcinoma cell (MDA-MB-231) remodeled its surrounding extracellular matrix (ECM) and the matrix remodeling was reversible. In this study we examined the matrix microstructure between cells and cellular interaction between them using quantitative confocal microscopy. The result shows that the matrix microstructure is the most significantly remodeled between cells consisting of aligned, and densified collagen fibers (collagen bundle)., the result shows that collagen bundle is irreversible and significantly change micromechanics of ECM around the bundle. We further examined cellular interaction with collagen bundle by analyzing dynamics of actin and talin formation along with the direction of bundle. Lastly, we analyzed dynamics of cellular protrusion and migrating direction of cells along the bundle.

  4. Tokyo Guidelines 2018: management bundles for acute cholangitis and cholecystitis.

    PubMed

    Mayumi, Toshihiko; Okamoto, Kohji; Takada, Tadahiro; Strasberg, Steven M; Solomkin, Joseph S; Schlossberg, David; Pitt, Henry A; Yoshida, Masahiro; Gomi, Harumi; Miura, Fumihiko; Garden, O James; Kiriyama, Seiki; Yokoe, Masamichi; Endo, Itaru; Asbun, Horacio J; Iwashita, Yukio; Hibi, Taizo; Umezawa, Akiko; Suzuki, Kenji; Itoi, Takao; Hata, Jiro; Han, Ho-Seong; Hwang, Tsann-Long; Dervenis, Christos; Asai, Koji; Mori, Yasuhisa; Huang, Wayne Shih-Wei; Belli, Giulio; Mukai, Shuntaro; Jagannath, Palepu; Cherqui, Daniel; Kozaka, Kazuto; Baron, Todd H; de Santibañes, Eduardo; Higuchi, Ryota; Wada, Keita; Gouma, Dirk J; Deziel, Daniel J; Liau, Kui-Hin; Wakabayashi, Go; Padbury, Robert; Jonas, Eduard; Supe, Avinash Nivritti; Singh, Harjit; Gabata, Toshifumi; Chan, Angus C W; Lau, Wan Yee; Fan, Sheung Tat; Chen, Miin-Fu; Ker, Chen-Guo; Yoon, Yoo-Seok; Choi, In-Seok; Kim, Myung-Hwan; Yoon, Dong-Sup; Kitano, Seigo; Inomata, Masafumi; Hirata, Koichi; Inui, Kazuo; Sumiyama, Yoshinobu; Yamamoto, Masakazu

    2018-01-01

    Management bundles that define items or procedures strongly recommended in clinical practice have been used in many guidelines in recent years. Application of these bundles facilitates the adaptation of guidelines and helps improve the prognosis of target diseases. In Tokyo Guidelines 2013 (TG13), we proposed management bundles for acute cholangitis and cholecystitis. Here, in Tokyo Guidelines 2018 (TG18), we redefine the management bundles for acute cholangitis and cholecystitis. Critical parts of the bundles in TG18 include the diagnostic process, severity assessment, transfer of patients if necessary, and therapeutic approach at each time point. Observance of these items and procedures should improve the prognosis of acute cholangitis and cholecystitis. Studies are now needed to evaluate the dissemination of these TG18 bundles and their effectiveness. Free full articles and mobile app of TG18 are available at: http://www.jshbps.jp/modules/en/index.php?content_id=47. Related clinical questions and references are also included. © 2017 Japanese Society of Hepato-Biliary-Pancreatic Surgery.

  5. Spontaneous Oscillation by Hair Bundles of the Bullfrog's Sacculus

    PubMed Central

    Martin, Pascal; Bozovic, D.; Choe, Y.; Hudspeth, A. J.

    2007-01-01

    One prominent manifestation of mechanical activity in hair cells is spontaneous otoacoustic emission, the unprovoked emanation of sound by an internal ear. Because active hair-bundle motility probably constitutes the active process of non-mammalian hair cells, we investigated the ability of hair bundles in the bullfrog's sacculus to produce oscillations that might underlie spontaneous otoacoustic emissions. When maintained in the ear's normal ionic milieu, many bundles oscillated spontaneously through distances as great as 80 nm at frequencies of 5-50 Hz. Whole-cell recording disclosed that the positive phase of movement was associated with the opening of transduction channels. Gentamicin, which blocks transduction channels, reversibly arrested oscillation; drugs that affect the cAMP phosphorylation pathway and might influence myosin's activity altered the rate of oscillation. Increasing the Ca2+ concentration rendered oscillations faster and smaller until they were suppressed; lowering the Ca2+ concentration moderately with chelators had the opposite effect. When a bundle was offset with a stimulus fiber, oscillations were transiently suppressed but gradually resumed. Loading a bundle by partial displacement clamping, which simulated the presence of the accessory structures to which a bundle is ordinarily attached, increased the frequency and diminished the magnitude of oscillation. These observations accord with a model in which oscillations arise from the interplay of the hair bundle's negative stiffness with the activity of adaptation motors and with Ca2+-dependent relaxation of gating springs. PMID:12805294

  6. Broadband and flexible acoustic focusing by metafiber bundles

    NASA Astrophysics Data System (ADS)

    Sun, Hong-Xiang; Chen, Jia-He; Ge, Yong; Yuan, Shou-Qi; Liu, Xiao-Jun

    2018-06-01

    We report a broadband and flexible acoustic focusing through metafiber bundles in air, in which each metafiber consists of eight circular and narrow rectangular cavities. The fractional bandwidth of the acoustic focusing could reach about 0.2, which arises from the eigenmodes of the metafiber structure. Besides, owing to the flexible characteristic of the metafibers, the focus position can be manipulated by bending the metafiber bundles, and the metafiber bundles could bypass rigid scatterers inside the lens structure. More interestingly, the acoustic propagation and focusing directions can be changed by using a designed right-angled direction converter fabricated by the metafibers, and a waveform converter and a focusing lens of the cylindrical acoustic source are realized based on the metafiber bundles. The proposed focusing lens has the advantages of broad bandwidth, flexible structure, and high focusing performance, showing great potentials in versatile applications.

  7. The flagellar master operon flhDC is a pleiotropic regulator involved in motility and virulence of the fish pathogen Yersinia ruckeri.

    PubMed

    Jozwick, A K S; Graf, J; Welch, T J

    2017-03-01

    To investigate the function of the master flagellar operon flhDC in the fish pathogen Yersinia ruckeri and compare the effect of a constructed flhD mutation to a naturally occurring fliR mutation causing loss-of-motility in emergent biotype 2 (BT2) strains. Yersinia ruckeri flhD and fliR mutants were constructed in a motile strain. Both mutations caused loss-of-motility, ablation of flagellin synthesis and phospholipase secretion, similar to naturally occurring BT2 strains. Transcriptome analysis confirmed flhDC regulation of flagellar, chemotaxis and phospholipase loci as well as other genes of diverse function. The flhD mutation confers a competitive advantage within the fish host when compared with its parent strain, while this advantage was not seen with the naturally occurring fliR mutation. An intact flhD is necessary for expression of the flagellar secretion system as well as other diverse loci, consistent with a role for flhD as a pleiotropic regulator. The maintenance of the flhD locus in Y. ruckeri strains suggests its importance for aspects of Y. ruckeri biology other than virulence, since the flhD mutation conferred a competitive advantage during experimental challenge of rainbow trout. Yersinia ruckeri is the causative agent of enteric red mouth disease, an invasive septicaemia that affects farmed salmonid fish species. Disease outbreaks can cause severe economic losses in aquaculture. BT2 variants, which have independently emerged worldwide, are an increasing threat to farmed fish production. Knowledge of mechanisms involved in virulence, conserved functions and gene regulation among strains may be exploited for the development of novel disease control strategies to prevent pathogen growth or virulence phenotypes within aquaculture. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  8. Fiber-bundle-basis sparse reconstruction for high resolution wide-field microendoscopy.

    PubMed

    Mekhail, Simon Peter; Abudukeyoumu, Nilupaer; Ward, Jonathan; Arbuthnott, Gordon; Chormaic, Síle Nic

    2018-04-01

    In order to observe deep regions of the brain, we propose the use of a fiber bundle for microendoscopy. Fiber bundles allow for the excitation and collection of fluorescence as well as wide field imaging while remaining largely impervious to image distortions brought on by bending. Furthermore, their thin diameter, from 200-500 µ m, means their impact on living tissue, though not absent, is minimal. Although wide field imaging with a bundle allows for a high temporal resolution since no scanning is involved, the largest criticism of bundle imaging is the drastically lowered spatial resolution. In this paper, we make use of sparsity in the object being imaged to up sample the low resolution images from the fiber bundle with compressive sensing. We take each image in a single shot by using a measurement basis dictated by the quasi-crystalline arrangement of the bundle's cores. We find that this technique allows us to increase the resolution of a typical image taken through a fiber bundle.

  9. Characterization of midrib vascular bundles of selected medicinal species in Rubiaceae

    NASA Astrophysics Data System (ADS)

    Nurul-Syahirah, M.; Noraini, T.; Latiff, A.

    2016-11-01

    An anatomical study was carried out on mature leaves of five selected medicinal species of Rubiaceae from Peninsular Malaysia. The chosen medicinal species were Aidia densiflora, Aidia racemosa, Chasallia chartacea, Hedyotis auricularia and Ixora grandifolia. The objective of this study is to determine the taxonomic value of midrib anatomical characteristics. Leaves samples were collected from Taman Paku Pakis, Universiti Kebangsaan Malaysia, Bangi, Selangor and Kledang Saiong Forest Reserve, Perak, Malaysia. Leaves samples then were fixed in spirit and acetic acid (3:1), the midrib parts then were sectioned using sliding microtome, cleared using Clorox, stained in Safranin and Alcian blue, mounted in Euparal and were observed under light microscope. Findings in this study have shown all species have collateral bundles. The midrib vascular bundles characteristics that can be used as tool to differentiate between species or genus are vascular bundles system (opened or closed), shape and arrangement of main vascular bundles, presence of both additional and medullary vascular bundles, position of additional vascular bundles, shape of medullary vascular bundles, presence of sclerenchyma cells ensheathed the vascular bundles. As a conclusion, midrib anatomical characteristics can be used to identify and discriminate medicinal plants species studied in the Rubiaceae.

  10. Cryopreservation of sperm bundles (spermatozeugmata) from endangered livebearing goodeids.

    PubMed

    Liu, Yue; Torres, Leticia; Tiersch, Terrence R

    2018-04-14

    More than half of fishes in the family Goodeidae are considered to be endangered, threatened, or vulnerable. Sperm cryopreservation is an effective tool for conserving genetic resources of imperiled populations, but development of protocols with livebearing fishes faces numerous challenges including the natural packaging of sperm into bundles. In this study the cryopreservation of sperm bundles (spermatozeugmata) of three goodeids species was evaluated. Sperm quality was evaluated by activation with NaCl-NaOH solution (at 300 mOsmol/kg and pH 11.8), and analysis of dissociable bundles and dissociation duration. Using Redtail Splitfin (Xenotoca eiseni) as a model, the effects of cryoprotectants (dimethyl sulfoxide, methanol, and glycerol) with different concentrations (5-15% v/v %), equilibration exposure times (1-60 min), cooling rates (5-40 °C/min), concentrations (4 × 10 4 -4 × 10 6 bundles/ml), buffers (HBSS, PBS and NaCl), and buffer osmolalities (200-400 mOsmol/kg) were investigated. After cooling and thawing, sperm bundles maintained their packed form. A specific protocol was developed (10% dimethyl sulfoxide, 20-min equilibration, 10 °C/min cooling rate, 4 × 10 6 bundles/ml, and 300 mOsmol/kg HBSS). This protocol yielded 89 ± 5% of post-thaw dissociable bundles with 209 ± 10 s of dissociation duration for X. eiseni, 96 ± 9% with 814 ± 14 s for Blackfin Goodea (Goodea atripinni), and 66 ± 2% with 726 ± 25 s for Striped Goodeid (Ataeniobius toweri). This is the first study of cryopreservation of sperm within bundles for livebearing fishes and provides a basis for establishment of germplasm repositories for goodeids and other livebearers. Copyright © 2018 Elsevier Inc. All rights reserved.

  11. Bundles over nearly-Kahler homogeneous spaces in heterotic string theory

    NASA Astrophysics Data System (ADS)

    Klaput, Michael; Lukas, Andre; Matti, Cyril

    2011-09-01

    We construct heterotic vacua based on six-dimensional nearly-Kahler homogeneous manifolds and non-trivial vector bundles thereon. Our examples are based on three specific group coset spaces. It is shown how to construct line bundles over these spaces, compute their properties and build up vector bundles consistent with supersymmetry and anomaly cancelation. It turns out that the most interesting coset is SU(3)/U(1)2. This space supports a large number of vector bundles which lead to consistent heterotic vacua, some of them with three chiral families.

  12. Mechanical Overstimulation of Hair Bundles: Suppression and Recovery of Active Motility

    PubMed Central

    Kao, Albert; Meenderink, Sebastiaan W. F.; Bozovic, Dolores

    2013-01-01

    We explore the effects of high-amplitude mechanical stimuli on hair bundles of the bullfrog sacculus. Under in vitro conditions, these bundles exhibit spontaneous limit cycle oscillations. Prolonged deflection exerted two effects. First, it induced an offset in the position of the bundle. Recovery to the original position displayed two distinct time scales, suggesting the existence of two adaptive mechanisms. Second, the stimulus suppressed spontaneous oscillations, indicating a change in the hair bundle’s dynamic state. After cessation of the stimulus, active bundle motility recovered with time. Both effects were dependent on the duration of the imposed stimulus. External calcium concentration also affected the recovery to the oscillatory state. Our results indicate that both offset in the bundle position and calcium concentration control the dynamic state of the bundle. PMID:23505461

  13. Local load-sharing fiber bundle model in higher dimensions.

    PubMed

    Sinha, Santanu; Kjellstadli, Jonas T; Hansen, Alex

    2015-08-01

    We consider the local load-sharing fiber bundle model in one to five dimensions. Depending on the breaking threshold distribution of the fibers, there is a transition where the fracture process becomes localized. In the localized phase, the model behaves as the invasion percolation model. The difference between the local load-sharing fiber bundle model and the equal load-sharing fiber bundle model vanishes with increasing dimensionality with the characteristics of a power law.

  14. Recognition of white matter bundles using local and global streamline-based registration and clustering.

    PubMed

    Garyfallidis, Eleftherios; Côté, Marc-Alexandre; Rheault, Francois; Sidhu, Jasmeen; Hau, Janice; Petit, Laurent; Fortin, David; Cunanne, Stephen; Descoteaux, Maxime

    2018-04-15

    Virtual dissection of diffusion MRI tractograms is cumbersome and needs extensive knowledge of white matter anatomy. This virtual dissection often requires several inclusion and exclusion regions-of-interest that make it a process that is very hard to reproduce across experts. Having automated tools that can extract white matter bundles for tract-based studies of large numbers of people is of great interest for neuroscience and neurosurgical planning. The purpose of our proposed method, named RecoBundles, is to segment white matter bundles and make virtual dissection easier to perform. This can help explore large tractograms from multiple persons directly in their native space. RecoBundles leverages latest state-of-the-art streamline-based registration and clustering to recognize and extract bundles using prior bundle models. RecoBundles uses bundle models as shape priors for detecting similar streamlines and bundles in tractograms. RecoBundles is 100% streamline-based, is efficient to work with millions of streamlines and, most importantly, is robust and adaptive to incomplete data and bundles with missing components. It is also robust to pathological brains with tumors and deformations. We evaluated our results using multiple bundles and showed that RecoBundles is in good agreement with the neuroanatomical experts and generally produced more dense bundles. Across all the different experiments reported in this paper, RecoBundles was able to identify the core parts of the bundles, independently from tractography type (deterministic or probabilistic) or size. Thus, RecoBundles can be a valuable method for exploring tractograms and facilitating tractometry studies. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Turkish and Native English Academic Writers' Use of Lexical Bundles

    ERIC Educational Resources Information Center

    Öztürk, Yusuf; Köse, Gül Durmusoglu

    2016-01-01

    Lexical bundles such as "on the other hand" and "as a result of" are extremely common and important in academic discourse. The appropriate use of lexical bundles typical of a specific academic discipline is important for writers and the absence of such bundles may not sound fluent and native-like. Recent studies (e.g. Adel…

  16. Hair-bundle proteomes of avian and mammalian inner-ear utricles

    PubMed Central

    Wilmarth, Phillip A.; Krey, Jocelyn F.; Shin, Jung-Bum; Choi, Dongseok; David, Larry L.; Barr-Gillespie, Peter G.

    2015-01-01

    Examination of multiple proteomics datasets within or between species increases the reliability of protein identification. We report here proteomes of inner-ear hair bundles from three species (chick, mouse, and rat), which were collected on LTQ or LTQ Velos ion-trap mass spectrometers; the constituent proteins were quantified using MS2 intensities, which are the summed intensities of all peptide fragmentation spectra matched to a protein. The data are available via ProteomeXchange with identifiers PXD002410 (chick LTQ), PXD002414 (chick Velos), PXD002415 (mouse Velos), and PXD002416 (rat LTQ). The two chick bundle datasets compared favourably to a third, already-described chick bundle dataset, which was quantified using MS1 peak intensities, the summed intensities of peptides identified by high-resolution mass spectrometry (PXD000104; updated analysis in PXD002445). The mouse bundle dataset described here was comparable to a different mouse bundle dataset quantified using MS1 intensities (PXD002167). These six datasets will be useful for identifying the core proteome of vestibular hair bundles. PMID:26645194

  17. Double-bundle anterior cruciate ligament reconstruction is superior to single-bundle reconstruction in terms of revision frequency: a study of 22,460 patients from the Swedish National Knee Ligament Register.

    PubMed

    Svantesson, Eleonor; Sundemo, David; Hamrin Senorski, Eric; Alentorn-Geli, Eduard; Musahl, Volker; Fu, Freddie H; Desai, Neel; Stålman, Anders; Samuelsson, Kristian

    2017-12-01

    Studies comparing single- and double-bundle anterior cruciate ligament (ACL) reconstructions often include a combined analysis of anatomic and non-anatomic techniques. The purpose of this study was to compare the revision rates between single- and double-bundle ACL reconstructions in the Swedish National Knee Ligament Register with regard to surgical variables as determined by the anatomic ACL reconstruction scoring checklist (AARSC). Patients from the Swedish National Knee Ligament Register who underwent either single- or double-bundle ACL reconstruction with hamstring tendon autograft during the period 2007-2014 were included. The follow-up period started with primary ACL reconstruction, and the outcome measure was set as revision surgery. An online questionnaire based on the items of the AARSC was used to determine the surgical technique implemented in the single-bundle procedures. These were organized into subgroups based on surgical variables, and the revision rates were compared with the double-bundle ACL reconstruction. Hazard ratios (HR) with 95% confidence interval (CI) was calculated and adjusted for confounders by Cox regression. A total of 22,460 patients were included in the study, of which 21,846 were single-bundle and 614 were double-bundle ACL reconstruction. Double-bundle ACL reconstruction had a revision frequency of 2.0% (n = 12) and single-bundle 3.2% (n = 689). Single-bundle reconstruction had an increased risk of revision surgery compared with double-bundle [adjusted HR 1.98 (95% CI 1.12-3.51), p = 0.019]. The subgroup analysis showed a significantly increased risk of revision surgery in patients undergoing single-bundle with anatomic technique using transportal drilling [adjusted HR 2.51 (95% CI 1.39-4.54), p = 0.002] compared with double-bundle ACL reconstruction. Utilizing a more complete anatomic technique according to the AARSC lowered the hazard rate considerably when transportal drilling was performed but still resulted in

  18. Improvements to Wire Bundle Thermal Modeling for Ampacity Determination

    NASA Technical Reports Server (NTRS)

    Rickman, Steve L.; Iannello, Christopher J.; Shariff, Khadijah

    2017-01-01

    Determining current carrying capacity (ampacity) of wire bundles in aerospace vehicles is critical not only to safety but also to efficient design. Published standards provide guidance on determining wire bundle ampacity but offer little flexibility for configurations where wire bundles of mixed gauges and currents are employed with varying external insulation jacket surface properties. Thermal modeling has been employed in an attempt to develop techniques to assist in ampacity determination for these complex configurations. Previous developments allowed analysis of wire bundle configurations but was constrained to configurations comprised of less than 50 elements. Additionally, for vacuum analyses, configurations with very low emittance external jackets suffered from numerical instability in the solution. A new thermal modeler is presented allowing for larger configurations and is not constrained for low bundle infrared emissivity calculations. Formulation of key internal radiation and interface conductance parameters is discussed including the effects of temperature and air pressure on wire to wire thermal conductance. Test cases comparing model-predicted ampacity and that calculated from standards documents are presented.

  19. Bundles of Norms About Teen Sex and Pregnancy.

    PubMed

    Mollborn, Stefanie; Sennott, Christie

    2015-09-01

    Teen pregnancy is a cultural battleground in struggles over morality, education, and family. At its heart are norms about teen sex, contraception, pregnancy, and abortion. Analyzing 57 interviews with college students, we found that "bundles" of related norms shaped the messages teens hear. Teens did not think their communities encouraged teen sex or pregnancy, but normative messages differed greatly, with either moral or practical rationalizations. Teens readily identified multiple norms intended to regulate teen sex, contraception, abortion, childbearing, and the sanctioning of teen parents. Beyond influencing teens' behavior, norms shaped teenagers' public portrayals and post hoc justifications of their behavior. Although norm bundles are complex to measure, participants could summarize them succinctly. These bundles and their conflicting behavioral prescriptions create space for human agency in negotiating normative pressures. The norm bundles concept has implications for teen pregnancy prevention policies and can help revitalize social norms for understanding health behaviors. © The Author(s) 2014.

  20. Deformation quantization with separation of variables of an endomorphism bundle

    NASA Astrophysics Data System (ADS)

    Karabegov, Alexander

    2014-01-01

    Given a holomorphic Hermitian vector bundle E and a star-product with separation of variables on a pseudo-Kähler manifold, we construct a star product on the sections of the endomorphism bundle of the dual bundle E∗ which also has the appropriately generalized property of separation of variables. For this star product we prove a generalization of Gammelgaard's graph-theoretic formula.

  1. Analytic convergence of harmonic metrics for parabolic Higgs bundles

    NASA Astrophysics Data System (ADS)

    Kim, Semin; Wilkin, Graeme

    2018-04-01

    In this paper we investigate the moduli space of parabolic Higgs bundles over a punctured Riemann surface with varying weights at the punctures. We show that the harmonic metric depends analytically on the weights and the stable Higgs bundle. This gives a Higgs bundle generalisation of a theorem of McOwen on the existence of hyperbolic cone metrics on a punctured surface within a given conformal class, and a generalisation of a theorem of Judge on the analytic parametrisation of these metrics.

  2. [A new type of flagellar structure. Type 9+n

    PubMed Central

    1977-01-01

    The ultrastructural study of the Eoacanthocephala sperm cell shows a variation from 0 to 5 in the number of the axial fibers in the axoneme. All the species of the order Eoacanthocephala available to us show this variation; moreover, every individual possesses simultaneously several different structural types. So, we are dealing with a new flagellar organization: 9+n, with 0 less than or equal to n less than or equal to 5. In the Quadrigyridae and the Tenuisentidae families, n varies from 0 to 4, with a maximum of 2 for most individuals, exceptionally at 1 for some individuals. In the Neoechinorhynchidae family, n varies from 0 to 5 with a conspicuous prevalence of 3 (from 84 to 99%, according to the individual). These results prompted us to reexamine the two other orders of Acanthocephala in which the structural types 9+2 or 9+0 have been considered as fixed. Indeed, we have found a few flagella the structure of which is different from the prevalent one. It seems, therefore, that the number of the central fibers of the axoneme in the Acanthocephala sperm cell is never absolutely fixed. PMID:557042

  3. High-order synchronization of hair cell bundles

    NASA Astrophysics Data System (ADS)

    Levy, Michael; Molzon, Adrian; Lee, Jae-Hyun; Kim, Ji-Wook; Cheon, Jinwoo; Bozovic, Dolores

    2016-12-01

    Auditory and vestibular hair cell bundles exhibit active mechanical oscillations at natural frequencies that are typically lower than the detection range of the corresponding end organs. We explore how these noisy nonlinear oscillators mode-lock to frequencies higher than their internal clocks. A nanomagnetic technique is used to stimulate the bundles without an imposed mechanical load. The evoked response shows regimes of high-order mode-locking. Exploring a broad range of stimulus frequencies and intensities, we observe regions of high-order synchronization, analogous to Arnold Tongues in dynamical systems literature. Significant areas of overlap occur between synchronization regimes, with the bundle intermittently flickering between different winding numbers. We demonstrate how an ensemble of these noisy spontaneous oscillators could be entrained to efficiently detect signals significantly above the characteristic frequencies of the individual cells.

  4. High-order synchronization of hair cell bundles

    PubMed Central

    Levy, Michael; Molzon, Adrian; Lee, Jae-Hyun; Kim, Ji-wook; Cheon, Jinwoo; Bozovic, Dolores

    2016-01-01

    Auditory and vestibular hair cell bundles exhibit active mechanical oscillations at natural frequencies that are typically lower than the detection range of the corresponding end organs. We explore how these noisy nonlinear oscillators mode-lock to frequencies higher than their internal clocks. A nanomagnetic technique is used to stimulate the bundles without an imposed mechanical load. The evoked response shows regimes of high-order mode-locking. Exploring a broad range of stimulus frequencies and intensities, we observe regions of high-order synchronization, analogous to Arnold Tongues in dynamical systems literature. Significant areas of overlap occur between synchronization regimes, with the bundle intermittently flickering between different winding numbers. We demonstrate how an ensemble of these noisy spontaneous oscillators could be entrained to efficiently detect signals significantly above the characteristic frequencies of the individual cells. PMID:27974743

  5. Igg Subclasses Targeting the Flagella of Salmonella enterica Serovar Typhimurium Can Mediate Phagocytosis and Bacterial Killing

    PubMed Central

    Goh, Yun Shan; Armour, Kathryn L; Clark, Michael R; Grant, Andrew J; Mastroeni, Pietro

    2016-01-01

    Invasive non-typhoidal Salmonella are a common cause of invasive disease in immuno-compromised individuals and in children. Multi-drug resistance poses challenges to disease control, with a critical need for effective vaccines. Flagellin is an attractive vaccine candidate due to surface exposure and high epitope copy number, but its potential as a target for opsonophacytic antibodies is unclear. We examined the effect of targeting flagella with different classes of IgG on the interaction between Salmonella Typhimurium and a human phagocyte-like cell line, THP-1. We tagged the FliC flagellar protein with a foreign CD52 mimotope (TSSPSAD) and bacteria were opsonized with a panel of humanised CD52 antibodies with the same antigen-binding V-region, but different constant regions. We found that IgG binding to flagella increases bacterial phagocytosis and reduces viable intracellular bacterial numbers. Opsonisation with IgG3, followed by IgG1, IgG4, and IgG2, resulted in the highest level of bacterial uptake and in the highest reduction in the intracellular load of viable bacteria. Taken together, our data provide proof-of-principle evidence that targeting flagella with antibodies can increase the antibacterial function of host cells, with IgG3 being the most potent subclass. These data will assist the rational design of urgently needed, optimised vaccines against iNTS disease. PMID:27366588

  6. Flagellar membrane fusion and protein exchange in trypanosomes; a new form of cell-cell communication?

    PubMed Central

    Imhof, Simon; Fragoso, Cristina; Hemphill, Andrew; von Schubert, Conrad; Li, Dong; Legant, Wesley; Betzig, Eric; Roditi, Isabel

    2016-01-01

    Diverse structures facilitate direct exchange of proteins between cells, including plasmadesmata in plants and tunnelling nanotubes in bacteria and higher eukaryotes.  Here we describe a new mechanism of protein transfer, flagellar membrane fusion, in the unicellular parasite Trypanosoma brucei. When fluorescently tagged trypanosomes were co-cultured, a small proportion of double-positive cells were observed. The formation of double-positive cells was dependent on the presence of extracellular calcium and was enhanced by placing cells in medium supplemented with fresh bovine serum. Time-lapse microscopy revealed that double-positive cells arose by bidirectional protein exchange in the absence of nuclear transfer.  Furthermore, super-resolution microscopy showed that this process occurred in ≤1 minute, the limit of temporal resolution in these experiments. Both cytoplasmic and membrane proteins could be transferred provided they gained access to the flagellum. Intriguingly, a component of the RNAi machinery (Argonaute) was able to move between cells, raising the possibility that small interfering RNAs are transported as cargo. Transmission electron microscopy showed that shared flagella contained two axonemes and two paraflagellar rods bounded by a single membrane. In some cases flagellar fusion was partial and interactions between cells were transient. In other cases fusion occurred along the entire length of the flagellum, was stable for several hours and might be irreversible. Fusion did not appear to be deleterious for cell function: paired cells were motile and could give rise to progeny while fused. The motile flagella of unicellular organisms are related to the sensory cilia of higher eukaryotes, raising the possibility that protein transfer between cells via cilia or flagella occurs more widely in nature. PMID:27239276

  7. Proteomics on the rims; insights into the biology of the nuclear envelope and flagellar pocket of trypanosomes

    PubMed Central

    Field, Mark C.; Adung’a, Vincent; Obado, Samson; Chait, Brian T.; Rout, Michael P.

    2014-01-01

    SUMMERY Trypanosomatids represent the causative agents of major diseases in humans, livestock and plants, with inevitable suffering and economic hardship as a result. They are also evolutionarily highly divergent organisms, and the many unique aspects of trypanosome biology provide opportunities in terms of identification of drug targets, the challenge of exploiting these putative targets, and at the same time significant scope for exploration of novel and divergent cell biology. We can estimate from genome sequences that the degree of divergence of trypanosomes from animals and fungi is extreme, with perhaps one third to one half of predicted trypanosome proteins having no known function based on homology or recognizable protein domains/architecture. Two highly important aspects of trypanosome biology are the flagellar pocket and the nuclear envelope, where in silico analysis clearly suggests great potential divergence in the proteome. The flagellar pocket is the sole site of endo- and exocytosis in trypanosomes and plays important roles in immune evasion via variant surface glycoprotein (VSG) trafficking and providing a location for sequestration of various invariant receptors. The trypanosome nuclear envelope has been largely unexplored, but by analogy with higher eukaryotes, roles in the regulation of chromatin and most significantly, in controlling VSG gene expression are expected. Here we discuss recent successful proteomics-based approaches towards characterization of the nuclear envelope and the endocytic apparatus, the identification of conserved and novel trypanosomatid-specific features, and the implications of these findings. PMID:22309600

  8. Heavy tailed bacterial motor switching statistics define macroscopic transport properties during upstream contamination by E. coli

    NASA Astrophysics Data System (ADS)

    Figueroa-Morales, N.; Rivera, A.; Altshuler, E.; Darnige, T.; Douarche, C.; Soto, R.; Lindner, A.; Clément, E.

    The motility of E. Coli bacteria is described as a run and tumble process. Changes of direction correspond to a switch in the flagellar motor rotation. The run time distribution is described as an exponential decay of characteristic time close to 1s. Remarkably, it has been demonstrated that the generic response for the distribution of run times is not exponential, but a heavy tailed power law decay, which is at odds with the motility findings. We investigate the consequences of the motor statistics in the macroscopic bacterial transport. During upstream contamination processes in very confined channels, we have identified very long contamination tongues. Using a stochastic model considering bacterial dwelling times on the surfaces related to the run times, we are able to reproduce qualitatively and quantitatively the evolution of the contamination profiles when considering the power law run time distribution. However, the model fails to reproduce the qualitative dynamics when the classical exponential run and tumble distribution is considered. Moreover, we have corroborated the existence of a power law run time distribution by means of 3D Lagrangian tracking. We then argue that the macroscopic transport of bacteria is essentially determined by the motor rotation statistics.

  9. Alpha 2 LASSO Data Bundles

    DOE Data Explorer

    Gustafson, William Jr; Vogelmann, Andrew; Endo, Satoshi; Toto, Tami; Xiao, Heng; Li, Zhijin; Cheng, Xiaoping; Kim, Jinwon; Krishna, Bhargavi

    2015-08-31

    The Alpha 2 release is the second release from the LASSO Pilot Phase that builds upon the Alpha 1 release. Alpha 2 contains additional diagnostics in the data bundles and focuses on cases from spring-summer 2016. A data bundle is a unified package consisting of LASSO LES input and output, observations, evaluation diagnostics, and model skill scores. LES input include model configuration information and forcing data. LES output includes profile statistics and full domain fields of cloud and environmental variables. Model evaluation data consists of LES output and ARM observations co-registered on the same grid and sampling frequency. Model performance is quantified by skill scores and diagnostics in terms of cloud and environmental variables.

  10. Lexical Bundles: Facilitating University "Talk" in Group Discussions

    ERIC Educational Resources Information Center

    Heng, Chan Swee; Kashiha, Hadi; Tan, Helen

    2014-01-01

    Group discussion forms an integral language experience for most language learners, providing them with an opportunity to express themselves in a naturalistic setting. Multi-word expressions are commonly used and one of them is lexical bundles. Lexical bundles are types of extended collocations that occur more commonly than we expect; they are…

  11. Risk Adjustment for Medicare Total Knee Arthroplasty Bundled Payments.

    PubMed

    Clement, R Carter; Derman, Peter B; Kheir, Michael M; Soo, Adrianne E; Flynn, David N; Levin, L Scott; Fleisher, Lee

    2016-09-01

    The use of bundled payments is growing because of their potential to align providers and hospitals on the goal of cost reduction. However, such gain sharing could incentivize providers to "cherry-pick" more profitable patients. Risk adjustment can prevent this unintended consequence, yet most bundling programs include minimal adjustment techniques. This study was conducted to determine how bundled payments for total knee arthroplasty (TKA) should be adjusted for risk. The authors collected financial data for all Medicare patients (age≥65 years) undergoing primary unilateral TKA at an academic center over a period of 2 years (n=941). Multivariate regression was performed to assess the effect of patient factors on the costs of acute inpatient care, including unplanned 30-day readmissions. This analysis mirrors a bundling model used in the Medicare Bundled Payments for Care Improvement initiative. Increased age, American Society of Anesthesiologists (ASA) class, and the presence of a Medicare Major Complications/Comorbid Conditions (MCC) modifier (typically representing major complications) were associated with increased costs (regression coefficients, $57 per year; $729 per ASA class beyond I; and $3122 for patients meeting MCC criteria; P=.003, P=.001, and P<.001, respectively). Differences in costs were not associated with body mass index, sex, or race. If the results are generalizable, Medicare bundled payments for TKA encompassing acute inpatient care should be adjusted upward by the stated amounts for older patients, those with elevated ASA class, and patients meeting MCC criteria. This is likely an underestimate for many bundling models, including the Comprehensive Care for Joint Replacement program, incorporating varying degrees of postacute care. Failure to adjust for factors that affect costs may create adverse incentives, creating barriers to care for certain patient populations. [Orthopedics. 2016; 39(5):e911-e916.]. Copyright 2016, SLACK Incorporated.

  12. A compressed sensing approach for resolution improvement in fiber-bundle based endomicroscopy

    NASA Astrophysics Data System (ADS)

    Dumas, John P.; Lodhi, Muhammad A.; Bajwa, Waheed U.; Pierce, Mark C.

    2018-02-01

    Endomicroscopy techniques such as confocal, multi-photon, and wide-field imaging have all been demonstrated using coherent fiber-optic imaging bundles. While the narrow diameter and flexibility of fiber bundles is clinically advantageous, the number of resolvable points in an image is conventionally limited to the number of individual fibers within the bundle. We are introducing concepts from the compressed sensing (CS) field to fiber bundle based endomicroscopy, to allow images to be recovered with more resolvable points than fibers in the bundle. The distal face of the fiber bundle is treated as a low-resolution sensor with circular pixels (fibers) arranged in a hexagonal lattice. A spatial light modulator is located conjugate to the object and distal face, applying multiple high resolution masks to the intermediate image prior to propagation through the bundle. We acquire images of the proximal end of the bundle for each (known) mask pattern and then apply CS inversion algorithms to recover a single high-resolution image. We first developed a theoretical forward model describing image formation through the mask and fiber bundle. We then imaged objects through a rigid fiber bundle and demonstrate that our CS endomicroscopy architecture can recover intra-fiber details while filling inter-fiber regions with interpolation. Finally, we examine the relationship between reconstruction quality and the ratio of the number of mask elements to the number of fiber cores, finding that images could be generated with approximately 28,900 resolvable points for a 1,000 fiber region in our platform.

  13. Ecosystem service bundles for analyzing tradeoffs in diverse landscapes

    PubMed Central

    Raudsepp-Hearne, C.; Peterson, G. D.; Bennett, E. M.

    2010-01-01

    A key challenge of ecosystem management is determining how to manage multiple ecosystem services across landscapes. Enhancing important provisioning ecosystem services, such as food and timber, often leads to tradeoffs between regulating and cultural ecosystem services, such as nutrient cycling, flood protection, and tourism. We developed a framework for analyzing the provision of multiple ecosystem services across landscapes and present an empirical demonstration of ecosystem service bundles, sets of services that appear together repeatedly. Ecosystem service bundles were identified by analyzing the spatial patterns of 12 ecosystem services in a mixed-use landscape consisting of 137 municipalities in Quebec, Canada. We identified six types of ecosystem service bundles and were able to link these bundles to areas on the landscape characterized by distinct social–ecological dynamics. Our results show landscape-scale tradeoffs between provisioning and almost all regulating and cultural ecosystem services, and they show that a greater diversity of ecosystem services is positively correlated with the provision of regulating ecosystem services. Ecosystem service-bundle analysis can identify areas on a landscape where ecosystem management has produced exceptionally desirable or undesirable sets of ecosystem services. PMID:20194739

  14. Darcy Permeability of Hollow Fiber Bundles Used in Blood Oxygenation Devices

    PubMed Central

    Pacella, Heather E.; Eash, Heidi J.; Federspiel, William J.

    2011-01-01

    Many industrial and biomedical devices (e.g. blood oxygenators and artificial lungs) use bundles of hollow fiber membranes for separation processes. Analyses of flow and mass transport within the shell-side of the fiber bundles most often model the bundle for simplicity as a packed bed or porous media, using a Darcy permeability coefficient estimated from the Blake-Kozeny equation to account for viscous drag from the fibers. In this study, we developed a simple method for measuring the Darcy permeability of hollow fiber membrane bundles and evaluated how well the Blake-Kozeny (BK) equation predicted the Darcy permeability for these bundles. Fiber bundles were fabricated from commercially available Celgard® ×30-240 fiber fabric (300 μm outer diameter fibers @ 35 and 54 fibers/inch) and from a fiber fabric with 193 μm fibers (61 fibers/inch). The fiber bundles were mounted to the bottom of an acrylic tube and Darcy permeability was determined by measuring the elapsed time for a column of glycerol solution to flow through a fiber bundle. The ratio of the measured Darcy permeability to that predicted from the BK equation varied from 1.09 to 0.56. A comprehensive literature review suggested a modified BK equation with the “constant” correlated to porosity. This modification improved the predictions of the BK equation, with the ratio of measured to predicted permeability varying from 1.13 to 0.84. PMID:22927706

  15. Characterization of a space orbited incoherent fiber optic bundle

    NASA Technical Reports Server (NTRS)

    Dewalt, Stephen A.; Taylor, Edward W.

    1993-01-01

    The results of a study performed to determine the effects of adverse space environments on a bundle of over 1800 optical fibers orbited for 69 months are reported. Experimental results are presented on an incoherent fiber optic bundle oriented in low Earth orbit aboard the Long Duration Exposure Facility (LDEF) satellite as part of the Space Environment Effects Experiment (M0006). Measurements were performed to determine if space induced radiation effects changed the fiber bundle characteristics. Data demonstrating the success of light transmitting fibers to withstand the adverse space environment are presented.

  16. Nurses' perceptions of a pressure ulcer prevention care bundle: a qualitative descriptive study.

    PubMed

    Roberts, Shelley; McInnes, Elizabeth; Wallis, Marianne; Bucknall, Tracey; Banks, Merrilyn; Chaboyer, Wendy

    2016-01-01

    Pressure ulcer prevention is a critical patient safety indicator for acute care hospitals. An innovative pressure ulcer prevention care bundle targeting patient participation in their care was recently tested in a cluster randomised trial in eight Australian hospitals. Understanding nurses' perspectives of such an intervention is imperative when interpreting results and translating evidence into practice. As part of a process evaluation for the main trial, this study assessed nurses' perceptions of the usefulness and impact of a pressure ulcer prevention care bundle intervention on clinical practice. This qualitative descriptive study involved semi-structured interviews with nursing staff at four Australian hospitals that were intervention sites for a cluster randomised trial testing a pressure ulcer prevention care bundle. Four to five participants were purposively sampled at each site. A trained interviewer used a semi-structured interview guide to question participants about their perceptions of the care bundle. Interviews were digitally recorded, transcribed and analysed using thematic analysis. Eighteen nurses from four hospitals participated in the study. Nurses' perceptions of the intervention are described in five themes: 1) Awareness of the pressure ulcer prevention care bundle and its similarity to current practice; 2) Improving awareness, communication and participation with the pressure ulcer prevention care bundle; 3) Appreciating the positive aspects of patient participation in care; 4) Perceived barriers to engaging patients in the pressure ulcer prevention care bundle; and 5) Partnering with nursing staff to facilitate pressure ulcer prevention care bundle implementation. Overall, nurses found the care bundle feasible and acceptable. They identified a number of benefits from the bundle, including improved communication, awareness and participation in pressure ulcer prevention care among patients and staff. However, nurses thought the care bundle was

  17. YeeU enhances the bundling of cytoskeletal polymers of MreB and FtsZ, antagonizing the CbtA (YeeV) toxicity in Escherichia coli.

    PubMed

    Masuda, Hisako; Tan, Qian; Awano, Naoki; Wu, Kuen-Phon; Inouye, Masayori

    2012-06-01

    All free-living bacteria carry the toxin-antitoxin (TA) systems controlling cell growth and death under stress conditions. YeeU-YeeV (CbtA) is one of the Escherichia coli TA systems, and the toxin, CbtA, has been reported to inhibit the polymerization of bacterial cytoskeletal proteins, MreB and FtsZ. Here, we demonstrate that the antitoxin, YeeU, is a novel type of antitoxin (type IV TA system), which does not form a complex with CbtA but functions as an antagonist for CbtA toxicity. Specifically, YeeU was found to directly interact with MreB and FtsZ, and enhance the bundling of their filamentous polymers in vitro. Surprisingly, YeeU neutralized not only the toxicity of CbtA but also the toxicity caused by other inhibitors of MreB and FtsZ, such as A22, SulA and MinC, indicating that YeeU-induced bundling of MreB and FtsZ has an intrinsic global stabilizing effect on their homeostasis. Here we propose to rename YeeU as CbeA for cytoskeleton bundling-enhancing factor A. © 2012 Blackwell Publishing Ltd.

  18. Effectiveness of a Model Bundle Payment Initiative for Femur Fracture Patients.

    PubMed

    Lott, Ariana; Belayneh, Rebekah; Haglin, Jack; Konda, Sanjit; Egol, Kenneth A

    2018-05-28

    Analyze the effectiveness of a BPCI (Bundle Payments for Care Improvement) initiative for patients who would be included in a future potential Surgical Hip and Femur Fracture Treatment (SHFFT) bundle. Retrospective cohort SETTING:: Single Academic Institution PATIENTS/PARTICIPANTS:: Patients discharged with operative fixation of a hip or femur fracture (DRG codes 480-482) between 1/2015-10/2016 were included. A BPCI initiative based upon an established program for BPCI Total Joint Arthroplasty (TJA) was initiated for patients with hip and femur fractures in January 2016. Patients were divided into non-bundle (care before initiative) and bundle (care with initiative) cohorts. Application of BPCI principles MAIN OUTCOME MEASURES:: Length of stay, location of discharge, readmissions RESULTS:: 116 patients participated in the "institutional bundle," and 126 received care prior to the initiative. There was a trend towards decreased mean length of stay, (7.3 ± 6.3 days vs. 6.8 ± 4.0 days, p=0.457) and decreased readmission within 90 days (22.2% vs. 18.1%, p=0.426). The number of patients discharged home doubled (30.2% vs. 14.3%, p=0.008). There was no difference in readmission rates in bundle vs. non-bundle patients based on discharged home status; however, bundle patients discharged to SNF trended towards less readmissions than non-bundle patients discharged to SNF (37.3% vs. 50.6%, p=0.402). Mean episode cost reduction due to initiative was estimated to be $6,450 using Medicare reimbursement data. This study demonstrates the potential success of a BPCI initiative at one institution in decreasing post-acute care facility utilization and cost of care when used for a hip and femur fracture population. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

  19. Pseudomonas aeruginosa Evasion of Phagocytosis Is Mediated by Loss of Swimming Motility and Is Independent of Flagellum Expression▿ †

    PubMed Central

    Amiel, Eyal; Lovewell, Rustin R.; O'Toole, George A.; Hogan, Deborah A.; Berwin, Brent

    2010-01-01

    Pseudomonas aeruginosa is a pathogenic Gram-negative bacterium that causes severe opportunistic infections in immunocompromised individuals; in particular, severity of infection with P. aeruginosa positively correlates with poor prognosis in cystic fibrosis (CF) patients. Establishment of chronic infection by this pathogen is associated with downregulation of flagellar expression and of other genes that regulate P. aeruginosa motility. The current paradigm is that loss of flagellar expression enables immune evasion by the bacteria due to loss of engagement by phagocytic receptors that recognize flagellar components and loss of immune activation through flagellin-mediated Toll-like receptor (TLR) signaling. In this work, we employ bacterial and mammalian genetic approaches to demonstrate that loss of motility, not the loss of the flagellum per se, is the critical factor in the development of resistance to phagocytosis by P. aeruginosa. We demonstrate that isogenic P. aeruginosa mutants deficient in flagellar function, but retaining an intact flagellum, are highly resistant to phagocytosis by both murine and human phagocytic cells at levels comparable to those of flagellum-deficient mutants. Furthermore, we show that loss of MyD88 signaling in murine phagocytes does not recapitulate the phagocytic deficit observed for either flagellum-deficient or motility-deficient P. aeruginosa mutants. Our data demonstrate that loss of bacterial motility confers a dramatic resistance to phagocytosis that is independent of both flagellar expression and TLR signaling. These findings provide an explanation for the well-documented observation of nonmotility in clinical P. aeruginosa isolates and for how this phenotype confers upon the bacteria an advantage in the context of immune evasion. PMID:20457788

  20. Pseudomonas aeruginosa evasion of phagocytosis is mediated by loss of swimming motility and is independent of flagellum expression.

    PubMed

    Amiel, Eyal; Lovewell, Rustin R; O'Toole, George A; Hogan, Deborah A; Berwin, Brent

    2010-07-01

    Pseudomonas aeruginosa is a pathogenic Gram-negative bacterium that causes severe opportunistic infections in immunocompromised individuals; in particular, severity of infection with P. aeruginosa positively correlates with poor prognosis in cystic fibrosis (CF) patients. Establishment of chronic infection by this pathogen is associated with downregulation of flagellar expression and of other genes that regulate P. aeruginosa motility. The current paradigm is that loss of flagellar expression enables immune evasion by the bacteria due to loss of engagement by phagocytic receptors that recognize flagellar components and loss of immune activation through flagellin-mediated Toll-like receptor (TLR) signaling. In this work, we employ bacterial and mammalian genetic approaches to demonstrate that loss of motility, not the loss of the flagellum per se, is the critical factor in the development of resistance to phagocytosis by P. aeruginosa. We demonstrate that isogenic P. aeruginosa mutants deficient in flagellar function, but retaining an intact flagellum, are highly resistant to phagocytosis by both murine and human phagocytic cells at levels comparable to those of flagellum-deficient mutants. Furthermore, we show that loss of MyD88 signaling in murine phagocytes does not recapitulate the phagocytic deficit observed for either flagellum-deficient or motility-deficient P. aeruginosa mutants. Our data demonstrate that loss of bacterial motility confers a dramatic resistance to phagocytosis that is independent of both flagellar expression and TLR signaling. These findings provide an explanation for the well-documented observation of nonmotility in clinical P. aeruginosa isolates and for how this phenotype confers upon the bacteria an advantage in the context of immune evasion.

  1. Management bundles for candidaemia: the impact of compliance on clinical outcomes

    PubMed Central

    Takesue, Yoshio; Ueda, Takashi; Mikamo, Hiroshige; Oda, Shigeto; Takakura, Shunji; Kitagawa, Yuko; Kohno, Shigeru; Masuda, A.; Yoshida, C.; Yasunaga, C.; Yamashita, C.; Nakataki, E.; Ohyagi, H.; Yagi, H.; Johnai, H.; Murai, H.; Hanamoto, H.; Nakamura, I.; Sanada, I.; Tandai, I.; Kuroki, J.; Ogawa, J.; Kawahara, K.; Amino, K.; Nakajima, K.; Yoshimoto, K.; Takeda, K.; Nakamura, K.; Suzuki, K.; Yamada, K.; Aizawa, M.; Hashimoto, M.; Ogata, M.; Shirano, M.; Kawada, M.; Kaneda, M.; Yoshioka, M.; Okuda, N.; Sugita, N.; Kikuchi, N.; Fuke, S.; Tsuchihashi, S.; Sugitani, S.; Ikuta, S.; Honda, S.; Nei, T.; Iwamura, T.; Yagi, T.; Kaji, T.; Ichimiya, Y.; Kobayashi, Y.; Minamishima, Y.; Goto, Y.; Hatano, Y.; Nagao, Y.; Yamagishi, Y.; Sashihara, J.; Tsukamoto, A.; Kawaoka, T.; Kobayashi, M.

    2015-01-01

    Objectives The Mycoses Forum in Japan has developed management bundles for candidaemia to incorporate into bedside practice. The aim of this study was to investigate nationwide compliance with the bundles and their impact on clinical outcomes. Methods Non-neutropenic patients treated with antifungals for candidaemia were surveyed. Bundles consist of nine items to complete. Data were sent to the central office between July 2011 and April 2012. Results Six hundred and eight patients were analysed. The compliance rate for achieving all elements was 6.9%, and it increased to 21.4% when compliance was analysed by the bundle except for oral switch. There was a significant difference in clinical success between patients with and without compliance [92.9% versus 75.8% (P = 0.011)]. Compliance with the bundles, however, failed to be an independent factor associated with favourable outcomes. When step-down oral therapy was excluded from the elements of compliance, compliance with the bundles was revealed to be an independent predictor of clinical success (OR 4.42, 95% CI 2.05–9.52) and mortality (OR 0.27, 95% CI 0.13–0.57). Independent individual elements contributing to clinical success were removal of central venous catheters within 24 h, assessment of clinical efficacy on the third to the fifth day and at least 2 weeks of therapy after clearance of candidaemia. Conclusions Compliance with the bundles for candidaemia had a beneficial effect on clinical outcomes. Promotion of the bundles approach may have the potential to narrow the gap between clinical evidence and bedside practice. PMID:25326087

  2. Revascularization of diaphyseal bone segments by vascular bundle implantation.

    PubMed

    Nagi, O N

    2005-11-01

    Vascularized bone transfer is an effective, established treatment for avascular necrosis and atrophic or infected nonunions. However, limited donor sites and technical difficulty limit its application. Vascular bundle transplantation may provide an alternative. However, even if vascular ingrowth is presumed to occur in such situations, its extent in aiding revascularization for ultimate graft incorporation is not well understood. A rabbit tibia model was used to study and compare vascularized, segmental, diaphyseal, nonvascularized conventional, and vascular bundle-implanted grafts with a combination of angiographic, radiographic, histopathologic, and bone scanning techniques. Complete graft incorporation in conventional grafts was observed at 6 months, whereas it was 8 to 12 weeks with either of the vascularized grafts. The pattern of radionuclide uptake and the duration of graft incorporation between vascular segmental bone grafts (with intact endosteal blood supply) and vascular bundle-implanted segmental grafts were similar. A vascular bundle implanted in the recipient bone was found to anastomose extensively with the intraosseous circulation at 6 weeks. Effective revascularization of bone could be seen when a simple vascular bundle was introduced into a segment of bone deprived of its normal blood supply. This simple technique offers promise for improvement of bone graft survival in clinical circumstances.

  3. Surgical Management of Neurovascular Bundle in Uterine Fibroid Pseudocapsule

    PubMed Central

    Malvasi, Antonio; Hurst, Brad S.; Tsin, Daniel A.; Davila, Fausto; Dominguez, Guillermo; Dell'edera, Domenico; Cavallotti, Carlo; Negro, Roberto; Gustapane, Sarah; Teigland, Chris M.; Mettler, Liselotte

    2012-01-01

    The uterine fibroid pseudocapsule is a fibro-neurovascular structure surrounding a leiomyoma, separating it from normal peripheral myometrium. The fibroid pseudocapsule is composed of a neurovascular network rich in neurofibers similar to the neurovascular bundle surrounding a prostate. The nerve-sparing radical prostatectomy has several intriguing parallels to myomectomy. It may serve either as a useful model in modern fibroid surgical removal, or it may accelerate our understanding of the role of the fibrovascular bundle and neurotransmitters in the healing and restoration of reproductive potential after intracapsular myomectomy. Surgical innovations, such as laparoscopic or robotic myomectomy applied to the intracapsular technique with magnification of the fibroid pseudocapsule surrounding a leiomyoma, originated from the radical prostatectomy method that highlighted a careful dissection of the neurovascular bundle to preserve sexual functioning after prostatectomy. Gentle uterine leiomyoma detachment from the pseudocapsule neurovascular bundle has allowed a reduction in uterine bleeding and uterine musculature trauma with sparing of the pseudocapsule neuropeptide fibers. This technique has had a favorable impact on functionality in reproduction and has improved fertility outcomes. Further research should determine the role of the myoma pseudocapsule neurovascular bundle in the formation, growth, and pathophysiological consequences of fibroids, including pain, infertility, and reproductive outcomes. PMID:22906340

  4. Forest Residues Bundling Project

    Treesearch

    U.S. Forest Service

    2007-01-01

    During the summer of 2003, the U.S. Forest Service conducted an evaluation of biomass bundling for forest residue extraction. This CD provides a report of the project results, a video documentary project record, and a collection of images from the project. Additional information is available at:

  5. Novel Membrane-Bound eIF2α Kinase in the Flagellar Pocket of Trypanosoma brucei▿

    PubMed Central

    Moraes, Maria Carolina S.; Jesus, Teresa C. L.; Hashimoto, Nilce N.; Dey, Madhusudan; Schwartz, Kevin J.; Alves, Viviane S.; Avila, Carla C.; Bangs, James D.; Dever, Thomas E.; Schenkman, Sergio; Castilho, Beatriz A.

    2007-01-01

    Translational control mediated by phosphorylation of the alpha subunit of the eukaryotic initiation factor 2 (eIF2α) is central to stress-induced programs of gene expression. Trypanosomatids, important human pathogens, display differentiation processes elicited by contact with the distinct physiological milieu found in their insect vectors and mammalian hosts, likely representing stress situations. Trypanosoma brucei, the agent of African trypanosomiasis, encodes three potential eIF2α kinases (TbeIF2K1 to -K3). We show here that TbeIF2K2 is a transmembrane glycoprotein expressed both in procyclic and in bloodstream forms. The catalytic domain of TbeIF2K2 phosphorylates yeast and mammalian eIF2α at Ser51. It also phosphorylates the highly unusual form of eIF2α found in trypanosomatids specifically at residue Thr169 that corresponds to Ser51 in other eukaryotes. T. brucei eIF2α, however, is not a substrate for GCN2 or PKR in vitro. The putative regulatory domain of TbeIF2K2 does not share any sequence similarity with known eIF2α kinases. In both procyclic and bloodstream forms TbeIF2K2 is mainly localized in the membrane of the flagellar pocket, an organelle that is the exclusive site of exo- and endocytosis in these parasites. It can also be detected in endocytic compartments but not in lysosomes, suggesting that it is recycled between endosomes and the flagellar pocket. TbeIF2K2 location suggests a relevance in sensing protein or nutrient transport in T. brucei, an organism that relies heavily on posttranscriptional regulatory mechanisms to control gene expression in different environmental conditions. This is the first membrane-associated eIF2α kinase described in unicellular eukaryotes. PMID:17873083

  6. Consensus Bundle on Maternal Mental Health: Perinatal Depression and Anxiety.

    PubMed

    Kendig, Susan; Keats, John P; Hoffman, M Camille; Kay, Lisa B; Miller, Emily S; Moore Simas, Tiffany A; Frieder, Ariela; Hackley, Barbara; Indman, Pec; Raines, Christena; Semenuk, Kisha; Wisner, Katherine L; Lemieux, Lauren A

    2017-03-01

    Perinatal mood and anxiety disorders are among the most common mental health conditions encountered by women of reproductive age. When left untreated, perinatal mood and anxiety disorders can have profound adverse effects on women and their children, ranging from increased risk of poor adherence to medical care, exacerbation of medical conditions, loss of interpersonal and financial resources, smoking and substance use, suicide, and infanticide. Perinatal mood and anxiety disorders are associated with increased risks of maternal and infant mortality and morbidity and are recognized as a significant patient safety issue. In 2015, the Council on Patient Safety in Women's Health Care convened an interdisciplinary workgroup to develop an evidence-based patient safety bundle to address maternal mental health. The focus of this bundle is perinatal mood and anxiety disorders. The bundle is modeled after other bundles released by the Council on Patient Safety in Women's Health Care and provides broad direction for incorporating perinatal mood and anxiety disorder screening, intervention, referral, and follow-up into maternity care practice across health care settings. This commentary provides information to assist with bundle implementation.

  7. Fabrication of Gate-Electrode Integrated Carbon-Nanotube Bundle Field Emitters

    NASA Technical Reports Server (NTRS)

    Toda, Risaku; Bronikowski, Michael; Luong, Edward; Manohara, Harish

    2008-01-01

    A continuing effort to develop carbon-nanotube-based field emitters (cold cathodes) as high-current-density electron sources has yielded an optimized device design and a fabrication scheme to implement the design. One major element of the device design is to use a planar array of bundles of carbon nanotubes as the field-emission tips and to optimize the critical dimensions of the array (principally, heights of bundles and distances between them) to obtain high area-averaged current density and high reliability over a long operational lifetime a concept that was discussed in more detail in Arrays of Bundles of Carbon Nanotubes as Field Emitters (NPO-40817), NASA Tech Briefs, Vol. 31, No. 2 (February 2007), page 58. Another major element of the design is to configure the gate electrodes (anodes used to extract, accelerate, and/or focus electrons) as a ring that overhangs a recess wherein the bundles of nanotubes are located, such that by virtue of the proximity between the ring and the bundles, a relatively low applied potential suffices to generate the large electric field needed for emission of electrons.

  8. The BID Domain of Type IV Secretion Substrates Forms a Conserved Four-Helix Bundle Topped with a Hook.

    PubMed

    Stanger, Frédéric V; de Beer, Tjaart A P; Dranow, David M; Schirmer, Tilman; Phan, Isabelle; Dehio, Christoph

    2017-01-03

    The BID (Bep intracellular delivery) domain functions as secretion signal in a subfamily of protein substrates of bacterial type IV secretion (T4S) systems. It mediates transfer of (1) relaxases and the attached DNA during bacterial conjugation, and (2) numerous Bartonella effector proteins (Beps) during protein transfer into host cells infected by pathogenic Bartonella species. Furthermore, BID domains of Beps have often evolved secondary effector functions within host cells. Here, we provide crystal structures for three representative BID domains and describe a novel conserved fold characterized by a compact, antiparallel four-helix bundle topped with a hook. The conserved hydrophobic core provides a rigid scaffold to a surface that, despite a few conserved exposed residues and similarities in charge distribution, displays significant variability. We propose that the genuine function of BID domains as T4S signal may primarily depend on their rigid structure, while the plasticity of their surface may facilitate adaptation to secondary effector functions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Bundled payment and enhanced recovery after surgery.

    PubMed

    Huang, Jeffrey

    2015-01-01

    Medicare's fee-for-service (FFS) payment model may contribute to unsustainable spending growth. Payers are turning to alternative payment methods. The leading alternative payment model to the FFS problem is bundled payment. The Centers for Medicare & Medicaid Services (CMS) is taking another step to improve healthcare quality at lower cost. The CMS's Center for Medicare and Medicaid Innovation developed four models of bundled payments and 48 discrete clinical condition episodes. Many surgical care procedures are included in the 48 different clinical condition episodes.

  10. 48 CFR 7.107 - Additional requirements for acquisitions involving bundling.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... the agency must conduct market research to determine whether bundling is necessary and justified (15 U.S.C. 644(e)(2)). Market research may indicate that bundling is necessary and justified if an agency...

  11. Quantitative study of bundle size effect on thermal conductivity of single-walled carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Feng, Ya; Inoue, Taiki; An, Hua; Xiang, Rong; Chiashi, Shohei; Maruyama, Shigeo

    2018-05-01

    Compared with isolated single-walled carbon nanotubes (SWNTs), thermal conductivity is greatly impeded in SWNT bundles; however, the measurement of the bundle size effect is difficult. In this study, the number of SWNTs in a bundle was determined based on the transferred horizontally aligned SWNTs on a suspended micro-thermometer to quantitatively study the effect of the bundle size on thermal conductivity. Increasing the bundle size significantly degraded the thermal conductivity. For isolated SWNTs, thermal conductivity was approximately 5000 ± 1000 W m-1 K-1 at room temperature, three times larger than that of the four-SWNT bundle. The logarithmical deterioration of thermal conductivity resulting from the increased bundle size can be attributed to the increased scattering rate with neighboring SWNTs based on the kinetic theory.

  12. Optical absorption and thermal transport of individual suspended carbon nanotube bundles.

    PubMed

    Hsu, I-Kai; Pettes, Michael T; Bushmaker, Adam; Aykol, Mehmet; Shi, Li; Cronin, Stephen B

    2009-02-01

    A focused laser beam is used to heat individual single-walled carbon nanotube bundles bridging two suspended microthermometers. By measurement of the temperature rise of the two thermometers, the optical absorption of 7.4-10.3 nm diameter bundles is found to be between 0.03 and 0.44% of the incident photons in the 0.4 microm diameter laser spot. The thermal conductance of the bundle is obtained with the additional measurement of the temperature rise of the nanotubes in the laser spot from shifts in the Raman G band frequency. According to the nanotube bundle diameter determined by transmission electron microscopy, the thermal conductivity is obtained.

  13. Adhering to a national surgical care bundle reduces the risk of surgical site infections

    PubMed Central

    Hopmans, Titia E. M.; Soetens, Loes C.; Wille, Jan C.; Geerlings, Suzanne E.; Vos, Margreet C.; van Benthem, Birgit H. B.; de Greeff, Sabine C.

    2017-01-01

    Background In 2008, a bundle of care to prevent Surgical Site Infections (SSIs) was introduced in the Netherlands. The bundle consisted of four elements: antibiotic prophylaxis according to local guidelines, no hair removal, normothermia and ‘hygiene discipline’ in the operating room (i.e. number of door movements). Dutch hospitals were advised to implement the bundle and to measure the outcome. This study’s goal was to assess how effective the bundle was in reducing SSI risk. Methods Hospitals assessed whether their staff complied with each of the bundle elements and voluntary reported compliance data to the national SSI surveillance network (PREZIES). From PREZIES data, we selected data from 2009 to 2014 relating to 13 types of surgical procedures. We excluded surgeries with missing (non)compliance data, and calculated for each remaining surgery with reported (non)compliance data the level of compliance with the bundle (that is, being compliant with 0, 1, 2, 3, or 4 of the elements). Subsequently, we used this level of compliance to assess the effect of bundle compliance on the SSI risk, using multilevel logistic regression techniques. Results 217 489 surgeries were included, of which 62 486 surgeries (29%) had complete bundle reporting. Within this group, the SSI risk was significantly lower for surgeries with complete bundle compliance compared to surgeries with lower compliance levels. Odds ratios ranged from 0.63 to 0.86 (risk reduction of 14% to 37%), while a 13% risk reduction was demonstrated for each point increase in compliance-level. Sensitivity analysis indicated that due to analysing reported bundles only, we probably underestimated the total effect of implementing the bundle. Conclusions This study demonstrated that adhering to a surgical care bundle significantly reduced the risk of SSIs. Reporting of and compliance with the bundle compliance can, however, still be improved. Therefore an even greater effect might be achieved. PMID:28877223

  14. Evidence for two populations of hair bundles in the sea anemone, Nematostella vectensis.

    PubMed

    Menard, Shelcie S; Watson, Glen M

    2017-06-01

    Cytochalasin D (CD) was employed to disrupt F-actin within stereocilia of anemone hair bundles. CD treatment decreases the abundance of hair bundles (by 85%) while significantly impairing predation. The remaining hair bundles are 'CD-resistant.' Surprisingly, the morphology and F-actin content of resistant hair bundles are comparable to those of untreated controls. However, the resistant hair bundles fail to respond normally to the N-acetylated sugar, NANA, by elongating. Instead, they remain at resting length. Immediately after CD treatment, when only CD-resistant hair bundles are present, nematocyst discharge is normal into targets touched to tentacles in the absence of vibrations (i.e., baseline) but fails to increase normally in the presence of nearby vibrations at 56Hz, a key frequency. After CD treatment, the abundance of hair bundles recovers to control levels within three hours. At 2h after CD treatment, when CD-resistant and CD-sensitive hair bundles are both present, but a full-recovery is not yet complete, somewhat enhanced discharge of nematocysts occurs into targets touched to tentacles in the presence of nearby vibrations at 56Hz (at least as compared to the response of CD-treated animals to contact with test probes in the absence of vibrations). Additionally, at 2h after CD-treatment, prey capture recovers to normal. Thus, two populations of hair bundles may be present on tentacles of sea anemones: those that are CD-resistant and those that are CD-sensitive. The functions of these hair bundles may be distinct. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Engineering biomimetic hair bundle sensors for underwater sensing applications

    NASA Astrophysics Data System (ADS)

    Kottapalli, Ajay Giri Prakash; Asadnia, Mohsen; Karavitaki, K. Domenica; Warkiani, Majid Ebrahimi; Miao, Jianmin; Corey, David P.; Triantafyllou, Michael

    2018-05-01

    We present the fabrication of an artificial MEMS hair bundle sensor designed to approximate the structural and functional principles of the flow-sensing bundles found in fish neuromast hair cells. The sensor consists of micro-pillars of graded height connected with piezoelectric nanofiber "tip-links" and encapsulated by a hydrogel cupula-like structure. Fluid drag force actuates the hydrogel cupula and deflects the micro-pillar bundle, stretching the nanofibers and generating electric charges. These biomimetic sensors achieve an ultrahigh sensitivity of 0.286 mV/(mm/s) and an extremely low threshold detection limit of 8.24 µm/s. A complete version of this paper has been published [1].

  16. Application of a transient heat transfer model for bundled, multiphase pipelines

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

    Brown, T.S.; Clapham, J.; Danielson, T.J.

    1996-12-31

    A computer model has been developed which accurately describes transient heat transfer in pipeline bundles. An arbitrary number of internal pipelines containing different fluids, flowing in either direction along with the input of heat to one or more of the fluids can be accommodated. The model is coupled to the transient, multiphase flow simulator OLGA. The lines containing the multiphase production fluids are modeled by OLGA, and the heat transfer between the internal lines, carrier pipe, and surroundings is handled by the bundle model. The model has been applied extensively to the design of a subsea, heated bundle system formore » the Britannia gas condensate field in the North Sea. The 15-km bundle system contains a 14{double_prime} production line, an 8{double_prime} test line, a 3{double_prime} methanol line, and a 12{double_prime} internal heating medium line within a 37.25{double_prime} carrier. The heating medium (water) flows in the internal heating medium line and in the annulus at 82,500 BPD. The primary purpose of the bundle system is to avoid the formation of hydrates. A secondary purpose is to avoid the deposition of paraffin. The bundle model was used to (1) compare the merits of two coaxial lines vs. a single bundle; (2) optimize the insulation levels on the carrier and internal lines; (3) determine the minimum time required to heat up the bundle; (4) determine heat input requirements to avoid hydrates throughout the field life, (5) determine temperature profiles along the lines for a range of production rates; (6) study ruptures of the production line into the bundle annulus; (7) determine minimum temperatures during depressurization; and (8) determine cool-down times. The results of these studies were used to size lines, select insulation levels, assess erosion potential, design for thermal expansion-induced stresses, and to select materials of construction.« less

  17. High thermodynamic stability of parametrically designed helical bundles

    DOE PAGES

    Huang, Po -Ssu; Oberdorfer, Gustav; Xu, Chunfu; ...

    2014-10-24

    Here we describe a procedure for designing proteins with backbones produced by varying the parameters in the Crick coiled coil–generating equations. Combinatorial design calculations identify low-energy sequences for alternative helix supercoil arrangements, and the helices in the lowest-energy arrangements are connected by loop building. We design an antiparallel monomeric untwisted three-helix bundle with 80-residue helices, an antiparallel monomeric right-handed four-helix bundle, and a pentameric parallel left-handed five-helix bundle. The designed proteins are extremely stable (extrapolated ΔG fold > 60 kilocalories per mole), and their crystal structures are close to those of the design models with nearly identical core packing betweenmore » the helices. The approach enables the custom design of hyperstable proteins with fine-tuned geometries for a wide range of applications.« less

  18. Fiber-optic-bundle-based optical coherence tomography.

    PubMed

    Xie, Tuqiang; Mukai, David; Guo, Shuguang; Brenner, Matthew; Chen, Zhongping

    2005-07-15

    A fiber-optic-bundle-based optical coherence tomography (OCT) probe method is presented. The experimental results demonstrate this multimode optical fiber-bundle-based OCT system can achieve a lateral resolution of 12 microm and an axial resolution of 10 microm with a superluminescent diode source. This novel OCT imaging approach eliminates any moving parts in the probe and has a primary advantage for use in extremely compact and safe OCT endoscopes for imaging internal organs and great potential to be combined with confocal endoscopic microscopy.

  19. Spinal surgery: variations in health care costs and implications for episode-based bundled payments.

    PubMed

    Ugiliweneza, Beatrice; Kong, Maiying; Nosova, Kristin; Huang, Kevin T; Babu, Ranjith; Lad, Shivanand P; Boakye, Maxwell

    2014-07-01

    Retrospective, observational. To simulate what episodes of care in spinal surgery might look like in a bundled payment system and to evaluate the associated costs and characteristics. Episode-based payment bundling has received considerable attention as a potential method to help curb the rise in health care spending and is being investigated as a new payment model as part of the Affordable Care Act. Although earlier studies investigated bundled payments in a number of surgical settings, very few focused on spine surgery, specifically. We analyzed data from MarketScan. Patients were included in the study if they underwent cervical or lumbar spinal surgery during 2000-2009, had at least 2-year preoperative and 90-day postoperative follow-up data. Patients were grouped on the basis of their diagnosis-related group (DRG) and then tracked in simulated episodes-of-care/payment bundles that lasted for the duration of 30, 60, and 90 days after the discharge from the index-surgical hospitalization. The total cost associated with each episode-of-care duration was measured and characterized. A total of 196,918 patients met our inclusion criteria. Significant variation existed between DRGs, ranging from $11,180 (30-day bundle, DRG 491) to $107,642 (30-day bundle, DRG 456). There were significant cost variations within each individual DRG. Postdischarge care accounted for a relatively small portion of overall bundle costs (range, 4%-8% in 90-day bundles). Total bundle costs remained relatively flat as bundle-length increased (total average cost of 30-day bundle: $33,522 vs. $35,165 for 90-day bundle). Payments to hospitals accounted for the largest portion of bundle costs (76%). There exists significant variation in total health care costs for patients who undergo spinal surgery, even within a given DRG. Better characterization of impacts of a bundled payment system in spine surgery is important for understanding the costs of index procedure hospital, physician services, and

  20. Steady-state stiffness of utricular hair cells depends on macular location and hair bundle structure.

    PubMed

    Spoon, Corrie; Moravec, W J; Rowe, M H; Grant, J W; Peterson, E H

    2011-12-01

    Spatial and temporal properties of head movement are encoded by vestibular hair cells in the inner ear. One of the most striking features of these receptors is the orderly structural variation in their mechanoreceptive hair bundles, but the functional significance of this diversity is poorly understood. We tested the hypothesis that hair bundle structure is a significant contributor to hair bundle mechanics by comparing structure and steady-state stiffness of 73 hair bundles at varying locations on the utricular macula. Our first major finding is that stiffness of utricular hair bundles varies systematically with macular locus. Stiffness values are highest in the striola, near the line of hair bundle polarity reversal, and decline exponentially toward the medial extrastriola. Striolar bundles are significantly more stiff than those in medial (median: 8.9 μN/m) and lateral (2.0 μN/m) extrastriolae. Within the striola, bundle stiffness is greatest in zone 2 (106.4 μN/m), a band of type II hair cells, and significantly less in zone 3 (30.6 μN/m), which contains the only type I hair cells in the macula. Bathing bundles in media that break interciliary links produced changes in bundle stiffness with predictable time course and magnitude, suggesting that links were intact in our standard media and contributed normally to bundle stiffness during measurements. Our second major finding is that bundle structure is a significant predictor of steady-state stiffness: the heights of kinocilia and the tallest stereocilia are the most important determinants of bundle stiffness. Our results suggest 1) a functional interpretation of bundle height variability in vertebrate vestibular organs, 2) a role for the striola in detecting onset of head movement, and 3) the hypothesis that differences in bundle stiffness contribute to diversity in afferent response dynamics.

  1. Steady-state stiffness of utricular hair cells depends on macular location and hair bundle structure

    PubMed Central

    Spoon, Corrie; Moravec, W. J.; Rowe, M. H.; Grant, J. W.

    2011-01-01

    Spatial and temporal properties of head movement are encoded by vestibular hair cells in the inner ear. One of the most striking features of these receptors is the orderly structural variation in their mechanoreceptive hair bundles, but the functional significance of this diversity is poorly understood. We tested the hypothesis that hair bundle structure is a significant contributor to hair bundle mechanics by comparing structure and steady-state stiffness of 73 hair bundles at varying locations on the utricular macula. Our first major finding is that stiffness of utricular hair bundles varies systematically with macular locus. Stiffness values are highest in the striola, near the line of hair bundle polarity reversal, and decline exponentially toward the medial extrastriola. Striolar bundles are significantly more stiff than those in medial (median: 8.9 μN/m) and lateral (2.0 μN/m) extrastriolae. Within the striola, bundle stiffness is greatest in zone 2 (106.4 μN/m), a band of type II hair cells, and significantly less in zone 3 (30.6 μN/m), which contains the only type I hair cells in the macula. Bathing bundles in media that break interciliary links produced changes in bundle stiffness with predictable time course and magnitude, suggesting that links were intact in our standard media and contributed normally to bundle stiffness during measurements. Our second major finding is that bundle structure is a significant predictor of steady-state stiffness: the heights of kinocilia and the tallest stereocilia are the most important determinants of bundle stiffness. Our results suggest 1) a functional interpretation of bundle height variability in vertebrate vestibular organs, 2) a role for the striola in detecting onset of head movement, and 3) the hypothesis that differences in bundle stiffness contribute to diversity in afferent response dynamics. PMID:21918003

  2. Conformational Spread in the Flagellar Motor Switch: A Model Study

    PubMed Central

    Maini, Philip K.; Berry, Richard M.; Bai, Fan

    2012-01-01

    The reliable response to weak biological signals requires that they be amplified with fidelity. In E. coli, the flagellar motors that control swimming can switch direction in response to very small changes in the concentration of the signaling protein CheY-P, but how this works is not well understood. A recently proposed allosteric model based on cooperative conformational spread in a ring of identical protomers seems promising as it is able to qualitatively reproduce switching, locked state behavior and Hill coefficient values measured for the rotary motor. In this paper we undertook a comprehensive simulation study to analyze the behavior of this model in detail and made predictions on three experimentally observable quantities: switch time distribution, locked state interval distribution, Hill coefficient of the switch response. We parameterized the model using experimental measurements, finding excellent agreement with published data on motor behavior. Analysis of the simulated switching dynamics revealed a mechanism for chemotactic ultrasensitivity, in which cooperativity is indispensable for realizing both coherent switching and effective amplification. These results showed how cells can combine elements of analog and digital control to produce switches that are simultaneously sensitive and reliable. PMID:22654654

  3. An evaluation of the impact of the ventilator care bundle.

    PubMed

    Crunden, Eddie; Boyce, Carolyn; Woodman, Helen; Bray, Barbara

    2005-01-01

    A number of interventions have been shown to improve the outcomes of patients who are invasively ventilated in intensive care units (ICUs). However, significant problems still exist in implementing research findings into clinical practice. The aim of this study was to assess whether the systematic and methodical implementation of evidence-based interventions encapsulated in a care bundle influenced length of ventilation and ICU length of stay (LOS). A ventilator care bundle was introduced within a general ICU and evaluated 1 year later. The care bundle was composed of four protocols that consisted of prophylaxis against peptic ulceration, prophylaxis against deep vein thrombosis, daily cessation of sedation and elevation of the patient's head and chest to at least 30 degrees to the horizontal. Compliance with the bundle was assessed, as was ICU LOS, ICU mortality and ICU/high-dependency unit patient throughput. Mean ICU LOS was reduced from 13-75 [standard deviation (SD) 19.11] days to 8.36 (SD 10.21) days (p<0.05). Mean ventilator days were reduced from 10.8 (SD 15.58) days to 6.1 (SD 8.88) days. Unit patient throughput increased by 30.1% and the number of invasively ventilated patients increased by 39.5%. Care bundles encourage the consistent and systematic application of evidence-based protocols used in particular treatment regimes. Since the introduction of the ventilator care bundle, length of ventilation and ICU LOS have reduced significantly.

  4. Bundles to prevent ventilator-associated pneumonia: how valuable are they?

    PubMed

    Wip, Charity; Napolitano, Lena

    2009-04-01

    To review the value of care bundles to prevent ventilator-associated pneumonia (VAP). The Ventilator Bundle contains four components, elevation of the head of the bed to 30-45 degrees, daily 'sedation vacation' and daily assessment of readiness to extubate, peptic ulcer disease prophylaxis, and deep venous thrombosis prophylaxis, aimed to improve outcome in mechanically ventilated patients, but not all are associated with VAP prevention. Daily spontaneous awakening and breathing trials are associated with early liberation from mechanical ventilation and VAP reduction. Although a small prospective, randomized clinical study documented that the semirecumbent position was associated with a significant reduction in VAP, more recent studies have documented that the semirecumbent position is difficult to maintain in mechanically ventilated patients and may not impact VAP reduction. Prophylaxis for peptic ulcer disease and deep venous thrombosis do not directly impact VAP reduction. Other methods to reduce VAP, such as oral care and hygiene, chlorhexidine in the posterior pharynx, and specialized endotracheal tubes (continuous aspiration of subglottic secretions, silver-coated), should be considered for inclusion in a revised Ventilator Bundle more specifically aimed at VAP prevention. The Ventilator Bundle is an effective method to reduce VAP rates in ICUs. The ventilator bundle should be modified and expanded to include specific processes of care that have been definitively demonstrated to be effective in VAP reduction or a specific VAP bundle created to focus on VAP prevention.

  5. Consensus Bundle on Maternal Mental Health: Perinatal Depression and Anxiety.

    PubMed

    Kendig, Susan; Keats, John P; Hoffman, M Camille; Kay, Lisa B; Miller, Emily S; Simas, Tiffany A Moore; Frieder, Ariela; Hackley, Barbara; Indman, Pec; Raines, Christena; Semenuk, Kisha; Wisner, Katherine L; Lemieux, Lauren A

    2017-03-01

    Perinatal mood and anxiety disorders are among the most common mental health conditions encountered by women of reproductive age. When left untreated, perinatal mood and anxiety disorders can have profound adverse effects on women and their children, ranging from increased risk of poor adherence to medical care, exacerbation of medical conditions, loss of interpersonal and financial resources, smoking and substance use, suicide, and infanticide. Perinatal mood and anxiety disorders are associated with increased risks of maternal and infant mortality and morbidity and are recognized as a significant patient safety issue. In 2015, the Council on Patient Safety in Women's Health Care convened an interdisciplinary work group to develop an evidence-based patient safety bundle to address maternal mental health. The focus of this bundle is perinatal mood and anxiety disorders. The bundle is modeled after other bundles released by the Council on Patient Safety in Women's Health Care and provides broad direction for incorporating perinatal mood and anxiety disorder screening, intervention, referral, and follow-up into maternity care practice across health care settings. This commentary provides information to assist with bundle implementation. © 2017 by the American College of Nurse-Midwives.

  6. Identifying the challenges and facilitators of implementing a COPD care bundle.

    PubMed

    Lennox, Laura; Green, Stuart; Howe, Cathy; Musgrave, Hannah; Bell, Derek; Elkin, Sarah

    2014-01-01

    Care bundles have been shown to improve outcomes, reduce hospital readmissions and reduce length of hospital stay; therefore increasing the speed of uptake and delivery of care bundles should be a priority in order to deliver more timely improvements and consistent high-quality care. Previous studies have detailed the difficulties of obtaining full compliance to bundle elements but few have described the underlying reasons for this. In order to improve future implementation this paper investigates the challenges encountered by clinical teams implementing a chronic obstructive pulmonary disease (COPD) care bundle and describes actions taken to overcome these challenges. An initial retrospective documentary analysis of data from seven clinical implementation teams was undertaken to review the challenges faced by the clinical teams. Three focus groups with healthcare professionals and managers explored solutions to these challenges developed during the project. Documentary analysis identified 28 challenges which directly impacted implementation of the COPD care bundle within five themes; staffing, infrastructure, process, use of improvement methodology and patient and public involvement. Focus groups revealed that the five most significant challenges for all groups were: staff too busy, staff shortages, lack of staff engagement, added workload of the bundle and patient coding issues. The participants shared facilitating factors used to overcome issues including: shifting perceptions to improve engagement, further education sessions to increase staff participation and gaining buy-in from managers through payment frameworks. Maximising the impact of a care bundle relies on its successful and timely implementation. Teams implementing the COPD care bundle encountered challenges that were common to all teams and sites. Understanding and learning from the challenges faced by previous endeavours and identifying the facilitators to overcoming these barriers provides an

  7. Identifying the challenges and facilitators of implementing a COPD care bundle

    PubMed Central

    Lennox, Laura; Green, Stuart; Howe, Cathy; Musgrave, Hannah; Bell, Derek; Elkin, Sarah

    2014-01-01

    Background Care bundles have been shown to improve outcomes, reduce hospital readmissions and reduce length of hospital stay; therefore increasing the speed of uptake and delivery of care bundles should be a priority in order to deliver more timely improvements and consistent high-quality care. Previous studies have detailed the difficulties of obtaining full compliance to bundle elements but few have described the underlying reasons for this. In order to improve future implementation this paper investigates the challenges encountered by clinical teams implementing a chronic obstructive pulmonary disease (COPD) care bundle and describes actions taken to overcome these challenges. Methods An initial retrospective documentary analysis of data from seven clinical implementation teams was undertaken to review the challenges faced by the clinical teams. Three focus groups with healthcare professionals and managers explored solutions to these challenges developed during the project. Results Documentary analysis identified 28 challenges which directly impacted implementation of the COPD care bundle within five themes; staffing, infrastructure, process, use of improvement methodology and patient and public involvement. Focus groups revealed that the five most significant challenges for all groups were: staff too busy, staff shortages, lack of staff engagement, added workload of the bundle and patient coding issues. The participants shared facilitating factors used to overcome issues including: shifting perceptions to improve engagement, further education sessions to increase staff participation and gaining buy-in from managers through payment frameworks. Conclusions Maximising the impact of a care bundle relies on its successful and timely implementation. Teams implementing the COPD care bundle encountered challenges that were common to all teams and sites. Understanding and learning from the challenges faced by previous endeavours and identifying the facilitators to

  8. Implementing a pressure ulcer prevention bundle in an adult intensive care.

    PubMed

    Tayyib, Nahla; Coyer, Fiona; Lewis, Peter A

    2016-12-01

    The incidence of pressure ulcers (PUs) in intensive care units (ICUs) is high and numerous strategies have been implemented to address this issue. One approach is the use of a PU prevention bundle. However, to ensure success care bundle implementation requires monitoring to evaluate the care bundle compliance rate, and to evaluate the effectiveness of implementation strategies in facilitating practice change. The aims of this study were to appraise the implementation of a series of high impact intervention care bundle components directed at preventing the development of PUs, within ICU, and to evaluate the effectiveness of strategies used to enhance the implementation compliance. An observational prospective study design was used. Implementation strategies included regular education, training, audit and feed-back and the presence of a champion in the ICU. Implementation compliance was measured along four time points using a compliance checklist. Of the 60 registered nurses (RNs) working in the critical care setting, 11 participated in this study. Study participants demonstrated a high level of compliance towards the PU prevention bundle implementation (78.1%), with 100% participant acceptance. No significant differences were found between participants' demographic characteristics and the compliance score. There was a significant effect for time in the implementation compliance (Wilks Lambda=0.29, F (3, 8)=6.35, p<0.016), indicating that RNs needed time to become familiar with the bundle and routinely implement it into their practice. PU incidence was not influenced by the compliance level of participants. The implementation strategies used showed a positive impact on compliance. Assessing and evaluating implementation compliance is critical to achieve a desired outcome (reduction in PU incidence). This study's findings also highlighted that while RNs needed time to familiarise themselves with the care bundle elements, their clinical practice was congruent with the

  9. Improved Conjugate Gradient Bundle Adjustment of Dunhuang Wall Painting Images

    NASA Astrophysics Data System (ADS)

    Hu, K.; Huang, X.; You, H.

    2017-09-01

    Bundle adjustment with additional parameters is identified as a critical step for precise orthoimage generation and 3D reconstruction of Dunhuang wall paintings. Due to the introduction of self-calibration parameters and quasi-planar constraints, the structure of coefficient matrix of the reduced normal equation is banded-bordered, making the solving process of bundle adjustment complex. In this paper, Conjugate Gradient Bundle Adjustment (CGBA) method is deduced by calculus of variations. A preconditioning method based on improved incomplete Cholesky factorization is adopt to reduce the condition number of coefficient matrix, as well as to accelerate the iteration rate of CGBA. Both theoretical analysis and experimental results comparison with conventional method indicate that, the proposed method can effectively conquer the ill-conditioned problem of normal equation and improve the calculation efficiency of bundle adjustment with additional parameters considerably, while maintaining the actual accuracy.

  10. Brill-Noether theory for vector bundles on projective curves

    NASA Astrophysics Data System (ADS)

    Ballico, E.

    1998-11-01

    In this paper we will study the Brill-Noether theory of vector bundles on a smooth projective curve X. As usual in papers on this topic we are mainly interested in stable or at least semistable bundles. Let Wkr, d(X) be the scheme of all stable vector bundles E on X with rank (E)=r, deg (E)=d and h0(X, E)[gt-or-equal, slanted]k+1. For a survey of the main known results, see the introduction of [6]. The referee has pointed out that the results in [6] were improved by V. Mercat in [14]; he proved that Wkr, d(X) is non-empty for d<2r if and only if k+1[less-than-or-eq, slant]r+(d[minus sign]r)/g. If X has general moduli the more interesting existence theorem was proved in [19]. However, in this paper we are mainly interested in very special curves X, e.g. the hyperelliptic or the bielliptic curves. We work over an algebraically closed base field K. In Section 5 we will assume char (K)=0. In Section 1 we will give some theorems of Clifford's type. In Section 2 we will construct several stable bundles with certain properties. Here the main tool is an operation (the +elementary transformation) which sends a vector bundle E on X to another vector bundle E[prime prime or minute] with rank (E[prime prime or minute])=rank (E) and deg (E[prime prime or minute])=deg (E)+1 (see Section 2 for its definition and its elementary properties). Using the +elementary transformations in Section 3 we will prove the following existence theorem which covers the case of a ‘small’ number of sections.

  11. Bundling Post-Acute Care Services into MS-DRG Payments

    PubMed Central

    Vertrees, James C.; Averill, Richard F.; Eisenhandler, Jon; Quain, Anthony; Switalski, James

    2013-01-01

    Objective A bundled hospital payment system that encompasses both acute and post-acute care has been proposed as a means of creating financial incentives in the Medicare fee-for-service system to foster care coordination and to improve the current disorganized system of post care. The objective of this study was to evaluate the statistical stability of alternative designs of a hospital payment system that includes post-acute care services to determine the feasibility of using a combined hospital and post-acute care bundle as a unit of payment. Methods The Medicare Severity-Diagnosis Related Groups (MS-DRGs) were subdivided into clinical subclasses that measured a patient's chronic illness burden to test whether a patient's chronic illness burden had a substantial impact on post-acute care expenditures. Using Medicare data the statistical performance of the MS-DRGs with and without the chronic illness subclasses was evaluated across a wide range of post-acute care windows and combinations of post-acute care service bundles using both submitted charges and Medicare payments. Results The statistical performance of the MS-DRGs as measured by R2 was consistently better when the chronic illness subclasses are included indicating that MS-DRGs by themselves are an inadequate unit of payment for post-acute care payment bundles. In general, R2 values increased as the post-acute care window length increased and decreased as more services were added to the post-acute care bundle. Discussion The study results suggest that it is feasible to develop a payment system that incorporates significant post-acute care services into the MS-DRG inpatient payment bundle. This expansion of the basic DRG payment approach can provide a strong financial incentive for providers to better coordinate care potentially leading to improved efficiency and outcome quality. PMID:24753970

  12. The impact of Lean bundles on hospital performance: does size matter?

    PubMed

    Al-Hyari, Khalil; Abu Hammour, Sewar; Abu Zaid, Mohammad Khair Saleem; Haffar, Mohamed

    2016-10-10

    Purpose The purpose of this paper is to study the effect of the implementation of Lean bundles on hospital performance in private hospitals in Jordan and evaluate how much the size of organization can affect the relationship between Lean bundles implementation and hospital performance. Design/methodology/approach The research is considered as quantitative method (descriptive and hypothesis testing). Three statistical techniques were adopted to analyse the data. Structural equation modeling techniques and multi-group analysis were used to examine the research's hypothesis, and to perform the required statistical analysis of the data from the survey. Reliability analysis and confirmatory factor analysis were used to test the construct validity, reliability and measurement loadings that were performed. Findings Lean bundles have been identified as an effective approach that can dramatically improve the organizational performance of private hospitals in Jordan. Main Lean bundles - just in time, human resource management, and total quality management are applicable to large, small and medium hospitals without significant differences in advantages that depend on size. Originality/value According to the researchers' best knowledge, this is the first research that studies the impact of Lean bundles implementation in healthcare sector in Jordan. This research also makes a significant contribution for decision makers in healthcare to increase their awareness of Lean bundles.

  13. Anatomic Double-Bundle Anterior Cruciate Ligament Reconstruction With a Free Quadriceps Tendon Autograft.

    PubMed

    Caterev, Sergiu; Nistor, Dan Viorel; Todor, Adrian

    2016-10-01

    Anatomic double-bundle anterior cruciate ligament (ACL) reconstruction aims to restore the 2 functional bundles of the ACL in an attempt to better reproduce the native biomechanics of the injured knee and promote long-term knee health. However, this concept is not fully accepted and is not performed on a standard basis. In addition, the superiority of this technique over the conventional single-bundle technique has been questioned, especially the long-term clinical results. One of the down sides of the double-bundle reconstruction is the complexity of the procedure, with increased risks, operative time, and costs compared with the single-bundle procedure. Also, the revision procedure, if necessary, is more challenging. We propose a technique that has some advantages over the traditional double-bundle procedure, using a single femoral tunnel, 2 tibial tunnels, and a free quadriceps tendon autograft.

  14. Phylogenetic and Metagenomic Analyses of Substrate-Dependent Bacterial Temporal Dynamics in Microbial Fuel Cells

    PubMed Central

    Zhang, Husen; Chen, Xi; Braithwaite, Daniel; He, Zhen

    2014-01-01

    Understanding the microbial community structure and genetic potential of anode biofilms is key to improve extracellular electron transfers in microbial fuel cells. We investigated effect of substrate and temporal dynamics of anodic biofilm communities using phylogenetic and metagenomic approaches in parallel with electrochemical characterizations. The startup non-steady state anodic bacterial structures were compared for a simple substrate, acetate, and for a complex substrate, landfill leachate, using a single-chamber air-cathode microbial fuel cell. Principal coordinate analysis showed that distinct community structures were formed with each substrate type. The bacterial diversity measured as Shannon index decreased with time in acetate cycles, and was restored with the introduction of leachate. The change of diversity was accompanied by an opposite trend in the relative abundance of Geobacter-affiliated phylotypes, which were acclimated to over 40% of total Bacteria at the end of acetate-fed conditions then declined in the leachate cycles. The transition from acetate to leachate caused a decrease in output power density from 243±13 mW/m2 to 140±11 mW/m2, accompanied by a decrease in Coulombic electron recovery from 18±3% to 9±3%. The leachate cycles selected protein-degrading phylotypes within phylum Synergistetes. Metagenomic shotgun sequencing showed that leachate-fed communities had higher cell motility genes including bacterial chemotaxis and flagellar assembly, and increased gene abundance related to metal resistance, antibiotic resistance, and quorum sensing. These differentially represented genes suggested an altered anodic biofilm community in response to additional substrates and stress from the complex landfill leachate. PMID:25202990

  15. Phylogenetic and metagenomic analyses of substrate-dependent bacterial temporal dynamics in microbial fuel cells.

    PubMed

    Zhang, Husen; Chen, Xi; Braithwaite, Daniel; He, Zhen

    2014-01-01

    Understanding the microbial community structure and genetic potential of anode biofilms is key to improve extracellular electron transfers in microbial fuel cells. We investigated effect of substrate and temporal dynamics of anodic biofilm communities using phylogenetic and metagenomic approaches in parallel with electrochemical characterizations. The startup non-steady state anodic bacterial structures were compared for a simple substrate, acetate, and for a complex substrate, landfill leachate, using a single-chamber air-cathode microbial fuel cell. Principal coordinate analysis showed that distinct community structures were formed with each substrate type. The bacterial diversity measured as Shannon index decreased with time in acetate cycles, and was restored with the introduction of leachate. The change of diversity was accompanied by an opposite trend in the relative abundance of Geobacter-affiliated phylotypes, which were acclimated to over 40% of total Bacteria at the end of acetate-fed conditions then declined in the leachate cycles. The transition from acetate to leachate caused a decrease in output power density from 243±13 mW/m2 to 140±11 mW/m2, accompanied by a decrease in Coulombic electron recovery from 18±3% to 9±3%. The leachate cycles selected protein-degrading phylotypes within phylum Synergistetes. Metagenomic shotgun sequencing showed that leachate-fed communities had higher cell motility genes including bacterial chemotaxis and flagellar assembly, and increased gene abundance related to metal resistance, antibiotic resistance, and quorum sensing. These differentially represented genes suggested an altered anodic biofilm community in response to additional substrates and stress from the complex landfill leachate.

  16. Field Emission Study of Carbon Nanotubes: High Current Density from Nanotube Bundle Arrays

    NASA Technical Reports Server (NTRS)

    Bronikowski, Micheal J.; Manohara, Harish M.; Siegel, Peter H.; Hunt, Brian D.

    2004-01-01

    We have investigated the field emission behavior of lithographically patterned bundles of multiwalled carbon nanotubes arranged in a variety of array geometries. Such arrays of nanotube bundles are found to perform significantly better in field emission than arrays of isolated nanotubes or dense, continuous mats of nanotubes, with the field emission performance depending on the bundle diameter and inter-bundle spacing. Arrays of 2-micrometers diameter nanotube bundles spaced 5 micrometers apart (edge-to-edge spacing) produced the largest emission densities, routinely giving 1.5 to 1.8 A/cm(sup 2) at approximately 4 V/micrometer electric field, and greater than 6 A/cm(sup 2) at 20 V/micrometers.

  17. Reduction of Surgical Site Infections after Implementation of a Bundle of Care

    PubMed Central

    Crolla, Rogier M. P. H.; van der Laan, Lijckle; Veen, Eelco J.; Hendriks, Yvonne; van Schendel, Caroline; Kluytmans, Jan

    2012-01-01

    Background Surgical Site Infections (SSI) are relatively frequent complications after colorectal surgery and are associated with substantial morbidity and mortality. Objective Implementing a bundle of care and measuring the effects on the SSI rate. Design Prospective quasi experimental cohort study. Methods A prospective surveillance for SSI after colorectal surgery was performed in the Amphia Hospital, Breda, from January 1, 2008 until January 1, 2012. As part of a National patient safety initiative, a bundle of care consisting of 4 elements covering the surgical process was introduced in 2009. The elements of the bundle were perioperative antibiotic prophylaxis, hair removal before surgery, perioperative normothermia and discipline in the operating room. Bundle compliance was measured every 3 months in a random sample of surgical procedures. Results Bundle compliance improved significantly from an average of 10% in 2009 to 60% in 2011. 1537 colorectal procedures were performed during the study period and 300 SSI (19.5%) occurred. SSI were associated with a prolonged length of stay (mean additional length of stay 18 days) and a significantly higher 6 months mortality (Adjusted OR: 2.71, 95% confidence interval 1.76–4.18). Logistic regression showed a significant decrease of the SSI rate that paralleled the introduction of the bundle. The adjusted Odds ratio of the SSI rate was 36% lower in 2011 compared to 2008. Conclusion The implementation of the bundle was associated with improved compliance over time and a 36% reduction of the SSI rate after adjustment for confounders. This makes the bundle an important tool to improve patient safety. PMID:22962619

  18. National Partnership for Maternal Safety: Consensus Bundle on Venous Thromboembolism.

    PubMed

    D'Alton, Mary E; Friedman, Alexander M; Smiley, Richard M; Montgomery, Douglas M; Paidas, Michael J; D'Oria, Robyn; Frost, Jennifer L; Hameed, Afshan B; Karsnitz, Deborah; Levy, Barbara S; Clark, Steven L

    2016-10-01

    Obstetric venous thromboembolism is a leading cause of severe maternal morbidity and mortality. Maternal death from thromboembolism is amenable to prevention, and thromboprophylaxis is the most readily implementable means of systematically reducing the maternal death rate. Observational data support the benefit of risk-factor-based prophylaxis in reducing obstetric thromboembolism. This bundle, developed by a multidisciplinary working group and published by the National Partnership for Maternal Safety under the guidance of the Council on Patient Safety in Women's Health Care, supports routine thromboembolism risk assessment for obstetric patients, with appropriate use of pharmacologic and mechanical thromboprophylaxis. Safety bundles outline critical clinical practices that should be implemented in every maternity unit. The safety bundle is organized into four domains: Readiness, Recognition, Response, and Reporting and Systems Learning. Although the bundle components may be adapted to meet the resources available in individual facilities, standardization within an institution is strongly encouraged.

  19. Mass Transport Through Carbon Nanotube-Polystyrene Bundles

    NASA Astrophysics Data System (ADS)

    Lin, Rongzhou; Tran, Tuan

    2016-05-01

    Carbon nanotubes have been widely used as test channels to study nanofluidic transport, which has been found to have distinctive properties compared to transport of fluids in macroscopic channels. A long-standing challenge in the study of mass transport through carbon nanotubes (CNTs) is the determination of flow enhancement. Various experimental investigations have been conducted to measure the flow rate through CNTs, mainly based on either vertically aligned CNT membranes or individual CNTs. Here, we proposed an alternative approach that can be used to quantify the mass transport through CNTs. This is a simple method relying on the use of carbon nanotube-polystyrene bundles, which are made of CNTs pulled out from a vertically aligned CNT array and glued together by polystyrene. We experimentally showed by using fluorescent tagging that the composite bundles allowed measureable and selective mass transport through CNTs. This type of composite bundle may be useful in various CNT research areas as they are simple to fabricate, less likely to form macroscopic cracks, and offer a high density of CNT pores while maintaining the aligned morphology of CNTs.

  20. Hexagonally Ordered Arrays of α-Helical Bundles Formed from Peptide-Dendron Hybrids

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

    Barkley, Deborah A.; Rokhlenko, Yekaterina; Marine, Jeannette E.

    Combining monodisperse building blocks that have distinct folding properties serves as a modular strategy for controlling structural complexity in hierarchically organized materials. We combine an α-helical bundle-forming peptide with self-assembling dendrons to better control the arrangement of functional groups within cylindrical nanostructures. Site-specific grafting of dendrons to amino acid residues on the exterior of the α-helical bundle yields monodisperse macromolecules with programmable folding and self-assembly properties. The resulting hybrid biomaterials form thermotropic columnar hexagonal mesophases in which the peptides adopt an α-helical conformation. Bundling of the α-helical peptides accompanies self-assembly of the peptide-dendron hybrids into cylindrical nanostructures. The bundle stoichiometrymore » in the mesophase agrees well with the size found in solution for α-helical bundles of peptides with a similar amino acid sequence.« less

  1. Right bundle branch block and anterior wall ST elevation myocardial infarction.

    PubMed

    Trofin, Monica; Israel, Carsten W; Barold, S Serge

    2017-09-01

    We report the case of an acute anterior wall ST elevation myocardial infarction with new left anterior fascicular block and pre-existing right bundle branch block. Due to a wide right bundle branch block, no ST segment elevation was visible in lead V1. The left anterior fascicular block was caused by proximal occlusion of the left artery descending and disappeared after acute revascularization. However, also the R' of the right bundle branch block became significantly shorter after revascularization, dismanteling a minor ST segment elevation. The ST elevation in lead V1 in anterior wall infarction and right bundle branch block may merge with the R' and cause a further QRS widening as an "equivalent" to the ST elevation.

  2. Telemetry in bundles: delay-tolerant networking for delay-challenged applications

    NASA Technical Reports Server (NTRS)

    Burleigh, S.

    2003-01-01

    This paper presents an overview of DTN concepts, including bundles and the Bundling overlay protocol. One possible scenario for the application of DTN to a telemetry return problem is described, and there is a brief discussion of the current state of DTN technology development.

  3. Accuracy of Shack-Hartmann wavefront sensor using a coherent wound fibre image bundle

    NASA Astrophysics Data System (ADS)

    Zheng, Jessica R.; Goodwin, Michael; Lawrence, Jon

    2018-03-01

    Shack-Hartmannwavefront sensors using wound fibre image bundles are desired for multi-object adaptive optical systems to provide large multiplex positioned by Starbugs. The use of a large-sized wound fibre image bundle provides the flexibility to use more sub-apertures wavefront sensor for ELTs. These compact wavefront sensors take advantage of large focal surfaces such as the Giant Magellan Telescope. The focus of this paper is to study the wound fibre image bundle structure defects effect on the centroid measurement accuracy of a Shack-Hartmann wavefront sensor. We use the first moment centroid method to estimate the centroid of a focused Gaussian beam sampled by a simulated bundle. Spot estimation accuracy with wound fibre image bundle and its structure impact on wavefront measurement accuracy statistics are addressed. Our results show that when the measurement signal-to-noise ratio is high, the centroid measurement accuracy is dominated by the wound fibre image bundle structure, e.g. tile angle and gap spacing. For the measurement with low signal-to-noise ratio, its accuracy is influenced by the read noise of the detector instead of the wound fibre image bundle structure defects. We demonstrate this both with simulation and experimentally. We provide a statistical model of the centroid and wavefront error of a wound fibre image bundle found through experiment.

  4. Viscous Dynamics of Lyme Disease and Syphilis Spirochetes Reveal Flagellar Torque and Drag

    PubMed Central

    Harman, Michael; Vig, Dhruv K.; Radolf, Justin D.; Wolgemuth, Charles W.

    2013-01-01

    The spirochetes that cause Lyme disease (Borrelia burgdorferi) and syphilis (Treponema pallidum) swim through viscous fluids, such as blood and interstitial fluid, by undulating their bodies as traveling, planar waves. These undulations are driven by rotation of the flagella within the periplasmic space, the narrow (∼20–40 nm in width) compartment between the inner and outer membranes. We show here that the swimming speeds of B. burgdorferi and T. pallidum decrease with increases in viscosity of the external aqueous milieu, even though the flagella are entirely intracellular. We then use mathematical modeling to show that the measured changes in speed are consistent with the exertion of constant torque by the spirochetal flagellar motors. Comparison of simulations, experiments, and a simple model for power dissipation allows us to estimate the torque and resistive drag that act on the flagella of these major spirochetal pathogens. PMID:24268139

  5. Consensus Bundle on Maternal Mental Health: Perinatal Depression and Anxiety.

    PubMed

    Kendig, Susan; Keats, John P; Hoffman, M Camille; Kay, Lisa B; Miller, Emily S; Moore Simas, Tiffany A; Frieder, Ariela; Hackley, Barbara; Indman, Pec; Raines, Christena; Semenuk, Kisha; Wisner, Katherine L; Lemieux, Lauren A

    Perinatal mood and anxiety disorders are among the most common mental health conditions encountered by women of reproductive age. When left untreated, perinatal mood and anxiety disorders can have profound adverse effects on women and their children, ranging from increased risk of poor adherence to medical care, exacerbation of medical conditions, loss of interpersonal and financial resources, smoking and substance use, suicide, and infanticide. Perinatal mood and anxiety disorders are associated with increased risks of maternal and infant mortality and morbidity and are recognized as a significant patient safety issue. In 2015, the Council on Patient Safety in Women's Health Care convened an interdisciplinary workgroup to develop an evidence-based patient safety bundle to address maternal mental health. The focus of this bundle is perinatal mood and anxiety disorders. The bundle is modeled after other bundles released by the Council on Patient Safety in Women's Health Care and provides broad direction for incorporating perinatal mood and anxiety disorder screening, intervention, referral, and follow-up into maternity care practice across health care settings. This commentary provides information to assist with bundle implementation. Copyright © 2017 AWHONN, the Association of Women’s Health, Obstetric and Neonatal Nurses. Published by Elsevier Inc. All rights reserved.

  6. Uncovering ecosystem service bundles through social preferences.

    PubMed

    Martín-López, Berta; Iniesta-Arandia, Irene; García-Llorente, Marina; Palomo, Ignacio; Casado-Arzuaga, Izaskun; Amo, David García Del; Gómez-Baggethun, Erik; Oteros-Rozas, Elisa; Palacios-Agundez, Igone; Willaarts, Bárbara; González, José A; Santos-Martín, Fernando; Onaindia, Miren; López-Santiago, Cesar; Montes, Carlos

    2012-01-01

    Ecosystem service assessments have increasingly been used to support environmental management policies, mainly based on biophysical and economic indicators. However, few studies have coped with the social-cultural dimension of ecosystem services, despite being considered a research priority. We examined how ecosystem service bundles and trade-offs emerge from diverging social preferences toward ecosystem services delivered by various types of ecosystems in Spain. We conducted 3,379 direct face-to-face questionnaires in eight different case study sites from 2007 to 2011. Overall, 90.5% of the sampled population recognized the ecosystem's capacity to deliver services. Formal studies, environmental behavior, and gender variables influenced the probability of people recognizing the ecosystem's capacity to provide services. The ecosystem services most frequently perceived by people were regulating services; of those, air purification held the greatest importance. However, statistical analysis showed that socio-cultural factors and the conservation management strategy of ecosystems (i.e., National Park, Natural Park, or a non-protected area) have an effect on social preferences toward ecosystem services. Ecosystem service trade-offs and bundles were identified by analyzing social preferences through multivariate analysis (redundancy analysis and hierarchical cluster analysis). We found a clear trade-off among provisioning services (and recreational hunting) versus regulating services and almost all cultural services. We identified three ecosystem service bundles associated with the conservation management strategy and the rural-urban gradient. We conclude that socio-cultural preferences toward ecosystem services can serve as a tool to identify relevant services for people, the factors underlying these social preferences, and emerging ecosystem service bundles and trade-offs.

  7. High-Resolution pH Imaging of Living Bacterial Cells To Detect Local pH Differences

    PubMed Central

    Morimoto, Yusuke V.; Kami-ike, Nobunori; Miyata, Tomoko; Kawamoto, Akihiro; Kato, Takayuki

    2016-01-01

    ABSTRACT Protons are utilized for various biological activities such as energy transduction and cell signaling. For construction of the bacterial flagellum, a type III export apparatus utilizes ATP and proton motive force to drive flagellar protein export, but the energy transduction mechanism remains unclear. Here, we have developed a high-resolution pH imaging system to measure local pH differences within living Salmonella enterica cells, especially in close proximity to the cytoplasmic membrane and the export apparatus. The local pH near the membrane was ca. 0.2 pH unit higher than the bulk cytoplasmic pH. However, the local pH near the export apparatus was ca. 0.1 pH unit lower than that near the membrane. This drop of local pH depended on the activities of both transmembrane export components and FliI ATPase. We propose that the export apparatus acts as an H+/protein antiporter to couple ATP hydrolysis with H+ flow to drive protein export. PMID:27923921

  8. Enhanced central venous catheter bundle for pediatric parenteral-dependent intestinal failure.

    PubMed

    Ormsby, Jennifer A; Bukoye, Bola; Lajoie, Debra; Shermont, Herminia; Martin, Lisa; Leger, Kierrah; Mahoney, Judy; Potter-Bynoe, Gail; Carpenter, Jane; Ozonoff, Al; Lee, Grace M

    2018-05-16

    Central line-associated bloodstream infections (CLABSIs) cause substantial morbidity and increase antimicrobial use and length of stay among hospitalized children in the United States. CLABSI occurs more frequently among high-risk pediatric patients, such as those with intestinal failure (IF) who are parenteral nutrition (PN) dependent. Following an increase in CLABSI rates, a quality improvement (QI) initiative was implemented. Using QI methodology, an enhanced central venous catheter (CVC) maintenance bundle was developed and implemented on 2 units for pediatric PN-dependent patients with IF. CLABSI rates were prospectively monitored pre- and postimplementation, and bundle element adherence was monitored. Enhanced bundle elements included chlorhexidine-impregnated patch, daily bathing, ethanol locks, 2 nurses for CVC care in a distraction-free zone, peripheral laboratory draws, bundling routine laboratory tests, and PN administration set changes every 24 hours. Adherence to enhanced bundle elements increased to >90% over 3 months. CLABSI rates averaged 1.41 per 1,000 central line days preimplementation compared with 0.40 per 1,000 device days postimplementation (P = .003), an 85% absolute reduction in CLABSI rates over 12 months. Patients with IF are at an increased risk for CLABSI. Enhanced CVC maintenance bundles that specifically target prevention practices in this population may be beneficial. Copyright © 2018 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  9. Lexical bundles in an advanced INTOCSU writing class and engineering texts: A functional analysis

    NASA Astrophysics Data System (ADS)

    Alquraishi, Mohammed Abdulrahman

    The purpose of this study is to investigate the functions of lexical bundles in two corpora: a corpus of engineering academic texts and a corpus of IEP advanced writing class texts. This study is concerned with the nature of formulaic language in Pathway IEPs and engineering texts, and whether those types of texts show similar or distinctive formulaic functions. Moreover, the study looked into lexical bundles found in an engineering 1.26 million-word corpus and an ESL 65000-word corpus using a concordancing program. The study then analyzed the functions of those lexical bundles and compared them statistically using chi-square tests. Additionally, the results of this investigation showed 236 unique frequent lexical bundles in the engineering corpus and 37 bundles in the pathway corpus. Also, the study identified several differences between the density and functions of lexical bundles in the two corpora. These differences were evident in the distribution of functions of lexical bundles and the minimal overlap of lexical bundles found in the two corpora. The results of this study call for more attention to formulaic language at ESP and EAP programs.

  10. Overhead electric power transmission line jumpering system for bundles of five or more subconductors

    DOEpatents

    Winkelman, Paul F.

    1982-01-01

    Jumpering of electric power transmission lines at a dead end tower. Two transmission line conductor bundles each contain five or more spaced apart subconductors (5) arranged in the shape of a cylinder having a circular cross section. The ends of each bundle of subconductors are attached with insulators to a dead end tower (1). Jumpering allows the electric current to flow between the two bundles of subconductors using jumper buses, internal jumper conductors, and external jumper conductors. One or more current collecting jumper buses (37) are located inside each bundle of subconductors with each jumper bus being attached to the end of a subconductor. Small-diameter internal jumper conductors (33) are located in the inherently electrically shielded area inside each bundle of subconductors with each subconductor (except ones having an attached jumper bus) having one internal jumper conductor connected between that subconductor's end and a jumper bus. Large-diameter external jumper conductors (9) are located outside each bundle of subconductors with one or more external jumper conductors being connected between the jumper buses in one bundle of subconductors and the jumper buses in the other bundle.

  11. Exploring Differential Bundle Functioning in Mathematics by Gender: The Effect of Hierarchical Modelling

    ERIC Educational Resources Information Center

    Ong, Yoke Mooi; Williams, Julian; Lamprianou, Iasonas

    2013-01-01

    Researchers interested in exploring substantive group differences are increasingly attending to bundles of items (or testlets): the aim is to understand how gender differences, for instance, are explained by differential performances on different types or bundles of items, hence differential bundle functioning (DBF). Some previous work has…

  12. Polyelectrolyte Bundles: Finite size at thermodynamic equilibrium?

    NASA Astrophysics Data System (ADS)

    Sayar, Mehmet

    2005-03-01

    Experimental observation of finite size aggregates formed by polyelectrolytes such as DNA and F-actin, as well as synthetic polymers like poly(p-phenylene), has created a lot of attention in recent years. Here, bundle formation in rigid rod-like polyelectrolytes is studied via computer simulations. For the case of hydrophobically modified polyelectrolytes finite size bundles are observed even in the presence of only monovalent counterions. Furthermore, in the absence of a hydrophobic backbone, we have also observed formation of finite size aggregates via multivalent counterion condensation. The size distribution of such aggregates and the stability is analyzed in this study.

  13. Sperm bundle and reproductive organs of carabid beetles tribe Pterostichini (Coleoptera: Carabidae)

    NASA Astrophysics Data System (ADS)

    Sasakawa, Kôji

    2007-05-01

    The morphological characteristics of sperm and reproductive organs may offer clues as to how reproductive systems have evolved. In this paper, the morphologies of the sperm and male reproductive organs of carabid beetles in the tribe Pterostichini (Coleoptera: Carabidae) are described, and the morphological associations among characters are examined. All species form sperm bundles in which the head of the sperm was embedded in a rod-shaped structure, i.e., spermatodesm. The spermatodesm shape (left-handed spiral, right-handed spiral, or without conspicuous spiral structure) and the condition of the sperm on the spermatodesm surface (with the tail free-moving or forming a thin, sheetlike structure) vary among species. In all species, the spiral directions of the convoluted seminal vesicles and vasa deferentia are the same on both sides of the body; that is, they show an asymmetric structure. The species in which the sperm bundle and the seminal vesicles both have a spiral structure could be classified into two types, with significant differences in sperm-bundle length between the two types. The species with a sperm-bundle spiral and seminal-vesicle spiral of almost the same diameter have longer sperm bundles than the species with a sperm-bundle spiral and seminal-vesicle tube of almost the same diameter. In the former type, the spiral directions of the sperm bundles and seminal vesicles are inevitably the same, whereas they differ in some species with the later type. Therefore, increased sperm bundle length appears to have been facilitated by the concordance of the sperm bundle’s coiling direction with the coiling direction of the seminal vesicle.

  14. Phase slips in oscillatory hair bundles.

    PubMed

    Roongthumskul, Yuttana; Shlomovitz, Roie; Bruinsma, Robijn; Bozovic, Dolores

    2013-04-05

    Hair cells of the inner ear contain an active amplifier that allows them to detect extremely weak signals. As one of the manifestations of an active process, spontaneous oscillations arise in fluid immersed hair bundles of in vitro preparations of selected auditory and vestibular organs. We measure the phase-locking dynamics of oscillatory bundles exposed to low-amplitude sinusoidal signals, a transition that can be described by a saddle-node bifurcation on an invariant circle. The transition is characterized by the occurrence of phase slips, at a rate that is dependent on the amplitude and detuning of the applied drive. The resultant staircase structure in the phase of the oscillation can be described by the stochastic Adler equation, which reproduces the statistics of phase slip production.

  15. Noncommutative Line Bundles and Gerbes

    NASA Astrophysics Data System (ADS)

    Jurčo, B.

    We introduce noncommutative line bundles and gerbes within the framework of deformation quantization. The Seiberg-Witten map is used to construct the corresponding noncommutative Čech cocycles. Morita equivalence of star products and quantization of twisted Poisson structures are discussed from this point of view.

  16. Use of the Delay-Tolerant Networking Bundle Protocol from Space

    NASA Technical Reports Server (NTRS)

    Wood, Lloyd; Ivancic, William D.; Eddy, Wesley M.; Stewart, Dave; Northam, James; Jackson, Chris; daSilvaCuriel, Alex

    2009-01-01

    The Disaster Monitoring Constellation (DMC), constructed by Survey Satellite Technology Ltd (SSTL), is a multisatellite Earth-imaging low-Earth-orbit sensor network where captured image swaths are stored onboard each satellite and later downloaded from the satellite payloads to a ground station. Store-and-forward of images with capture and later download gives each satellite the characteristics of a node in a Delay/Disruption Tolerant Network (DTN). Originally developed for the Interplanetary Internet, DTNs are now under investigation in an Internet Research Task Force (IRTF) DTN research group (RG), which has developed a bundle architecture and protocol. The DMC is currently unique in its adoption of the Internet Protocol (IP) for its imaging payloads and for satellite command and control, based around reuse of commercial networking and link protocols. These satellites use of IP has enabled earlier experiments with the Cisco router in Low Earth Orbit (CLEO) onboard the constellation's UK-DMC satellite. Earth images are downloaded from the satellites using a custom IPbased high-speed transfer protocol developed by SSTL, Saratoga, which tolerates unusual link environments. Saratoga has been documented in the Internet Engineering Task Force (IETF) for wider adoption. We experiment with use of DTNRG bundle concepts onboard the UKDMC satellite, by examining how Saratoga can be used as a DTN convergence layer to carry the DTNRG Bundle Protocol, so that sensor images can be delivered to ground stations and beyond as bundles. This is the first successful use of the DTNRG Bundle Protocol in a space environment. We use our practical experience to examine the strengths and weaknesses of the Bundle Protocol for DTN use, paying attention to fragmentation, custody transfer, and reliability issues.

  17. Presenting Lexical Bundles for Explicit Noticing with Schematic Linguistic Representation

    ERIC Educational Resources Information Center

    Thomson, Haidee Elizabeth

    2016-01-01

    Lexical bundles are essential for fluency, but their incompleteness is a stumbling block for learners. In this study, two presentation methods to increase awareness of lexical bundles through explicit noticing are explored and compared with incidental exposure. The three conditions in this study were as follows: noticing with schematic linguistic…

  18. Medicare's bundling pilot: including post-acute care services.

    PubMed

    Dummit, Laura A

    2011-03-28

    Fee-for-service Medicare, in which a separate payment is made for each service, rewards health care providers for delivering more services, but not necessarily coordinating those services over time or across settings. To help address these concerns, the Patient Protection and Affordable Care Act of 2010 requires Medicare to experiment with making a bundled payment for a hospitalization plus post-acute care, that is, the recuperative or rehabilitative care following a hospital discharge. This bundled payment approach is intended to promote more efficient care across the acute/post-acute episode because the entity that receives the payment has financial incentives to keep episode costs below the payment. Although the entity is expected to control costs through improved care coordination and efficiency, it could stint on care or avoid expensive patients instead. This issue brief focuses on the unique challenges posed by the inclusion of post-acute care services in a payment bundle and special considerations in implementing and evaluating the episode payment approach.

  19. Increased reliability of nuclear magnetic resonance protein structures by consensus structure bundles.

    PubMed

    Buchner, Lena; Güntert, Peter

    2015-02-03

    Nuclear magnetic resonance (NMR) structures are represented by bundles of conformers calculated from different randomized initial structures using identical experimental input data. The spread among these conformers indicates the precision of the atomic coordinates. However, there is as yet no reliable measure of structural accuracy, i.e., how close NMR conformers are to the "true" structure. Instead, the precision of structure bundles is widely (mis)interpreted as a measure of structural quality. Attempts to increase precision often overestimate accuracy by tight bundles of high precision but much lower accuracy. To overcome this problem, we introduce a protocol for NMR structure determination with the software package CYANA, which produces, like the traditional method, bundles of conformers in agreement with a common set of conformational restraints but with a realistic precision that is, throughout a variety of proteins and NMR data sets, a much better estimate of structural accuracy than the precision of conventional structure bundles. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Consensus Bundle on Prevention of Surgical Site Infections After Major Gynecologic Surgery.

    PubMed

    Pellegrini, Joseph E; Toledo, Paloma; Soper, David E; Bradford, William C; Cruz, Deborah A; Levy, Barbara S; Lemieux, Lauren A

    2017-01-01

    Surgical site infections are the most common complication of surgery in the United States. Of surgeries in women of reproductive age, hysterectomy is one of the most frequently performed, second only to cesarean birth. Therefore, prevention of surgical site infections in women undergoing gynecologic surgery is an ideal topic for a patient safety bundle. The primary purpose of this safety bundle is to provide recommendations that can be implemented into any surgical environment in an effort to reduce the incidence of surgical site infection. This bundle was developed by a multidisciplinary team convened by the Council on Patient Safety in Women's Health Care. The bundle is organized into four domains: Readiness, Recognition and Prevention, Response, and Reporting and Systems Learning. In addition to recommendations for practice, each of the domains stresses communication and teamwork between all members of the surgical team. Although the bundle components are designed to be adaptable to work in a variety of clinical settings, standardization within institutions is encouraged.

  1. Elevated temperature triggers human respiratory syncytial virus F protein six-helix bundle formation

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

    Yunus, Abdul S.; Jackson, Trent P.; Crisafi, Katherine

    2010-01-20

    Human respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract infection in infants, immunocompromised patients, and the elderly. The RSV fusion (F) protein mediates fusion of the viral envelope with the target cell membrane during virus entry and is a primary target for antiviral drug and vaccine development. The F protein contains two heptad repeat regions, HR1 and HR2. Peptides corresponding to these regions form a six-helix bundle structure that is thought to play a critical role in membrane fusion. However, characterization of six-helix bundle formation in native RSV F protein has been hindered by themore » fact that a trigger for F protein conformational change has yet to be identified. Here we demonstrate that RSV F protein on the surface of infected cells undergoes a conformational change following exposure to elevated temperature, resulting in the formation of the six-helix bundle structure. We first generated and characterized six-helix bundle-specific antibodies raised against recombinant peptides modeling the RSV F protein six-helix bundle structure. We then used these antibodies as probes to monitor RSV F protein six-helix bundle formation in response to a diverse array of potential triggers of conformational changes. We found that exposure of 'membrane-anchored' RSV F protein to elevated temperature (45-55 deg. C) was sufficient to trigger six-helix bundle formation. Antibody binding to the six-helix bundle conformation was detected by both flow cytometry and cell-surface immunoprecipitation of the RSV F protein. None of the other treatments, including interaction with a number of potential receptors, resulted in significant binding by six-helix bundle-specific antibodies. We conclude that native, untriggered RSV F protein exists in a metastable state that can be converted in vitro to the more stable, fusogenic six-helix bundle conformation by an increase in thermal energy. These findings help to better define the

  2. Bundling in Place: Translating the NGSS into Place-Based Earth-System Science Curricula

    NASA Astrophysics Data System (ADS)

    Semken, S. C.

    2016-12-01

    Bundling is the process of grouping Performance Expectations (PEs) from the Next Generation Science Standards (NGSS) into coherent units based on a defined topic, idea, question, or phenomenon. Bundling sorts the PEs for a given grade or grade band into a teachable narrative: a key stage in building curriculum, instruction, and assessment from the NGSS. To encourage and facilitate this, bundling guidelines have recently been released on the NGSS website (nextgenscience.org/glossary/bundlesbundling), and example bundles for different grade bands and disciplines are also being developed and posted there. According to these guidelines the iterative process of bundling begins with organization of PEs according to natural connections among them, and alignment of the three NGSS dimensions (Disciplinary Core Ideas, Cross-Cutting Concepts, and Science and Engineering Practices) that underpin each PE. Bundles are grouped by coherence and increasing complexity into courses, and courses into course sets that should encompass all PEs for a grade band. Bundling offers a natural way to translate the NGSS into highly contextualized curricula such as place-based (PB) teaching, which is situated in specific places or regions and focused on natural and cultural features, processes, phenomena, history, and challenges to sustainability therein. Attributes of place and our individual and collective connections to place (sense of place) directly inform PB curriculum, pedagogy, and assessment. PEs can be bundled by their relevance to these themes. Following the NGSS guidelines, I model the process for PB instruction by bundling PEs around the themes of Paleozoic geology and carbonate deposition and their relationships to mining and calcining of limestone in Anthropocene cement production for developing communities. The bundles integrate aspects of Earth history, the carbon cycle, mineral resources, climate change, and sustainability using specific local examples and narratives. They are

  3. Negative hair-bundle stiffness betrays a mechanism for mechanical amplification by the hair cell.

    PubMed

    Martin, P; Mehta, A D; Hudspeth, A J

    2000-10-24

    Hearing and balance rely on the ability of hair cells in the inner ear to sense miniscule mechanical stimuli. In each cell, sound or acceleration deflects the mechanosensitive hair bundle, a tuft of rigid stereocilia protruding from the cell's apical surface. By altering the tension in gating springs linked to mechanically sensitive transduction channels, this deflection changes the channels' open probability and elicits an electrical response. To detect weak stimuli despite energy losses caused by viscous dissipation, a hair cell can use active hair-bundle movement to amplify its mechanical inputs. This amplificatory process also yields spontaneous bundle oscillations. Using a displacement-clamp system to measure the mechanical properties of individual hair bundles from the bullfrog's ear, we found that an oscillatory bundle displays negative slope stiffness at the heart of its region of mechanosensitivity. Offsetting the hair bundle's position activates an adaptation process that shifts the region of negative stiffness along the displacement axis. Modeling indicates that the interplay between negative bundle stiffness and the motor responsible for mechanical adaptation produces bundle oscillation similar to that observed. Just as the negative resistance of electrically excitable cells and of tunnel diodes can be embedded in a biasing circuit to amplify electrical signals, negative stiffness can be harnessed to amplify mechanical stimuli in the ear.

  4. Uncovering Ecosystem Service Bundles through Social Preferences

    PubMed Central

    Martín-López, Berta; Iniesta-Arandia, Irene; García-Llorente, Marina; Palomo, Ignacio; Casado-Arzuaga, Izaskun; Amo, David García Del; Gómez-Baggethun, Erik; Oteros-Rozas, Elisa; Palacios-Agundez, Igone; Willaarts, Bárbara; González, José A.; Santos-Martín, Fernando; Onaindia, Miren; López-Santiago, Cesar; Montes, Carlos

    2012-01-01

    Ecosystem service assessments have increasingly been used to support environmental management policies, mainly based on biophysical and economic indicators. However, few studies have coped with the social-cultural dimension of ecosystem services, despite being considered a research priority. We examined how ecosystem service bundles and trade-offs emerge from diverging social preferences toward ecosystem services delivered by various types of ecosystems in Spain. We conducted 3,379 direct face-to-face questionnaires in eight different case study sites from 2007 to 2011. Overall, 90.5% of the sampled population recognized the ecosystem’s capacity to deliver services. Formal studies, environmental behavior, and gender variables influenced the probability of people recognizing the ecosystem’s capacity to provide services. The ecosystem services most frequently perceived by people were regulating services; of those, air purification held the greatest importance. However, statistical analysis showed that socio-cultural factors and the conservation management strategy of ecosystems (i.e., National Park, Natural Park, or a non-protected area) have an effect on social preferences toward ecosystem services. Ecosystem service trade-offs and bundles were identified by analyzing social preferences through multivariate analysis (redundancy analysis and hierarchical cluster analysis). We found a clear trade-off among provisioning services (and recreational hunting) versus regulating services and almost all cultural services. We identified three ecosystem service bundles associated with the conservation management strategy and the rural-urban gradient. We conclude that socio-cultural preferences toward ecosystem services can serve as a tool to identify relevant services for people, the factors underlying these social preferences, and emerging ecosystem service bundles and trade-offs. PMID:22720006

  5. Heterotic line bundle models on elliptically fibered Calabi-Yau three-folds

    NASA Astrophysics Data System (ADS)

    Braun, Andreas P.; Brodie, Callum R.; Lukas, Andre

    2018-04-01

    We analyze heterotic line bundle models on elliptically fibered Calabi-Yau three-folds over weak Fano bases. In order to facilitate Wilson line breaking to the standard model group, we focus on elliptically fibered three-folds with a second section and a freely-acting involution. Specifically, we consider toric weak Fano surfaces as base manifolds and identify six such manifolds with the required properties. The requisite mathematical tools for the construction of line bundle models on these spaces, including the calculation of line bundle cohomology, are developed. A computer scan leads to more than 400 line bundle models with the right number of families and an SU(5) GUT group which could descend to standard-like models after taking the ℤ2 quotient. A common and surprising feature of these models is the presence of a large number of vector-like states.

  6. Direct His bundle pacing post AVN ablation.

    PubMed

    Lakshmanadoss, Umashankar; Aggarwal, Ashim; Huang, David T; Daubert, James P; Shah, Abrar

    2009-08-01

    Atrioventricular nodal (AVN) ablation with concomitant pacemaker implantation is one of the strategies that reduce symptoms in patients with atrial fibrillation (AF). However, the long-term adverse effects of right ventricular (RV) apical pacing have led to the search for alternating sites of pacing. Biventricular pacing produces a significant improvement in functional capacity over RV pacing in patients undergoing AVN ablation. Another alternative site for pacing is direct His bundle to reduce the adverse outcome of RV pacing. Here, we present a case of direct His bundle pacing using steerable lead delivery system in a patient with symptomatic paroxysmal AF with concurrent AVN ablation.

  7. bop5 mutations reveal new roles for the IC138 phosphoprotein in the regulation of flagellar motility and asymmetric waveforms

    PubMed Central

    VanderWaal, Kristyn E.; Yamamoto, Ryosuke; Wakabayashi, Ken-ichi; Fox, Laura; Kamiya, Ritsu; Dutcher, Susan K.; Bayly, Phillip V.; Sale, Winfield S.; Porter, Mary E.

    2011-01-01

    I1 dynein, or dynein f, is a highly conserved inner arm isoform that plays a key role in the regulation of flagellar motility. To understand how the IC138 IC/LC subcomplex modulates I1 activity, we characterized the molecular lesions and motility phenotypes of several bop5 alleles. bop5-3, bop5-4, and bop5-5 are null alleles, whereas bop5-6 is an intron mutation that reduces IC138 expression. I1 dynein assembles into the axoneme, but the IC138 IC/LC subcomplex is missing. bop5 strains, like other I1 mutants, swim forward with reduced swimming velocities and display an impaired reversal response during photoshock. Unlike mutants lacking the entire I1 dynein, however, bop5 strains exhibit normal phototaxis. bop5 defects are rescued by transformation with the wild-type IC138 gene. Analysis of flagellar waveforms reveals that loss of the IC138 subcomplex reduces shear amplitude, sliding velocities, and the speed of bend propagation in vivo, consistent with the reduction in microtubule sliding velocities observed in vitro. The results indicate that the IC138 IC/LC subcomplex is necessary to generate an efficient waveform for optimal motility, but it is not essential for phototaxis. These findings have significant implications for the mechanisms by which IC/LC complexes regulate dynein motor activity independent of effects on cargo binding or complex stability. PMID:21697502

  8. SO(32) heterotic line bundle models

    NASA Astrophysics Data System (ADS)

    Otsuka, Hajime

    2018-05-01

    We search for the three-generation standard-like and/or Pati-Salam models from the SO(32) heterotic string theory on smooth, quotient complete intersection Calabi-Yau threefolds with multiple line bundles, each with structure group U(1). These models are S- and T-dual to intersecting D-brane models in type IIA string theory. We find that the stable line bundles and Wilson lines lead to the standard model gauge group with an extra U(1) B-L via a Pati-Salam-like symmetry and the obtained spectrum consists of three chiral generations of quarks and leptons, and vector-like particles. Green-Schwarz anomalous U(1) symmetries control not only the Yukawa couplings of the quarks and leptons but also the higher-dimensional operators causing the proton decay.

  9. Sequence Bundles: a novel method for visualising, discovering and exploring sequence motifs

    PubMed Central

    2014-01-01

    Background We introduce Sequence Bundles--a novel data visualisation method for representing multiple sequence alignments (MSAs). We identify and address key limitations of the existing bioinformatics data visualisation methods (i.e. the Sequence Logo) by enabling Sequence Bundles to give salient visual expression to sequence motifs and other data features, which would otherwise remain hidden. Methods For the development of Sequence Bundles we employed research-led information design methodologies. Sequences are encoded as uninterrupted, semi-opaque lines plotted on a 2-dimensional reconfigurable grid. Each line represents a single sequence. The thickness and opacity of the stack at each residue in each position indicates the level of conservation and the lines' curved paths expose patterns in correlation and functionality. Several MSAs can be visualised in a composite image. The Sequence Bundles method is designed to favour a tangible, continuous and intuitive display of information. Results We have developed a software demonstration application for generating a Sequence Bundles visualisation of MSAs provided for the BioVis 2013 redesign contest. A subsequent exploration of the visualised line patterns allowed for the discovery of a number of interesting features in the dataset. Reported features include the extreme conservation of sequences displaying a specific residue and bifurcations of the consensus sequence. Conclusions Sequence Bundles is a novel method for visualisation of MSAs and the discovery of sequence motifs. It can aid in generating new insight and hypothesis making. Sequence Bundles is well disposed for future implementation as an interactive visual analytics software, which can complement existing visualisation tools. PMID:25237395

  10. Frequent pauses in Escherichia coli flagella elongation revealed by single cell real-time fluorescence imaging.

    PubMed

    Zhao, Ziyi; Zhao, Yifan; Zhuang, Xiang-Yu; Lo, Wei-Chang; Baker, Matthew A B; Lo, Chien-Jung; Bai, Fan

    2018-05-14

    The bacterial flagellum is a large extracellular protein organelle that extrudes from the cell surface. The flagellar filament is assembled from tens of thousands of flagellin subunits that are exported through the flagellar type III secretion system. Here, we measure the growth of Escherichia coli flagella in real time and find that, although the growth rate displays large variations at similar lengths, it decays on average as flagella lengthen. By tracking single flagella, we show that the large variations in growth rate occur as a result of frequent pauses. Furthermore, different flagella on the same cell show variable growth rates with correlation. Our observations are consistent with an injection-diffusion model, and we propose that an insufficient cytoplasmic flagellin supply is responsible for the pauses in flagellar growth in E. coli.

  11. Spin transport in carbon nanotubes bundles: An ab-initio study

    NASA Astrophysics Data System (ADS)

    Meena, Shweta; Choudhary, Sudhanshu

    2017-10-01

    First principles investigations are performed on understanding the spin-polarized transport in carbon nanotubes and carbon nanotube bundles consisting of (8 , 0) and (17 , 0) SWCNTs kept in vertical (out-of-plane) arrangement and contacted by two CrO2 Half-Metallic-Ferromagnetic (HMF) electrodes. On comparison of the results for all the structures, it is observed that carbon nanotube bundle consisting of (17 , 0) CNT offers high TMR ∼100% and the transport phenomenon is tunneling, since there are no transmission states near Fermi level. However, in individual (8 , 0) and (17 , 0) CNT the transport is not because of tunneling, since there are significant number of transmission states near Fermi level. High Magneto Resistance (MR) 96% and 99% is observed in individual (8 , 0) and (17 , 0) CNTs respectively. Both TMR and Spin Injection Efficiency η (Spin-Filtration) are higher in (17 , 0) carbon nanotube bundle structure, which is due to carbon nanotube bundle acting as a perfect barrier in vertical (out-of-plane) arrangement resulting in negligible spin-down current (I↓) in both Parallel Configuration (PC) and Antiparallel Configuration (APC).

  12. Compressive force generation by a bundle of living biofilaments

    NASA Astrophysics Data System (ADS)

    Ramachandran, Sanoop; Ryckaert, Jean-Paul

    2013-08-01

    To study the compressional forces exerted by a bundle of living stiff filaments pressing on a surface, akin to the case of an actin bundle in filopodia structures, we have performed particulate molecular dynamics simulations of a grafted bundle of parallel living (self-assembling) filaments, in chemical equilibrium with a solution of their constitutive monomers. Equilibrium is established as these filaments, grafted at one end to a wall of the simulation box, grow at their chemically active free end, and encounter the opposite confining wall of the simulation box. Further growth of filaments requires bending and thus energy, which automatically limit the populations of longer filaments. The resulting filament sizes distribution and the force exerted by the bundle on the obstacle are analyzed for different grafting densities and different sub- or supercritical conditions, these properties being compared with the predictions of the corresponding ideal confined bundle model. In this analysis, non-ideal effects due to interactions between filaments and confinement effects are singled out. For all state points considered at the same temperature and at the same gap width between the two surfaces, the force per filament exerted on the opposite wall appears to be a function of a rescaled free monomer density hat{ρ }_1^eff. This quantity can be estimated directly from the characteristic length of the exponential filament size distribution P observed in the size domain where these grafted filaments are not in direct contact with the wall. We also analyze the dynamics of the filament contour length fluctuations in terms of effective polymerization (U) and depolymerization (W) rates, where again it is possible to disentangle non-ideal and confinement effects.

  13. The Actions of Calcium on Hair Bundle Mechanics in Mammalian Cochlear Hair Cells

    PubMed Central

    Beurg, Maryline; Nam, Jong-Hoon; Crawford, Andrew; Fettiplace, Robert

    2008-01-01

    Sound stimuli excite cochlear hair cells by vibration of each hair bundle, which opens mechanotransducer (MT) channels. We have measured hair-bundle mechanics in isolated rat cochleas by stimulation with flexible glass fibers and simultaneous recording of the MT current. Both inner and outer hair-cell bundles exhibited force-displacement relationships with a nonlinearity that reflects a time-dependent reduction in stiffness. The nonlinearity was abolished, and hair-bundle stiffness increased, by maneuvers that diminished calcium influx through the MT channels: lowering extracellular calcium, blocking the MT current with dihydrostreptomycin, or depolarizing to positive potentials. To simulate the effects of Ca2+, we constructed a finite-element model of the outer hair cell bundle that incorporates the gating-spring hypothesis for MT channel activation. Four calcium ions were assumed to bind to the MT channel, making it harder to open, and, in addition, Ca2+ was posited to cause either a channel release or a decrease in the gating-spring stiffness. Both mechanisms produced Ca2+ effects on adaptation and bundle mechanics comparable to those measured experimentally. We suggest that fast adaptation and force generation by the hair bundle may stem from the action of Ca2+ on the channel complex and do not necessarily require the direct involvement of a myosin motor. The significance of these results for cochlear transduction and amplification are discussed. PMID:18178649

  14. The impact of a ventilator bundle on preventing ventilator-associated pneumonia: a multicenter study.

    PubMed

    Eom, Joong Sik; Lee, Mi-Suk; Chun, Hee-Kyung; Choi, Hee Jung; Jung, Sun-Young; Kim, Yeon-Sook; Yoon, Seon Jin; Kwak, Yee Gyung; Oh, Gang-Bok; Jeon, Min-Hyok; Park, Sun-Young; Koo, Hyun-Sook; Ju, Young-Su; Lee, Jin Seo

    2014-01-01

    For prevention of ventilator-associated pneumonia (VAP), a bundle approach was applied to patients receiving mechanical ventilation in intensive care units. The incidence of VAP and the preventive efficacy of the VAP bundle were investigated. A quasi-experimental study was conducted in adult intensive care units of 6 university hospitals with similar VAP rates. We implemented the VAP bundle between March 2011 and June 2011, then compared the rate of VAP after implementation of the VAP bundle with the rate in the previous 8 months. Our ventilator bundle included head of bed elevation, peptic ulcer disease prophylaxis, deep venous thrombosis prophylaxis, and oral decontamination with chlorhexidine 0.12%. Continuous aspiration of subglottic secretions was an option. Implementation of the VAP bundle reduced the VAP rate from a mean of 4.08 cases per 1,000 ventilator-days to 1.16 cases per 1,000 ventilator-days. The incidence density ratio (rate) was 0.28 (95% confidence interval, 0.275-0.292). Implementing the appropriate VAP bundle significantly decreased the incidence of VAP in patients with mechanical ventilation. Copyright © 2014 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

  15. Multi-scale fluorescence imaging of bacterial infections in animal models

    NASA Astrophysics Data System (ADS)

    Bixler, Joel N.; Kong, Ying; Cirillo, Jeffrey D.; Maitland, Kristen C.

    2013-03-01

    Tuberculosis, caused by Mycobacterium tuberculosis (Mtb), currently affects roughly one-third of the world's population. Drug resistant strains of Mtb decrease the effectiveness of current therapeutics and demand the development of new antimicrobial therapies. In addition, the current vaccine, Bacille Calmette Guérin (BCG), has variable efficacy for disease prevention in different populations. Animal studies are often limited by the need to sacrifice at discrete time points for pathology and tissue homogenization, which greatly reduces spatial and temporal resolution. Optical imaging offers the potential for a minimally-invasive solution to imaging on a macroscopic and microscopic scale, allowing for high resolution study of infection. We have integrated a fluorescence microendoscope into a whole-animal optical imaging system, allowing for simultaneous microscopic and macroscopic imaging of tdTomato expressing BCG in vivo. A 535 nm LED was collimated and launched into a 10,000 element fiber bundle with an outer diameter of 0.66 mm. The fiber bundle can be inserted through an intra-tracheal catheter into the lung of a mouse. Fluorescence emission can either be (1) collected by the bundle and imaged onto the surface of a CCD camera for localized detection or (2) the fluorescence can be imaged by the whole animal imaging system providing macroscopic information. Results from internal localized excitation and external whole body detection indicate the potential for imaging bacterial infections down to 100 colony forming units. This novel imaging technique has the potential to allow for functional studies, enhancing the ability to assess new therapeutic agents.

  16. Adsorption site analysis of impurity embedded single-walled carbon nanotube bundles

    USGS Publications Warehouse

    Agnihotri, S.; Mota, J.P.B.; Rostam-Abadi, M.; Rood, M.J.

    2006-01-01

    Bundle morphology and adsorptive contributions from nanotubes and impurities are studied both experimentally and by simulation using a computer-aided methodology, which employs a small physisorbed probe molecule to explore the porosity of nanotube samples. Grand canonical Monte Carlo simulation of nitrogen adsorption on localized sites of a bundle is carried out to predict adsorption in its accessible internal pore volume and on its external surface as a function of tube diameter. External adsorption is split into the contributions from the clean surface of the outermost nanotubes of the bundle and from the surface of the impurities. The site-specific isotherms are then combined into a global isotherm for a given sample using knowledge of its tube-diameter distribution obtained by Raman spectroscopy. The structural parameters of the sample, such as the fraction of open-ended nanotubes and the contributions from impurities and nanotube bundles to total external surface area, are determined by fitting the experimental nitrogen adsorption data to the simulated isotherm. The degree of closure between experimental and calculated adsorption isotherms for samples manufactured by two different methods, to provide different nanotube morphology and contamination level, further strengthens the validity and resulting interpretations based on the proposed approach. The average number of nanotubes per bundle and average bundle size, within a sample, are also quantified. The proposed method allows for extrapolation of adsorption properties to conditions where the purification process is 100% effective at removing all impurities and opening access to all intrabundle adsorption sites. ?? 2006 Elsevier Ltd. All rights reserved.

  17. Formation and structural organization of the egg-sperm bundle of the scleractinian coral Montipora capitata

    NASA Astrophysics Data System (ADS)

    Padilla-Gamiño, J. L.; Weatherby, T. M.; Waller, R. G.; Gates, R. D.

    2011-06-01

    The majority of scleractinian corals are hermaphrodites that broadcast spawn their gametes separately or packaged as egg-sperm bundles during spawning events that are timed to the lunar cycle. The egg-sperm bundle is an efficient way of transporting gametes to the ocean surface where fertilization takes place, while minimizing sperm dilution and maximizing the opportunity for gamete encounters during a spawning event. To date, there are few studies that focus on the formation and structure of egg-sperm bundle. This study explores formation, ultrastructure, and longevity of the egg-sperm bundle in Montipora capitata, a major reef building coral in Hawai`i. Our results show that the egg-sperm bundle is formed by a mucus layer secreted by the oocytes. The sperm package is located at the center of each bundle, possibly reflecting the development of male and female gametes in different mesenteries. Once the egg-sperm bundle has reached the ocean surface, it breaks open within 10-35 min, depending on the environmental conditions (i.e., wind, water turbulence). Although the bundle has an ephemeral life span, the formation of an egg-sperm bundle is a fundamental part of the reproductive process that could be strongly influenced by climate change and deterioration of water quality (due to anthropogenic effects) and thus requires further investigation.

  18. Effects of Mechanical Loading on the Dynamics of Hair-Cell Stereociliary Bundles

    NASA Astrophysics Data System (ADS)

    Fredrickson, Lea

    Hearing is remarkably sensitive and still not entirely understood. Hair cells of the inner ear are the mechano-electrical transducers of sound and understanding how they function is essential to the understanding of hearing in general. Spontaneous oscillations exhibited by stereociliary bundles of the bullfrog sacculus provide a useful probe for the study of the hair cells' internal dynamic state. In this work we study the effects of mechanical loading on these hair-cell bundles in order to study their dynamics. When applying stiffness loads, we find that the spontaneous oscillation profile changes from multimode to single mode with light loading, and decreases in amplitude and increases in frequency with stiffer loads. We also find that tuning decreases with increasing load such that at loads comparable to in vivo conditions the tuning is flat. We further explore loading via deflections to hair cell bundles, both in the form of steady-state offsets and slow ramps. We find that steady state offsets lead to significant modulation of the characteristic frequency of response, decreasing the frequency in the channels closed direction (negative) and increasing it in the channels open direction (positive). Attachment to the overlying membrane was found, in vitro, to affect bundle offset position in hair cells of the bullfrog sacculus. Application of similar offsets on free-standing, spontaneously oscillating hair bundles shows modulation of their dynamic state, i.e. oscillation profile, characteristic frequency, and response to stimulus. Large offsets are found to arrest spontaneous oscillations, which recover upon reversal of the stimulus. The dynamical state of the hair bundle is dependent on both the history and direction of the offset stimulus. Oscillation suppression occurs much more readily in the negative direction and the bundle behavior approaching quiescence is distinct from that in the positive direction. With the change in spontaneous oscillation frequency

  19. Calculation of Non-Bonded Forces Due to Sliding of Bundled Carbon Nanotubes

    NASA Technical Reports Server (NTRS)

    Frankland, S. J. V.; Bandorawalla, T.; Gates, T. S.

    2003-01-01

    An important consideration for load transfer in bundles of single-walled carbon nanotubes is the nonbonded (van der Waals) forces between the nanotubes and their effect on axial sliding of the nanotubes relative to each other. In this research, the non-bonded forces in a bundle of seven hexagonally packed (10,10) single-walled carbon nanotubes are represented as an axial force applied to the central nanotube. A simple model, based on momentum balance, is developed to describe the velocity response of the central nanotube to the applied force. The model is verified by comparing its velocity predictions with molecular dynamics simulations that were performed on the bundle with different force histories applied to the central nanotube. The model was found to quantitatively predict the nanotube velocities obtained from the molecular dynamics simulations. Both the model and the simulations predict a threshold force at which the nanotube releases from the bundle. This force converts to a shear yield strength of 10.5-11.0 MPa for (10,10) nanotubes in a bundle.

  20. Consensus Bundle on Prevention of Surgical Site Infections After Major Gynecologic Surgery.

    PubMed

    Pellegrini, Joseph E; Toledo, Paloma; Soper, David E; Bradford, William C; Cruz, Deborah A; Levy, Barbara S; Lemieux, Lauren A

    2017-02-06

    Surgical site infections are the most common complication of surgery in the United states. Of surgeries in women of reproductive age, hysterectomy is one of the most frequently performed, second only to cesarean birth. Therefore, prevention of surgical site infections in women undergoing gynecologic surgery is an ideal topic for a patient safety bundle. The primary purpose of this safety bundle is to provide recommendations that can be implemented into any surgical environment in an effot to reduce the incidence of surgical site infection. This bundle was developed by a multidisciplinary team convened by the Council on Patient Safety in Women's Health Care. The bundle is organized into four domains: Readiness, Recognition and Prevention, Response, and Reporting and Systems Learning. In addition to recommendations for practice, each of the domains stresses communication and teamwork between all members of the surgical team. Although the bundle components are designed to be adaptable to work in a variety of clinical settings, standardization within institutions is encouraged. Copyright ©2016 American College of Obstetricians and Gynecologists.

  1. Infinitesimal moduli of G2 holonomy manifolds with instanton bundles

    NASA Astrophysics Data System (ADS)

    de la Ossa, Xenia; Larfors, Magdalena; Svanes, Eirik E.

    2016-11-01

    We describe the infinitesimal moduli space of pairs ( Y, V) where Y is a manifold with G 2 holonomy, and V is a vector bundle on Y with an instanton connection. These structures arise in connection to the moduli space of heterotic string compactifications on compact and non-compact seven dimensional spaces, e.g. domain walls. Employing the canonical G 2 cohomology developed by Reyes-Carrión and Fernández and Ugarte, we show that the moduli space decomposes into the sum of the bundle moduli {H}_{{overset{ěe }{d}}_A}^1(Y,End(V)) plus the moduli of the G 2 structure preserving the instanton condition. The latter piece is contained in {H}_{overset{ěe }{d}θ}^1(Y,TY) , and is given by the kernel of a map overset{ěe }{F} which generalises the concept of the Atiyah map for holomorphic bundles on complex manifolds to the case at hand. In fact, the map overset{ěe }{F} is given in terms of the curvature of the bundle and maps {H}_{overset{ěe }{d}θ}^1(Y,TY) into {H}_{{overset{ěe }{d}}_A}^2(Y,End(V)) , and moreover can be used to define a cohomology on an extension bundle of TY by End( V). We comment further on the resemblance with the holomorphic Atiyah algebroid and connect the story to physics, in particular to heterotic compactifications on ( Y, V) when α' = 0.

  2. Femoral tunnel placement in single-bundle anterior cruciate ligament reconstruction: a cadaveric study relating transtibial lateralized femoral tunnel position to the anteromedial and posterolateral bundle femoral origins of the anterior cruciate ligament.

    PubMed

    Rue, John-Paul H; Ghodadra, Neil; Bach, Bernard R

    2008-01-01

    There is controversy regarding the necessity of reconstructing both the posterolateral and anteromedial bundles of the anterior cruciate ligament. A laterally oriented transtibial drilled femoral tunnel replaces portions of the femoral footprints of the anteromedial and posterolateral bundles of the anterior cruciate ligament. Descriptive laboratory study. Footprints of the anteromedial and posterolateral bundles of the anterior cruciate ligament were preserved on 7 matched pairs (5 female, 2 male) of fresh-frozen human cadaveric femurs (14 femurs total). Each femur was anatomically oriented and secured in a custom size-appropriate, side-matched replica tibia model to simulate transtibial retrograde drilling of a 10-mm femoral tunnel in each specimen. The relationship of the tunnel relative to footprints of both bundles of the anterior cruciate ligament was recorded using a Microscribe MX digitizer. The angle of the femoral tunnel relative to the vertical 12-o'clock position was recorded for all 14 specimens; only 10 specimens were used for footprint measurements. On average, the 10-mm femoral tunnel overlapped 50% of the anteromedial bundle (range, 2%-83%) and 51% of the posterolateral bundle (range, 16%-97%). The footprint of the anteromedial bundle occupied 32% (range, 3%-49%) of the area of the tunnel; the footprint of the posterolateral bundle contributed 26% (range, 7%-41%). The remainder of the area of the 10-mm tunnel did not overlap with the anterior cruciate ligament footprint. The mean absolute angle of the femoral tunnel as measured directly on the specimen was 48 degrees (range, 42 degrees-53 degrees) from vertical, corresponding to approximately a 10:30 clock face position on a right knee. Anterior cruciate ligament reconstruction using a laterally oriented transtibial drilled femoral tunnel incorporates portions of the anteromedial and posterolateral bundle origins of the native anterior cruciate ligament. A laterally oriented transtibial drilled

  3. Fabrication of bundle-structured tube-leaky optical fibers for infrared thermal imaging

    NASA Astrophysics Data System (ADS)

    Kobayashi, T.; Katagiri, T.; Matsuura, Y.

    2017-02-01

    Bundled glass tubular fibers were fabricated by glass drawing technique for endoscopic infrared-thermal imaging. The bundle fibers were made of borosilicate glass and have a structure like a photonic crystal fiber having multiple hollow cores. Fabricated fibers have a length of 90 cm and each pixel sizes are less than 80 μm. By setting the thickness of glass wall to a quarter-wavelength optical thickness, light is confined in the air core as a leaky mode with a low loss owing to the interference effect of the thin glass wall and this type of hollow-core fibers is known as tube leaky fibers. The transmission losses of bundled fibers were firstly measured and it was found that bundled tube-leaky fibers have reasonably low transmission losses in spite of the small pixel size. Then thermal images were delivered by the bundled fibers combining with an InSb infrared camera. Considering applications with rigid endoscopes, an imaging system composed of a 30-cm long fiber bundle and a half-ball lens with a diameter of 2 mm was fabricated. By using this imaging system, a metal wire with a thickness of 200 μm was successfully observed and another test showed that the minimum detected temperature was 32.0 °C and the temperature resolution of the system was around 0.7 °C.

  4. Analysis of multimode fiber bundles for endoscopic spectral-domain optical coherence tomography

    PubMed Central

    Risi, Matthew D.; Makhlouf, Houssine; Rouse, Andrew R.; Gmitro, Arthur F.

    2016-01-01

    A theoretical analysis of the use of a fiber bundle in spectral-domain optical coherence tomography (OCT) systems is presented. The fiber bundle enables a flexible endoscopic design and provides fast, parallelized acquisition of the OCT data. However, the multimode characteristic of the fibers in the fiber bundle affects the depth sensitivity of the imaging system. A description of light interference in a multimode fiber is presented along with numerical simulations and experimental studies to illustrate the theoretical analysis. PMID:25967012

  5. The MIMIC Model as a Tool for Differential Bundle Functioning Detection

    ERIC Educational Resources Information Center

    Finch, W. Holmes

    2012-01-01

    Increasingly, researchers interested in identifying potentially biased test items are encouraged to use a confirmatory, rather than exploratory, approach. One such method for confirmatory testing is rooted in differential bundle functioning (DBF), where hypotheses regarding potential differential item functioning (DIF) for sets of items (bundles)…

  6. Design and impact of bundled payment for detox and follow-up care.

    PubMed

    Quinn, Amity E; Hodgkin, Dominic; Perloff, Jennifer N; Stewart, Maureen T; Brolin, Mary; Lane, Nancy; Horgan, Constance M

    2017-11-01

    Recent payment reforms promote movement from fee-for-service to alternative payment models that shift financial risk from payers to providers, incentivizing providers to manage patients' utilization. Bundled payment, an episode-based fixed payment that includes the prices of a group of services that would typically treat an episode of care, is expanding in the United States. Bundled payment has been recommended as a way to pay for comprehensive SUD treatment and has the potential to improve treatment engagement after detox, which could reduce detox readmissions, improve health outcomes, and reduce medical care costs. However, if moving to bundled payment creates large losses for some providers, it may not be sustainable. The objective of this study was to design the first bundled payment for detox and follow-up care and to estimate its impact on provider revenues. Massachusetts Medicaid beneficiaries' behavioral health, medical, and pharmacy claims from July 2010-April 2013 were used to build and test a detox bundled payment for continuously enrolled adults (N=5521). A risk adjustment model was developed using general linear modeling to predict beneficiaries' episode costs. The projected payments to each provider from the risk adjustment analysis were compared to the observed baseline costs to determine the potential impact of a detox bundled payment reform on organizational revenues. This was modeled in two ways: first assuming no change in behavior and then assuming a supply-side cost sharing behavioral response of a 10% reduction in detox readmissions and an increase of one individual counseling and one group counseling session. The mean total 90-day detox episode cost was $3743. Nearly 70% of the total mean cost consists of the index detox, psychiatric inpatient care, and short-term residential care. Risk mitigation, including risk adjustment, substantially reduced the variation of the mean episode cost. There are opportunities for organizations to gain revenue

  7. Lagrangians and Euler morphisms from connections on the frame bundle

    NASA Astrophysics Data System (ADS)

    Kurek, J.; Mikulski, W. M.

    2011-07-01

    We classify all natural operators transforming torsion free classical linear connections ∇ on m-dimensional manifolds M into r-th order Lagrangians λ(∇) and Euler morphisms E(∇) on the linear frame bundle P1M. We also briefly write how this classification result can be generalized on higher order frame bundles PkM instead of P1M.

  8. Muon Bundles as a Sign of Strangelets from the Universe

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

    Kankiewicz, P.; Rybczyński, M.; Włodarczyk, Z.

    Recently, the CERN ALICE experiment observed muon bundles of very high multiplicities in its dedicated cosmic ray (CR) run, thereby confirming similar findings from the LEP era at CERN (in the CosmoLEP project). Originally, it was argued that they apparently stem from the primary CRs with a heavy masses. We propose an alternative possibility arguing that muonic bundles of highest multiplicity are produced by strangelets, hypothetical stable lumps of strange quark matter infiltrating our universe. We also address the possibility of additionally deducing their directionality which could be of astrophysical interest. Significant evidence for anisotropy of arrival directions of themore » observed high-multiplicity muonic bundles is found. Estimated directionality suggests their possible extragalactic provenance.« less

  9. Microtubule bundling plays a role in ethylene-mediated cortical microtubule reorientation in etiolated Arabidopsis hypocotyls.

    PubMed

    Ma, Qianqian; Sun, Jingbo; Mao, Tonglin

    2016-05-15

    The gaseous hormone ethylene is known to regulate plant growth under etiolated conditions (the 'triple response'). Although organization of cortical microtubules is essential for cell elongation, the underlying mechanisms that regulate microtubule organization by hormone signaling, including ethylene, are ambiguous. In the present study, we demonstrate that ethylene signaling participates in regulation of cortical microtubule reorientation. In particular, regulation of microtubule bundling is important for this process in etiolated hypocotyls. Time-lapse analysis indicated that selective stabilization of microtubule-bundling structures formed in various arrays is related to ethylene-mediated microtubule orientation. Bundling events and bundle growth lifetimes were significantly increased in oblique and longitudinal arrays, but decreased in transverse arrays in wild-type cells in response to ethylene. However, the effects of ethylene on microtubule bundling were partially suppressed in a microtubule-bundling protein WDL5 knockout mutant (wdl5-1). This study suggests that modulation of microtubule bundles that have formed in certain orientations plays a role in reorienting microtubule arrays in response to ethylene-mediated etiolated hypocotyl cell elongation. © 2016. Published by The Company of Biologists Ltd.

  10. Enhanced field emission properties of carbon nanotube bundles confined in SiO2 pits

    NASA Astrophysics Data System (ADS)

    Lim, Yu Dian; Grapov, Dmitry; Hu, Liangxing; Kong, Qinyu; Tay, Beng Kang; Labunov, Vladimir; Miao, Jianmin; Coquet, Philippe; Aditya, Sheel

    2018-02-01

    It has been widely reported that carbon nanotubes (CNTs) exhibit superior field emission (FE) properties due to their high aspect ratios and unique structural properties. Among the various types of CNTs, random growth CNTs exhibit promising FE properties due to their reduced inter-tube screening effect. However, growing random growth CNTs on individual catalyst islands often results in spread out CNT bundles, which reduces overall field enhancement. In this study, significant improvement in FE properties in CNT bundles is demonstrated by confining them in microfabricated SiO2 pits. Growing CNT bundles in narrow (0.5 μm diameter and 2 μm height) SiO2 pits achieves FE current density of 1-1.4 A cm-2, which is much higher than for freestanding CNT bundles (76.9 mA cm-2). From the Fowler Nordheim plots, confined CNT bundles show a higher field enhancement factor. This improvement can be attributed to the reduced bundle diameter by SiO2 pit confinement, which yields bundles with higher aspect ratios. Combining the obtained outcomes, it can be conclusively summarized that confining CNTs in SiO2 pits yields higher FE current density due to the higher field enhancement of confined CNTs.

  11. 47 CFR 76.1514 - Bundling of video and local exchange services.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 4 2014-10-01 2014-10-01 false Bundling of video and local exchange services... RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Open Video Systems § 76.1514 Bundling of video and local exchange services. An open video system operator may offer video and local exchange...

  12. 47 CFR 76.1514 - Bundling of video and local exchange services.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 4 2013-10-01 2013-10-01 false Bundling of video and local exchange services... RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Open Video Systems § 76.1514 Bundling of video and local exchange services. An open video system operator may offer video and local exchange...

  13. 47 CFR 76.1514 - Bundling of video and local exchange services.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 4 2012-10-01 2012-10-01 false Bundling of video and local exchange services... RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Open Video Systems § 76.1514 Bundling of video and local exchange services. An open video system operator may offer video and local exchange...

  14. 47 CFR 76.1514 - Bundling of video and local exchange services.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 4 2010-10-01 2010-10-01 false Bundling of video and local exchange services... RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Open Video Systems § 76.1514 Bundling of video and local exchange services. An open video system operator may offer video and local exchange...

  15. 47 CFR 76.1514 - Bundling of video and local exchange services.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 4 2011-10-01 2011-10-01 false Bundling of video and local exchange services... RADIO SERVICES MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Open Video Systems § 76.1514 Bundling of video and local exchange services. An open video system operator may offer video and local exchange...

  16. Ab initio density functional theory investigation of electronic properties of semiconducting single-walled carbon nanotube bundles

    NASA Astrophysics Data System (ADS)

    Moradian, Rostam; Behzad, Somayeh; Azadi, Sam

    2008-09-01

    By using ab initio density functional theory we investigated the structural and electronic properties of semiconducting (7, 0), (8, 0) and (10, 0) carbon nanotube bundles. The energetic and electronic evolutions of nanotubes in the bundling process are also studied. The effects of inter-tube coupling on the electronic dispersions of semiconducting carbon nanotube bundles are demonstrated. Our results show that the inter-tube coupling decreases the energy gap in semiconducting nanotubes. We found that bundles of (7, 0) and (8, 0) carbon nanotubes have metallic feature, while (10, 0) bundle is a semiconductor with an energy gap of 0.22 eV. To clarify our results the band structures of isolated and bundled nanotubes are compared.

  17. Structured Post-IQ Domain Governs Selectivity of Myosin X for Fascin-Actin Bundles*

    PubMed Central

    Nagy, Stanislav; Rock, Ronald S.

    2010-01-01

    Without guidance cues, cytoskeletal motors would traffic components to the wrong destination with disastrous consequences for the cell. Recently, we identified a motor protein, myosin X, that identifies bundled actin filaments for transport. These bundles direct myosin X to a unique destination, the tips of cellular filopodia. Because the structural and kinetic features that drive bundle selection are unknown, we employed a domain-swapping approach with the nonselective myosin V to identify the selectivity module of myosin X. We found a surprising role of the myosin X tail region (post-IQ) in supporting long runs on bundles. Moreover, the myosin X head is adapted for initiating processive runs on bundles. We found that the tail is structured and biases the orientation of the two myosin X heads because a targeted insertion that introduces flexibility in the tail abolishes selectivity. Together, these results suggest how myosin motors may manage to read cellular addresses. PMID:20538587

  18. Bacterial Community Sstructure and Novel Species of Magnetotactic Bacteria in Sediments from a Seamount in the Mariana Volcanic Arc

    NASA Astrophysics Data System (ADS)

    PAN, H.; LIU, J.; Zhang, W.; Xiao, T.; Wu, L. F.

    2017-12-01

    Seamounts are unique ecosystems where undersea mountains rise abruptly from the sea floor and interact dynamically with underwater currents, creating peculiar biological habitats with various microbial community structures. Certain bacteria associated with seamounts form conspicuous extracellular iron oxide structures, including encrusted stalks, flattened bifurcating tubes, and filamentous sheaths. To extend knowledge of seamount microorganisms we performed a systematic analysis of the population composition and occurrence of live magnetotactic bacteria (MTB) in sediments of a seamount in the Mariana volcanic arc. Proteobacteria dominated at 13 stations, and were the second in abundance to members of the Firmicutes at a deep station on a steep slope facing the Yap-Mariana trench. We found MTB that synthesize intracellular iron-oxide nanocrystals in biogenic sediments at all 14 stations, at seawater depths ranging from 238 to 2023 m. A novel flagellar apparatus, and the most complex yet reported, was observed in magnetotactic cocci; it comprises one or two bundles of 19 flagella arranged in a 3:4:5:4:3 array. Phylogenetic analysis of 16S rRNA gene sequences identified 16 novel species of MTB specific to this seamount. The geographic properties at the various stations on the seamount appear to be important in shaping the microbial community structure.

  19. Understanding nurses' views on a pressure ulcer prevention care bundle: a first step towards successful implementation.

    PubMed

    Chaboyer, Wendy; Gillespie, Brigid M

    2014-12-01

    To explore nurses' views of the barriers and facilitators to the use of a newly devised patient-centred pressure ulcer prevention care bundle. Given pressure ulcer prevention strategies are not implemented consistently, the use of a pressure ulcer care bundle may improve implementation given bundles generally assist in standardising care. A quality improvement project was undertaken after a pressure ulcer prevention care bundle was developed and pilot-tested. Short, conversational interviews with nurse explored their views of a patient-centred pressure ulcer care bundle. Interviews were audio-taped and transcribed. Inductive content analysis was used to analyse the transcripts. A total of 20 nurses were interviewed. Five categories with corresponding subcategories emerged from the analysis. They were increasing awareness of pressure ulcer prevention, prompting pressure ulcer prevention activities, promoting active patient participation, barriers to using a pressure ulcer prevention care bundle and enabling integration of the pressure ulcer prevention care bundle into routine practice. Benefits of using a patient-centred pressure ulcer prevention care bundle may include prompting patients and staff to implement prevention strategies and promote active patient participation in care. The success of the care bundle relied on both patients' willingness to participate and nurses' willingness to incorporate it into their routine work. A patient-centred pressure ulcer prevention care bundle may facilitate more consistent implementation of pressure ulcer prevention strategies and active patient participation in care. © 2014 John Wiley & Sons Ltd.

  20. Management of acute Achilles tendon rupture with tendon-bundle technique

    PubMed Central

    Li, Chun-Guang; Li, Bing

    2017-01-01

    Objective *These authors contributed equally to this work.To explore tendon-bundle technique for treating Achilles tendon rupture with no defects. Methods Patients with full unilateral Achilles tendon rupture with no defects were included. The Achilles tendon medial edge surgical repair approach was used, revealing horsetail-like rupture bundles. Tendon bundles were anatomically realigned and repaired end-to-end using 5-0 sutures. Patients were followed-up for 1 year, and assessed for differences between the repaired versus healthy limb. Results Out of 24 patients (18 male, 6 female; aged 19–56 years) at 1 year following surgery, mean American Orthopaedic Foot and Ankle Society score was 92.4 ± 5.9; mean differences between the surgically repaired versus contralateral side in dorsiflexion and plantarflexion angle were 3.5 ± 2.3° and 5.6 ± 3.2°, respectively; mean difference in calf circumference between the two sides was 0.9 ± 0.5 cm; and mean increase in Achilles tendon width versus the healthy side was 0.8 ± 0.2 cm. By 1 year post-surgery, there were no significant between-side differences in dorsiflexion and plantarflexion angle, or calf circumference. Conclusions Tendon-bundle surgery resulted in good ankle function restoration and low complication rates. Tendon-bundle surgery may reduce blood supply destruction and maximally preserve Achilles tendon length, and may be effective for treating Achilles tendon rupture with no defects. PMID:28222622

  1. Hemlock woolly adelgid (Homoptera: Adelgidae): stylet bundle insertion and feeding sites

    Treesearch

    Rebecca F. Young; Kathleen S. Shields; Graeme P. Berlyn

    1995-01-01

    Stylet bundle insertion site, path traveled, and feeding site were examined for the hemlock woolly adelgid, Adelges tsugae Annand, on needles from current and previous years of eastern hemlock, Tsuga canadensis Carriere. The stylet bundle is composed of 4 individual stylets--2 outer mandibular stylets and 2 inner maxillary stylets...

  2. In Helicobacter pylori auto-inducer-2, but not LuxS/MccAB catalysed reverse transsulphuration, regulates motility through modulation of flagellar gene transcription

    PubMed Central

    2010-01-01

    Background LuxS may function as a metabolic enzyme or as the synthase of a quorum sensing signalling molecule, auto-inducer-2 (AI-2); hence, the mechanism underlying phenotypic changes upon luxS inactivation is not always clear. In Helicobacter pylori, we have recently shown that, rather than functioning in recycling methionine as in most bacteria, LuxS (along with newly-characterised MccA and MccB), synthesises cysteine via reverse transsulphuration. In this study, we investigated whether and how LuxS controls motility of H. pylori, specifically if it has its effects via luxS-required cysteine metabolism or via AI-2 synthesis only. Results We report that disruption of luxS renders H. pylori non-motile in soft agar and by microscopy, whereas disruption of mccAHp or mccBHp (other genes in the cysteine provision pathway) does not, implying that the lost phenotype is not due to disrupted cysteine provision. The motility defect of the ΔluxSHp mutant was complemented genetically by luxSHp and also by addition of in vitro synthesised AI-2 or 4, 5-dihydroxy-2, 3-pentanedione (DPD, the precursor of AI-2). In contrast, exogenously added cysteine could not restore motility to the ΔluxSHp mutant, confirming that AI-2 synthesis, but not the metabolic effect of LuxS was important. Microscopy showed reduced number and length of flagella in the ΔluxSHp mutant. Immunoblotting identified decreased levels of FlaA and FlgE but not FlaB in the ΔluxSHp mutant, and RT-PCR showed that the expression of flaA, flgE, motA, motB, flhA and fliI but not flaB was reduced. Addition of DPD but not cysteine to the ΔluxSHp mutant restored flagellar gene transcription, and the number and length of flagella. Conclusions Our data show that as well as being a metabolic enzyme, H. pylori LuxS has an alternative role in regulation of motility by modulating flagellar transcripts and flagellar biosynthesis through production of the signalling molecule AI-2. PMID:20691071

  3. Consensus Bundle on Prevention of Surgical Site Infections After Major Gynecologic Surgery.

    PubMed

    Pellegrini, Joseph E; Toledo, Paloma; Soper, David E; Bradford, William C; Cruz, Deborah A; Levy, Barbara S; Lemieux, Lauren A

    Surgical site infections are the most common complications of surgery in the United States. Of surgeries in women of reproductive age, hysterectomy is one of the most frequently performed, second only to cesarean birth. Therefore, prevention of surgical site infections in women undergoing gynecologic surgery is an ideal topic for a patient safety bundle. The primary purpose of this safety bundle is to provide recommendations that can be implemented into any surgical environment in an effort to reduce the incidence of surgical site infection. This bundle was developed by a multidisciplinary team convened by the Council on Patient Safety in Women's Health Care. The bundle is organized into four domains: Readiness, Recognition and Prevention, Response, and Reporting and Systems Learning. In addition to recommendations for practice, each of the domains stresses communication and teamwork between all members of the surgical team. Although the bundle components are designed to be adaptable to work in a variety of clinical settings, standardization within institutions is encouraged. Copyright © 2017 AWHONN, the Association of Women’s Health, Obstetric and Neonatal Nurses. Published by Elsevier Inc. All rights reserved.

  4. Big things come in bundled packages: implications of bundled payment systems in health care reimbursement reform.

    PubMed

    Delisle, Dennis R

    2013-01-01

    With passage of the Affordable Care Act, the ever-evolving landscape of health care braces for another shift in the reimbursement paradigm. As health care costs continue to rise, providers are pressed to deliver efficient, high-quality care at flat to minimally increasing rates. Inherent systemwide inefficiencies between payers and providers at various clinical settings pose a daunting task for enhancing collaboration and care coordination. A change from Medicare's fee-for-service reimbursement model to bundled payments offers one avenue for resolution. Pilots using such payment models have realized varying degrees of success, leading to the development and upcoming implementation of a bundled payment initiative led by the Center for Medicare and Medicaid Innovation. Delivery integration is critical to ensure high-quality care at affordable costs across the system. Providers and payers able to adapt to the newly proposed models of payment will benefit from achieving cost reductions and improved patient outcomes and realize a competitive advantage.

  5. Study on galloping behavior of iced eight bundle conductor transmission lines

    NASA Astrophysics Data System (ADS)

    Zhou, Linshu; Yan, Bo; Zhang, Liang; Zhou, Song

    2016-02-01

    Wind tunnel test was carried out to obtain the aerodynamic coefficients of an eight bundle conductor accreted with crescent-shaped ice. A user-defined cable element with torsional degree of freedom is developed in ABAQUS software to capture the torsional deformation of the iced conductors during galloping. By means of the user-defined cable element, different damping ratios in in-plane, out-of-plane and torsional directions of the conductors can be defined and the aerodynamic forces varying with their motion status can be exerted on the conductors conveniently when ABAQUS is used to simulate galloping of transmission lines. A wind tunnel test to model galloping of an iced eight bundle conductor segment was carried out, and the validity of the numerical simulation method is demonstrated by the agreement of the galloping orbit of the bundle conductor segment model recorded in the test and that by the numerical simulation. Furthermore, galloping behavior, including dynamic responses, galloping orbits, frequencies, vibration modes and amplitudes, of typical iced eight bundle conductor transmission lines in the cases of different span lengths, initial tensions in sub-conductors, wind velocities, angles of wind attack and damping ratios is studied, and the galloping behavior of the lines with internal resonance conditions is discussed. The obtained results may provide a fundamental tool for the development of anti-galloping techniques of eight bundle conductor transmission lines.

  6. Limiting (zero-load) speed of the rotary motor of Escherichia coli is independent of the number of torque-generating units

    PubMed Central

    Wang, Bin; Zhang, Rongjing; Yuan, Junhua

    2017-01-01

    Rotation of the bacterial flagellar motor is driven by multiple torque-generating units (stator elements). The torque-generating dynamics can be understood in terms of the “duty ratio” of the stator elements, that is, the fraction of time a stator element engages with the rotor during its mechanochemical cycle. The dependence of the limiting speed (zero-load speed) of the motor on the number of stator elements is the determining test of the duty ratio, which has been controversial experimentally and theoretically over the past decade. Here, we developed a method combining laser dark-field microscopy and optical trapping to resolve this controversy. We found that the zero-load speed is independent of the number of stator elements for the bacterial flagellar motor in Escherichia coli, demonstrating that these elements have a duty ratio close to 1. PMID:29109285

  7. Limiting (zero-load) speed of the rotary motor of Escherichia coli is independent of the number of torque-generating units.

    PubMed

    Wang, Bin; Zhang, Rongjing; Yuan, Junhua

    2017-11-21

    Rotation of the bacterial flagellar motor is driven by multiple torque-generating units (stator elements). The torque-generating dynamics can be understood in terms of the "duty ratio" of the stator elements, that is, the fraction of time a stator element engages with the rotor during its mechanochemical cycle. The dependence of the limiting speed (zero-load speed) of the motor on the number of stator elements is the determining test of the duty ratio, which has been controversial experimentally and theoretically over the past decade. Here, we developed a method combining laser dark-field microscopy and optical trapping to resolve this controversy. We found that the zero-load speed is independent of the number of stator elements for the bacterial flagellar motor in Escherichia coli , demonstrating that these elements have a duty ratio close to 1.

  8. Exploring Bundling Theory with Geometry

    ERIC Educational Resources Information Center

    Eckalbar, John C.

    2006-01-01

    The author shows how instructors might successfully introduce students in principles and intermediate microeconomic theory classes to the topic of bundling (i.e., the selling of two or more goods as a package, rather than separately). It is surprising how much students can learn using only the tools of high school geometry. To be specific, one can…

  9. Betti numbers of graded modules and cohomology of vector bundles

    NASA Astrophysics Data System (ADS)

    Eisenbud, David; Schreyer, Frank-Olaf

    2009-07-01

    In the remarkable paper Graded Betti numbers of Cohen-Macaulay modules and the multiplicity conjecture, Mats Boij and Jonas Soederberg conjectured that the Betti table of a Cohen-Macaulay module over a polynomial ring is a positive linear combination of Betti tables of modules with pure resolutions. We prove a strengthened form of their conjectures. Applications include a proof of the Multiplicity Conjecture of Huneke and Srinivasan and a proof of the convexity of a fan naturally associated to the Young lattice. With the same tools we show that the cohomology table of any vector bundle on projective space is a positive rational linear combination of the cohomology tables of what we call supernatural vector bundles. Using this result we give new bounds on the slope of a vector bundle in terms of its cohomology.

  10. Development and pilot testing of a patient-participatory pressure ulcer prevention care bundle.

    PubMed

    Gillespie, Brigid M; Chaboyer, Wendy; Sykes, Mark; O'Brien, Jennifer; Brandis, Susan

    2014-01-01

    This study developed and piloted a patient-centered pressure ulcer prevention care bundle for adult hospitalized patients to promote patient participation in prevention. The care bundle had 3 core messages: (1) keep moving, (2) care for your skin, and (3) ensure a good diet. A brief video, combined brochure/checklist, and poster were developed as training resources. Patient evaluation identified benefits of the care bundle; however, the combined checklist/brochure was rarely used.

  11. Broadcast Spawning Coral Mussismilia hispida Can Vertically Transfer its Associated Bacterial Core

    PubMed Central

    Leite, Deborah C. A.; Leão, Pedro; Garrido, Amana G.; Lins, Ulysses; Santos, Henrique F.; Pires, Débora O.; Castro, Clovis B.; van Elsas, Jan D.; Zilberberg, Carla; Rosado, Alexandre S.; Peixoto, Raquel S.

    2017-01-01

    The hologenome theory of evolution (HTE), which is under fierce debate, presupposes that parts of the microbiome are transmitted from one generation to the next [vertical transmission (VT)], which may also influence the evolution of the holobiont. Even though bacteria have previously been described in early life stages of corals, these early life stages (larvae) could have been inoculated in the water and not inside the parental colony (through gametes) carrying the parental microbiome. How Symbiodinium is transmitted to offspring is also not clear, as only one study has described this mechanism in spawners. All other studies refer to incubators. To explore the VT hypothesis and the key components being transferred, colonies of the broadcast spawner species Mussismilia hispida were kept in nurseries until spawning. Gamete bundles, larvae and adult corals were analyzed to identify their associated microbiota with respect to composition and location. Symbiodinium and bacteria were detected by sequencing in gametes and coral planula larvae. However, no cells were detected using microscopy at the gamete stage, which could be related to the absence of those cells inside the oocytes/dispersed in the mucus or to a low resolution of our approach. A preliminary survey of Symbiodinium diversity indicated that parental colonies harbored Symbiodinium clades B, C and G, whereas only clade B was found in oocytes and planula larvae [5 days after fertilization (a.f.)]. The core bacterial populations found in the bundles, planula larvae and parental colonies were identified as members of the genera Burkholderia, Pseudomonas, Acinetobacter, Ralstonia, Inquilinus and Bacillus, suggesting that these populations could be vertically transferred through the mucus. The collective data suggest that spawner corals, such as M. hispida, can transmit Symbiodinium cells and the bacterial core to their offspring by a coral gamete (and that this gamete, with its bacterial load, is released into

  12. Designing cooperatively folded abiotic uni- and multimolecular helix bundles

    NASA Astrophysics Data System (ADS)

    de, Soumen; Chi, Bo; Granier, Thierry; Qi, Ting; Maurizot, Victor; Huc, Ivan

    2018-01-01

    Abiotic foldamers, that is foldamers that have backbones chemically remote from peptidic and nucleotidic skeletons, may give access to shapes and functions different to those of peptides and nucleotides. However, design methodologies towards abiotic tertiary and quaternary structures are yet to be developed. Here we report rationally designed interactional patterns to guide the folding and assembly of abiotic helix bundles. Computational design facilitated the introduction of hydrogen-bonding functionalities at defined locations on the aromatic amide backbones that promote cooperative folding into helix-turn-helix motifs in organic solvents. The hydrogen-bond-directed aggregation of helices not linked by a turn unit produced several thermodynamically and kinetically stable homochiral dimeric and trimeric bundles with structures that are distinct from the designed helix-turn-helix. Relative helix orientation within the bundles may be changed from parallel to tilted on subtle solvent variations. Altogether, these results prefigure the richness and uniqueness of abiotic tertiary structure behaviour.

  13. Coulomb explosion: a novel approach to separate single-walled carbon nanotubes from their bundle.

    PubMed

    Liu, Guangtong; Zhao, Yuanchun; Zheng, Kaihong; Liu, Zheng; Ma, Wenjun; Ren, Yan; Xie, Sishen; Sun, Lianfeng

    2009-01-01

    A novel approach based on Coulomb explosion has been developed to separate single-walled carbon nanotubes (SWNTs) from their bundle. With this technique, we can readily separate a bundle of SWNTs into smaller bundles with uniform diameter as well as some individual SWNTs. The separated SWNTs have a typical length of several microns and form a nanotree at one end of the original bundle. More importantly, this separating procedure involves no surfactant and includes only one-step physical process. The separation method offers great conveniences for the subsequent individual SWNT or multiterminal SWNTs device fabrication and their physical properties studies.

  14. Optical design of endoscopic shape-tracker using quantum dots embedded in fiber bundles

    NASA Astrophysics Data System (ADS)

    Eisenstein, Jessica; Gavalis, Robb; Wong, Peter Y.; Cao, Caroline G. L.

    2009-08-01

    Colonoscopy is the current gold standard for colon cancer screening and diagnosis. However, the near-blind navigation process employed during colonoscopy results in endoscopist disorientation and scope looping, leading to missed detection of tumors, incorrect localization, and pain for the patient. A fiber optic bend sensor, which would fit into the working channel of a colonoscope, is developed to aid navigation through the colon during colonoscopy. The bend sensor is comprised of a bundle of seven fibers doped with quantum dots (QDs). Each fiber within the bundle contains a unique region made up of three zones with differently-colored QDs, spaced 120° apart circumferentially on the fiber. During bending at the QD region, light lost from the fiber's core is coupled into one of the QD zones, inducing fluorescence of the corresponding color whose intensity is proportional to the degree of bending. A complementary metal oxide semiconductor camera is used to obtain an image of the fluorescing end faces of the fiber bundle. The location of the fiber within the bundle, the color of fluorescence, and the fluorescence intensity are used to determine the bundle's bending location, direction, and degree of curvature, respectively. Preliminary results obtained using a single fiber with three QD zones and a seven-fiber bundle containing one active fiber with two QDs (180° apart) demonstrate the feasibility of the concept. Further developments on fiber orientation during bundling and the design of a graphical user interface to communicate bending information are also discussed.

  15. Functional Activity of Antibodies Directed towards Flagellin Proteins of Non-Typhoidal Salmonella

    PubMed Central

    Boyd, Mary A.; Wang, Jin Y.; Tulapurkar, Mohan E.; Pasetti, Marcela F.; Levine, Myron M.; Simon, Raphael

    2016-01-01

    Non-typhoidal Salmonella (NTS) serovars Typhimurium and Enteritidis are major causes of invasive bacterial infections in children under 5 years old in sub-Saharan Africa, with case fatality rates of ~20%. There are no licensed NTS vaccines for humans. Vaccines that induce antibodies against a Salmonella Typhi surface antigen, Vi polysaccharide, significantly protect humans against typhoid fever, establishing that immune responses to Salmonella surface antigens can be protective. Flagella proteins, abundant surface antigens in Salmonella serovars that cause human disease, are also powerful immunogens, but the functional capacity of elicited anti-flagellar antibodies and their role in facilitating bacterial clearance has been unclear. We examined the ability of anti-flagellar antibodies to mediate microbial killing by immune system components in-vitro and assessed their role in protecting mice against invasive Salmonella infection. Polyclonal (hyperimmune sera) and monoclonal antibodies raised against phase 1 flagellin proteins of S. Enteritidis and S. Typhimurium facilitated bacterial uptake and killing of the homologous serovar pathogen by phagocytes. Polyclonal anti-flagellar antibodies accompanied by complement also achieved direct bacterial killing. Serum bactericidal activity was restricted to Salmonella serovars expressing the same flagellin used as immunogen. Notably, individual anti-flagellin monoclonal antibodies with complement were not bactericidal, but this biological activity was restored when different monoclonal anti-flagellin antibodies were combined. Passive transfer immunization with a monoclonal IgG antibody specific for phase 1 flagellin from S. Typhimurium protected mice against lethal challenge with a representative African invasive S. Typhimurium strain. These findings have relevance for the use of flagellin proteins in NTS vaccines, and confirm the role of anti-flagellin antibodies as mediators of protective immunity. PMID:26998925

  16. Historical dynamics in ecosystem service bundles.

    PubMed

    Renard, Delphine; Rhemtulla, Jeanine M; Bennett, Elena M

    2015-10-27

    Managing multiple ecosystem services (ES), including addressing trade-offs between services and preventing ecological surprises, is among the most pressing areas for sustainability research. These challenges require ES research to go beyond the currently common approach of snapshot studies limited to one or two services at a single point in time. We used a spatiotemporal approach to examine changes in nine ES and their relationships from 1971 to 2006 across 131 municipalities in a mixed-use landscape in Quebec, Canada. We show how an approach that incorporates time and space can improve our understanding of ES dynamics. We found an increase in the provision of most services through time; however, provision of ES was not uniformly enhanced at all locations. Instead, each municipality specialized in providing a bundle (set of positively correlated ES) dominated by just a few services. The trajectory of bundle formation was related to changes in agricultural policy and global trends; local biophysical and socioeconomic characteristics explained the bundles' increasing spatial clustering. Relationships between services varied through time, with some provisioning and cultural services shifting from a trade-off or no relationship in 1971 to an apparent synergistic relationship by 2006. By implementing a spatiotemporal perspective on multiple services, we provide clear evidence of the dynamic nature of ES interactions and contribute to identifying processes and drivers behind these changing relationships. Our study raises questions about using snapshots of ES provision at a single point in time to build our understanding of ES relationships in complex and dynamic social-ecological systems.

  17. Two Types of Genetic Interaction Implicate the Whirligig Gene of Drosophila Melanogaster in Microtubule Organization in the Flagellar Axoneme

    PubMed Central

    Green, L. L.; Wolf, N.; McDonald, K. L.; Fuller, M. T.

    1990-01-01

    The mutant nc4 allele of whirligig (3-54.4) of Drosophila melanogaster fails to complement mutations in an α-tubulin locus, α1t, mutations in a β-tubulin locus, B2t, or a mutation in the haywire locus. However, wrl fails to map to any of the known α- or β-tubulin genes. The extragenic failure to complement could indicate that the wrl product participates in structural interactions with microtubule proteins. The whirligig locus appears to be haploinsufficient for male fertility. Both a deficiency of wrl and possible loss of function alleles obtained by reverting the failure to complement between wrl(nc4) and B2t(n) are dominant male sterile in a genetic background wild type for tubulin. The dominant male sterility of the revertant alleles is suppressed if the flies are also heterozygous for B2t(n), for a deficiency of α1t, or for the hay(nc2) allele. These results suggest that it is not the absolute level of wrl gene product but its level relative to tubulin or microtubule function that is important for normal spermatogenesis. The phenotype of homozygous wrl mutants suggests that the whirligig product plays a role in postmeiotic spermatid differentiation, possibly in organizing the microtubules of the sperm flagellar axoneme. Flies homozygous for either wrl(nc4) or revertant alleles are viable and female fertile but male sterile. Premeiotic and meiotic stages of spermatogenesis appear normal. However, in post-meiotic stages, flagellar axonemes show loss of the accessory microtubule on the B-subfiber of outer doublet microtubules, outer triplet instead of outer doublet microtubules, and missing central pair microtubules. PMID:2127579

  18. Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression.

    PubMed

    Chung, Jeong Min; Lee, Sangmin; Jung, Hyun Suk

    2017-05-01

    Bacterial expression is commonly used to produce recombinant and truncated mutant eukaryotic proteins. However, heterologous protein expression may render synthesized proteins insoluble. The conventional method used to express a poorly soluble protein, which involves denaturation and refolding, is time-consuming and inefficient. There are several non-denaturing approaches that can increase the solubility of recombinant proteins that include using different bacterial cell strains, altering the time of induction, lowering the incubation temperature, and employing different detergents for purification. In this study, we compared several non-denaturing protocols to express and purify two insoluble 34 kDa actin-bundling protein mutants. The solubility of the mutant proteins was not affected by any of the approaches except for treatment with the detergent sarkosyl. These results indicate that sarkosyl can effectively improve the solubility of insoluble proteins during bacterial expression. Copyright © 2016. Published by Elsevier Inc.

  19. Crystallographic and Molecular Dynamics Analysis of Loop Motions Unmasking the Peptidoglycan-Binding Site in Stator Protein MotB of Flagellar Motor

    PubMed Central

    Nahar, Musammat F.; Buckle, Ashley M.; Roujeinikova, Anna

    2011-01-01

    Background The C-terminal domain of MotB (MotB-C) shows high sequence similarity to outer membrane protein A and related peptidoglycan (PG)-binding proteins. It is believed to anchor the power-generating MotA/MotB stator unit of the bacterial flagellar motor to the peptidoglycan layer of the cell wall. We previously reported the first crystal structure of this domain and made a puzzling observation that all conserved residues that are thought to be essential for PG recognition are buried and inaccessible in the crystal structure. In this study, we tested a hypothesis that peptidoglycan binding is preceded by, or accompanied by, some structural reorganization that exposes the key conserved residues. Methodology/Principal Findings We determined the structure of a new crystalline form (Form B) of Helicobacter pylori MotB-C. Comparisons with the existing Form A revealed conformational variations in the petal-like loops around the carbohydrate binding site near one end of the β-sheet. These variations are thought to reflect natural flexibility at this site required for insertion into the peptidoglycan mesh. In order to understand the nature of this flexibility we have performed molecular dynamics simulations of the MotB-C dimer. The results are consistent with the crystallographic data and provide evidence that the three loops move in a concerted fashion, exposing conserved MotB residues that have previously been implicated in binding of the peptide moiety of peptidoglycan. Conclusion/Significance Our structural analysis provides a new insight into the mechanism by which MotB inserts into the peptidoglycan mesh, thus anchoring the power-generating complex to the cell wall. PMID:21533052

  20. Lexical Bundles in the Academic Writing of Advanced Chinese EFL Learners

    ERIC Educational Resources Information Center

    Wei, Yaoyu; Lei, Lei

    2011-01-01

    The present study investigated the use of lexical bundles in the academic writing of advanced Chinese EFL learners. A corpus of doctoral dissertations by the learners and a corpus of published journal articles by professional writers were collected for the study. Four-word lexical bundles in the two corpora were identified and analysed. Results…

  1. Lexical Bundle Analysis in Mathematics Classroom Discourse: The Significance of Stance

    ERIC Educational Resources Information Center

    Herbel-Eisenmann, Beth; Wagner, David; Cortes, Viviana

    2010-01-01

    In this article, we introduce the lexical bundle, defined by corpus linguists as a group of three or more words that frequently recur together, in a single group, in a particular register (Biber, Johansson, Leech, Conrad, & Finegan, 2006; Cortes, "English for Specific Purposes" 23:397-423, 2004). Attention to lexical bundles helps to explore…

  2. The effects of fee bundling on dental utilization.

    PubMed Central

    Porter, J; Coyte, P C; Barnsley, J; Croxford, R

    1999-01-01

    OBJECTIVE: To examine dental utilization following an adjustment to the provincial fee schedule in which preventive maintenance (recall) services were bundled at lower fees. DATA SOURCES/STUDY SETTING: Blue Cross dental insurance claims for claimants associated with four major Ontario employers using a common insurance plan over the period 1987-1990. STUDY DESIGN: This before-and-after design analyzes the dental claims experience over a four-year period for 4,455 individuals 18 years of age and older one year prior to the bundling of services, one year concurrent with the change, and two years after the introduction of bundling. The dependent variable is the annual adjusted payment per user. DATA COLLECTION/EXTRACTION METHODS: The analysis was based on all claims submitted by adult users for services received at recall visits and who reported at least one visit of this type between 1987 and 1990. In these data, 26,177 services were provided by 1,214 dentists and represent 41 percent of all adult service claims submitted over the four years of observation. PRINCIPAL FINDINGS: Real per capita payment for adult recall services decreased by 0.3 percent in the year bundling was implemented (1988), but by the end of the study period such payments had increased 4.8 percent relative to pre-bundling levels. Multiple regression analysis assessed the role of patient and provider variables in the upward trend of per capita payments. The following variables were significant in explaining 37 percent of the variation in utilization over the period of observation: subscriber employment location; ever having received periodontal scaling or ever having received restorative services; regular user; dentist's school of graduation; and interactions involving year, service type, and regular user status. CONCLUSIONS: The volume and intensity of services received by adult patients increased when fee constraints were imposed on dentists. Future efforts to contain dental expenditures through

  3. Managing bundled payments.

    PubMed

    Draper, Andrew

    2011-04-01

    Results of Medicare's ACE demonstration project and Geisinger Health System's ProvenCare initiative provide insight into the challenges hospitals will face as bundled payment proliferates. An early analysis of these results suggests that hospitals would benefit from bringing full automation using clinical IT tools to bear in their efforts to meet these challenges. Other important factors contributing to success include board and physician leadership, organizational structure, pricing methodology for bidding, evidence-based medical practice guidelines, supply cost management, process efficiency management, proactive and aggressive case management, business development and marketing strategy, and the financial management system.

  4. Distributed temperature sensing inside a 19-rod bundle

    DOE PAGES

    Lomperski, S.; Bremer, N.; Gerardi, C.

    2017-05-23

    The temperature field within a model of a sodium-cooled fast reactor fuel rod bundle was measured using Ø155 μm fiber optic distributed temperature sensors (DTS). The bundle consists of 19 electrically-heated rods Ø6.3 mm and 865 mm long. Working fluids were argon and air at atmospheric pressure and Reynolds numbers up to 300. A 20 m-long DTS was threaded through Ø1 mm capillaries wound around rods as wire-wraps. The sensor generated 173 measurements along each rod at 5 mm resolution for a total of 3300 data locations. A second DTS, 58 m long, was suspended between rods to provide 9300more » fluid temperature measurements at 20 mm resolution. Such data density makes it possible to construct 3D maps of the temperature field that are beyond the reach of traditional sensors such as thermocouples. This is illustrated through a series of steady-state and transient tests. As a result, the work demonstrates the feasibility of mapping temperature within the close confines of a rod bundle at resolutions suitable for validation of computational fluid dynamics codes.« less

  5. Coupling and Elastic Loading Affect the Active Response by the Inner Ear Hair Cell Bundles

    PubMed Central

    Strimbu, Clark Elliott; Fredrickson-Hemsing, Lea; Bozovic, Dolores

    2012-01-01

    Active hair bundle motility has been proposed to underlie the amplification mechanism in the auditory endorgans of non-mammals and in the vestibular systems of all vertebrates, and to constitute a crucial component of cochlear amplification in mammals. We used semi-intact in vitro preparations of the bullfrog sacculus to study the effects of elastic mechanical loading on both natively coupled and freely oscillating hair bundles. For the latter, we attached glass fibers of different stiffness to the stereocilia and observed the induced changes in the spontaneous bundle movement. When driven with sinusoidal deflections, hair bundles displayed phase-locked response indicative of an Arnold Tongue, with the frequency selectivity highest at low amplitudes and decreasing under stronger stimulation. A striking broadening of the mode-locked response was seen with increasing stiffness of the load, until approximate impedance matching, where the phase-locked response remained flat over the physiological range of frequencies. When the otolithic membrane was left intact atop the preparation, the natural loading of the bundles likewise decreased their frequency selectivity with respect to that observed in freely oscillating bundles. To probe for signatures of the active process under natural loading and coupling conditions, we applied transient mechanical stimuli to the otolithic membrane. Following the pulses, the underlying bundles displayed active movement in the opposite direction, analogous to the twitches observed in individual cells. Tracking features in the otolithic membrane indicated that it moved in phase with the bundles. Hence, synchronous active motility evoked in the system of coupled hair bundles by external input is sufficient to displace large overlying structures. PMID:22479461

  6. Class III myosins shape the auditory hair bundles by limiting microvilli and stereocilia growth

    PubMed Central

    Lelli, Andrea; Michel, Vincent; Boutet de Monvel, Jacques; Cortese, Matteo; Bosch-Grau, Montserrat; Aghaie, Asadollah; Perfettini, Isabelle; Dupont, Typhaine; Avan, Paul

    2016-01-01

    The precise architecture of hair bundles, the arrays of mechanosensitive microvilli-like stereocilia crowning the auditory hair cells, is essential to hearing. Myosin IIIa, defective in the late-onset deafness form DFNB30, has been proposed to transport espin-1 to the tips of stereocilia, thereby promoting their elongation. We show that Myo3a−/−Myo3b−/− mice lacking myosin IIIa and myosin IIIb are profoundly deaf, whereas Myo3a-cKO Myo3b−/− mice lacking myosin IIIb and losing myosin IIIa postnatally have normal hearing. Myo3a−/−Myo3b−/− cochlear hair bundles display robust mechanoelectrical transduction currents with normal kinetics but show severe embryonic abnormalities whose features rapidly change. These include abnormally tall and numerous microvilli or stereocilia, ungraded stereocilia bundles, and bundle rounding and closure. Surprisingly, espin-1 is properly targeted to Myo3a−/−Myo3b−/− stereocilia tips. Our results uncover the critical role that class III myosins play redundantly in hair-bundle morphogenesis; they unexpectedly limit the elongation of stereocilia and of subsequently regressing microvilli, thus contributing to the early hair bundle shaping. PMID:26754646

  7. Effect of dielectrophoretic force on swimming bacteria.

    PubMed

    Tran, Ngoc Phu; Marcos

    2015-07-01

    Dielectrophoresis (DEP) has been applied widely in bacterial manipulation such as separating, concentrating, and focusing. Previous studies primarily focused on the collective effects of DEP force on the bacterial population. However, the influence of DEP force on the swimming of a single bacterium had not been investigated. In this study, we present a model to analyze the effect of DEP force on a swimming helically flagellated bacterium, particularly on its swimming direction and velocity. We consider a simple DEP force that acts along the X-direction, and its strength as well as direction varies with the X- and Y-positions. Resistive force theory is employed to compute the hydrodynamic force on the bacterium's flagellar bundle, and the effects of both DEP force and rotational diffusion on the swimming of the bacterium are simultaneously taken into consideration using the Fokker-Planck equation. We show the mechanism of how DEP force alters the orientation and velocity of the bacterium. In most cases, the DEP force dominantly influences the orientation of the swimming bacterium; however, when the DEP force strongly varies along the Y-direction, the rotational diffusion is also responsible for determining the bacterium's reorientation. More interestingly, the variance of DEP force along the Y-direction causes the bacterium to experience a translational velocity perpendicular to its primary axis, and this phenomenon could be utilized to focus the bacteria. Finally, we show the feasibility of applying our findings to achieve bacterial focusing. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Antiprotozoal glutathione derivatives with flagellar membrane binding activity against T. brucei rhodesiense.

    PubMed

    Daunes, Sylvie; Yardley, Vanessa; Croft, Simon L; D'Silva, Claudius

    2017-02-15

    A new series of N-substituted S-(2,4-dinitrophenyl)glutathione dibutyl diesters were synthesized to improve in vitro anti-protozoal activity against the pathogenic parasites Trypanosoma brucei rhodesiense, Trypanosoma cruzi and Leishmania donovani. The results obtained indicate that N-substituents enhance the inhibitory properties of glutathione diesters whilst showing reduced toxicity against KB cells as in the cases of compounds 5, 9, 10, 16, 18 and 19. We suggest that the interaction of N-substituted S-(2,4-dinitrophenyl) glutathione dibutyl diesters with T. b. brucei occurs mainly by weak hydrophobic interactions such as London and van der Waals forces. A QSAR study indicated that the inhibitory activity of the peptide is associated negatively with the average number of C atoms, N C and positively to S ZX, the ZX shadow a geometric descriptor related to molecular size and orientation of the compound. HPLC-UV studies in conjunction with optical microscopy indicate that the observed selectivity of inhibition of these compounds against bloodstream form T. b. brucei parasites in comparison to L. donovani under the same conditions is due to intracellular uptake via endocytosis in the flagellar pocket. Copyright © 2016. Published by Elsevier Ltd.

  9. Project IMPACT Pilot Report: Feasibility of Implementing a Hospital-to-Home Transition Bundle.

    PubMed

    Mallory, Leah A; Osorio, Snezana Nena; Prato, B Stephen; DiPace, Jennifer; Schmutter, Lisa; Soung, Paula; Rogers, Amanda; Woodall, William J; Burley, Kayla; Gage, Sandra; Cooperberg, David

    2017-03-01

    To improve hospital to home transitions, a 4-element pediatric patient-centered transition bundle was developed, including: a transition readiness checklist; predischarge teach-back education; timely and complete written handoff to the primary care provider; and a postdischarge phone call. The objective of this study was to demonstrate the feasibility of bundle implementation and report initial outcomes at 4 pilot sites. Outcome measures included postdischarge caregiver ability to teach-back key home management information and 30-day reuse rates. A multisite, observational time series using multiple planned sequential interventions to implement bundle components with non-technology-supported and technology-supported patients. Data were collected via electronic health record reviews and during postdischarge phone calls. Statistical process control charts were used to assess outcomes. Four pilot sites implemented the bundle between January 2014 and May 2015 for 2601 patients, of whom 1394 had postdischarge telephone encounters. Improvement was noted in the implementation of all bundle elements with the transitions readiness checklist posing the greatest feasibility challenge. Phone contact connection rates were 69%. Caregiver ability to teach-back essential home management information postdischarge improved from 18% to 82%. No improvement was noted in reuse rates, which differed dramatically between technology-supported and non-technology-supported patients. A pediatric care transition bundle was successfully tested and implemented, as demonstrated by improvement in all process measures, as well as caregiver home management skills. Important considerations for successful implementation and evaluation of the discharge bundle include the role of local context, electronic health record integration, and subgroup analysis for technology-supported patients. Copyright © 2017 by the American Academy of Pediatrics.

  10. Decreased Surgical Site Infection Rate in Hysterectomy: Effect of a Gynecology-Specific Bundle.

    PubMed

    Andiman, Sarah E; Xu, Xiao; Boyce, John M; Ludwig, Elizabeth M; Rillstone, Heidi R W; Desai, Vrunda B; Fan, Linda L

    2018-06-01

    We implemented a hysterectomy-specific surgical site infection prevention bundle after a higher-than-expected surgical site infection rate was identified at our institution. We evaluate how this bundle affected the surgical site infection rate, length of hospital stay, and 30-day postoperative readmission rate. This is a quality improvement study featuring retrospective analysis of a prospectively implemented, multidisciplinary team-designed surgical site infection prevention bundle that consisted of chlorhexidine-impregnated preoperative wipes, standardized aseptic surgical preparation, standardized antibiotic dosing, perioperative normothermia, surgical dressing maintenance, and direct feedback to clinicians when the protocol was breached. There were 2,099 hysterectomies completed during the 33-month study period. There were 61 surgical site infections (4.51%) in the pre-full bundle implementation period and 14 (1.87%) in the post-full bundle implementation period; we found a sustained reduction in the proportion of patients experiencing surgical site infection during the last 8 months of the study period. After adjusting for clinical characteristics, patients who underwent surgery after full implementation were less likely to develop a surgical site infection (adjusted odds ratio [OR] 0.46, P=.01) than those undergoing surgery before full implementation. Multivariable regression analysis showed no statistically significant difference in postoperative days of hospital stay (adjusted mean ratio 0.95, P=.09) or rate of readmission for surgical site infection-specific indication (adjusted OR 2.65, P=.08) between the before and after full-bundle implementation periods. The multidisciplinary implementation of a gynecologic perioperative surgical site infection prevention bundle was associated with a significant reduction in surgical site infection rate in patients undergoing hysterectomy.

  11. High-resolution imaging of retinal nerve fiber bundles in glaucoma using adaptive optics scanning laser ophthalmoscopy.

    PubMed

    Takayama, Kohei; Ooto, Sotaro; Hangai, Masanori; Ueda-Arakawa, Naoko; Yoshida, Sachiko; Akagi, Tadamichi; Ikeda, Hanako Ohashi; Nonaka, Atsushi; Hanebuchi, Masaaki; Inoue, Takashi; Yoshimura, Nagahisa

    2013-05-01

    To detect pathologic changes in retinal nerve fiber bundles in glaucomatous eyes seen on images obtained by adaptive optics (AO) scanning laser ophthalmoscopy (AO SLO). Prospective cross-sectional study. Twenty-eight eyes of 28 patients with open-angle glaucoma and 21 normal eyes of 21 volunteer subjects underwent a full ophthalmologic examination, visual field testing using a Humphrey Field Analyzer, fundus photography, red-free SLO imaging, spectral-domain optical coherence tomography, and imaging with an original prototype AO SLO system. The AO SLO images showed many hyperreflective bundles suggesting nerve fiber bundles. In glaucomatous eyes, the nerve fiber bundles were narrower than in normal eyes, and the nerve fiber layer thickness was correlated with the nerve fiber bundle widths on AO SLO (P < .001). In the nerve fiber layer defect area on fundus photography, the nerve fiber bundles on AO SLO were narrower compared with those in normal eyes (P < .001). At 60 degrees on the inferior temporal side of the optic disc, the nerve fiber bundle width was significantly lower, even in areas without nerve fiber layer defect, in eyes with glaucomatous eyes compared with normal eyes (P = .026). The mean deviations of each cluster in visual field testing were correlated with the corresponding nerve fiber bundle widths (P = .017). AO SLO images showed reduced nerve fiber bundle widths both in clinically normal and abnormal areas of glaucomatous eyes, and these abnormalities were associated with visual field defects, suggesting that AO SLO may be useful for detecting early nerve fiber bundle abnormalities associated with loss of visual function. Copyright © 2013 Elsevier Inc. All rights reserved.

  12. Fiber Bundle Model Under Heterogeneous Loading

    NASA Astrophysics Data System (ADS)

    Roy, Subhadeep; Goswami, Sanchari

    2018-03-01

    The present work deals with the behavior of fiber bundle model under heterogeneous loading condition. The model is explored both in the mean-field limit as well as with local stress concentration. In the mean field limit, the failure abruptness decreases with increasing order k of heterogeneous loading. In this limit, a brittle to quasi-brittle transition is observed at a particular strength of disorder which changes with k. On the other hand, the model is hardly affected by such heterogeneity in the limit where local stress concentration plays a crucial role. The continuous limit of the heterogeneous loading is also studied and discussed in this paper. Some of the important results related to fiber bundle model are reviewed and their responses to our new scheme of heterogeneous loading are studied in details. Our findings are universal with respect to the nature of the threshold distribution adopted to assign strength to an individual fiber.

  13. National Partnership for Maternal Safety: Consensus Bundle on Severe Hypertension During Pregnancy and the Postpartum Period.

    PubMed

    Bernstein, Peter S; Martin, James N; Barton, John R; Shields, Laurence E; Druzin, Maurice L; Scavone, Barbara M; Frost, Jennifer; Morton, Christine H; Ruhl, Catherine; Slager, Joan; Tsigas, Eleni Z; Jaffer, Sara; Menard, M Kathryn

    2017-08-01

    Complications arising from hypertensive disorders of pregnancy are among the leading causes of preventable severe maternal morbidity and mortality. Timely and appropriate treatment has the potential to significantly reduce hypertension-related complications. To assist health care providers in achieving this goal, this patient safety bundle provides guidance to coordinate and standardize the care provided to women with severe hypertension during pregnancy and the postpartum period. This is one of several patient safety bundles developed by multidisciplinary work groups of the National Partnership for Maternal Safety under the guidance of the Council on Patient Safety in Women's Health Care. These safety bundles outline critical clinical practices that should be implemented in every maternity care setting. Similar to other bundles that have been developed and promoted by the Partnership, the hypertension safety bundle is organized into four domains: Readiness, Recognition and Prevention, Response, and Reporting and Systems Learning. Although the bundle components may be adapted to meet the resources available in individual facilities, standardization within an institution is strongly encouraged. This commentary provides information to assist with bundle implementation.

  14. Transduction channels' gating can control friction on vibrating hair-cell bundles in the ear.

    PubMed

    Bormuth, Volker; Barral, Jérémie; Joanny, Jean-François; Jülicher, Frank; Martin, Pascal

    2014-05-20

    Hearing starts when sound-evoked mechanical vibrations of the hair-cell bundle activate mechanosensitive ion channels, giving birth to an electrical signal. As for any mechanical system, friction impedes movements of the hair bundle and thus constrains the sensitivity and frequency selectivity of auditory transduction. Friction is generally thought to result mainly from viscous drag by the surrounding fluid. We demonstrate here that the opening and closing of the transduction channels produce internal frictional forces that can dominate viscous drag on the micrometer-sized hair bundle. We characterized friction by analyzing hysteresis in the force-displacement relation of single hair-cell bundles in response to periodic triangular stimuli. For bundle velocities high enough to outrun adaptation, we found that frictional forces were maximal within the narrow region of deflections that elicited significant channel gating, plummeted upon application of a channel blocker, and displayed a sublinear growth for increasing bundle velocity. At low velocity, the slope of the relation between the frictional force and velocity was nearly fivefold larger than the hydrodynamic friction coefficient that was measured when the transduction machinery was decoupled from bundle motion by severing tip links. A theoretical analysis reveals that channel friction arises from coupling the dynamics of the conformational change associated with channel gating to tip-link tension. Varying channel properties affects friction, with faster channels producing smaller friction. We propose that this intrinsic source of friction may contribute to the process that sets the hair cell's characteristic frequency of responsiveness.

  15. Soluble components of the flagellar export apparatus, FliI, FliJ, and FliH, do not deliver flagellin, the major filament protein, from the cytosol to the export gate.

    PubMed

    Sajó, Ráchel; Liliom, Károly; Muskotál, Adél; Klein, Agnes; Závodszky, Péter; Vonderviszt, Ferenc; Dobó, József

    2014-11-01

    Flagella, the locomotion organelles of bacteria, extend from the cytoplasm to the cell exterior. External flagellar proteins are synthesized in the cytoplasm and exported by the flagellar type III secretion system. Soluble components of the flagellar export apparatus, FliI, FliH, and FliJ, have been implicated to carry late export substrates in complex with their cognate chaperones from the cytoplasm to the export gate. The importance of the soluble components in the delivery of the three minor late substrates FlgK, FlgL (hook-filament junction) and FliD (filament-cap) has been convincingly demonstrated, but their role in the transport of the major filament component flagellin (FliC) is still unclear. We have used continuous ATPase activity measurements and quartz crystal microbalance (QCM) studies to characterize interactions between the soluble export components and flagellin or the FliC:FliS substrate-chaperone complex. As controls, interactions between soluble export component pairs were characterized providing Kd values. FliC or FliC:FliS did not influence the ATPase activity of FliI alone or in complex with FliH and/or FliJ suggesting lack of interaction in solution. Immobilized FliI, FliH, or FliJ did not interact with FliC or FliC:FliS detected by QCM. The lack of interaction in the fluid phase between FliC or FliC:FliS and the soluble export components, in particular with the ATPase FliI, suggests that cells use different mechanisms for the export of late minor substrates, and the major substrate, FliC. It seems that the abundantly produced flagellin does not require the assistance of the soluble export components to efficiently reach the export gate. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Cell shape can mediate the spatial organization of the bacterial cytoskeleton

    NASA Astrophysics Data System (ADS)

    Wang, Siyuan; Wingreen, Ned

    2013-03-01

    The bacterial cytoskeleton guides the synthesis of cell wall and thus regulates cell shape. Since spatial patterning of the bacterial cytoskeleton is critical to the proper control of cell shape, it is important to ask how the cytoskeleton spatially self-organizes in the first place. In this work, we develop a quantitative model to account for the various spatial patterns adopted by bacterial cytoskeletal proteins, especially the orientation and length of cytoskeletal filaments such as FtsZ and MreB in rod-shaped cells. We show that the combined mechanical energy of membrane bending, membrane pinning, and filament bending of a membrane-attached cytoskeletal filament can be sufficient to prescribe orientation, e.g. circumferential for FtsZ or helical for MreB, with the accuracy of orientation increasing with the length of the cytoskeletal filament. Moreover, the mechanical energy can compete with the chemical energy of cytoskeletal polymerization to regulate filament length. Notably, we predict a conformational transition with increasing polymer length from smoothly curved to end-bent polymers. Finally, the mechanical energy also results in a mutual attraction among polymers on the same membrane, which could facilitate tight polymer spacing or bundling. The predictions of the model can be verified through genetic, microscopic, and microfluidic approaches.

  17. An Exploration of Human Well-Being Bundles as Identifiers of Ecosystem Service Use Patterns

    PubMed Central

    Biggs, Reinette; Reyers, Belinda

    2016-01-01

    We take a social-ecological systems perspective to investigate the linkages between ecosystem services and human well-being in South Africa. A recent paper identified different types of social-ecological systems in the country, based on distinct bundles of ecosystem service use. These system types were found to represent increasingly weak direct feedbacks between nature and people, from rural “green-loop” communities to urban “red-loop” societies. Here we construct human well-being bundles and explore whether the well-being bundles can be used to identify the same social-ecological system types that were identified using bundles of ecosystem service use. Based on national census data, we found three distinct well-being bundle types that are mainly characterized by differences in income, unemployment and property ownership. The distribution of these well-being bundles approximates the distribution of ecosystem service use bundles to a substantial degree: High levels of income and education generally coincided with areas characterised by low levels of direct ecosystem service use (or red-loop systems), while the majority of low well-being areas coincided with medium and high levels of direct ecosystem service use (or transition and green-loop systems). However, our results indicate that transformations from green-loop to red-loop systems do not always entail an immediate improvement in well-being, which we suggest may be due to a time lag between changes in the different system components. Using human well-being bundles as an indicator of social-ecological dynamics may be useful in other contexts since it is based on socio-economic data commonly collected by governments, and provides important insights into the connections between ecosystem services and human well-being at policy-relevant sub-national scales. PMID:27695120

  18. An Exploration of Human Well-Being Bundles as Identifiers of Ecosystem Service Use Patterns.

    PubMed

    Hamann, Maike; Biggs, Reinette; Reyers, Belinda

    2016-01-01

    We take a social-ecological systems perspective to investigate the linkages between ecosystem services and human well-being in South Africa. A recent paper identified different types of social-ecological systems in the country, based on distinct bundles of ecosystem service use. These system types were found to represent increasingly weak direct feedbacks between nature and people, from rural "green-loop" communities to urban "red-loop" societies. Here we construct human well-being bundles and explore whether the well-being bundles can be used to identify the same social-ecological system types that were identified using bundles of ecosystem service use. Based on national census data, we found three distinct well-being bundle types that are mainly characterized by differences in income, unemployment and property ownership. The distribution of these well-being bundles approximates the distribution of ecosystem service use bundles to a substantial degree: High levels of income and education generally coincided with areas characterised by low levels of direct ecosystem service use (or red-loop systems), while the majority of low well-being areas coincided with medium and high levels of direct ecosystem service use (or transition and green-loop systems). However, our results indicate that transformations from green-loop to red-loop systems do not always entail an immediate improvement in well-being, which we suggest may be due to a time lag between changes in the different system components. Using human well-being bundles as an indicator of social-ecological dynamics may be useful in other contexts since it is based on socio-economic data commonly collected by governments, and provides important insights into the connections between ecosystem services and human well-being at policy-relevant sub-national scales.

  19. Computer-Aided Resolution of an Experimental Paradox in Bacterial Chemotaxis

    PubMed Central

    Abouhamad, Walid N.; Bray, Dennis; Schuster, Martin; Boesch, Kristin C.; Silversmith, Ruth E.; Bourret, Robert B.

    1998-01-01

    Escherichia coli responds to its environment by means of a network of intracellular reactions which process signals from membrane-bound receptors and relay them to the flagellar motors. Although characterization of the reactions in the chemotaxis signaling pathway is sufficiently complete to construct computer simulations that predict the phenotypes of mutant strains with a high degree of accuracy, two previous experimental investigations of the activity remaining upon genetic deletion of multiple signaling components yielded several contradictory results (M. P. Conley, A. J. Wolfe, D. F. Blair, and H. C. Berg, J. Bacteriol. 171:5190–5193, 1989; J. D. Liu and J. S. Parkinson, Proc. Natl. Acad. Sci. USA 86:8703–8707, 1989). For example, “building up” the pathway by adding back CheA and CheY to a gutted strain lacking chemotaxis genes resulted in counterclockwise flagellar rotation whereas “breaking down” the pathway by deleting chemotaxis genes except cheA and cheY resulted in alternating episodes of clockwise and counterclockwise flagellar rotation. Our computer simulation predicts that trace amounts of CheZ expressed in the gutted strain could account for this difference. We tested this explanation experimentally by constructing a mutant containing a new deletion of the che genes that cannot express CheZ and verified that the behavior of strains built up from the new deletion does in fact conform to both the phenotypes observed for breakdown strains and computer-generated predictions. Our findings consolidate the present view of the chemotaxis signaling pathway and highlight the utility of molecularly based computer models in the analysis of complex biochemical networks. PMID:9683468

  20. Experimental observation of spatially resolved photo-luminescence intensity distribution in dual mode upconverting nanorod bundles

    PubMed Central

    Kumar, Pawan; Singh, Satbir; Singh, V. N.; Singh, Nidhi; Gupta, R. K.; Gupta, Bipin Kumar

    2017-01-01

    A novel method for demonstration of photoluminescence intensity distribution in upconverting nanorod bundles using confocal microscopy is reported. Herein, a strategy for the synthesis of highly luminescent dual mode upconverting/downshift Y1.94O3:Ho3+0.02/Yb3+0.04 nanorod bundles by a facile hydrothermal route has been introduced. These luminescent nanorod bundles exhibit strong green emission at 549 nm upon excitations at 449 nm and 980 nm with quantum efficiencies of ~6.3% and ~1.1%, respectively. The TEM/HRTEM results confirm that these bundles are composed of several individual nanorods with diameter of ~100 nm and length in the range of 1–3 μm. Furthermore, two dimensional spatially resolved photoluminescence intensity distribution study has been carried out using confocal photoluminescence microscope throughout the nanorod bundles. This study provides a new direction for the potential use of such emerging dual mode nanorod bundles as photon sources for next generation flat panel optical display devices, bio-medical applications, luminescent security ink and enhanced energy harvesting in photovoltaic applications. PMID:28211891

  1. Exploring the membrane fusion mechanism through force-induced disassembly of HIV-1 six-helix bundle

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

    Gao, Kai; Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101; University of Chinese Academy of Sciences, Beijing 100049

    Enveloped virus, such as HIV-1, employs membrane fusion mechanism to invade into host cell. HIV-1 gp41 ectodomain uses six-helix bundle configuration to accomplish this process. Using molecular dynamic simulations, we confirmed the stability of this six-helix bundle by showing high occupancy of hydrogen bonds and hydrophobic interactions. Key residues and interactions important for the bundle integration were characterized by force-induced unfolding simulations of six-helix bundle, exhibiting the collapse order of these groups of interactions. Moreover, our results in some way concerted with a previous theory that the formation of coiled-coil choose a route which involved cooperative interactions between the N-terminalmore » and C-terminal helix. -- Highlights: •Unfolding of HIV-1 gp41 six-helix bundle is studied by molecular dynamics simulations. •Specific interactions responsible for the stability of HIV-1 envelope post-fusion conformation were identified. •The gp41 six-helix bundle transition inducing membrane fusion might be a cooperative process of the three subunits.« less

  2. Reinforcement of single-walled carbon nanotube bundles by intertube bridging

    NASA Astrophysics Data System (ADS)

    Kis, A.; Csányi, G.; Salvetat, J.-P.; Lee, Thien-Nga; Couteau, E.; Kulik, A. J.; Benoit, W.; Brugger, J.; Forró, L.

    2004-03-01

    During their production, single-walled carbon nanotubes form bundles. Owing to the weak van der Waals interaction that holds them together in the bundle, the tubes can easily slide on each other, resulting in a shear modulus comparable to that of graphite. This low shear modulus is also a major obstacle in the fabrication of macroscopic fibres composed of carbon nanotubes. Here, we have introduced stable links between neighbouring carbon nanotubes within bundles, using moderate electron-beam irradiation inside a transmission electron microscope. Concurrent measurements of the mechanical properties using an atomic force microscope show a 30-fold increase of the bending modulus, due to the formation of stable crosslinks that effectively eliminate sliding between the nanotubes. Crosslinks were modelled using first-principles calculations, showing that interstitial carbon atoms formed during irradiation in addition to carboxyl groups, can independently lead to bridge formation between neighbouring nanotubes.

  3. Muon and Gamma Bundles tracing Up-going Tau Neutrino Astronomy

    NASA Astrophysics Data System (ADS)

    Fargion, D.; de Santis, M.; de Sanctis Lucentini, P. G.; Grossi, M.

    2004-11-01

    Up-going and Horizontal Tau Air-Showers, UpTaus and HorTaus, may trace Ultra High Energy Neutrino Tau Earth Skimming at the edge of the horizons. Their secondaries (μ± and γ bundles with e± pair flashes) might trace their nature over UHECR secondaries in horizontal showers. Indeed the atmosphere act as a perfect amplifier as well as a filter for showers: down-ward and horizontal μ bundles may still be originated by far Ultra High Energy Cosmic Rays skimming the terrestrial atmosphere but their rich gamma component will be exponentially suppressed. At large zenith angles after crossing a large slant depth (Xmax > 3 × 103 g cm-2) the number of μ± and secondary γ's (produced by the e± pair from μ decay in flight) is comparable. On the other hand, up-ward muon bundles from UpTaus and HorTaus may arise within a young shower with a larger gamma-muon ratio (˜ 102), leaving its characteristic imprint. We estimate the UpTaus and HorTaus rate from the Earth and we evaluate the consequent event rate of μ±, e± and γ bundles. We show that such events even for minimal GZK neutrino fluxes could be detected by scintillator arrays placed on mountains at 1 -5 km and pointing to the horizon. The required array areas are within tens-hundreds of square meters. An optimal structure is an array of crown-like twin detectors facing the horizons. We argue that such detectors will be able to detect both muonic bundles at a minimal average flux of 10-11 cm-2 s-1 sr-1 and electromagnetic particles (γ, e±) at 3 × 10-9 cm-2 s-1 sr-1, a few times each year, even for the minimal GZK ν flux.

  4. Risk of Revision Was Not Reduced by a Double-bundle ACL Reconstruction Technique: Results From the Scandinavian Registers.

    PubMed

    Aga, Cathrine; Kartus, Jüri-Tomas; Lind, Martin; Lygre, Stein Håkon Låstad; Granan, Lars-Petter; Engebretsen, Lars

    2017-10-01

    Double-bundle anterior cruciate ligament (ACL) reconstruction has demonstrated improved biomechanical properties and moderately better objective outcomes compared with single-bundle reconstructions. This could make an impact on the rerupture rate and reduce the risk of revisions in patients undergoing double-bundle ACL reconstruction compared with patients reconstructed with a traditional single-bundle technique. The National Knee Ligament Registers in Scandinavia provide information that can be used to evaluate the revision outcome after ACL reconstructions. The purposes of the study were (1) to compare the risk of revision between double-bundle and single-bundle reconstructions, reconstructed with autologous hamstring tendon grafts; (2) to compare the risk of revision between double-bundle hamstring tendon and single-bundle bone-patellar tendon-bone autografts; and (3) to compare the hazard ratios for the same two research questions after Cox regression analysis was performed. Data collection of primary ACL reconstructions from the National Knee Ligament Registers in Denmark, Norway, and Sweden from July 1, 2005, to December 31, 2014, was retrospectively analyzed. A total of 60,775 patients were included in the study; 994 patients were reconstructed with double-bundle hamstring tendon grafts, 51,991 with single-bundle hamstring tendon grafts, and 7790 with single-bundle bone-patellar tendon-bone grafts. The double-bundle ACL-reconstructed patients were compared with the two other groups. The risk of revision for each research question was detected by the risk ratio, hazard ratio, and the corresponding 95% confidence intervals. Kaplan-Meier analysis was used to estimate survival at 1, 2, and 5 years for the three different groups. Furthermore, a Cox proportional hazard regression model was applied and the hazard ratios were adjusted for country, age, sex, meniscal or chondral injury, and utilized fixation devices on the femoral and tibial sides. There were no

  5. Differential geometric invariants for time-reversal symmetric Bloch-bundles: The “Real” case

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

    De Nittis, Giuseppe, E-mail: gidenittis@mat.puc.cl; Gomi, Kiyonori, E-mail: kgomi@math.shinshu-u.ac.jp

    2016-05-15

    Topological quantum systems subjected to an even (resp. odd) time-reversal symmetry can be classified by looking at the related “Real” (resp. “Quaternionic”) Bloch-bundles. If from one side the topological classification of these time-reversal vector bundle theories has been completely described in De Nittis and Gomi [J. Geom. Phys. 86, 303–338 (2014)] for the “Real” case and in De Nittis and Gomi [Commun. Math. Phys. 339, 1–55 (2015)] for the “Quaternionic” case, from the other side it seems that a classification in terms of differential geometric invariants is still missing in the literature. With this article and its companion [G. Demore » Nittis and K. Gomi (unpublished)] we want to cover this gap. More precisely, we extend in an equivariant way the theory of connections on principal bundles and vector bundles endowed with a time-reversal symmetry. In the “Real” case we generalize the Chern-Weil theory and we show that the assignment of a “Real” connection, along with the related differential Chern class and its holonomy, suffices for the classification of “Real” vector bundles in low dimensions.« less

  6. Unwrapping the Bundle: An Examination of Research Libraries and the "Big Deal"

    ERIC Educational Resources Information Center

    Strieb, Karla L.; Blixrud, Julia C.

    2014-01-01

    This study presents and analyzes the findings of a 2012 survey of member libraries belonging to the Association of Research Libraries (ARL) about publishers' large journal bundles and compares the results to earlier surveys. The data illuminate five research questions: market penetration, journal bundle construction, collection format shifts,…

  7. Transduction channels’ gating can control friction on vibrating hair-cell bundles in the ear

    PubMed Central

    Bormuth, Volker; Barral, Jérémie; Joanny, Jean-François; Jülicher, Frank; Martin, Pascal

    2014-01-01

    Hearing starts when sound-evoked mechanical vibrations of the hair-cell bundle activate mechanosensitive ion channels, giving birth to an electrical signal. As for any mechanical system, friction impedes movements of the hair bundle and thus constrains the sensitivity and frequency selectivity of auditory transduction. Friction is generally thought to result mainly from viscous drag by the surrounding fluid. We demonstrate here that the opening and closing of the transduction channels produce internal frictional forces that can dominate viscous drag on the micrometer-sized hair bundle. We characterized friction by analyzing hysteresis in the force–displacement relation of single hair-cell bundles in response to periodic triangular stimuli. For bundle velocities high enough to outrun adaptation, we found that frictional forces were maximal within the narrow region of deflections that elicited significant channel gating, plummeted upon application of a channel blocker, and displayed a sublinear growth for increasing bundle velocity. At low velocity, the slope of the relation between the frictional force and velocity was nearly fivefold larger than the hydrodynamic friction coefficient that was measured when the transduction machinery was decoupled from bundle motion by severing tip links. A theoretical analysis reveals that channel friction arises from coupling the dynamics of the conformational change associated with channel gating to tip-link tension. Varying channel properties affects friction, with faster channels producing smaller friction. We propose that this intrinsic source of friction may contribute to the process that sets the hair cell’s characteristic frequency of responsiveness. PMID:24799674

  8. Using the HELIOS facility for assessment of bundle-jacket thermal coupling in a CICC

    NASA Astrophysics Data System (ADS)

    Lacroix, B.; Rousset, B.; Cloez, H.; Decool, P.; Duchateau, J. L.; Hoa, C.; Luchier, N.; Nicollet, S.; Topin, F.

    2016-12-01

    In a Cable In Conduit Conductor (CICC) cooled by forced circulation of supercritical helium, the heat exchange in the bundle region can play a significant role for conductor safe operation, while remaining a quite uncertain parameter. Heat exchange between bundle and jacket depends on the relative contributions of convective heat transfer due to the helium flow inside the bundle and of thermal resistance due to the wrappings between the cable and the conduit. In order to qualify this thermal coupling at realistic operating conditions, a dedicated experiment on a 1.2 m sample of ITER Toroidal Field (TF) dummy conductor was designed and performed in the HELIOS test facility at CEA Grenoble. Several methods were envisaged, and the choice was made to assess bundle-jacket heat transfer coefficient by measuring the temperature of a solid copper cylinder inserted over the conductor jacket and submitted to heat deposition on its outer surface. The mock-up was manufactured and tested in spring 2015. Bundle-jacket heat transfer coefficient was found in the range 300-500 W m-2 K-1. Results analysis suggests that the order of magnitude of convective heat transfer coefficient inside bundle is closer to Colburn-Reynolds analogy than to Dittus-Boelter correlation, and that bundle-jacket thermal coupling is mainly limited by thermal resistance due to wrappings. A model based on an equivalent layer of stagnant helium between wraps and jacket was proposed and showed a good consistency with the experiment, with relevant values for the helium layer thickness.

  9. Incremental Implicit Learning of Bundles of Statistical Patterns

    PubMed Central

    Qian, Ting; Jaeger, T. Florian; Aslin, Richard N.

    2016-01-01

    Forming an accurate representation of a task environment often takes place incrementally as the information relevant to learning the representation only unfolds over time. This incremental nature of learning poses an important problem: it is usually unclear whether a sequence of stimuli consists of only a single pattern, or multiple patterns that are spliced together. In the former case, the learner can directly use each observed stimulus to continuously revise its representation of the task environment. In the latter case, however, the learner must first parse the sequence of stimuli into different bundles, so as to not conflate the multiple patterns. We created a video-game statistical learning paradigm and investigated 1) whether learners without prior knowledge of the existence of multiple “stimulus bundles” — subsequences of stimuli that define locally coherent statistical patterns — could detect their presence in the input, and 2) whether learners are capable of constructing a rich representation that encodes the various statistical patterns associated with bundles. By comparing human learning behavior to the predictions of three computational models, we find evidence that learners can handle both tasks successfully. In addition, we discuss the underlying reasons for why the learning of stimulus bundles occurs even when such behavior may seem irrational. PMID:27639552

  10. Development of a blunt chest injury care bundle: An integrative review.

    PubMed

    Kourouche, Sarah; Buckley, Thomas; Munroe, Belinda; Curtis, Kate

    2018-06-01

    Blunt chest injuries (BCI) are associated with high rates of morbidity and mortality. There are many interventions for BCI which may be able to be combined as a care bundle for improved and more consistent outcomes. To review and integrate the BCI management interventions to inform the development of a BCI care bundle. A structured search of the literature was conducted to identify studies evaluating interventions for patients with BCI. Databases MEDLINE, CINAHL, PubMed and Scopus were searched from 1990-April 2017. A two-step data extraction process was conducted using pre-defined data fields, including research quality indicators. Each study was appraised using a quality assessment tool, scored for level of evidence, then data collated into categories. Interventions were also assessed using the APEASE criteria then integrated to develop a BCI care bundle. Eighty-one articles were included in the final analysis. Interventions that improved BCI outcomes were grouped into three categories; respiratory intervention, analgesia and surgical intervention. Respiratory interventions included continuous positive airway pressure and high flow nasal oxygen. Analgesia interventions included regular multi-modal analgesia and paravertebral or epidural analgesia. Surgical fixation was supported for use in moderate to severe rib fractures/BCI. Interventions supported by evidence and that met APEASE criteria were combined into a BCI care bundle with four components: respiratory adjuncts, analgesia, complication prevention, and surgical fixation. The key components of a BCI care bundle are respiratory support, analgesia, complication prevention including chest physiotherapy and surgical fixation. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Food bundling as a health nudge: Investigating consumer fruit and vegetable selection using behavioral economics.

    PubMed

    Carroll, Kathryn A; Samek, Anya; Zepeda, Lydia

    2018-02-01

    Displaying bundles of healthy foods at the grocery store is a health nudge that simplifies shopping and may have the potential for increasing fruit and vegetable (F&V) purchasing. To evaluate the impact of food bundling, we conduct an artefactual field experiment with community participants in a laboratory set up as a grocery store. Dual-self theory suggests that food choices may differ depending on whether shoppers are under cognitive load - in our experiment, we exogenously vary whether bundles are displayed (with and without a price discount) and whether shoppers are under cognitive load. Our findings align with prior studies that suggest unhealthy options are more likely to be selected when cognitive resources are constrained. When bundles are displayed, we observe increased F&V purchasing. We also observe a significant interaction between cognitive load and price discounting. We find discounted bundles are more effective in the absence of cognitive load, but non-discounted bundles are more effective when shoppers are under cognitive load. Although more research is warranted, our findings suggest that when shopping under cognitive load, it is possible that discounts impose additional cognitive strain on the shopping experience. For retailers and policymakers, our results point to the potential power of bundling as a strategy for increasing healthy food purchasing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. The bundling of actin with polyethylene glycol 8000 in the presence and absence of gelsolin.

    PubMed Central

    Goverman, J; Schick, L A; Newman, J

    1996-01-01

    Actin filament and bundle formation occur in the cytosol under conditions of very high total macromolecular concentration. In this study we have utilized the inert molecule polyethylene glycol 8000 (PEG) as a means of simulating crowded conditions in vitro. Column-purified Ca-actin was polymerized in the absence and presence of gelsolin (to regulate mean filament lengths between 50 and 5000 mers) and PEG (2-8%) using various concentrations of KCl and/or 2 mM divalent cations. Bundling was characterized by the scattered light intensity and mean diffusion coefficients obtained from dynamic light scattering, as well as by fluorescence and phase-contrast microscopy. The minimum concentration of KCl required for bundling decreases both with increasing concentration of PEG at a fixed mean filament length, and with decreasing filament length at a fixed concentration of PEG. In the absence of divalent cation, bundling is reversible on dilution, as determined by intensity levels, diffusion coefficients, and microscopy. However, with either 2 mM Mg2+ or Ca2+ added, bundling is irreversible under conditions of higher PEG concentrations or longer filaments, indicating that osmotic pressure effects cannot fully explain actin bundling with PEG. Weaker divalent cation-binding sites on actin as well as disulfide bonds appear to be involved in the irreversible bundling. Images FIGURE 7 PMID:8874022

  13. Covariance and the hierarchy of frame bundles

    NASA Technical Reports Server (NTRS)

    Estabrook, Frank B.

    1987-01-01

    This is an essay on the general concept of covariance, and its connection with the structure of the nested set of higher frame bundles over a differentiable manifold. Examples of covariant geometric objects include not only linear tensor fields, densities and forms, but affinity fields, sectors and sector forms, higher order frame fields, etc., often having nonlinear transformation rules and Lie derivatives. The intrinsic, or invariant, sets of forms that arise on frame bundles satisfy the graded Cartan-Maurer structure equations of an infinite Lie algebra. Reduction of these gives invariant structure equations for Lie pseudogroups, and for G-structures of various orders. Some new results are introduced for prolongation of structure equations, and for treatment of Riemannian geometry with higher-order moving frames. The use of invariant form equations for nonlinear field physics is implicitly advocated.

  14. Assessing the Effect of Language Demand in Bundles of Math Word Problems

    ERIC Educational Resources Information Center

    Banks, Kathleen; Jeddeeni, Ahmad; Walker, Cindy M.

    2016-01-01

    Differential bundle functioning (DBF) analyses were conducted to determine whether seventh and eighth grade second language learners (SLLs) had lower probabilities of answering bundles of math word problems correctly that had heavy language demands, when compared to non-SLLs of equal math proficiency. Math word problems on each of four test forms…

  15. Constructed microhabitat bundles for sampling fishes and crayfishes in coastal plain streams

    Treesearch

    Melvin L. Warren; A.L. Sheldon; W.R. Haag

    2009-01-01

    We investigated fish and crayfish use of standardized, constructed microhabitats (bundles) in three northern Mississippi streams. Cypress Creek and the Little Tallahatchie Canal were channelized and incised and had little woody cover; Puskus Creek was unchannelized and unincised and had abundant woody cover. We constructed three types of bundles (cane, leaf, and string...

  16. Simulations of resonant Raman response in bundles of semiconductor carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Roslyak, Oleksiy; Piryatinski, Andrei; Doorn, Stephen; Haroz, Erik; Telg, Hagen; Duque, Juan; Crochet, Jared; Simpson, J. R.; Hight Walker, A. R.; LANL Collaboration; Fordham Collaboration; NIST Collaboration

    This work is motivated by an experimental study of resonant Raman spectroscopy under E22 excitation, which shows a new, sharp feature associated with bundling in (6,5) semiconductor carbon nanotubes. In order to provide an insight into the experimental data, we model Raman excitation spectra using our modified discrete dipole approximation (DDA) method. The calculations account for the exciton states polarized along and across the nanotube axis that are characterized by a small energy splitting. Strong polarization of the nanotubes forming the bundle results in the exciton state mixing whose spectroscopic signatures such as peaks positions, line widths, and depolarization ratio are calculated and compared to the experiment. Furthermore, the effects of the energy and structural disorder, as well as structural defects within the bundle are also examined and compared with the experimental data.

  17. Simulations of resonant Raman response in bundles of semiconductor carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Roslyak, Oleksiy; Doorn, Stephen; Haroz, Erik; Duque, Juan; Crochet, Jared; Telg, Hagen; Hight Walker, Angela; Simpson, Jeffrey; Piryatinski, Andrei

    This work is motivated by experimental study of resonant Raman response associated with E22 exciton state coupled to G+-mode vibrational mode in bundles of (6,5) semiconductor carbon nanotubes. In order to provide an insight into experimental data, we model Raman excitation spectra using our modified discrete dipole approximation (DDA) method. The calculations account for the exciton states polarized along and across the nanotube axis that are characterized by a small energy splitting. Strong polarization of the nanotubes forming the bundle results in the exciton state mixing whose spectroscopic signatures such as peaks positions, line widths, and depolarization ratio are calculated and compared to the experiment. Furthermore, the effects of the energy and structural disorder, as well as structural defects within the bundle are also examined and compared with the experimental data.

  18. Ab initio density functional theory investigation of structural and electronic properties of silicon carbide nanotube bundles

    NASA Astrophysics Data System (ADS)

    Moradian, Rostam; Behzad, Somayeh; Chegel, Raad

    2008-10-01

    By using ab initio density functional theory the structural and electronic properties of isolated and bundled (8,0) and (6,6) silicon carbide nanotubes (SiCNTs) are investigated. Our results show that for such small diameter nanotubes the inter-tube interaction causes a very small radial deformation, while band splitting and reduction of the semiconducting energy band gap are significant. We compared the equilibrium interaction energy and inter-tube separation distance of (8,0) SiCNT bundle with (10,0) carbon nanotube (CNT) bundle where they have the same radius. We found that there is a larger inter-tube separation and weaker inter-tube interaction in the (8,0) SiCNT bundle with respect to (10,0) CNT bundle, although they have the same radius.

  19. Swimming Motility Mediates the Formation of Neutrophil Extracellular Traps Induced by Flagellated Pseudomonas aeruginosa

    PubMed Central

    Sil, Payel; Chassaing, Benoit; Yoo, Dae-goon; Gewirtz, Andrew T.; Goldberg, Joanna B.; McCarter, Linda L.; Rada, Balázs

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen causing severe infections often characterized by robust neutrophilic infiltration. Neutrophils provide the first line of defense against P. aeruginosa. Aside from their defense conferred by phagocytic activity, neutrophils also release neutrophil extracellular traps (NETs) to immobilize bacteria. Although NET formation is an important antimicrobial process, the details of its mechanism are largely unknown. The identity of the main components of P. aeruginosa responsible for triggering NET formation is unclear. In this study, our focus was to identify the main bacterial factors mediating NET formation and to gain insight into the underlying mechanism. We found that P. aeruginosa in its exponential growth phase promoted strong NET formation in human neutrophils while its NET-inducing ability dramatically decreased at later stages of bacterial growth. We identified the flagellum as the primary component of P. aeruginosa responsible for inducing NET extrusion as flagellum-deficient bacteria remained seriously impaired in triggering NET formation. Purified P. aeruginosa flagellin, the monomeric component of the flagellum, does not stimulate NET formation in human neutrophils. P. aeruginosa-induced NET formation is independent of the flagellum-sensing receptors TLR5 and NLRC4 in both human and mouse neutrophils. Interestingly, we found that flagellar motility, not flagellum binding to neutrophils per se, mediates NET release induced by flagellated bacteria. Immotile, flagellar motor-deficient bacterial strains producing paralyzed flagella did not induce NET formation. Forced contact between immotile P. aeruginosa and neutrophils restored their NET-inducing ability. Both the motAB and motCD genetic loci encoding flagellar motor genes contribute to maximal NET release; however the motCD genes play a more important role. Phagocytosis of P. aeruginosa and superoxide production by neutrophils were also largely dependent upon

  20. Ab initio density functional theory investigation of Li-intercalated silicon carbide nanotube bundles

    NASA Astrophysics Data System (ADS)

    Moradian, Rostam; Behzad, Somayeh; Chegel, Raad

    2009-06-01

    We present the results of ab initio density functional theory calculations on the energetic, and geometric and electronic structure of Li-intercalated ( 6,6) silicon carbide nanotube (SiCNT) bundles. Our results show that intercalation of lithium leads to the significant changes in the geometrical structure. The most prominent effect of Li intercalation on the electronic band structure is a shift of the Fermi energy which occurs as a result of charge transfer from lithium to the SiCNTs. All the Li-intercalated ( 6,6) SiCNT bundles are predicted to be metallic representing a substantial change in electronic properties relative to the undoped bundle, which is a wide band gap semiconductor. Both inside of the nanotube and the interstitial space are susceptible for intercalation. The present calculations suggest that the SiCNT bundle is a promising candidate for the anode material in battery applications.

  1. The Shape of a Ponytail and the Statistical Physics of Hair Fiber Bundles

    NASA Astrophysics Data System (ADS)

    Goldstein, Raymond E.; Warren, Patrick B.; Ball, Robin C.

    2012-02-01

    From Leonardo to the Brothers Grimm our fascination with hair has endured in art and science. Yet, a quantitative understanding of the shapes of a hair bundles has been lacking. Here we combine experiment and theory to propose an answer to the most basic question: What is the shape of a ponytail? A model for the shape of hair bundles is developed from the perspective of statistical physics, treating individual fibers as elastic filaments with random intrinsic curvatures. The combined effects of bending elasticity, gravity, and bundle compressibility are recast as a differential equation for the envelope of a bundle, in which the compressibility enters through an ``equation of state.'' From this, we identify the balance of forces in various regions of the ponytail, extract the equation of state from analysis of ponytail shapes, and relate the observed pressure to the measured random curvatures of individual hairs.

  2. Implementing a Daily Maintenance Care Bundle to Prevent Central Line-Associated Bloodstream Infections in Pediatric Oncology Patients.

    PubMed

    Duffy, Elizabeth A; Rodgers, Cheryl C; Shever, Leah L; Hockenberry, Marilyn J

    2015-01-01

    Eliminating central line-associated bloodstream infection (CLABSI) is a national priority. Central venous catheter (CVC) care bundles are composed of a series of interventions that, when used together, are effective in preventing CLABSI. A CVC daily maintenance care bundle includes procedural guidelines for hygiene, dressing changes, and access as well as specific timeframes. Failure to complete one of the components of the care bundle predisposes the patient to a bloodstream infection. A nurse-led multidisciplinary team implemented and, for six months, sustained a daily maintenance care bundle for pediatric oncology patients. This quality improvement project focused on nursing staffs' implementation of the daily maintenance care bundle and the sustainment of the intervention. The project used a pre-post program design to evaluate outcomes of CVC daily maintenance care bundle compliancy and CLABSI. A statistically significant increase between the pre- and post-assessments of the compliance was noted with the CVC daily maintenance care bundle. CLABSI infection rates decreased during the intervention. Strategies to implement practice change and promote sustainability are discussed. © 2015 by Association of Pediatric Hematology/Oncology Nurses.

  3. Modular Bundle Adjustment for Photogrammetric Computations

    NASA Astrophysics Data System (ADS)

    Börlin, N.; Murtiyoso, A.; Grussenmeyer, P.; Menna, F.; Nocerino, E.

    2018-05-01

    In this paper we investigate how the residuals in bundle adjustment can be split into a composition of simple functions. According to the chain rule, the Jacobian (linearisation) of the residual can be formed as a product of the Jacobians of the individual steps. When implemented, this enables a modularisation of the computation of the bundle adjustment residuals and Jacobians where each component has limited responsibility. This enables simple replacement of components to e.g. implement different projection or rotation models by exchanging a module. The technique has previously been used to implement bundle adjustment in the open-source package DBAT (Börlin and Grussenmeyer, 2013) based on the Photogrammetric and Computer Vision interpretations of Brown (1971) lens distortion model. In this paper, we applied the technique to investigate how affine distortions can be used to model the projection of a tilt-shift lens. Two extended distortion models were implemented to test the hypothesis that the ordering of the affine and lens distortion steps can be changed to reduce the size of the residuals of a tilt-shift lens calibration. Results on synthetic data confirm that the ordering of the affine and lens distortion steps matter and is detectable by DBAT. However, when applied to a real camera calibration data set of a tilt-shift lens, no difference between the extended models was seen. This suggests that the tested hypothesis is false and that other effects need to be modelled to better explain the projection. The relatively low implementation effort that was needed to generate the models suggest that the technique can be used to investigate other novel projection models in photogrammetry, including modelling changes in the 3D geometry to better understand the tilt-shift lens.

  4. ACL double-bundle reconstruction with one tibial tunnel provides equal stability compared to two tibial tunnels.

    PubMed

    Drews, Björn Holger; Seitz, Andreas Martin; Huth, Jochen; Bauer, Gerhard; Ignatius, Anita; Dürselen, Lutz

    2017-05-01

    The purpose of this study was to investigate whether an anterior cruciate ligament (ACL) double-bundle reconstruction with one tibial tunnel displays the same in vitro stability as a conventional double-bundle reconstruction with two tibial tunnels when using the same tensioning protocol. In 11 fresh-frozen cadaveric knees, ACL double-bundle reconstruction with one and two tibial tunnels was performed. The two grafts were tightened using 80 N in different flexion angles (anteromedial-bundle at 60° and posterolateral-bundle at 15°). Anterior tibial translation (134 N) and translation with combined rotatory and valgus loads (10 Nm valgus stress and 4 Nm internal tibial torque) were determined at 0°, 30°, 60° and 90° flexion. Measurements were taken in intact ACL, resected ACL, three-tunnel reconstruction and four-tunnel reconstruction. Additionally, the tension on the grafts was determined. Student's t test was performed for statistical analysis of the related samples. Significance was set at p < 0.017 according to Bonferroni correction. The two reconstructive techniques displayed no significant differences in comparison with the intact ACL in anterior tibial translation at 0°, 60° and 90° of flexion. The same results were obtained for the anterior tibial translation with a combined rotatory load at 60° and 90°. When directly comparing both reconstructive techniques, there were no significant differences for the anterior tibial translation and combined rotatory load at all flexion angles. The measured tension on grafts displayed similar load sharing between both bundles. Except at full extension, both grafts displayed a significantly different tension increase under anterior tibial translation for both techniques (p = 0.0086). Tightening both bundles in ACL double-bundle reconstruction with one or two tibial tunnels in different flexion angles achieved comparable restoration of stability, although there was different load sharing on the bundles

  5. Electrocardiogram: his bundle potentials can be recorded noninvasively beat by beat on surface electrocardiogram.

    PubMed

    Wang, Gaopin; Liu, Renguang; Chang, Qinghua; Xu, Zhaolong; Zhang, Yingjie; Pan, Dianzhu

    2017-03-15

    The micro waveform of His bundle potential can't be recorded beat-to-beat on surface electrocardiogram yet. We have found that the micro-wavelets before QRS complex may be related to atrioventricular conduction system potentials. This study is to explore the possibility of His bundle potential can be noninvasively recorded on surface electrocardiogram. We randomized 65 patients undergoing radiofrequency catheter ablation of paroxysmal superventricular tachycardia (exclude overt Wolff-Parkinson-White syndrome) to receive "conventional electrocardiogram" and "new electrocardiogram" before the procedure. His bundle electrogram was collected during the procedure. Comparative analysis of PA s (PA interval recorded on surface electrocardiogram), AH s (AH interval recorded on surface electrocardiogram) and HV s (HV interval recorded on surface electrocardiogram) interval recorded on surface "new electrocardiogram" and PA, AH, HV interval recorded on His bundle electrogram was investigated. There was no difference (P > 0.05) between groups in HV s interval (49.63 ± 6.19 ms) and HV interval (49.35 ± 6.49 ms). Results of correlational analysis found that HV S interval was significantly positively associated with HV interval (r = 0.929; P < 0.01). His bundle potentials can be noninvasively recorded on surface electrocardiogram. Noninvasive His bundle potential tracing might represent a new method for locating the site of atrioventricular block and identifying the origin of a wide QRS complex.

  6. Cross-Linker Unbinding and Self-Similarity in Bundled Cytoskeletal Networks

    NASA Astrophysics Data System (ADS)

    Lieleg, O.; Bausch, A. R.

    2007-10-01

    The macromechanical properties of purely bundled in vitro actin networks are not only determined by the micromechanical properties of individual bundles but also by molecular unbinding events of the actin-binding protein (ABP) fascin. Under high mechanical load the network elasticity depends on the forced unbinding of individual ABPs in a rate dependent manner. Cross-linker unbinding in combination with the structural self-similarity of the network enables the introduction of a concentration-time superposition principle—broadening the mechanically accessible frequency range over 8 orders of magnitude.

  7. Mutational Analyses of HAMP Helices Suggest a Dynamic Bundle Model of Input-Output Signaling in Chemoreceptors

    PubMed Central

    Zhou, Qin; Ames, Peter; Parkinson, John S.

    2009-01-01

    SUMMARY To test the gearbox model of HAMP signaling in the E. coli serine receptor, Tsr, we generated a series of amino acid replacements at each residue of the AS1 and AS2 helices. The residues most critical for Tsr function defined hydrophobic packing faces consistent with a 4-helix bundle. Suppression patterns of helix lesions conformed to the the predicted packing layers in the bundle. Although the properties and patterns of most AS1 and AS2 lesions were consistent with both proposed gearbox structures, some mutational features specifically indicate the functional importance of an x-da bundle over an alternative a-d bundle. These genetic data suggest that HAMP signaling could simply involve changes in the stability of its x-da bundle. We propose that Tsr HAMP controls output signals by modulating destabilizing phase clashes between the AS2 helices and the adjoining kinase control helices. Our model further proposes that chemoeffectors regulate HAMP bundle stability through a control cable connection between the transmembrane segments and AS1 helices. Attractant stimuli, which cause inward piston displacements in chemoreceptors, should reduce cable tension, thereby stabilizing the HAMP bundle. This study shows how transmembrane signaling and HAMP input-output control could occur without the helix rotations central to the gearbox model. PMID:19656294

  8. EsxB, a secreted protein from Bacillus anthracis forms two distinct helical bundles

    DOE PAGES

    Fan, Yao; Tan, Kemin; Chhor, Gekleng; ...

    2015-07-03

    The EsxB protein from Bacillus anthracis belongs to the WXG100 family, a group of proteins secreted by a specialized secretion system. We have determined the crystal structures of recombinant EsxB and discovered that the small protein (~10 kDa), comprised of a helix-loop-helix (HLH) hairpin, is capable of associating into two different helical bundles. The two basic quaternary assemblies of EsxB are an antiparallel (AP) dimer and a rarely observed bisecting U (BU) dimer. This structural duality of EsxB is believed to originate from the heptad repeat sequence diversity of the first helix of its HLH hairpin, which allows for twomore » alternative helix packing. The flexibility of EsxB and the ability to form alternative helical bundles underscore the possibility that this protein can serve as an adaptor in secretion and can form hetero-oligomeric helix bundle(s) with other secreted members of the WXG100 family, such as EsxW. The highly conserved WXG motif is located within the loop of the HLH hairpin and is mostly buried within the helix bundle suggesting that its role is mainly structural. The exact functions of the motif, including a proposed role as a secretion signal, remain unknown.« less

  9. Biomechanical Comparison: Single-Bundle versus Double-Bundle Posterior Cruciate Ligament Reconstruction Techniques.

    PubMed

    Milles, Jeffrey L; Nuelle, Clayton W; Pfeiffer, Ferris; Stannard, James P; Smith, Patrick; Kfuri, Mauricio; Cook, James L

    2017-05-01

    Controversy exists regarding double-bundle (DB) versus single-bundle (SB) posterior cruciate ligament (PCL) reconstruction, with differences in multiple variables affecting biomechanical and clinical results. Our objective was to compare immediate postimplantation biomechanics of SB versus DB reconstructions to determine the relative importance of restoring both PCL bundles versus total graft volume. Twenty knees were randomly assigned to five techniques ( n  = 4 knees/technique), performed by three surgeons experienced in their technique(s), three SB techniques ( n  = 12; all-inside arthroscopic inlay, all-inside suspensory fixation, and arthroscopic-assisted open onlay), and two DB techniques ( n  = 8; arthroscopic-assisted open inlay and all-inside suspensory fixation). Each knee was tested in three conditions: PCL-intact, PCL-deficient, and post-PCL reconstruction. Testing consisted of a posterior-directed force at four knee flexion angles, 10, 30, 60, and 90 degrees, to measure load to 5 mm of posterior displacement, maximum displacement (at 100 N load), and stiffness. Data for each knee were normalized, combined into two groups (SB and DB), and then compared using one-way analysis of variance. Graft volumes were calculated and analyzed to determine if differences significantly influenced the biomechanical results. Intact knees were stiffer than both groups at most angles ( p  < 0.02; p  < 0.05). DB was stiffer than SB at all angles except 30 degrees ( p  < 0.05). Intact knees had less laxity than SB ( p  < 0.03) and DB ( p  < 0.05) at 60 and 90 degrees. DB had less laxity than SB at all angles except 60 degrees ( p  < 0.05). Intact knees required more load than SB at 30, 60, and 90 degrees ( p  < 0.01) and more than DB at 60 and 90 degrees ( p  < 0.05). DB required more load than SB at 30, 60, and 90 degrees ( p  < 0.01). Graft volumes did not have strong correlations ( r  = 0.13-0.37) to any

  10. Transcriptional Analysis of the MrpJ Network: Modulation of Diverse Virulence-Associated Genes and Direct Regulation of mrp Fimbrial and flhDC Flagellar Operons in Proteus mirabilis

    PubMed Central

    Bode, Nadine J.; Debnath, Irina; Kuan, Lisa; Schulfer, Anjelique; Ty, Maureen

    2015-01-01

    The enteric bacterium Proteus mirabilis is associated with a significant number of catheter-associated urinary tract infections (UTIs). Strict regulation of the antagonistic processes of adhesion and motility, mediated by fimbriae and flagella, respectively, is essential for disease progression. Previously, the transcriptional regulator MrpJ, which is encoded by the mrp fimbrial operon, has been shown to repress both swimming and swarming motility. Here we show that MrpJ affects an array of cellular processes beyond adherence and motility. Microarray analysis found that expression of mrpJ mimicking levels observed during UTIs leads to differential expression of 217 genes related to, among other functions, bacterial virulence, type VI secretion, and metabolism. We probed the molecular mechanism of transcriptional regulation by MrpJ using transcriptional reporters and chromatin immunoprecipitation (ChIP). Binding of MrpJ to two virulence-associated target gene promoters, the promoters of the flagellar master regulator flhDC and mrp itself, appears to be affected by the condensation state of the native chromosome, although both targets share a direct MrpJ binding site proximal to the transcriptional start. Furthermore, an mrpJ deletion mutant colonized the bladders of mice at significantly lower levels in a transurethral model of infection. Additionally, we observed that mrpJ is widely conserved in a collection of recent clinical isolates. Altogether, these findings support a role of MrpJ as a global regulator of P. mirabilis virulence. PMID:25847961

  11. Transcriptional analysis of the MrpJ network: modulation of diverse virulence-associated genes and direct regulation of mrp fimbrial and flhDC flagellar operons in Proteus mirabilis.

    PubMed

    Bode, Nadine J; Debnath, Irina; Kuan, Lisa; Schulfer, Anjelique; Ty, Maureen; Pearson, Melanie M

    2015-06-01

    The enteric bacterium Proteus mirabilis is associated with a significant number of catheter-associated urinary tract infections (UTIs). Strict regulation of the antagonistic processes of adhesion and motility, mediated by fimbriae and flagella, respectively, is essential for disease progression. Previously, the transcriptional regulator MrpJ, which is encoded by the mrp fimbrial operon, has been shown to repress both swimming and swarming motility. Here we show that MrpJ affects an array of cellular processes beyond adherence and motility. Microarray analysis found that expression of mrpJ mimicking levels observed during UTIs leads to differential expression of 217 genes related to, among other functions, bacterial virulence, type VI secretion, and metabolism. We probed the molecular mechanism of transcriptional regulation by MrpJ using transcriptional reporters and chromatin immunoprecipitation (ChIP). Binding of MrpJ to two virulence-associated target gene promoters, the promoters of the flagellar master regulator flhDC and mrp itself, appears to be affected by the condensation state of the native chromosome, although both targets share a direct MrpJ binding site proximal to the transcriptional start. Furthermore, an mrpJ deletion mutant colonized the bladders of mice at significantly lower levels in a transurethral model of infection. Additionally, we observed that mrpJ is widely conserved in a collection of recent clinical isolates. Altogether, these findings support a role of MrpJ as a global regulator of P. mirabilis virulence. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. Creep rupture of fiber bundles: A molecular dynamics investigation

    NASA Astrophysics Data System (ADS)

    Linga, G.; Ballone, P.; Hansen, Alex

    2015-08-01

    The creep deformation and eventual breaking of polymeric samples under a constant tensile load F is investigated by molecular dynamics based on a particle representation of the fiber bundle model. The results of the virtual testing of fibrous samples consisting of 40 000 particles arranged on Nc=400 chains reproduce characteristic stages seen in the experimental investigations of creep in polymeric materials. A logarithmic plot of the bundle lifetime τ versus load F displays a marked curvature, ruling out a simple power-law dependence of τ on F . A power law τ ˜F-4 , however, is recovered at high load. We discuss the role of reversible bond breaking and formation on the eventual fate of the sample and simulate a different type of creep testing, imposing a constant stress rate on the sample up to its breaking point. Our simulations, relying on a coarse-grained representation of the polymer structure, introduce new features into the standard fiber bundle model, such as real-time dynamics, inertia, and entropy, and open the way to more detailed models, aiming at material science aspects of polymeric fibers, investigated within a sound statistical mechanics framework.

  13. System for supporting a bundled tube fuel injector within a combustor

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

    LeBegue, Jeffrey Scott; Melton, Patrick Benedict; Westmoreland, III, James Harold

    A combustor includes an end cover having an outer side and an inner side, an outer barrel having a forward end that is adjacent to the inner side of the end cover and an aft end that is axially spaced from the forward end. An inner barrel is at least partially disposed concentrically within the outer barrel and is fixedly connected to the outer barrel. A fluid conduit extends downstream from the end cover. A first bundled tube fuel injector segment is disposed concentrically within the inner barrel. The bundled tube fuel injector segment includes a fuel plenum that ismore » in fluid communication with the fluid conduit and a plurality of parallel tubes that extend axially through the fuel plenum. The bundled tube fuel injector segment is fixedly connected to the inner barrel.« less

  14. The flagellar motor of Caulobacter crescentus generates more torque when a cell swims backward

    PubMed Central

    Lele, Pushkar P.; Roland, Thibault; Shrivastava, Abhishek; Chen, Yihao; Berg, Howard C.

    2016-01-01

    Caulobacter crescentus, a monotrichous bacterium, swims by rotating a single right-handed helical filament. CW motor rotation thrusts the cell forward 1, a mode of motility known as the pusher mode; CCW motor rotation pulls the cell backward, a mode of motility referred to as the puller mode 2. The situation is opposite in E. coli, a peritrichous bacterium, where CCW rotation of multiple left-handed filaments drives the cell forward. The flagellar motor in E. coli generates more torque in the CCW direction than the CW direction in swimming cells 3,4. However, monotrichous bacteria including C. crescentus swim forward and backward at similar speeds, prompting the assumption that motor torques in the two modes are the same 5,6. Here, we present evidence that motors in C. crescentus develop higher torques in the puller mode than in the pusher mode, and suggest that the anisotropy in torque-generation is similar in two species, despite the differences in filament handedness and motor bias (probability of CW rotation). PMID:27499800

  15. New Feature Observed in the Raman Resonance Excitation Profiles of (6 , 5) -Enriched, Selectively Bundled SWCNTs

    NASA Astrophysics Data System (ADS)

    Hight Walker, A. R.; Simpson, J. R.; Roslyak, O.; Haroz, E.; Telg, H.; Duque, J. G.; Crochet, J. J.; Piryatinski, A.; Doorn, S. K.

    Understanding the photophysics of exciton behavior in single wall carbon nanotube (SWCNT) bundles remains important for opto-electronic device applications. We report resonance Raman spectroscopy (RRS) measurements on (6 , 5) -enriched SWCNTs, dispersed in aqueous solutions and separated using density gradient ultracentrifugation into fractions of increasing bundling. Near-IR to UV absorption spectroscopy shows a redshift and broadening of the main excitonic transitions with increasing bundling. A continuously tunable dye laser coupled to a triple-grating spectrometer affords measurement of Raman resonance excitation profiles (REPs) over a range of wavelengths covering the (6 , 5) -E22 range (505 to 585) nm. REPs of both the radial breathing mode (RBM) and G-band reveal a redshifting and broadening of the (6 , 5) E22 transition energy with increasing bundling. Additionally, we observe an unexpected peak in the REP of bundled SWCNTs, which is shifted lower in energy than the main E22 and is anomalously narrow. We compare these observations to a theoretical model that examines the origin of this peak in relation to bundle polarization-enhanced exciton response.

  16. Textbook-Bundled Metacognitive Tools: A Study of LearnSmart's Efficacy in General Chemistry

    ERIC Educational Resources Information Center

    Thadani, Vandana; Bouvier-Brown, Nicole C.

    2016-01-01

    College textbook publishers increasingly bundle sophisticated technology-based study tools with their texts. These tools appear promising, but empirical work on their efficacy is needed. We examined whether LearnSmart, a study tool bundled with McGraw-Hill's textbook "Chemistry" (Chang & Goldsby, 2013), improved learning in an…

  17. The Mr 140,000 Intermediate Chain of Chlamydomonas Flagellar Inner Arm Dynein Is a WD-Repeat Protein Implicated in Dynein Arm Anchoring

    PubMed Central

    Yang, Pinfen; Sale, Winfield S.

    1998-01-01

    Previous structural and biochemical studies have revealed that the inner arm dynein I1 is targeted and anchored to a unique site located proximal to the first radial spoke in each 96-nm axoneme repeat on flagellar doublet microtubules. To determine whether intermediate chains mediate the positioning and docking of dynein complexes, we cloned and characterized the 140-kDa intermediate chain (IC140) of the I1 complex. Sequence and secondary structural analysis, with particular emphasis on β-sheet organization, predicted that IC140 contains seven WD repeats. Reexamination of other members of the dynein intermediate chain family of WD proteins indicated that these polypeptides also bear seven WD/β-sheet repeats arranged in the same pattern along each intermediate chain protein. A polyclonal antibody was raised against a 53-kDa fusion protein derived from the C-terminal third of IC140. The antibody is highly specific for IC140 and does not bind to other dynein intermediate chains or proteins in Chlamydomonas flagella. Immunofluorescent microscopy of Chlamydomonas cells confirmed that IC140 is distributed along the length of both flagellar axonemes. In vitro reconstitution experiments demonstrated that the 53-kDa C-terminal fusion protein binds specifically to axonemes lacking the I1 complex. Chemical cross-linking indicated that IC140 is closely associated with a second intermediate chain in the I1 complex. These data suggest that IC140 contains domains responsible for the assembly and docking of the I1 complex to the doublet microtubule cargo. PMID:9843573

  18. Prevalence of Split Nerve Fiber Layer Bundles in Healthy People Imaged with Spectral Domain Optical Coherence Tomography.

    PubMed

    Gür Güngör, Sirel; Akman, Ahmet; Sarıgül Sezenöz, Almila; Tanrıaşıkı, Gülşah

    2016-12-01

    The presence of retinal nerve fiber layer (RNFL) split bundles was recently described in normal eyes scanned using scanning laser polarimetry and by histologic studies. Split bundles may resemble RNFL loss in healthy eyes. The aim of our study was to determine the prevalence of nerve fiber layer split bundles in healthy people. We imaged 718 eyes of 359 healthy persons with the spectral domain optical coherence tomography in this cross-sectional study. All eyes had intraocular pressure of 21 mmHg or less, normal appearance of the optic nerve head, and normal visual fields (Humphrey Field Analyzer 24-2 full threshold program). In our study, a bundle was defined as 'split' when there is localized defect not resembling a wedge defect in the RNFL deviation map with a symmetrically divided RNFL appearance on the RNFL thickness map. The classification was performed by two independent observers who used an identical set of reference examples to standardize the classification. Inter-observer consensus was reached in all cases. Bilateral superior split bundles were seen in 19 cases (5.29%) and unilateral superior split was observed in 15 cases (4.16%). In 325 cases (90.52%) there was no split bundle. Split nerve fiber layer bundles, in contrast to single nerve fiber layer bundles, are not common findings in healthy eyes. In eyes with normal optic disc appearance, especially when a superior RNFL defect is observed in RNFL deviation map, the RNLF thickness map and graphs should also be examined for split nerve fiber layer bundles.

  19. First genomic insights into members of a candidate bacterial phylum responsible for wastewater bulking

    PubMed Central

    Ohashi, Akiko; Parks, Donovan H.; Yamauchi, Toshihiro; Tyson, Gene W.

    2015-01-01

    Filamentous cells belonging to the candidate bacterial phylum KSB3 were previously identified as the causative agent of fatal filament overgrowth (bulking) in a high-rate industrial anaerobic wastewater treatment bioreactor. Here, we obtained near complete genomes from two KSB3 populations in the bioreactor, including the dominant bulking filament, using differential coverage binning of metagenomic data. Fluorescence in situ hybridization with 16S rRNA-targeted probes specific for the two populations confirmed that both are filamentous organisms. Genome-based metabolic reconstruction and microscopic observation of the KSB3 filaments in the presence of sugar gradients indicate that both filament types are Gram-negative, strictly anaerobic fermenters capable of non-flagellar based gliding motility, and have a strikingly large number of sensory and response regulator genes. We propose that the KSB3 filaments are highly sensitive to their surroundings and that cellular processes, including those causing bulking, are controlled by external stimuli. The obtained genomes lay the foundation for a more detailed understanding of environmental cues used by KSB3 filaments, which may lead to more robust treatment options to prevent bulking. PMID:25650158

  20. Coupling biochemistry and hydrodynamics captures hyperactivated sperm motility in a simple flagellar model

    PubMed Central

    Olson, Sarah D.; Suarez, Susan S.; Fauci, Lisa J.

    2011-01-01

    Hyperactivation in mammalian sperm is characterized by highly asymmetrical waveforms and an increase in the amplitude of flagellar bends. It is important for the sperm to be able to achieve hyperactivated motility in order to reach and fertilize the egg. Calcium (Ca2+) dynamics are known to play a large role in the initiation and maintenance of hyperactivated motility. Here we present an integrative model that couples the CatSper channel mediated Ca2+ dynamics of hyperactivation to a mechanical model of an idealized sperm flagellum in a 3-d viscous, incompressible fluid. The mechanical forces are due to passive stiffness properties and active bending moments that are a function of the local Ca2+ concentration along the length of the flagellum. By including an asymmetry in bending moments to reflect an asymmetry in the axoneme’s response to Ca2+, we capture the transition from activated motility to hyperactivated motility. We examine the effects of elastic properties of the flagellum and the Ca2+ dynamics on the overall swimming patterns. The swimming velocities of the model flagellum compare well with data for hyperactivated mouse sperm. PMID:21669209

  1. Bundle-branch reentry ventricular tachycardia after transcatheter aortic valve replacement

    PubMed Central

    de la Rosa Riestra, Adriana; Rubio Caballero, José Amador; Freites Estévez, Alfonso; Alonso Belló, Javier; Botas Rodríguez, Javier

    2016-01-01

    An 83-year-old male suffering from severe symptomatic aortic valve stenosis received an implant of a biological aortic prosthesis through the femoral artery without complications. Seven days after dischargement he experienced a syncope. The patient was wearing an ECG holter monitor that day, which showed a wide QRS complex tachycardia of 300 beats per minute. The electrophysiological study revealed a bundle-branch reentry ventricular tachycardia as the cause of the syncope. Radio-frequency was applied on the right-bundle branch. Twelve months later, the patient has remained asymptomatic. PMID:27134443

  2. Association of a Bundled-Payment Program With Cost and Outcomes in Full-Cycle Breast Cancer Care.

    PubMed

    Wang, C Jason; Cheng, Skye H; Wu, Jen-You; Lin, Yi-Ping; Kao, Wen-Hsin; Lin, Chia-Li; Chen, Yin-Jou; Tsai, Shu-Ling; Kao, Feng-Yu; Huang, Andrew T

    2017-03-01

    Value-driven payment system reform is a potential tool for aligning economic incentives with the improvement of quality and efficiency of health care and containment of cost. Such a payment system has not been researched satisfactorily in full-cycle cancer care. To examine the association of outcomes and medical expenditures with a bundled-payment pay-for-performance program for breast cancer in Taiwan compared with a fee-for-service (FFS) program. Data were obtained from the Taiwan Cancer Database, National Health Insurance Claims Data, the National Death Registry, and the bundled-payment enrollment file. Women with newly diagnosed breast cancer and a documented first cancer treatment from January 1, 2004, to December 31, 2008, were selected from the Taiwan Cancer Database and followed up for 5 years, with the last follow-up data available on December 31, 2013. Patients in the bundled-payment program were matched at a ratio of 1:3 with control individuals in an FFS program using a propensity score method. The final sample of 17 940 patients included 4485 (25%) in the bundled-payment group and 13 455 (75%) in the FFS group. Rates of adherence to quality indicators, survival rates, and medical payments (excluding bonuses paid in the bundled-payment group). The Kaplan-Meier method was used to calculate 5-year overall and event-free survival rates by cancer stage, and the Cox proportional hazards regression model was used to examine the effect of the bundled-payment program on overall and event-free survival. Sensitivity analysis for bonus payments in the bundled-payment group was also performed. The study population included 17 940 women (mean [SD] age, 52.2 [10.3] years). In the bundled-payment group, 1473 of 4215 patients (34.9%) with applicable quality indicators had full (100%) adherence to quality indicators compared with 3438 of 12 506 patients (27.5%) with applicable quality indicators in the FFS group (P < .001). The 5-year event-free survival

  3. Experience with Designing and Implementing a Bundled Payment Program for Total Hip Replacement.

    PubMed

    Whitcomb, Winthrop F; Lagu, Tara; Krushell, Robert J; Lehman, Andrew P; Greenbaum, Jordan; McGirr, Joan; Pekow, Penelope S; Calcasola, Stephanie; Benjamin, Evan; Mayforth, Janice; Lindenauer, Peter K

    2015-09-01

    Bundled payments, also known as episode-based payments, are intended to contain health care costs and promote quality. In 2011 a bundled payment pilot program for total hip replacement was implemented by an integrated health care delivery system in conjunction with a commercial health plan subsidiary. In July 2015 the Centers for Medicare & Medicaid Services (CMS) proposed the Comprehensive Care for Joint Replacement Model to test bundled payment for hip and knee replacement. Stakeholders were identified and a structure for program development and implementation was created. An Oversight Committee provided governance over a Clinical Model Subgroup and a Financial Model Subgroup. The pilot program included (1) a clinical model of care encompassing the period from the preoperative evaluation through the third postoperative visit, (2) a pricing model, (3) a program to share savings, and (4) a patient engagement and expectation strategy. Compared to 32 historical controls-patients treated before bundle implementation-45 post-bundle-implementation patients with total hip replacement had a similar length of hospital stay (3.0 versus 3.4 days, p=.24), higher rates of discharge to home or home with services than to a rehabilitation facility (87% versus 63%), similar adjusted median total payments ($22,272 versus $22,567, p=.43), and lower median posthospital payments ($704 versus $1,121, p=.002), and were more likely to receive guideline-consistent care (99% versus 95%, p=.05). The bundled payment pilot program was associated with similar total costs, decreased posthospital costs, fewer discharges to rehabilitation facilities, and improved quality. Successful implementation of the program hinged on buy-in from stakeholders and close collaboration between stakeholders and the clinical and financial teams.

  4. Thick Filament Length and Isoform Composition Determine Self-Organized Contractile Units in Actomyosin Bundles

    PubMed Central

    Thoresen, Todd; Lenz, Martin; Gardel, Margaret L.

    2013-01-01

    Diverse myosin II isoforms regulate contractility of actomyosin bundles in disparate physiological processes by variations in both motor mechanochemistry and the extent to which motors are clustered into thick filaments. Although the role of mechanochemistry is well appreciated, the extent to which thick filament length regulates actomyosin contractility is unknown. Here, we study the contractility of minimal actomyosin bundles formed in vitro by mixtures of F-actin and thick filaments of nonmuscle, smooth, and skeletal muscle myosin isoforms with varied length. Diverse myosin II isoforms guide the self-organization of distinct contractile units within in vitro bundles with shortening rates similar to those of in vivo myofibrils and stress fibers. The tendency to form contractile units increases with the thick filament length, resulting in a bundle shortening rate proportional to the length of constituent myosin thick filament. We develop a model that describes our data, providing a framework in which to understand how diverse myosin II isoforms regulate the contractile behaviors of disordered actomyosin bundles found in muscle and nonmuscle cells. These experiments provide insight into physiological processes that use dynamic regulation of thick filament length, such as smooth muscle contraction. PMID:23442916

  5. Probing sequence dependence of folding pathway of α-helix bundle proteins through free energy landscape analysis.

    PubMed

    Shao, Qiang

    2014-06-05

    A comparative study on the folding of multiple three-α-helix bundle proteins including α3D, α3W, and the B domain of protein A (BdpA) is presented. The use of integrated-tempering-sampling molecular dynamics simulations achieves reversible folding and unfolding events in individual short trajectories, which thus provides an efficient approach to sufficiently sample the configuration space of protein and delineate the folding pathway of α-helix bundle. The detailed free energy landscape analyses indicate that the folding mechanism of α-helix bundle is not uniform but sequence dependent. A simple model is then proposed to predict folding mechanism of α-helix bundle on the basis of amino acid composition: α-helical proteins containing higher percentage of hydrophobic residues than charged ones fold via nucleation-condensation mechanism (e.g., α3D and BdpA) whereas proteins having opposite tendency in amino acid composition more likely fold via the framework mechanism (e.g., α3W). The model is tested on various α-helix bundle proteins, and the predicted mechanism is similar to the most approved one for each protein. In addition, the common features in the folding pathway of α-helix bundle protein are also deduced. In summary, the present study provides comprehensive, atomic-level picture of the folding of α-helix bundle proteins.

  6. The 5 Clinical Pillars of Value for Total Joint Arthroplasty in a Bundled Payment Paradigm.

    PubMed

    Kim, Kelvin; Iorio, Richard

    2017-06-01

    Our large, urban, tertiary, university-based institution reflects on its 4-year experience with Bundled Payments for Care Improvement. We will describe the importance of 5 clinical pillars that have contributed to the early success of our bundled payment initiative. We are convinced that value-based care delivered through bundled payment initiatives is the best method to optimize patient outcomes while rewarding surgeons and hospitals for adapting to the evolving healthcare reforms. We summarize a number of experiences and lessons learned since the implementation of Bundled Payments for Care Improvement at our institution. Our experience has led to the development of more refined clinical pathways and coordination of care through evidence-based approaches. We have established that the success of the bundled payment program rests on the following 5 main clinical pillars: (1) optimizing patient selection and comorbidities; (2) optimizing care coordination, patient education, shared decision making, and patient expectations; (3) using a multimodal pain management protocol and minimizing narcotic use to facilitate rapid rehabilitation; (4) optimizing blood management, and standardizing venous thromboembolic disease prophylaxis treatment by risk standardizing patients and minimizing the use of aggressive anticoagulation; and (5) minimizing post-acute facility and resource utilization, and maximizing home resources for patient recovery. From our extensive experience with bundled payment models, we have established 5 clinical pillars of value for bundled payments. Our hope is that these principles will help ease the transition to value-based care for less-experienced healthcare systems. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Electronic structure and optical properties of boron nitride nanotube bundles from first principles

    NASA Astrophysics Data System (ADS)

    Behzad, Somayeh

    2015-06-01

    The electronic and optical properties of bundled armchair and zigzag boron nitride nanotubes (BNNTs) are investigated by using density functional theory. Owing to the inter-tube coupling, the dispersions along the tube axis and in the plane perpendicular to the tube axis of BNNT bundles are significantly varied, which are characterized by the decrease of band gap, the splitting of the doubly degenerated states, the expansions of valence and conduction bands. The calculated dielectric functions of the armchair and zigzag bundles are similar to that of the isolated tubes, except for the appearance of broadened peaks, small shifts of peak positions about 0.1 eV and increasing of peak intensities.

  8. Structure and Characterization of Vertically Aligned Single-Walled Carbon Nanotube Bundles

    DOE PAGES

    Márquez, Francisco; López, Vicente; Morant, Carmen; ...

    2010-01-01

    Arrmore » ays of vertically aligned single-walled carbon nanotube bundles, SWCNTs, have been synthesized by simple alcohol catalytic chemical vapor deposition process, carried out at 800 ° C . The formed SWCNTs are organized in small groups perpendicularly aligned and attached to the substrate. These small bundles show a constant diameter of ca. 30 nm and are formed by the adhesion of no more than twenty individual SWCNTs perfectly aligned along their length.« less

  9. Mapping Flagellar Genes in Chlamydomonas Using Restriction Fragment Length Polymorphisms

    PubMed Central

    Ranum, LPW.; Thompson, M. D.; Schloss, J. A.; Lefebvre, P. A.; Silflow, C. D.

    1988-01-01

    To correlate cloned nuclear DNA sequences with previously characterized mutations in Chlamydomonas and, to gain insight into the organization of its nuclear genome, we have begun to map molecular markers using restriction fragment length polymorphisms (RFLPs). A Chlamydomonas reinhardtii strain (CC-29) containing phenotypic markers on nine of the 19 linkage groups was crossed to the interfertile species Chlamydomonas smithii. DNA from each member of 22 randomly selected tetrads was analyzed for the segregation of RFLPs associated with cloned genes detected by hybridization with radioactive DNA probes. The current set of markers allows the detection of linkage to new molecular markers over approximately 54% of the existing genetic map. This study focused on mapping cloned flagellar genes and genes whose transcripts accumulate after deflagellation. Twelve different molecular clones have been assigned to seven linkage groups. The α-1 tubulin gene maps to linkage group III and is linked to the genomic sequence homologous to pcf6-100, a cDNA clone whose corresponding transcript accumulates after deflagellation. The α-2 tubulin gene maps to linkage group IV. The two β-tubulin genes are linked, with the β-1 gene being approximately 12 cM more distal from the centromere than the β-2 gene. A clone corresponding to a 73-kD dynein protein maps to the opposite arm of the same linkage group. The gene corresponding to the cDNA clone pcf6-187, whose mRNA accumulates after deflagellation, maps very close to the tightly linked pf-26 and pf-1 mutations on linkage group V. PMID:2906025

  10. Species-dependent hydrodynamics of flagellum-tethered bacteria in early biofilm development.

    PubMed

    Bennett, Rachel R; Lee, Calvin K; De Anda, Jaime; Nealson, Kenneth H; Yildiz, Fitnat H; O'Toole, George A; Wong, Gerard C L; Golestanian, Ramin

    2016-02-01

    Monotrichous bacteria on surfaces exhibit complex spinning movements. Such spinning motility is often a part of the surface detachment launch sequence of these cells. To understand the impact of spinning motility on bacterial surface interactions, we develop a hydrodynamic model of a surface-bound bacterium, which reproduces behaviours that we observe in Pseudomonas aeruginosa, Shewanella oneidensis and Vibrio cholerae, and provides a detailed dictionary for connecting observed spinning behaviour to bacteria-surface interactions. Our findings indicate that the fraction of the flagellar filament adhered to the surface, the rotation torque of this appendage, the flexibility of the flagellar hook and the shape of the bacterial cell dictate the likelihood that a microbe will detach and the optimum orientation that it should have during detachment. These findings are important for understanding species-specific reversible attachment, the key transition event between the planktonic and biofilm lifestyle for motile, rod-shaped organisms. © 2016 The Author(s).

  11. Constrained ripple optimization of Tokamak bundle divertors

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

    Hively, L.M.; Rome, J.A.; Lynch, V.E.

    1983-02-01

    Magnetic field ripple from a tokamak bundle divertor is localized to a small toroidal sector and must be treated differently from the usual (distributed) toroidal field (TF) coil ripple. Generally, in a tokamak with an unoptimized divertor design, all of the banana-trapped fast ions are quickly lost due to banana drift diffusion or to trapping between the 1/R variation in absolute value vector B ..xi.. B and local field maxima due to the divertor. A computer code has been written to optimize automatically on-axis ripple subject to these constraints, while varying up to nine design parameters. Optimum configurations have lowmore » on-axis ripple (<0.2%) so that, now, most banana-trapped fast ions are confined. Only those ions with banana tips near the outside region (absolute value theta < or equal to 45/sup 0/) are lost. However, because finite-sized TF coils have not been used in this study, the flux bundle is not expanded.« less

  12. Reactor application of an improved bundle divertor

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

    Yang, T.F.; Ruck, G.W.; Lee, A.Y.

    1978-11-01

    A Bundle Divertor was chosen as the impurity control and plasma exhaust system for the beam driven Demonstration Tokamak Hybrid Reactor - DTHR. In the context of a preconceptual design study of the reactor and associated facility a bundle divertor concept was developed and integrated into the reactor system. The overall system was found feasible and scalable for reactors with intermediate torodial field strengths on axis. The important design characteristics are: the overall average current density of the divertor coils is 0.73 kA for each tesla of toroidal field on axis; the divertor windings are made from super-conducting cables supportedmore » by steel structures and are designed to be maintainable; the particle collection assembly and auxiliary cryosorption vacuum pump are dual systems designed such that they can be reactivated alterntively to allow for continuous reactor operation; and the power requirement for energizing and operating the divertor is about 5 MW.« less

  13. Material distribution in light water reactor-type bundles tested under severe accident conditions

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

    Noack, V.; Hagen, S.J.L.; Hofmann, P.

    1997-02-01

    Severe fuel damage experiments simulating small-break loss-of-coolant accidents have been carried out in the CORA out-of-pile test facility at Forschungszentrum Karlsruhe. Rod bundles with electrically heated fuel rod simulators containing annular UO{sub 2} pellets, UO{sub 2} full pellet rods, and absorber rods of two kinds (Ag/In/Cd to represent pressurized water reactor conditions and B{sub 4}C to represent boiling water reactor and VVER-1000 fuel elements) were subjected to temperature transients up to 2,300 K. A special method was applied to determine the axial mass distribution of bundle materials. The low-temperature melt formation by various interactions between zirconium and components of absorbermore » and spacer grids strongly influences the bundle degradation and material relocation. Absorber materials can separate from the fuel by a noncoherent relocation of the materials at different temperatures. The distributions of solidified materials in the different test bundles show a clear dependence on the axial temperature profile. Coolant channel blockages are observed mainly at the lower end of the bundle, i.e., near the lowest elevation at which an oxidation excursion resulting from the highly exothermic zirconium-steam reaction had been experienced. This elevation corresponds with a steep axial temperature gradient in the maximum temperature attained. Oxide layers on Zircaloy result in reduced melt formation.« less

  14. Laughter-induced left bundle branch block.

    PubMed

    Chow, Grant V; Desai, Dipan; Spragg, David D; Zakaria, Sammy

    2012-10-01

    We present the case of a patient with ischemic heart disease and intermittent left bundle branch block, reproducibly induced by laughter. Following treatment of ischemia with successful deployment of a drug-eluting stent, no further episodes of inducible LBBB were seen. Transient ischemia, exacerbated by elevated intrathoracic pressure during laughter, may have contributed to onset of this phenomenon. © 2012 Wiley Periodicals, Inc.

  15. Study of imaging fiber bundle coupling technique in IR system

    NASA Astrophysics Data System (ADS)

    Chen, Guoqing; Yang, Jianfeng; Yan, Xingtao; Song, Yansong

    2017-02-01

    Due to its advantageous imaging characteristic and banding flexibility, imaging fiber bundle can be used for line-plane-switching push-broom infrared imaging. How to precisely couple the fiber bundle in the optics system is the key to get excellent image for transmission. After introducing the basic system composition and structural characteristics of the infrared systems coupled with imaging fiber bundle, this article analysis the coupling efficiency and the design requirements of its relay lenses with the angle of the numerical aperture selecting in the system and cold stop matching of the refrigerant infrared detector. For an actual need, one relay coupling system has been designed with the magnification is -0.6, field of objective height is 4mm, objective numerical aperture is 0.15, which has excellent image quality and enough coupling efficiency. In the end, the push broom imaging experiment is carried out. The results show that the design meets the requirements of light energy efficiency and image quality. This design has a certain reference value for the design of the infrared fiber optical system.

  16. Sustained Reduction of Ventilator-Associated Pneumonia Rates Using Real-Time Course Correction With a Ventilator Bundle Compliance Dashboard.

    PubMed

    Talbot, Thomas R; Carr, Devin; Parmley, C Lee; Martin, Barbara J; Gray, Barbara; Ambrose, Anna; Starmer, Jack

    2015-11-01

    The effectiveness of practice bundles on reducing ventilator-associated pneumonia (VAP) has been questioned. To implement a comprehensive program that included a real-time bundle compliance dashboard to improve compliance and reduce ventilator-associated complications. DESIGN Before-and-after quasi-experimental study with interrupted time-series analysis. SETTING Academic medical center. In 2007 a comprehensive institutional ventilator bundle program was developed. To assess bundle compliance and stimulate instant course correction of noncompliant parameters, a real-time computerized dashboard was developed. Program impact in 6 adult intensive care units (ICUs) was assessed. Bundle compliance was noted as an overall cumulative bundle adherence assessment, reflecting the percentage of time all elements were concurrently in compliance for all patients. The VAP rate in all ICUs combined decreased from 19.5 to 9.2 VAPs per 1,000 ventilator-days following program implementation (P<.001). Bundle compliance significantly increased (Z100 score of 23% in August 2007 to 83% in June 2011 [P<.001]). The implementation resulted in a significant monthly decrease in the overall ICU VAP rate of 3.28/1,000 ventilator-days (95% CI, 2.64-3.92/1,000 ventilator-days). Following the intervention, the VAP rate decreased significantly at a rate of 0.20/1,000 ventilator-days per month (95% CI, 0.14-0.30/1,000 ventilator-days per month). Among all adult ICUs combined, improved bundle compliance was moderately correlated with monthly VAP rate reductions (Pearson correlation coefficient, -0.32). A prevention program using a real-time bundle adherence dashboard was associated with significant sustained decreases in VAP rates and an increase in bundle compliance among adult ICU patients.

  17. Wolff-Parkinson-White syndrome type B and left bundle-branch block: electrophysiologic and radionuclide study

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

    Rakovec, P.; Kranjec, I.; Fettich, J.J.

    1985-01-01

    Coinciding left bundle-branch block and Wolff-Parkinson-White syndrome type B, a very rare electrocardiographic occurrence, was found in a patient with dilated cardiomyopathy. Electrophysiologic study revealed eccentric retrograde atrial activation during ventricular pacing, suggesting right-sided accessory pathway. At programmed atrial pacing, effective refractory period of the accessory pathway was 310 ms; at shorter pacing coupling intervals, normal atrioventricular conduction with left bundle-branch block was seen. Left bundle-branch block was seen also with His bundle pacing. Radionuclide phase imaging demonstrated right ventricular phase advance and left ventricular phase delay; both right and left ventricular phase images revealed broad phase distribution histograms. Combinedmore » electrophysiologic and radionuclide investigations are useful to disclose complex conduction abnormalities and their mechanical correlates.« less

  18. Implementation of a Preventive Services Bundle in Academic Pediatric Primary Care Centers.

    PubMed

    Samaan, Zeina Marcho; Brown, Courtney M; Morehous, John; Perkins, Alison A; Kahn, Robert S; Mansour, Mona E

    2016-03-01

    Previous studies have documented poor rates of delivery of preventive services, 1 of the core services provided in the primary care medical home setting. We aimed to increase the reliability of delivering a bundle of preventive services to patients 0 to 14 months of age from 58% of patient visits to 95% of visits. The bundle includes administration of routine vaccinations, offering influenza vaccination, completed lead screening, completed developmental screening tool, screening for maternal depression and food insecurity, and documentation of gestational age. The setting was 3 academic pediatric primary care clinics that serve 31,000 patients (>90% Medicaid). Quality improvement methodology was used and key driver diagram was determined. Patient "Ideal Visit Flow" and the Responsible, Accountable, Support, Consulted, and Informed Matrix were developed to drive accountability for components of the ideal flow. Plan, Do, Study, Act cycles were used to develop successful interventions. The percent of patients seen who received all bundle elements for which they were eligible was plotted weekly on a run chart, and statistical process control methods were used to determine a significant change in performance. The preintervention percentage of patient visits ages 0 to 14 months receiving all preventive service bundle elements was 58%. The postintervention percentage is 92%. Innovative redesign led to improvement in percentage of patients age 0 to 14 months who received the entire preventive services bundle. Key elements for success were multidisciplinary site-specific teams, redesigned visit flow, effective communication, and resources for data and project management. Copyright © 2016 by the American Academy of Pediatrics.

  19. Bundle Data Approach at GES DISC Targeting Natural Hazards

    NASA Technical Reports Server (NTRS)

    Shie, Chung-Lin; Shen, Suhung; Kempler, Steven J.

    2015-01-01

    Severe natural phenomena such as hurricane, volcano, blizzard, flood and drought have the potential to cause immeasurable property damages, great socioeconomic impact, and tragic loss of human life. From searching to assessing the Big, i.e., massive and heterogeneous scientific data (particularly, satellite and model products) in order to investigate those natural hazards, it has, however, become a daunting task for Earth scientists and applications researchers, especially during recent decades. The NASA Goddard Earth Sciences Data and Information Service Center (GES DISC) has served Big Earth science data, and the pertinent valuable information and services to the aforementioned users of diverse communities for years. In order to help and guide our users to online readily (i.e., with a minimum effort) acquire their requested data from our enormous resource at GES DISC for studying their targeted hazard event, we have thus initiated a Bundle Data approach in 2014, first targeting the hurricane event topic. We have recently worked on new topics such as volcano and blizzard. The bundle data of a specific hazard event is basically a sophisticated integrated data package consisting of a series of proper datasets containing a group of relevant (knowledge--based) data variables readily accessible to users via a system-prearranged table linking those data variables to the proper datasets (URLs). This online approach has been developed by utilizing a few existing data services such as Mirador as search engine; Giovanni for visualization; and OPeNDAP for data access, etc. The online Data Cookbook site at GES DISC is the current host for the bundle data. We are now also planning on developing an Automated Virtual Collection Framework that shall eventually accommodate the bundle data, as well as further improve our management in Big Data.

  20. Arrays of Bundles of Carbon Nanotubes as Field Emitters

    NASA Technical Reports Server (NTRS)

    Manohara, Harish; Bronkowski, Michael

    2007-01-01

    Experiments have shown that with suitable choices of critical dimensions, planar arrays of bundles of carbon nanotubes (see figure) can serve as high-current-density field emitter (cold-cathode) electron sources. Whereas some hot-cathode electron sources must be operated at supply potentials of thousands of volts, these cold-cathode sources generate comparable current densities when operated at tens of volts. Consequently, arrays of bundles of carbon nanotubes might prove useful as cold-cathode sources in miniature, lightweight electron-beam devices (e.g., nanoklystrons) soon to be developed. Prior to the experiments, all reported efforts to develop carbon-nanotube-based field-emission sources had yielded low current densities from a few hundred microamperes to a few hundred milliamperes per square centimeter. An electrostatic screening effect, in which taller nanotubes screen the shorter ones from participating in field emission, was conjectured to be what restricts the emission of electrons to such low levels. It was further conjectured that the screening effect could be reduced and thus emission levels increased by increasing the spacing between nanotubes to at least by a factor of one to two times the height of the nanotubes. While this change might increase the emission from individual nanotubes, it would decrease the number of nanotubes per unit area and thereby reduce the total possible emission current. Therefore, to maximize the area-averaged current density, it would be necessary to find an optimum combination of nanotube spacing and nanotube height. The present concept of using an array of bundles of nanotubes arises partly from the concept of optimizing the spacing and height of field emitters. It also arises partly from the idea that single nanotubes may have short lifetimes as field emitters, whereas bundles of nanotubes could afford redundancy so that the loss of a single nanotube would not significantly reduce the overall field emission.