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Sample records for activating protein flap

  1. A Single Amino Acid Difference between Mouse and Human 5-Lipoxygenase Activating Protein (FLAP) Explains the Speciation and Differential Pharmacology of Novel FLAP Inhibitors.

    PubMed

    Blevitt, Jonathan M; Hack, Michael D; Herman, Krystal; Chang, Leon; Keith, John M; Mirzadegan, Tara; Rao, Navin L; Lebsack, Alec D; Milla, Marcos E

    2016-06-10

    5-Lipoxygenase activating protein (FLAP) plays a critical role in the metabolism of arachidonic acid to leukotriene A4, the precursor to the potent pro-inflammatory mediators leukotriene B4 and leukotriene C4 Studies with small molecule inhibitors of FLAP have led to the discovery of a drug binding pocket on the protein surface, and several pharmaceutical companies have developed compounds and performed clinical trials. Crystallographic studies and mutational analyses have contributed to a general understanding of compound binding modes. During our own efforts, we identified two unique chemical series. One series demonstrated strong inhibition of human FLAP but differential pharmacology across species and was completely inactive in assays with mouse or rat FLAP. The other series was active across rodent FLAP, as well as human and dog FLAP. Comparison of rodent and human FLAP amino acid sequences together with an analysis of a published crystal structure led to the identification of amino acid residue 24 in the floor of the putative binding pocket as a likely candidate for the observed speciation. On that basis, we tested compounds for binding to human G24A and mouse A24G FLAP mutant variants and compared the data to that generated for wild type human and mouse FLAP. These studies confirmed that a single amino acid mutation was sufficient to reverse the speciation observed in wild type FLAP. In addition, a PK/PD method was established in canines to enable preclinical profiling of mouse-inactive compounds. PMID:27129215

  2. A Single Amino Acid Difference between Mouse and Human 5-Lipoxygenase Activating Protein (FLAP) Explains the Speciation and Differential Pharmacology of Novel FLAP Inhibitors.

    PubMed

    Blevitt, Jonathan M; Hack, Michael D; Herman, Krystal; Chang, Leon; Keith, John M; Mirzadegan, Tara; Rao, Navin L; Lebsack, Alec D; Milla, Marcos E

    2016-06-10

    5-Lipoxygenase activating protein (FLAP) plays a critical role in the metabolism of arachidonic acid to leukotriene A4, the precursor to the potent pro-inflammatory mediators leukotriene B4 and leukotriene C4 Studies with small molecule inhibitors of FLAP have led to the discovery of a drug binding pocket on the protein surface, and several pharmaceutical companies have developed compounds and performed clinical trials. Crystallographic studies and mutational analyses have contributed to a general understanding of compound binding modes. During our own efforts, we identified two unique chemical series. One series demonstrated strong inhibition of human FLAP but differential pharmacology across species and was completely inactive in assays with mouse or rat FLAP. The other series was active across rodent FLAP, as well as human and dog FLAP. Comparison of rodent and human FLAP amino acid sequences together with an analysis of a published crystal structure led to the identification of amino acid residue 24 in the floor of the putative binding pocket as a likely candidate for the observed speciation. On that basis, we tested compounds for binding to human G24A and mouse A24G FLAP mutant variants and compared the data to that generated for wild type human and mouse FLAP. These studies confirmed that a single amino acid mutation was sufficient to reverse the speciation observed in wild type FLAP. In addition, a PK/PD method was established in canines to enable preclinical profiling of mouse-inactive compounds.

  3. Granulocyte/macrophage colony-stimulating factor stimulates the expression of the 5-lipoxygenase-activating protein (FLAP) in human neutrophils

    PubMed Central

    1994-01-01

    The synthesis of leukotrienes in human blood neutrophils chiefly relies on the activity of two enzymes, phospholipase A2 and 5-lipoxygenase (5- LO). In turn, the activation of the 5-LO requires the participation of a recently characterized membrane-bound protein, the 5-LO-activating protein (FLAP). In this study, we have investigated conditions under which FLAP expression in neutrophils may be modulated. Of several cytokines tested, only granulocyte/macrophage colony-stimulating factor (GM-CSF) (and to a lesser extent tumor necrosis factor alpha) significantly increased expression of FLAP. GM-CSF increased FLAP mRNA steady-state levels in a time- and dose-dependent manner. The stimulatory effect of GM-CSF on FLAP mRNA was inhibited by prior treatment of the cells with the transcription inhibitor, actinomycin D, and pretreatment of the cells with the protein synthesis inhibitor, cycloheximide, failed to prevent the increase in FLAP mRNA induced by GM-CSF. The accumulation of newly synthesized FLAP, as determined by immunoprecipitation after incorporation of 35S-labeled amino acids, was also increased after incubation of neutrophils with GM-CSF. In addition, the total level of FLAP protein was increased in GM-CSF- treated neutrophils, as determined by two-dimensional gel electrophoresis, followed by Western blot. GM-CSF did not alter the stability of the FLAP protein, indicating that the effect of GM-CSF on FLAP accumulation was the consequence of increased de novo synthesis as opposed to decreased degradation of FLAP. Finally, incubation of neutrophils with the synthetic glucocorticoid dexamethasone directly stimulated the upregulation of FLAP mRNA and protein, and enhanced the effect of GM-CSF. Taken together, these data demonstrate that FLAP expression may be upmodulated after appropriate stimulation of neutrophils. The increase in FLAP expression induced by GM-CSF in inflammatory conditions could confer upon neutrophils a prolonged capacity to synthesize

  4. 4,5-Diarylisoxazol-3-carboxylic acids: A new class of leukotriene biosynthesis inhibitors potentially targeting 5-lipoxygenase-activating protein (FLAP).

    PubMed

    Banoglu, Erden; Çelikoğlu, Erşan; Völker, Susanna; Olgaç, Abdurrahman; Gerstmeier, Jana; Garscha, Ulrike; Çalışkan, Burcu; Schubert, Ulrich S; Carotti, Andrea; Macchiarulo, Antonio; Werz, Oliver

    2016-05-01

    In this article, we report novel leukotriene (LT) biosynthesis inhibitors that may target 5-lipoxygenase-activating protein (FLAP) based on the previously identified isoxazole derivative (8). The design and synthesis was directed towards a subset of 4,5-diaryl-isoxazole-3-carboxylic acid derivatives as LT biosynthesis inhibitors. Biological evaluation disclosed a new skeleton of potential anti-inflammatory agents, exemplified by 39 and 40, which potently inhibit cellular 5-LO product synthesis (IC50 = 0.24 μM, each) seemingly by targeting FLAP with weak inhibition on 5-LO (IC50 ≥ 8 μM). Docking studies and molecular dynamic simulations with 5-LO and FLAP provide valuable insights into potential binding modes of the inhibitors. Together, these diaryl-isoxazol-3-carboxylic acids may possess potential as leads for development of effective anti-inflammatory drugs through inhibition of LT biosynthesis. PMID:26922224

  5. DNA and Protein Requirements for Substrate Conformational Changes Necessary for Human Flap Endonuclease-1-catalyzed Reaction.

    PubMed

    Algasaier, Sana I; Exell, Jack C; Bennet, Ian A; Thompson, Mark J; Gotham, Victoria J B; Shaw, Steven J; Craggs, Timothy D; Finger, L David; Grasby, Jane A

    2016-04-01

    Human flap endonuclease-1 (hFEN1) catalyzes the essential removal of single-stranded flaps arising at DNA junctions during replication and repair processes. hFEN1 biological function must be precisely controlled, and consequently, the protein relies on a combination of protein and substrate conformational changes as a prerequisite for reaction. These include substrate bending at the duplex-duplex junction and transfer of unpaired reacting duplex end into the active site. When present, 5'-flaps are thought to thread under the helical cap, limiting reaction to flaps with free 5'-terminiin vivo Here we monitored DNA bending by FRET and DNA unpairing using 2-aminopurine exciton pair CD to determine the DNA and protein requirements for these substrate conformational changes. Binding of DNA to hFEN1 in a bent conformation occurred independently of 5'-flap accommodation and did not require active site metal ions or the presence of conserved active site residues. More stringent requirements exist for transfer of the substrate to the active site. Placement of the scissile phosphate diester in the active site required the presence of divalent metal ions, a free 5'-flap (if present), a Watson-Crick base pair at the terminus of the reacting duplex, and the intact secondary structure of the enzyme helical cap. Optimal positioning of the scissile phosphate additionally required active site conserved residues Tyr(40), Asp(181), and Arg(100)and a reacting duplex 5'-phosphate. These studies suggest a FEN1 reaction mechanism where junctions are bound and 5'-flaps are threaded (when present), and finally the substrate is transferred onto active site metals initiating cleavage.

  6. DNA and Protein Requirements for Substrate Conformational Changes Necessary for Human Flap Endonuclease-1-catalyzed Reaction*

    PubMed Central

    Algasaier, Sana I.; Exell, Jack C.; Bennet, Ian A.; Thompson, Mark J.; Gotham, Victoria J. B.; Shaw, Steven J.; Craggs, Timothy D.; Finger, L. David; Grasby, Jane A.

    2016-01-01

    Human flap endonuclease-1 (hFEN1) catalyzes the essential removal of single-stranded flaps arising at DNA junctions during replication and repair processes. hFEN1 biological function must be precisely controlled, and consequently, the protein relies on a combination of protein and substrate conformational changes as a prerequisite for reaction. These include substrate bending at the duplex-duplex junction and transfer of unpaired reacting duplex end into the active site. When present, 5′-flaps are thought to thread under the helical cap, limiting reaction to flaps with free 5′-termini in vivo. Here we monitored DNA bending by FRET and DNA unpairing using 2-aminopurine exciton pair CD to determine the DNA and protein requirements for these substrate conformational changes. Binding of DNA to hFEN1 in a bent conformation occurred independently of 5′-flap accommodation and did not require active site metal ions or the presence of conserved active site residues. More stringent requirements exist for transfer of the substrate to the active site. Placement of the scissile phosphate diester in the active site required the presence of divalent metal ions, a free 5′-flap (if present), a Watson-Crick base pair at the terminus of the reacting duplex, and the intact secondary structure of the enzyme helical cap. Optimal positioning of the scissile phosphate additionally required active site conserved residues Tyr40, Asp181, and Arg100 and a reacting duplex 5′-phosphate. These studies suggest a FEN1 reaction mechanism where junctions are bound and 5′-flaps are threaded (when present), and finally the substrate is transferred onto active site metals initiating cleavage. PMID:26884332

  7. Power performance optimization and loads alleviation with active flaps using individual flap control

    NASA Astrophysics Data System (ADS)

    Pettas, Vasilis; Barlas, Thanasis; Gertz, Drew; Madsen, Helge A.

    2016-09-01

    The present article investigates the potential of Active Trailing Edge Flaps (ATEF) in terms of increase in annual energy production (AEP) as well as reduction of fatigue loads. The basis for this study is the DTU 10 MW Reference Wind Turbine (RWT) simulated using the aeroelastic code HAWC2. In an industrial-oriented manner the baseline rotor is upscaled by 5% and the ATEFs are implemented in the outer 30% of the blades. The flap system is kept simple and robust with a single flap section and control with wind speed, rotor azimuth, root bending moments and angle of attack in flap's mid-section being the sensor inputs. The AEP is increased due to the upscaling but also further due to the flap system while the fatigue loads in components of interest (blade, tower, nacelle and main bearing) are reduced close to the level of the original turbine. The aim of this study is to demonstrate a simple and applicable method that can be a technology enabler for rotor upscaling and lowering cost of energy.

  8. Active Control of a Flapping Wing in a Gust Setup

    NASA Astrophysics Data System (ADS)

    Wallace, Ryan; Anderson, Mark; Glauser, Mark

    2006-11-01

    The aim of this experiment is to determine the response of a flapping Micro Air Vehicle wing to a wind gust while in forward and hovering flight and apply an active control to respond to the wind gust. The flapping wing is driven by a DC brushless motor which is geared to allow for flapping at frequencies up to 3 Hz. The wing is set up vertically in the wind tunnel, and can flap up to angles of 120 degrees. To simulate a wind gust perpendicular to the free stream flow a diffuser is set up on top of the wind tunnel. Strain gages are attached to the wing. It has been shown while simultaneously measuring the dynamical strain and the velocity field with a PIV system, that a realistic estimate of the wake flow field can obtained using low dimensional tools (POD, mLSE). The wing and the flapping mechanism are mounted directly on a force balance to calculate the lift being produced. In order to prevent flow separation on the wing during a sudden wind gust the wing is actively deformed by an attached piezoelectric actuator. The end result is to have closed loop control to produce stable hovering and forward flight.

  9. Active Flow Control of Lifting Surface With Flap-Current Activities and Future Directions

    NASA Technical Reports Server (NTRS)

    Ahmadi, G.; Marzocca, P.; Jha, R.; Alstorm, B.; Obied, S.; Kabir, P.; Shahrabi, A.

    2010-01-01

    The main objective is to develop effective control strategies for separation control of an airfoil with a single hinge flap. The specific objectives are: Develop an active control architecture for flow control around an airfoil with flap. Design, fabricate, a wind tunnel test of a high lift wing (with flap) with integrated actuators and sensors. Design, development and fabrication of synthetic jet actuators. Develop appropriate control strategy for application to the airfoil. Wind tunnel testing of the high lift wing at various angles of attack and flap positions with closed loop control.

  10. Localized, Non-Harmonic Active Flap Motions for Low Frequency In-Plane Rotor Noise Reduction

    NASA Technical Reports Server (NTRS)

    Sim, Ben W.; Potsdam, Mark; Kitaplioglu, Cahit; LeMasurier, Philip; Lorber, Peter; Andrews, Joseph

    2012-01-01

    A first-of-its-kind demonstration of the use of localized, non-harmonic active flap motions, for suppressing low frequency, in-plane rotor noise, is reported in this paper. Operational feasibility is verified via testing of the full-scale AATD/Sikorsky/UTRC active flap demonstration rotor in the NFAC's 40- by 80-Foot anechoic wind tunnel. Effectiveness of using localized, non-harmonic active flap motions are compared to conventional four-per-rev harmonic flap motions, and also active flap motions derived from closed-loop acoustics implementations. All three approaches resulted in approximately the same noise reductions over an in-plane three-by-three microphone array installed forward and near in-plane of the rotor in the nearfield. It is also reported that using an active flap in this localized, non-harmonic manner, resulted in no more that 2% rotor performance penalty, but had the tendency to incur higher hub vibration levels.

  11. Dynamics of an Active-Site Flap Contributes to Catalysis in a JAMM Family Metallo Deubiquitinase.

    PubMed

    Bueno, Amy N; Shrestha, Rashmi K; Ronau, Judith A; Babar, Aditya; Sheedlo, Michael J; Fuchs, Julian E; Paul, Lake N; Das, Chittaranjan

    2015-10-01

    The endosome-associated deubiquitinase (DUB) AMSH is a member of the JAMM family of zinc-dependent metallo isopeptidases with high selectivity for Lys63-linked polyubiquitin chains, which play a key role in endosomal-lysosomal sorting of activated cell surface receptors. The catalytic domain of the enzyme features a flexible flap near the active site that opens and closes during its catalytic cycle. Structural analysis of its homologues, AMSH-LP (AMSH-like protein) and the fission yeast counterpart, Sst2, suggests that a conserved Phe residue in the flap may be critical for substrate binding and/or catalysis. To gain insight into the contribution of this flap in substrate recognition and catalysis, we generated mutants of Sst2 and characterized them using a combination of enzyme kinetics, X-ray crystallography, molecular dynamics simulations, and isothermal titration calorimetry (ITC). Our analysis shows that the Phe residue in the flap contributes key interactions during the rate-limiting step but not to substrate binding, since mutants of Phe403 exhibit a defect only in kcat but not in KM. Moreover, ITC studies show Phe403 mutants have similar KD for ubiquitin compared to the wild-type enzyme. The X-ray structures of both Phe403Ala and the Phe403Trp, in both the free and ubiquitin bound form, reveal no appreciable structural change that might impair substrate or alter product binding. We observed that the side chain of the Trp residue is oriented identically with respect to the isopeptide moiety of the substrate as the Phe residue in the wild-type enzyme, so the loss of activity seen in this mutant cannot be explained by the absence of a group with the ability to provide van der Waals interactions that facilitate the hyrdolysis of the Lys63-linked diubiquitin. Molecular dynamics simulations indicate that the flap in the Trp mutant is quite flexible, allowing almost free rotation of the indole side chain. Therefore, it is possible that these different dynamic

  12. Enhanced Correlation of SMART Active Flap Rotor Loads

    NASA Technical Reports Server (NTRS)

    Kottapalli, Sesi

    2011-01-01

    This is a follow-on study to a 2010 correlation effort. Measured data from the SMART rotor test in the NASA Ames 40- by 80- Foot Wind Tunnel are compared with CAMRAD II calculations. As background, during the wind tunnel test, unexpectedly high inboard loads were encountered, and it was hypothesized at that time that due to changes in the flexbeams over the years, the flexbeam properties used in the analysis needed updating. Boeing Mesa, recently updated these properties. This correlation study uses the updated flexbeam properties. Compared to earlier studies, the following two enhancements are implemented: i) the inboard loads (pitchcase and flexbeam loads) correlation is included for the first time (reliable prediction of the inboard loads is a prerequisite for any future anticipated flight-testing); ii) the number of blade modes is increased to better capture the flap dynamics and the pitchcase-flexbeam dynamics. Also, aerodynamically, both the rolled-up wake model and the more complex, multiple trailer wake model are used, with the latter slightly improving the blade chordwise moment correlation. This sensitivity to the wake model indicates that CFD is needed. Three high-speed experimental cases, one uncontrolled free flap case and two commanded flap cases, are considered. The two commanded flap cases include a 2o flap deflection at 5P case and a 0o flap deflection case. For the free flap case, selected modifications to the HH-06 section flap airfoil pitching moment table are implemented. For the commanded 2o flap case, the experimental flap variation is approximately matched by increasing the analytical flap hinge stiffness. This increased flap hinge stiffness is retained for the commanded 0o flap case also, which is treated as a free flap case, but with larger flap hinge stiffness. The change in the mid-span and outboard loads correlation due to the updating of the flexbeam properties is not significant. Increasing the number of blade modes results in an

  13. Prefabrication of bone by use of a vascularized periosteal flap and bone morphogenetic protein.

    PubMed

    Vögelin M D, E; Jones, N F; Lieberman, J R; Baker, J M; Tsingotjidou, A S; Brekke, J H

    2002-01-01

    The purpose of this pilot study was to prefabricate a vascularized bone graft by using a vascularized periosteal flap containing osteoprogenitor cells, a structural matrix, and recombinant human bone morphogenetic protein-2 (rhBMP-2). In a rat model, a periosteal flap vascularized by the saphenous artery and vein was dissected off the medial surface of the tibia. This flap consisted of three layers-periosteum, muscle, and fascia-and was tubed on itself to form a watertight chamber that was then transferred on its vascular pedicle to the groin. A total of 78 vascularized periosteal chambers were constructed in 39 animals and divided into 10 groups. In group 1, the periosteal chamber was left empty. Groups 2, 3, and 4 consisted of the periosteal flap and rhBMP-2, but in group 3, the proximal vascular pedicle was ligated, and in group 4, the flap was harvested without the periosteal layer and turned inside out. Groups 5 through 10 consisted of the vascularized periosteal flap containing several different structural matrices (calcium alginate spheres, polylactic acid, or demineralized bone matrix) with or without rhBMP-2. Animals were killed at 2, 4, or 8 weeks in each group. The presence and density of any new bone formation was evaluated both radiologically and histologically. Significant bone formation was seen only in those periosteal flaps containing rhBMP-2 and either the calcium alginate or polylactic acid matrix. New bone formation increased both radiologically and histologically from 2 weeks to 8 weeks only in the periosteal flaps containing the polylactic acid matrix and rhBMP-2. This preliminary study therefore suggests that four factors-blood supply, osteoprogenitor cells in the periosteal layer, a biodegradable matrix, and rhBMP-2-are required for optimal prefabrication of a vascularized bone graft.

  14. Active Control of Separation From the Flap of a Supercritical Airfoil

    NASA Technical Reports Server (NTRS)

    Melton, La Tunia Pack; Yao, Chung-Sheng; Seifert, Avi

    2003-01-01

    Active flow control in the form of periodic zero-mass-flux excitation was applied at several regions on the leading edge and trailing edge flaps of a simplified high-lift system t o delay flow separation. The NASA Energy Efficient Transport (EET) supercritical airfoil was equipped with a 15% chord simply hinged leading edge flap and a 25% chord simply hinged trailing edge flap. Detailed flow features were measured in an attempt to identify optimal actuator placement. The measurements included steady and unsteady model and tunnel wall pressures, wake surveys, arrays of surface hot-films, flow visualization, and particle image velocimetry (PIV). The current paper describes the application of active separation control at several locations on the deflected trailing edge flap. High frequency (F(+) approx.= 10) and low frequency amplitude modulation (F(+)AM approx.= 1) of the high frequency excitation were used for control. Preliminary efforts to combine leading and trailing edge flap excitations are also reported.

  15. N protein from lambdoid phages transforms NusA into an antiterminator by modulating NusA-RNA polymerase flap domain interactions

    PubMed Central

    Mishra, Saurabh; Sen, Ranjan

    2015-01-01

    Interaction of the lambdoid phage N protein with the bacterial transcription elongation factor NusA is the key component in the process of transcription antitermination. A convex surface of E. coli NusA-NTD, located opposite to its RNA polymerase-binding domain (the β-flap domain), directly interacts with N in the antitermination complex. We hypothesized that this N-NusA interaction induces allosteric effects on the NusA-RNAP interaction leading to transformation of NusA into a facilitator of the antitermination process. Here we showed that mutations in β-flap domain specifically defective for N antitermination exhibited altered NusA-nascent RNA interaction and have widened RNA exit channel indicating an intricate role of flap domain in the antitermination. The presence of N reoriented the RNAP binding surface of NusA-NTD, which changed its interaction pattern with the flap domain. These changes caused significant spatial rearrangement of the β-flap as well as the β′ dock domains to form a more constricted RNA exit channel in the N-modified elongation complex (EC), which might play key role in converting NusA into a facilitator of the N antitermination. We propose that in addition to affecting the RNA exit channel and the active center of the EC, β-flap domain rearrangement is also a mechanistic component in the N antitermination process. PMID:25990722

  16. Human exonuclease 1 (EXO1) activity characterization and its function on flap structures

    PubMed Central

    Keijzers, Guido; Bohr, Vilhelm A.; Rasmussen, Lene Juel

    2015-01-01

    Human exonuclease 1 (EXO1) is involved in multiple DNA metabolism processes, including DNA repair and replication. Most of the fundamental roles of EXO1 have been described in yeast. Here, we report a biochemical characterization of human full-length EXO1. Prior to assay EXO1 on different DNA flap structures, we determined factors essential for the thermodynamic stability of EXO1. We show that enzymatic activity and stability of EXO1 on DNA is modulated by temperature. By characterization of EXO1 flap activity using various DNA flap substrates, we show that EXO1 has a strong capacity for degrading double stranded DNA and has a modest endonuclease or 5′ flap activity. Furthermore, we report novel mechanistic insights into the processing of flap structures, showing that EXO1 preferentially cleaves one nucleotide inwards in a double stranded region of a forked and nicked DNA flap substrates, suggesting a possible role of EXO1 in strand displacement. PMID:26182368

  17. Active Management of Flap-Edge Trailing Vortices

    NASA Technical Reports Server (NTRS)

    Greenblatt, David; Yao, Chung-Sheng; Vey, Stefan; Paschereit, Oliver C.; Meyer, Robert

    2008-01-01

    The vortex hazard produced by large airliners and increasingly larger airliners entering service, combined with projected rapid increases in the demand for air transportation, is expected to act as a major impediment to increased air traffic capacity. Significant reduction in the vortex hazard is possible, however, by employing active vortex alleviation techniques that reduce the wake severity by dynamically modifying its vortex characteristics, providing that the techniques do not degrade performance or compromise safety and ride quality. With this as background, a series of experiments were performed, initially at NASA Langley Research Center and subsequently at the Berlin University of Technology in collaboration with the German Aerospace Center. The investigations demonstrated the basic mechanism for managing trailing vortices using retrofitted devices that are decoupled from conventional control surfaces. The basic premise for managing vortices advanced here is rooted in the erstwhile forgotten hypothesis of Albert Betz, as extended and verified ingeniously by Coleman duPont Donaldson and his collaborators. Using these devices, vortices may be perturbed at arbitrarily long wavelengths down to wavelengths less than a typical airliner wingspan and the oscillatory loads on the wings, and hence the vehicle, are small. Significant flexibility in the specific device has been demonstrated using local passive and active separation control as well as local circulation control via Gurney flaps. The method is now in a position to be tested in a wind tunnel with a longer test section on a scaled airliner configuration. Alternatively, the method can be tested directly in a towing tank, on a model aircraft, a light aircraft or a full-scale airliner. The authors believed that this method will have significant appeal from an industry perspective due to its retrofit potential with little to no impact on cruise (devices tucked away in the cove or retracted); low operating power

  18. Flap Endonuclease 1

    PubMed Central

    Balakrishnan, Lata; Bambara, Robert A.

    2013-01-01

    First discovered as a structure-specific endonuclease that evolved to cut at the base of single-stranded flaps, flap endonuclease (FEN1) is now recognized as a central component of cellular DNA metabolism. Substrate specificity allows FEN1 to process intermediates of Okazaki fragment maturation, long-patch base excision repair, telomere maintenance, and stalled replication fork rescue. For Okazaki fragments, the RNA primer is displaced into a 5′ flap and then cleaved off. FEN1 binds to the flap base and then threads the 5′ end of the flap through its helical arch and active site to create a configuration for cleavage. The threading requirement prevents this active nuclease from cutting the single-stranded template between Okazaki fragments. FEN1 efficiency and specificity are critical to the maintenance of genome fidelity. Overall, recent advances in our knowledge of FEN1 suggest that it was an ancient protein that has been fine-tuned over eons to coordinate many essential DNA transactions. PMID:23451868

  19. Role of induced vortex interaction in a semi-active flapping foil based energy harvester

    NASA Astrophysics Data System (ADS)

    Wu, J.; Chen, Y. L.; Zhao, N.

    2015-09-01

    The role of induced vortex interaction in a semi-active flapping foil based energy harvester is numerically examined in this work. A NACA0015 airfoil, which acts as an energy harvester, is placed in a two-dimensional laminar flow. It performs an imposed pitching motion that subsequently leads to a plunging motion. Two auxiliary smaller foils, which rotate about their centers, are arranged above and below the flapping foil, respectively. As a consequence, the vortex interaction between the flapping foil and the rotating foil is induced. At a Reynolds number of 1100 and the position of the pitching axis at one-third chord, the effects of the distance between two auxiliary foils, the phase difference between the rotating motion and the pitching motion as well as the frequency of pitching motion on the power extraction performance are systematically investigated. It is found that compared to the single flapping foil, the efficiency improvement of overall power extraction for the flapping foil with two auxiliary foils can be achieved. Based on the numerical analysis, it is indicated that the enhanced power extraction, which is caused by the increased lift force, thanks to the induced vortex interaction, directly benefits the efficiency enhancement.

  20. Structural integrity design for an active helicopter rotor blade with piezoelectric flap actuators

    NASA Astrophysics Data System (ADS)

    Lee, Jaehwan; Shin, SangJoon

    2011-04-01

    Helicopter uses a rotor system to generate lift, thrust and forces, and its aerodynamic environment is generally complex. Unsteady aerodynamic environment arises such as blade vortex interaction. This unsteady aerodynamic environment induces vibratory aerodynamic loads and high aeroacoustic noise. The aerodynamic load and aeroacoustic noise is at N times the rotor blade revolutions (N/rev). But conventional rotor control system composed of pitch links and swash plate is not capable of adjusting such vibratory loads because its control is restricted to 1/rev. Many active control methodologies have been examined to alleviate the problem. The blade using active control device manipulates the blade pitch angle with N/rev. In this paper, Active Trailing-edge Flap blade, which is one of the active control methods, is designed to reduce the unsteady aerodynamic loads. Active Trailing-edge Flap blade uses a trailing edge flap manipulated by an actuator to change camber line of the airfoil. Piezoelectric actuators are installed inside the blade to manipulate the trailing edge flap.

  1. Pivoting output unit control systems activated by jacks. [for controlling aircraft flaps

    NASA Technical Reports Server (NTRS)

    Belliere, P.

    1978-01-01

    An invention to be used for controlling aircraft flaps is described. It is applicable to control systems with two coaxial output units which pivot simultaneously with respect to two fixed units and which are activated by two opposed, straight coaxial jacks.

  2. The reduction of rotorcraft power and vibration using optimally controlled active gurney flap

    NASA Astrophysics Data System (ADS)

    Bae, Eui Sung

    The main topic of the present study is the application of active control scheme for the reduction of rotorcraft main rotor power reduction and vibratory load. When the helicopter is operated near its flight boundary, the required power and vibratory loads rapidly increases which impose a limit on the helicopter operation. Various methods were proposed and studied in order to achieve performance improvement under such operating condition. The effect of active control scheme was examined for its impact on the performance improvement and vibration reduction in the present study. Numerical simulations are based on the UH-60A Blackhawk helicopter with an active Gurney flap spanning from 70%R to 80%R of the main rotor. For obtaining the aeroelastic response of the rotor blade, finite element method was used to represent elastic blade. The aerodynamic loads acting on the blade are provided by CFD based 2D lookup table. Prescribed wake model was used to resolve the induced inflow over the rotor disk. The unsteady aerodynamic behavior due to the higher harmonic actuation of active Gurney flap was resolved by the time-domain unsteady aerodynamic model. The first part of preliminary study covers parametric study using Gurney flap. Starting with simple rigid blade representation of the rotor blade, the effect of 1/rev Gurney flap actuation was examined on three different gross weights. The effect of active Gurney flap width, the chordwise location of active Gurney flap, the effect of unsteady aerodynamic model, and the effect of 2/rev actuation frequency were examined. The second part of preliminary study was conducted with the elastic blade model to include the effect of torsion dynamics. Performance improvement using active Gurney flap was examined for maximizing thrust capability at two flight speeds. 1/rev Gurney flap actuation increased the gross weight capability up to 1,000 lbs. Also, 1/rev actuation of Gurney flap increased maximum altitude limit of baseline rotor by 1

  3. Application of Sequential Quadratic Programming to Minimize Smart Active Flap Rotor Hub Loads

    NASA Technical Reports Server (NTRS)

    Kottapalli, Sesi; Leyland, Jane

    2014-01-01

    In an analytical study, SMART active flap rotor hub loads have been minimized using nonlinear programming constrained optimization methodology. The recently developed NLPQLP system (Schittkowski, 2010) that employs Sequential Quadratic Programming (SQP) as its core algorithm was embedded into a driver code (NLP10x10) specifically designed to minimize active flap rotor hub loads (Leyland, 2014). Three types of practical constraints on the flap deflections have been considered. To validate the current application, two other optimization methods have been used: i) the standard, linear unconstrained method, and ii) the nonlinear Generalized Reduced Gradient (GRG) method with constraints. The new software code NLP10x10 has been systematically checked out. It has been verified that NLP10x10 is functioning as desired. The following are briefly covered in this paper: relevant optimization theory; implementation of the capability of minimizing a metric of all, or a subset, of the hub loads as well as the capability of using all, or a subset, of the flap harmonics; and finally, solutions for the SMART rotor. The eventual goal is to implement NLP10x10 in a real-time wind tunnel environment.

  4. Design of flapping wings for application to single active degree of freedom micro air vehicles

    NASA Astrophysics Data System (ADS)

    Chang, Kelvin Thomas

    This dissertation covers an experimental program to understand how wing compliance influences the performance of flapping micro air vehicle wings. The focus is the design of a membraned flapping wing for a single active degree of freedom mechanism, looking to maximize thrust performance in hover conditions. The optimization approach is unique in that experiments were the chosen engine as opposed to a computation model; this is because of the complexity involved in hover-mode flapping aerodynamics. The flapping mechanism and manufacturing process for fabricating the wings were carefully developed. The uncertainty in the thrust measurement was identified and reduced by implementing precision machining and repeatable techniques for fabrication. This resulted in a reduction of the manufacturing coefficient of variation from 16.8% to 2.6%. Optimization was then conducted for a single objective (Maximize thrust), using a three parameter design space, finding the highest thrust performance in wings with high aspect ratio; then, a multi-objective optimization was conducted with two objectives (Thrust and Power) and a four parameter space. The research then shifted focus to identifying the stiffness and deformation characteristics of high performance wing designs. Static stiffness measurements with a simple line load suggested that high chordwise stiffness or lower spanwise stiffness would be favorable for aerodynamic performance. To explore more components of the deformation, a full-field imaging technique was used and a uniform load was substituted to engage with the membrane. It was found that there is a range of torsional compliance where the wing is most efficient especially at higher flapping frequencies. The final component of the study was the dynamic deformation measurement. The two system, four camera digital image correlation setup uses stroboscopic measurement to capture the wing deformation. The phase shift between the twist and stroke, and the tip deflection

  5. Reduced In-Plane, Low Frequency Helicopter Noise of an Active Flap Rotor

    NASA Technical Reports Server (NTRS)

    Sim, Ben W.; Janakiram, Ram D.; Barbely, Natasha L.; Solis, Eduardo

    2009-01-01

    Results from a recent joint DARPA/Boeing/NASA/Army wind tunnel test demonstrated the ability to reduce in-plane, low frequency noise of the full-scale Boeing-SMART rotor using active flaps. Test data reported in this paper illustrated that acoustic energy in the first six blade-passing harmonics could be reduced by up to 6 decibels at a moderate airspeed, level flight condition corresponding to advance ratio of 0.30. Reduced noise levels were attributed to selective active flap schedules that modified in-plane blade airloads on the advancing side of the rotor, in a manner, which generated counteracting acoustic pulses that partially offset the negative pressure peaks associated with in-plane, steady thickness noise. These favorable reduced-noise operating states are a strong function of the active flap actuation amplitude, frequency and phase. The associated noise reductions resulted in reduced aural detection distance by up to 18%, but incurred significant vibratory load penalties due to increased hub shear forces. Small reductions in rotor lift-to-drag ratios, of no more than 3%, were also measured

  6. Active Control of Separation From the Flap of a Supercritical Airfoil

    NASA Technical Reports Server (NTRS)

    Melton, LaTunia Pack; Yao, Chung-Sheng; Seifert, Avi

    2006-01-01

    Zero-mass-flux periodic excitation was applied at several regions on a simplified high-lift system to delay the occurrence of flow separation. The NASA Energy Efficient Transport (EET) supercritical airfoil was equipped with a 15% chord simply hinged leading edge flap and a 25% chord simply hinged trailing edge flap. Detailed flow features were measured in an attempt to identify optimal actuator placement. The measurements included steady and unsteady model and tunnel wall pressures, wake surveys, arrays of surface hot-films, flow visualization, and particle image velocimetry (PIV). The current paper describes the application of active separation control at several locations on the deflected trailing edge flap. High frequency (F(+) approximately equal to 10) and low frequency amplitude modulation (F(+) sub AM approximately equal to 1) of the high frequency excitation were used for control. It was noted that the same performance gains were obtained with amplitude modulation and required only 30% of the momentum input required by pure sine excitation.

  7. Pharmacologic partial salvage of a failing free flap with recombinant tissue plasminogen activator (rt-PA).

    PubMed

    Atiyeh, B S; Fuleihan, N S; Musharafieh, R S

    1999-11-01

    Despite all the technical improvements in microvascular surgery and the experience gained in clinical practice, thrombosis at the site of microanastomosis remains a significant problem and a continuous source of frustration to most microsurgeons. Early recognition of vascular complications and prompt reexploration with vascular revision remain an essential and standard conduct for salvage. However, in situations where conditions for no-reflow have been established due to severe vasospasm or prolonged ischemia time, it becomes obvious that surgical reexploration alone is not enough to salvage a failing flap or a replanted limb. In such situations, the loss of the revascularized tissues seems to be inevitable. The authors describe their experience in partially salvaging a failing free flap with recombinant tissue plasminogen activator (rt-PA), reversing an established state of no-reflow. Pharmacologic manipulation of the complex and variable factors influencing anastomotic patency in microvascular tissue transfer seems to offer a new hope for preventing failures, as well as for salvaging failing flaps. It appears also that free-tissue transfer failure is not an all-or-none phenomenon.

  8. Effects of vascular endothelial growth factor on survival of surgical flaps: a review of experimental studies.

    PubMed

    Fang, Taolin; Lineaweaver, William C; Chen, Michael B; Kisner, Carson; Zhang, Feng

    2014-01-01

    Partial or complete necrosis of skin flaps remains a significant problem in plastic and reconstructive surgery. Growth factors have shown promise in improving flap survival through increased angiogenesis and blood supply to the flap. Vascular endothelial growth factor (VEGF) is the most widely investigated and successful one. But the mechanisms of the effects are still not very clear. In the course of a series of experiments, we indicated that tissue survival of surgical flaps could be improved by both preoperative (sustained phase effect) and intraoperative (acute phase effect) application of VEGF. We reviewed both experimental and clinical investigations on the use of VEGF with surgical flaps to summarize the evidence of both phases of VEGF activity in promotion of flaps survival in detail. With the combinations of acute and sustained phases of effects, VEGF protein and gene, VEGF morphologic actions, and VEGF histochemical modulations suggest a pattern of VEGF activity that can be superimposed on classic descriptive mechanisms of tissue survival of flaps.

  9. Active interrogation of helicopter main rotor faults using trailing edge flap actuation

    NASA Astrophysics Data System (ADS)

    Stevens, Patricia Lynn

    Over the past decade, the helicopter community has become increasingly interested in health monitoring. The rotor system, however, is not sufficiently covered in the current Health and Usage Monitoring Systems (HUMS). This dissertation describes the development and evaluation of a new approach for detecting helicopter rotor faults in which active trailing edge flaps are used to interrogate the system. This work is based on the presumption that trailing edge flaps would be installed for the primary purpose of vibration and/or noise control; health monitoring is a secondary use. Using this approach, the blade is excited by an interrogation signal, which is a low amplitude oscillation at a few discrete frequencies. The blade response is measured and the health of the system is determined using a frequency domain damage identification algorithm. Damage detection and location are achieved via the residual force vector. The residual force vector, coupled with an understanding of the system physics, also provides nature characterization. Quantification of damage extent is achieved via a frequency domain adaptation of the Asymmetric Minimum Rank Perturbation Theory. The active interrogation system is evaluated using an aeroelastic finite element model of the rotor system in hover, including an advanced unsteady aerodynamic model to predict the trailing edge flap loads. Realistic damage models, including distributed bending stiffness damage, torsional stiffness damage, control system stiffness damage, cracks and ballistic damage, are seeded in the rotor system model. Results demonstrate detection, location and quantification of extent of all of the faults tested. The effects of noise and modeling errors are discussed and mitigation techniques are developed. Additionally, a measurability study is included. Benefits of this work include both improved health monitoring for rotorcraft as well as insights into the application of structural damage detection algorithms to a

  10. Numerical analysis of active chordwise flexibility on the performance of non-symmetrical flapping airfoils

    NASA Astrophysics Data System (ADS)

    Tay, W. B.; Lim, K. B.

    2010-01-01

    This paper investigates the effect of active chordwise flexing on the lift, thrust and propulsive efficiency of three types of airfoils. The factors studied are the flexing center location, standard two-sided flexing as well as a type of single-sided flexing. The airfoils are simulated to flap with four configurations, and the effects of flexing under these configurations are investigated. Results show that flexing is not necessarily beneficial for the performance of the airfoils. However, with the correct parameters, efficiency is as high as 0.76 by placing the flexing centre at the trailing edge. The average thrust coefficient is more than twice as high, from 1.63 to 3.57 with flapping and flexing under the right conditions. Moreover, the single-sided flexing also gives an average lift coefficient as high as 4.61 for the S1020 airfoil. The shape of the airfoil does alter the effect of flexing too. Deviating the flexing phase angle away from 90° does not give a significant improvement to the airfoil’s performance. These results greatly enhance the design of a better performing ornithopter wing.

  11. Complementary non-radioactive assays for investigation of human flap endonuclease 1 activity

    PubMed Central

    Dorjsuren, Dorjbal; Kim, Daemyung; Maloney, David J.; Wilson, David M.; Simeonov, Anton

    2011-01-01

    FEN1, a key participant in DNA replication and repair, is the major human flap endonuclease that recognizes and cleaves flap DNA structures. Deficiencies in FEN1 function or deletion of the fen1 gene have profound biological effects, including the suppression of repair of DNA damage incurred from the action of various genotoxic agents. Given the importance of FEN1 in resolving abnormal DNA structures, inhibitors of the enzyme carry a potential as enhancers of DNA-interactive anticancer drugs. To facilitate the studies of FEN1 activity and the search for novel inhibitors, we developed a pair of complementary-readout homogeneous assays utilizing fluorogenic donor/quencher and AlphaScreen chemiluminescence strategies. A previously reported FEN1 inhibitor 3-hydroxy-5-methyl-1-phenylthieno[2,3-d]pyrimidine-2,4(1H,3H)-dione displayed equal potency in the new assays, in agreement with its published IC50. The assays were optimized to a low 4 µl volume and used to investigate a set of small molecules, leading to the identification of previously-unreported FEN1 inhibitors, among which aurintricarboxylic acid and NSC-13755 (an arylstibonic derivative) displayed submicromolar potency (average IC50 of 0.59 and 0.93 µM, respectively). The availability of these simple complementary assays obviates the need for undesirable radiotracer-based assays and should facilitate efforts to develop novel inhibitors for this key biological target. PMID:21062821

  12. Thermostable flap endonuclease from the archaeon, Pyrococcus horikoshii, cleaves the replication fork-like structure endo/exonucleolytically.

    PubMed

    Matsui, E; Kawasaki, S; Ishida, H; Ishikawa, K; Kosugi, Y; Kikuchi, H; Kawarabayashi, Y; Matsui, I

    1999-06-25

    The flap endonuclease gene homologue from the hyperthermophilic archaeon, Pyrococcus horikoshii, was overexpressed in Escherichia coli and purified. The results of gel filtration indicated that this protein was a 41-kDa monomer. P. horikoshii flap endonuclease (phFEN) cleaves replication fork-like substrates (RF) and 5' double-strand flap structures (DF) using both flap endonuclease and 5'-3'-exonuclease activities. The mammalian flap endonuclease (mFEN) is a single-strand flap-specific endonuclease (Harrington, J. J., and Lieber, M. R. (1994) EMBO J. 13, 1235-1246), but the action patterns of phFEN appear to be quite different from those of mFEN at this point. The DF-specific flap endonuclease and 5'-exonuclease activities have not yet been reported. Therefore, this is the first report of the specific endo/exonuclease activities of phFEN. The DF-specific 5'-exonuclease activity degraded the downstream primer of 3' single-flap structure and was 15 times higher than the activities against nicked substrates without 3' flap strand. DF-specific flap endonuclease cleaved the 5' double-flap strand in DF and the lagging strand in RF at the junction portion. Because the RF appears to be the intermediate structure, due to the arrest of the replication fork, the double strand breaks after the arrests of the replication forks are probably caused by phFEN.

  13. Expression, purification and crystallization of human 5-lipoxygenase-activating protein with leukotriene-biosynthesis inhibitors

    SciTech Connect

    Xu, Shihua; McKeever, Brian M.; Wisniewski, Douglas; Miller, Douglas K.; Spencer, Robert H.; Chu, Lin; Ujjainwalla, Feroze; Yamin, Ting-Ting; Evans, Jilly F.; Becker, Joseph W.; Ferguson, Andrew D.

    2007-12-01

    The expression, purification and crystallization of human 5-lipoxygenase-activating protein in complex with two leukotriene-biosynthesis inhibitors is decribed. The processes that were used to generate diffraction quality crystals are presented in detail. The nuclear membrane protein 5-lipoxygenase-activating protein (FLAP) plays an essential role in leukotriene synthesis. Recombinant full-length human FLAP with a C-terminal hexahistidine tag has been expressed and purified from the cytoplasmic membrane of Escherichia coli. Diffraction-quality crystals of FLAP in complex with leukotriene-synthesis inhibitor MK-591 and with an iodinated analogue of MK-591 have been grown using the sitting-drop vapor-diffusion method. The crystals exhibit tetragonal symmetry (P42{sub 1}2) and diffracted to a resolution limit of 4 Å.

  14. Foot and ankle reconstruction: an experience on the use of 14 different flaps in 226 cases.

    PubMed

    Zhu, Yue-Liang; Wang, Yi; He, Xiao-Qing; Zhu, Min; Li, Fu-Bin; Xu, Yong-Qing

    2013-11-01

    The aim of this report was to present our experience on the use of different flaps for soft tissue reconstruction of the foot and ankle. From 2007 to 2012, the soft tissue defects of traumatic injuries of the foot and ankle were reconstructed using 14 different flaps in 226 cases (162 male and 64 female). There were 62 pedicled flaps and 164 free flaps used in reconstruction. The pedicled flaps included sural flap, saphenous flap, dorsal pedal neurocutaneous flap, pedicled peroneal artery perforator flap, pedicled tibial artery perforator flap, and medial plantar flap. The free flaps were latissimus musculocutaneous flap, anterolateral thigh musculocutaneous flap, groin flap, lateral arm flap, anterolateral thigh perforator flap, peroneal artery perforator flap, thoracdorsal artery perforator flap, medial arm perforator flap. The sensory nerve coaptation was not performed for all of flaps. One hundred and ninety-four cases were combined with open fractures. One hundred and sixty-two cases had tendon. Among 164 free flaps, 8 flaps were completely lost, in which the defects were managed by the secondary procedures. Among the 57 flaps for plantar foot coverage (25 pedicled flaps and 32 free flaps), ulcers were developed in 5 pedicled flaps and 6 free flaps after weight bearing, and infection was found in 14 flaps. The donor site complications were seen in 3 cases with the free anterolateral thigh perforator flap transfer. All of limbs were preserved and the patients regained walking and daily activities. All of patients except for one regained protective sensation from 3 to 12 months postoperatively. Our experience showed that the sural flap and saphenous flap could be good options for the coverage of the defects at malleolus, dorsal hindfoot and midfoot. Plantar foot, forefoot and large size defects could be reconstructed with free anterolateral thigh perforator flap. For the infected wounds with dead spce, the free latissimus dorsi musculocutaneous flap remained to

  15. Tissue expansion using osmotically active hydrogel systems for direct closure of the donor defect of the radial forearm flap.

    PubMed

    Bergé, S J; Wiese, K G; von Lindern, J J; Niederhagen, B; Appel, T; Reich, R H

    2001-07-01

    Although widely used, the radial forearm flap has been criticized for the poor quality of its donor site. Attempts to avoid donor-site problems have concentrated on the elaboration of the split-thickness and full-thickness skin graft methods of reconstruction. Skin grafts frequently fail over the flexor carpi radialis tendon, leading to chronic skin breakdown or, at best, tendon adhesion. Tissue expansion appears to be a good alternative that allows the use of local tissues to ultimately improve the forearm donor-site appearance. To avoid the disadvantages of traditional silicone balloon expanders (such as pressure peaks, infection, the valve at a distance from the expander, postoperative fillings), an osmotically active system was used. In an 18-month prospective study, 10 osmotically active hydrogel tissue expanders were placed on the forearms of 10 patients. The radial forearm flap was performed for intraoral reconstruction after surgical resection of oral cavity malignancies. The study showed that, in nine out of 10 patients, the expanded skin achieved was sufficient to cover the donor site after raising the forearm flap. Additionally, the expansion-related swelling pressure was well tolerated by the patients, the cosmetic results were very satisfactory, and the incidence of complications was very low. By using osmotically active hydrogel tissue expanders, there is no postoperative filling and no risk of complications arising from defective balloon expanders, filling valves, or missing ports.

  16. Control of Flap Vortices

    NASA Technical Reports Server (NTRS)

    Greenblatt, David

    2005-01-01

    A wind tunnel investigation was carried out on a semi-span wing model to assess the feasibility of controlling vortices emanating from outboard flaps and tip-flaps by actively varying the degree of boundary layer separation. Separation was varied by means of perturbations produced from segmented zero-efflux oscillatory blowing slots, while estimates of span loadings and vortex sheet strengths were obtained by integrating wing surface pressures. These estimates were used as input to inviscid rollup relations as a means of predicting changes to the vortex characteristics resulting from the perturbations. Surveys of flow in the wake of the outboard and tip-flaps were made using a seven-hole probe, from which the vortex characteristics were directly deduced. Varying the degree of separation had a marked effect on vortex location, strength, tangential velocity, axial velocity and size for both outboard and tip-flaps. Qualitative changes in vortex characteristics were well predicted by the inviscid rollup relations, while the failure to account for viscosity was presumed to be the main reason for observed discrepancies. Introducing perturbations near the outboard flap-edges or on the tip-flap exerted significant control over vortices while producing negligible lift excursions.

  17. 2D CFD Analysis of an Airfoil with Active Continuous Trailing Edge Flap

    NASA Astrophysics Data System (ADS)

    Jaksich, Dylan; Shen, Jinwei

    2014-11-01

    Efficient and quieter helicopter rotors can be achieved through on-blade control devices, such as active Continuous Trailing-Edge Flaps driven by embedded piezoelectric material. This project aims to develop a CFD simulation tool to predict the aerodynamic characteristics of an airfoil with CTEF using open source code: OpenFOAM. Airfoil meshes used by OpenFOAM are obtained with MATLAB scripts. Once created it is possible to rotate the airfoil to various angles of attack. When the airfoil is properly set up various OpenFOAM properties, such as kinematic viscosity and flow velocity, are altered to achieve the desired testing conditions. Upon completion of a simulation, the program gives the lift, drag, and moment coefficients as well as the pressure and velocity around the airfoil. The simulation is then repeated across multiple angles of attack to give full lift and drag curves. The results are then compared to previous test data and other CFD predictions. This research will lead to further work involving quasi-steady 2D simulations incorporating NASTRAN to model aeroelastic deformation and eventually to 3D aeroelastic simulations. NSF ECE Grant #1358991 supported the first author as an REU student.

  18. Vibration reduction in helicopter rotors using an actively controlled partial span trailing edge flap located on the blade

    NASA Technical Reports Server (NTRS)

    Millott, T. A.; Friedmann, P. P.

    1994-01-01

    This report describes an analytical study of vibration reduction in a four-bladed helicopter rotor using an actively controlled, partial span, trailing edge flap located on the blade. The vibration reduction produced by the actively controlled flap (ACF) is compared with that obtained using individual blade control (IBC), in which the entire blade is oscillated in pitch. For both cases a deterministic feedback controller is implemented to reduce the 4/rev hub loads. For all cases considered, the ACF produced vibration reduction comparable with that obtained using IBC, but consumed only 10-30% of the power required to implement IBC. A careful parametric study is conducted to determine the influence of blade torsional stiffness, spanwise location of the control flap, and hinge moment correction on the vibration reduction characteristics of the ACF. The results clearly demonstrate the feasibility of this new approach to vibration reduction. It should be emphasized than the ACF, used together with a conventional swashplate, is completely decoupled from the primary flight control system and thus it has no influence on the airworthiness of the helicopter. This attribute is potentially a significant advantage when compared to IBC.

  19. Low Speed and High Speed Correlation of SMART Active Flap Rotor Loads

    NASA Technical Reports Server (NTRS)

    Kottapalli, Sesi B. R.

    2010-01-01

    Measured, open loop and closed loop data from the SMART rotor test in the NASA Ames 40- by 80- Foot Wind Tunnel are compared with CAMRAD II calculations. One open loop high-speed case and four closed loop cases are considered. The closed loop cases include three high-speed cases and one low-speed case. Two of these high-speed cases include a 2 deg flap deflection at 5P case and a test maximum-airspeed case. This study follows a recent, open loop correlation effort that used a simple correction factor for the airfoil pitching moment Mach number. Compared to the earlier effort, the current open loop study considers more fundamental corrections based on advancing blade aerodynamic conditions. The airfoil tables themselves have been studied. Selected modifications to the HH-06 section flap airfoil pitching moment table are implemented. For the closed loop condition, the effect of the flap actuator is modeled by increased flap hinge stiffness. Overall, the open loop correlation is reasonable, thus confirming the basic correctness of the current semi-empirical modifications; the closed loop correlation is also reasonable considering that the current flap model is a first generation model. Detailed correlation results are given in the paper.

  20. Simultaneous BVI noise and vibration reduction in rotorcraft using actively-controlled flaps and including performance considerations

    NASA Astrophysics Data System (ADS)

    Patt, Daniel A.

    This work presents the development and application of an active control approach for reduction of both vibration and noise induced by helicopter rotor blade vortex interaction (BVI). Control is implemented through single or dual actively controlled flaps (ACFs) on each blade. Low-speed helicopter flight is prone to severe BVI, resulting in elevated vibration and noise levels. Existing research has suggested that when some form of active control is used to reduce vibration, noise will increase and vice versa. The present research achieves simultaneous reduction of noise and vibration, and also investigates the physical sources of the observed reduction. The initial portion of this work focused on developing a tool for simulating helicopter noise and vibrations in the BVI flight regime. A method for predicting compressible unsteady blade surface pressure distribution on rotor blades was developed and combined with an enhanced free-wake model and an acoustic prediction tool with provisions for blade flexibility. These elements were incorporated within an aeroelastic analysis featuring fully coupled flap-lag-torsional blade dynamics. Subsequently, control algorithms were developed that were effective for reducing noise and vibration even in the nonlinear BVI flight regime; saturation limits were incorporated constraining flap deflections to specified limits. The resulting simulation was also validated with a wide range of experimental data, achieving excellent correlation. Finally, a number of active control studies were performed. Multi-component vibration reductions of 40--80% could be achieved, while incurring a small noise penalty. Noise was reduced using an onboard feedback microphone; reductions of 4--10 dB on the advancing side were observed on a plane beneath the rotor when using dual flaps. Finally, simultaneous noise and vibration reduction was studied. A reduction of about 5 dB in noise on the advancing side combined with a 60% reduction in vibration was

  1. Effects of calcitriol on random skin flap survival in rats.

    PubMed

    Zhou, Kai-liang; Zhang, Yi-hui; Lin, Ding-sheng; Tao, Xian-yao; Xu, Hua-zi

    2016-01-01

    Calcitriol, a metabolite of vitamin D, is often used in osteoporosis clinics. However, the material has other bioactivities; for example, it accelerates angiogenesis, has anti-inflammatory properties, and inhibits oxidative stress. We investigated the effects of calcitriol in a random skin flap rat model. "McFarlane flap" models were established in 84 male Sprague Dawley rats, divided into two groups. One group received intraperitoneal injections of calcitriol (2 μg/kg/day) whereas control rats received intraperitoneal injections of saline. The percentage flap survival area and tissue water content were measured 7 days later, which showed that calcitriol improved flap survival area and reduced tissue edema. It also increased the mean vessel density and upregulated levels of VEGF mRNA/protein, both of which promote flap angiogenesis. Moreover, it decreased leukocyte and macrophage infiltration, reduced the inflammatory proteins IL1β and IL6, increased SOD activity, decreased MDA content, and upregulated the level of autophagy. Overall, our results suggest that calcitriol promotes skin flap survival by accelerating angiogenesis, having anti-inflammatory effects, reducing oxidative stress, and promoting autophagy. PMID:26732750

  2. Reduction of Urease Activity by Interaction with the Flap Covering the Active Site

    PubMed Central

    Macomber, Lee; Minkara, Mona S.; Hausinger, Robert P.; Merz, Kenneth M.

    2015-01-01

    With the increasing appreciation for the human microbiome coupled with the global rise of antibiotic resistant organisms, it is imperative that new methods be developed to specifically target pathogens. To that end, a novel computational approach was devised to identify compounds that reduce the activity of urease, a medically important enzyme of Helicobacter pylori, Proteus mirabilis, and many other microorganisms. Urease contains a flexible loop that covers its active site; Glide was used to identify small molecules predicted to lock this loop in an open conformation. These compounds were screened against the model urease from Klebsiella aerogenes and the natural products epigallocatechin and quercetin were shown to inhibit at low and high micromolar concentrations, respectively. These molecules exhibit a strong time-dependent inactivation of urease that was not due to their oxygen sensitivity. Rather, these compounds appear to inactivate urease by reacting with a specific Cys residue located on the flexible loop. Substitution of this cysteine by alanine in the C319A variant increased the urease resistance to both epigallocatechin and quercetin, as predicted by the computational studies. Protein dynamics are integral to the function of many enzymes; thus, identification of compounds that lock an enzyme into a single conformation presents a useful approach to define potential inhibitors. PMID:25594724

  3. Stress optimization of leaf-spring crossed flexure pivots for an active Gurney flap mechanism

    NASA Astrophysics Data System (ADS)

    Freire Gómez, Jon; Booker, Julian D.; Mellor, Phil H.

    2015-04-01

    The EU's Green Rotorcraft programme is pursuing the development of a functional and airworthy Active Gurney Flap (AGF) for a full-scale helicopter rotor blade. Interest in the development of this `smart adaptive rotor blade' technology lies in its potential to provide a number of aerodynamic benefits, which would in turn translate into a reduction in fuel consumption and noise levels. The AGF mechanism selected employs leaf-spring crossed flexure pivots. These provide important advantages over bearings as they are not susceptible to seizing and do not require maintenance (i.e. lubrication or cleaning). A baseline design of this mechanism was successfully tested both in a fatigue rig and in a 2D wind tunnel environment at flight-representative deployment schedules. For full validation, a flight test would also be required. However, the severity of the in-flight loading conditions would likely compromise the mechanical integrity of the pivots' leaf-springs in their current form. This paper investigates the scope for stress reduction through three-dimensional shape optimization of the leaf-springs of a generic crossed flexure pivot. To this end, a procedure combining a linear strain energy formulation, a parametric leaf-spring profile definition and a series of optimization algorithms is employed. The resulting optimized leaf-springs are proven to be not only independent of the angular rotation at which the pivot operates, but also linearly scalable to leaf-springs of any length, minimum thickness and width. Validated using non-linear finite element analysis, the results show very significant stress reductions relative to pivots with constant cross section leaf-springs, of up to as much as 30% for the specific pivot configuration employed in the AGF mechanism. It is concluded that shape optimization offers great potential for reducing stress in crossed flexure pivots and, consequently, for extending their fatigue life and/or rotational range.

  4. Evidence That the DNA Mismatch Repair System Removes 1-Nucleotide Okazaki Fragment Flaps*♦

    PubMed Central

    Kadyrova, Lyudmila Y.; Dahal, Basanta K.; Kadyrov, Farid A.

    2015-01-01

    The DNA mismatch repair (MMR) system plays a major role in promoting genome stability and suppressing carcinogenesis. In this work, we investigated whether the MMR system is involved in Okazaki fragment maturation. We found that in the yeast Saccharomyces cerevisiae, the MMR system and the flap endonuclease Rad27 act in overlapping pathways that protect the nuclear genome from 1-bp insertions. In addition, we determined that purified yeast and human MutSα proteins recognize 1-nucleotide DNA and RNA flaps. In reconstituted human systems, MutSα, proliferating cell nuclear antigen, and replication factor C activate MutLα endonuclease to remove the flaps. ATPase and endonuclease mutants of MutLα are defective in the flap removal. These results suggest that the MMR system contributes to the removal of 1-nucleotide Okazaki fragment flaps. PMID:26224637

  5. Contractile activity of the pectoralis in the zebra finch according to mode and velocity of flap-bounding flight.

    PubMed

    Tobalske, Bret W; Puccinelli, Lisa A; Sheridan, David C

    2005-08-01

    We studied flying zebra finch (Taeniopygia guttata, N = 12), to provide a new test of a long-standing ;fixed-gear' hypothesis that flap-bounding birds use only intermittent non-flapping phases, instead of variation in muscle activity, to vary mechanical power output in flight. Using sonomicrometry and electromyography, we measured in vivo fascicle length and neuromuscular recruitment in the pectoralis as the birds flew in different flight modes (level, ascending, descending; mean velocity 1.6+/-0.3 m s(-1)) and across velocities in a new, variable-speed wind tunnel (0-12 m s(-1)). Synchronized high-speed digital video (250 Hz) provided a record of wing kinematics. Flight mode had a significant effect upon pectoralis strain, strain rate, fractional shortening and the relative timing of muscle activity (onset, offset and duration). Among flight velocities, we observed significant variation in pectoralis strain, fractional lengthening and shortening, strain rate, relative electromyographic (EMG) amplitude, and EMG duration and offset. In particular, variation in strain rate and relative EMG amplitude indicates that the fixed-gear hypothesis should be rejected. Instead, it appears that zebra finch vary work and power output within wingbeats by modulating muscle contractile behavior and between wingbeats using intermittent bounds. Muscle activity patterns and wing kinematics were similar between free flight and wind tunnel flight at similar speeds. Comparing flights with and without surgically implanted transducers and electrodes, zebra finch exhibited a reduction in maximum velocity (from 14 to 12 m s(-1)) and a significant increase in wingbeat frequency and percent time flapping. This identifies a potential limitation of in vivo flight measurements, and similar studies of bird flight should, therefore, include measurements of the extent to which flight performance is compromised by experimental protocol.

  6. HIGH EFFICIENCY STRUCTURAL FLOWTHROUGH ROTOR WITH ACTIVE FLAP CONTROL: VOLUME ZERO: OVERVIEW AND COMMERCIAL PATH

    SciTech Connect

    Zuteck, Michael D.; Jackson, Kevin L.; Santos, Richard A.

    2015-05-16

    The Zimitar one-piece rotor primary structure is integrated, so balanced thrust and gravity loads flow through the hub region without transferring out of its composite material. Large inner rotor geometry is used since there is no need to neck down to a blade root region and pitch bearing. Rotor control is provided by a highly redundant, five flap system on each blade, sized so that easily handled standard electric linear actuators are sufficient.

  7. HIGH EFFICIENCY STRUCTURAL FLOWTHROUGH ROTOR WITH ACTIVE FLAP CONTROL: VOLUME TWO: INNOVATION & COST OF ENERGY

    SciTech Connect

    Zuteck, Michael D.; Jackson, Kevin L.; Santos, Richard A.

    2015-05-16

    The Zimitar one-piece rotor primary structure is integrated, so balanced thrust and gravity loads flow through the hub region without transferring out of its composite material. Large inner rotor geometry is used since there is no need to neck down to a blade root region and pitch bearing. Rotor control is provided by a highly redundant, five flap system on each blade, sized so that easily handled standard electric linear actuators are sufficient.

  8. HIGH EFFICIENCY STRUCTURAL FLOWTHROUGH ROTOR WITH ACTIVE FLAP CONTROL: VOLUME THREE: MARKET & TEAM

    SciTech Connect

    Zuteck, Michael D.; Jackson, Kevin L.; Santos, Richard A.

    2015-05-16

    The Zimitar one-piece rotor primary structure is integrated, so balanced thrust and gravity loads flow through the hub region without transferring out of its composite material. Large inner rotor geometry is used since there is no need to neck down to a blade root region and pitch bearing. Rotor control is provided by a highly redundant, five flap system on each blade, sized so that easily handled standard electric linear actuators are sufficient.

  9. HIGH EFFICIENCY STRUCTURAL FLOWTHROUGH ROTOR WITH ACTIVE FLAP CONTROL: VOLUME ONE: PRELIMINARY DESIGN REPORT

    SciTech Connect

    Zuteck, Michael D.; Jackson, Kevin L.; Santos, Richard A.; Chow, Ray; Nordenholz, Thomas R.; Wamble, John Lee

    2015-05-16

    The Zimitar one-piece rotor primary structure is integrated, so balanced thrust and gravity loads flow through the hub region without transferring out of its composite material. Large inner rotor geometry is used since there is no need to neck down to a blade root region and pitch bearing. Rotor control is provided by a highly redundant, five flap system on each blade, sized so that easily handled standard electric linear actuators are sufficient.

  10. Distinct parts of leukotriene C{sub 4} synthase interact with 5-lipoxygenase and 5-lipoxygenase activating protein

    SciTech Connect

    Strid, Tobias; Svartz, Jesper; Franck, Niclas; Hallin, Elisabeth; Ingelsson, Bjoern; Soederstroem, Mats; Hammarstroem, Sven

    2009-04-17

    Leukotriene C{sub 4} is a potent inflammatory mediator formed from arachidonic acid and glutathione. 5-Lipoxygenase (5-LO), 5-lipoxygenase activating protein (FLAP) and leukotriene C{sub 4} synthase (LTC{sub 4}S) participate in its biosynthesis. We report evidence that LTC{sub 4}S interacts in vitro with both FLAP and 5-LO and that these interactions involve distinct parts of LTC{sub 4}S. FLAP bound to the N-terminal part/first hydrophobic region of LTC{sub 4}S. This part did not bind 5-LO which bound to the second hydrophilic loop of LTC{sub 4}S. Fluorescent FLAP- and LTC{sub 4}S-fusion proteins co-localized at the nuclear envelope. Furthermore, GFP-FLAP and GFP-LTC{sub 4}S co-localized with a fluorescent ER marker. In resting HEK293/T or COS-7 cells GFP-5-LO was found mainly in the nuclear matrix. Upon stimulation with calcium ionophore, GFP-5-LO translocated to the nuclear envelope allowing it to interact with FLAP and LTC{sub 4}S. Direct interaction of 5-LO and LTC{sub 4}S in ionophore-stimulated (but not un-stimulated) cells was demonstrated by BRET using GFP-5-LO and Rluc-LTC{sub 4}S.

  11. 5-Lipoxygenase Activating Protein Reduction Ameliorates Cognitive Deficit, Synaptic Dysfunction, and Neuropathology in a Mouse Model of Alzheimer’s Disease

    PubMed Central

    Giannopoulos, Phillip F.; Chu, Jin; Joshi, Yash B.; Sperow, Margaret; Li, Jin-Guo; Kirby, Lynn G.; Praticò, Domenico

    2013-01-01

    Background 5-lipoxygenase activating protein (FLAP) is abundantly present in the central nervous system. Although its function has been extensively interrogated in the context of peripheral inflammation, novel roles for this protein are emerging in the central nervous system. The objective of our study was to investigate the functional role that FLAP plays in a mouse model of Alzheimer’s disease (AD) with plaques and tangles (i.e., 3×Tg mice). Methods By implementing a genetic knockout of FLAP and pharmacologic inhibition with a FLAP inhibitor (MK-591), we evaluated the effect on the AD-like neuropathology, cognition, and synaptic plasticity in the 3×Tg mice. Results We show that reduction of FLAP leads to amelioration of cognition and memory along with the rescuing of synaptic dysfunction at an early age before the development of overt neuropathology. Genetic knockout and pharmacologic inhibition of FLAP also yielded an improvement in AD pathology through a reduction in Aβ via the γ-secretase pathway and a decrease in tau phosphorylation through the cdk5 pathway. Conclusions Our studies identify a novel functional role for FLAP in regulating memory and synaptic plasticity. They establish this protein at the crossroad of multiple pathways that ultimately contribute to the development of the entire AD-like phenotype, making it a viable therapeutic target with disease-modifying capacity for the treatment of this disease. PMID:23683389

  12. Benefits of using omental pedicle flap over muscle flap for closure of open window thoracotomy

    PubMed Central

    Chikaishi, Yasuhiro; Kuwata, Taiji; Takenaka, Masaru; Oka, Soichi; Hirai, Ayako; Imanishi, Naoko; Kuroda, Koji; Tanaka, Fumihiro

    2016-01-01

    Background Open window thoracotomy (OWT) as well as its closure are challenging. Transposition of omental pedicle and muscle flaps is often performed for OWT closure; however, the better technique among the two is unknown. The purpose of this series was to evaluate the outcomes of using both omental pedicle and muscle flaps for the aforementioned closure. Methods This was an observational retrospective cohort study on 27 consecutive patients who underwent OWT closure at a single institution between January 2005 and December 2014. The operation was performed using either omental pedicle or muscle flap with thoracoplasty. We compared both techniques in terms of the patient background [sex, age, body mass index (BMI) and C-reactive protein (CRP) before OWT and serum albumin levels before OWT closure], presence of methicillin-resistant Staphylococcus aureus (MRSA) infection, rate of bronchopleural fistula (BPF), duration of OWT, recurrence of local infection, morbidity, duration of indwelling drainage after operation, success, mortality and postoperative hospital stay. Results There were 9 (33.3%) omental pedicle flap procedures and 18 (66.7%) muscle flap procedures. The rate of local recurrence after closure of OWT was significantly higher with muscle flap than with omental pedicle flap (0% vs. 50.0%, P=0.012). The median duration of postoperative hospital stay was significantly shorter with omental pedicle flap than that with muscle flap (16.0 vs. 41.5 days, P=0.037). Mortality was observed in 2 patients (11.2%) in the muscle flap group and no patient in the omental pedicle flap group. Success rate was similar between the two groups (100% for omental pedicle flap vs. 83.3% for muscle flap). Conclusions Omental pedicle flap was superior to muscle flap in terms of reducing local recurrence and shortening postoperative hospital stay. However, mortality, morbidity and success rates were not affected by the choice of flap. PMID:27499959

  13. Effects of calcitriol on random skin flap survival in rats

    PubMed Central

    Zhou, Kai-liang; Zhang, Yi-hui; Lin, Ding-sheng; Tao, Xian-yao; Xu, Hua-zi

    2016-01-01

    Calcitriol, a metabolite of vitamin D, is often used in osteoporosis clinics. However, the material has other bioactivities; for example, it accelerates angiogenesis, has anti-inflammatory properties, and inhibits oxidative stress. We investigated the effects of calcitriol in a random skin flap rat model. “McFarlane flap” models were established in 84 male Sprague Dawley rats, divided into two groups. One group received intraperitoneal injections of calcitriol (2 μg/kg/day) whereas control rats received intraperitoneal injections of saline. The percentage flap survival area and tissue water content were measured 7 days later, which showed that calcitriol improved flap survival area and reduced tissue edema. It also increased the mean vessel density and upregulated levels of VEGF mRNA/protein, both of which promote flap angiogenesis. Moreover, it decreased leukocyte and macrophage infiltration, reduced the inflammatory proteins IL1β and IL6, increased SOD activity, decreased MDA content, and upregulated the level of autophagy. Overall, our results suggest that calcitriol promotes skin flap survival by accelerating angiogenesis, having anti-inflammatory effects, reducing oxidative stress, and promoting autophagy. PMID:26732750

  14. Regulation of yeast DNA polymerase δ-mediated strand displacement synthesis by 5'-flaps.

    PubMed

    Koc, Katrina N; Stodola, Joseph L; Burgers, Peter M; Galletto, Roberto

    2015-04-30

    The strand displacement activity of DNA polymerase δ is strongly stimulated by its interaction with proliferating cell nuclear antigen (PCNA). However, inactivation of the 3'-5' exonuclease activity is sufficient to allow the polymerase to carry out strand displacement even in the absence of PCNA. We have examined in vitro the basic biochemical properties that allow Pol δ-exo(-) to carry out strand displacement synthesis and discovered that it is regulated by the 5'-flaps in the DNA strand to be displaced. Under conditions where Pol δ carries out strand displacement synthesis, the presence of long 5'-flaps or addition in trans of ssDNA suppress this activity. This suggests the presence of a secondary DNA binding site on the enzyme that is responsible for modulation of strand displacement activity. The inhibitory effect of a long 5'-flap can be suppressed by its interaction with single-stranded DNA binding proteins. However, this relief of flap-inhibition does not simply originate from binding of Replication Protein A to the flap and sequestering it. Interaction of Pol δ with PCNA eliminates flap-mediated inhibition of strand displacement synthesis by masking the secondary DNA site on the polymerase. These data suggest that in addition to enhancing the processivity of the polymerase PCNA is an allosteric modulator of other Pol δ activities.

  15. Effect of microbial transglutaminase and non-meat proteins on functional properties of low NaCl, phosphate-free patties made from channel catfish (Ictalurus punctatus) belly flap meat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was aimed at developing value-added low sodium chloride (NaCl), phosphate-free restructured patties using minced channel catfish (Ictalurus punctatus) belly flap meat. The effect of microbial transglutaminase (MTGase, 0.7%) and non-meat proteins (isolated soy protein, ISP and whey protein...

  16. An Active Flow Circulation Controlled Flap Concept for General Aviation Aircraft Applications

    NASA Technical Reports Server (NTRS)

    Jones, Gregory S.; Viken, Sally A.; Washburn, Anthony E.; Jenkins, Luther N.; Cagle, C. Mark

    2002-01-01

    A recent focus on revolutionary aerodynamic concepts has highlighted the technology needs of general aviation and personal aircraft. New and stringent restrictions on these types of aircraft have placed high demands on aerodynamic performance, noise, and environmental issues. Improved high lift performance of these aircraft can lead to slower takeoff and landing speeds that can be related to reduced noise and crash survivability issues. Circulation Control technologies have been around for 65 years, yet have been avoided due to trade offs of mass flow, pitching moment, perceived noise etc. The need to improve the circulation control technology for general aviation and personal air-vehicle applications is the focus of this paper. This report will describe the development of a 2-D General Aviation Circulation Control (GACC) wing concept that utilizes a pulsed pneumatic flap.

  17. Repositioning free laser in situ keratomileusis flaps.

    PubMed

    Todani, Amit; Al-Arfaj, Khalid; Melki, Samir A

    2010-02-01

    We describe a protocol for adequate repositioning of free laser in situ keratomileusis (LASIK) corneal flaps created by a Moria M2 microkeratome even in the absence of fiduciary marks. In an enucleated porcine globe, a free flap was created by initially placing a longitudinal incision at the proposed hinge site followed by activating the forward pass of the automated microkeratome. A protocol was devised based on placement of a positioning dot on the free flap before the flap is retrieved from the microkeratome head. Preplaced surgical landmarks were used as a guide to determine the correct alignment of the free flap. Adequate orientation of the free flap to the stromal bed was achieved in 9 porcine eyes using the positioning dot method. The technique is applicable to the Moria M2 microkeratome only and must be validated for other types of keratomes.

  18. Separating proteins with activated carbon.

    PubMed

    Stone, Matthew T; Kozlov, Mikhail

    2014-07-15

    Activated carbon is applied to separate proteins based on differences in their size and effective charge. Three guidelines are suggested for the efficient separation of proteins with activated carbon. (1) Activated carbon can be used to efficiently remove smaller proteinaceous impurities from larger proteins. (2) Smaller proteinaceous impurities are most efficiently removed at a solution pH close to the impurity's isoelectric point, where they have a minimal effective charge. (3) The most efficient recovery of a small protein from activated carbon occurs at a solution pH further away from the protein's isoelectric point, where it is strongly charged. Studies measuring the binding capacities of individual polymers and proteins were used to develop these three guidelines, and they were then applied to the separation of several different protein mixtures. The ability of activated carbon to separate proteins was demonstrated to be broadly applicable with three different types of activated carbon by both static treatment and by flowing through a packed column of activated carbon. PMID:24898563

  19. The racquet conjunctival flap.

    PubMed

    Cies, W A; Odeh-Nasrala, N

    1976-01-01

    In situations requiring a conjunctival flap to cover a corneal defect, the racquet flap technique is a simple relatively noninvasive method. Less conjunctiva is sacrificed and yet the flap is fully maintained by the one pedicle. The bulbar conjunctiva is maintained in an essentially normal state.

  20. Keyhole Flap Nipple Reconstruction.

    PubMed

    Chen, Joseph I; Cash, Camille G; Iman, Al-Haj; Spiegel, Aldona J; Cronin, Ernest D

    2016-05-01

    Nipple-areola reconstruction is often one of the final but most challenging aspects of breast reconstruction. However, it is an integral and important component of breast reconstruction because it transforms the mound into a breast. We performed 133 nipple-areola reconstructions during a period of 4 years. Of these reconstructions, 76 of 133 nipple-areola complexes were reconstructed using the keyhole flap technique. The tissue used for the keyhole dermoadipose flap technique include transverse rectus abdominus myocutaneous flaps (60/76), latissimus dorsi flaps (15/76), or mastectomy skin flaps after tissue expanders (1/76). The average patient follow-up was 17 months. The design of the flap is based on a keyhole configuration. The base of the flap determines the width of the future nipple, whereas the length of the flap determines the projection. We try to match the projection of the contralateral nipple if present. The keyhole flap is simple to construct yet reliable. It provides good symmetry and projection and avoids the creation of new scars. The areola is then tattooed approximately 3 months after the nipple reconstruction. PMID:27579228

  1. Flow Over Swept Flaps and Flap Tips

    NASA Technical Reports Server (NTRS)

    Bradshaw, Peter; Buice, Carl U.

    1995-01-01

    Preliminary measurements have been made of the flow over the tip of an unswept wing flap. To achieve an acceptable Reynolds number based on flap chord, the flap chord was chosen equal to the chord of the main airfoil (c = 19 in. approx. 0.48 m). The model was mounted in a 30 in. x 30 in. wind tunnel running at up to 100 ft/sec. (30 m/s): severe wind-tunnel interference was accepted, and any computations would be done using the tunnel walls as the boundaries of the computational domain. Maximum Reynolds number based on flap chord and tunnel speed was about 1.O x lO(exp 6). The grant ended before a full set of measurements could be made, but the work done so far yields a useful picture of the flow. The vortex originates at about mid-chord on the flap and rises rapidly above the chord line. It has a concentrated core, with total pressure lower than the ambient static pressure, and there is no evidence of large-scale wandering. A simple method of model construction, giving light weight and excellent surface finish, was developed.

  2. Facial flap complications.

    PubMed

    Zoumalan, Richard A; Murakami, Craig S

    2012-06-01

    Knowledge of fundamental concepts can help decrease the chance of complications in plastic surgery. Local flap reconstruction for facial defects has many pitfalls. This article describes common complications in local flap reconstruction of the face and describes strategies that prevent problems.

  3. Allostery in BAX protein activation.

    PubMed

    Jiang, Zhenyan; Zhang, Hansi; Böckmann, Rainer A

    2016-11-01

    BAX is a member of the proapoptotic BCL-2 family of proteins, which is involved in the regulation of the intrinsic pathway of apoptosis. In the process of apoptosis, BH3-only molecules activate cytosolic BAX. Activated BAX molecules insert into the mitochondrial outer membrane with their [Formula: see text]-helix and form oligomers that lead to membrane poration, resulting in the release of apoptogenic factors including cytochrome c. Recently, a novel interaction site for the binding of the BIM SAHB ligand to BAX was reported. BIM SAHB binding was shown to invoke the exposure of the 6A7 epitope (amino acids 13-19) and of the BH3 domain of BAX, followed by mobilization of the BAX [Formula: see text]-helix. However, the intramolecular pathway for signal transmission in BAX, from BIM SAHB binding to mobilization of the [Formula: see text]-helix largely remained elusive. For a molecular understanding of the activation of BAX, and thus the first steps in apoptosis, we performed microsecond atomistic molecular dynamics simulations both of the BAX protein and of the BAX:BIM SAHB complex in aqueous solution. In agreement with experiment, the 6A7 and BH3 domains adopt a more solvent-exposed conformation within the BAX:BIM SAHB complex. BIM SAHB binding was found to stabilize the secondary structure of the [Formula: see text]9-helix. A force distribution analysis revealed a force network of residue-residue interactions responsible for signal transmission from the BIM SAHB binding site predominantly via the [Formula: see text]4- and [Formula: see text]6-helices to the [Formula: see text]9-helix on the opposite site of the protein.

  4. [The abdominal drop flap].

    PubMed

    Bodin, F; Liverneaux, P; Seigle-Murandi, F; Facca, S; Bruant-Rodier, C; Dissaux, C; Chaput, B

    2015-08-01

    The skin between the mastectomy scar and the future infra-mammary fold may be managed in different ways in delayed breast reconstruction using a DIEP (deep inferior epigastric perforator). Conserving this skin and positioning the flap skin paddle in the middle of the breast usually highlights skin color disparity because of two visible transition zones. Resection of the entire skin under the scar may be more aesthetic but limits direct closure possibility in case of flap failure. In order to benefit from both aesthetic result and safe surgical method, we propose the abdominal drop flap. The inferior thoracic skin flap is detached from the thoracic wall beyond the future infra-mammary fold, preserved and pushed under the breast.

  5. Total arm flap.

    PubMed

    Becker, D W

    1987-11-01

    The development of an unusual and rarely indicated total arm flap is described in the context of widely indicated and automatically used principles passed down by the recognized father of plastic surgery, Sir Harold G. Gillies.

  6. Flapping of Insectile Wings

    NASA Astrophysics Data System (ADS)

    Huang, Yangyang; Kanso, Eva

    2015-11-01

    Insects use flight muscles attached at the base of the wings to produce impressive wing flapping frequencies. Yet the effects of muscle stiffness on the performance of insect wings remain unclear. Here, we construct an insectile wing model, consisting of two rigid wings connected at their base by an elastic torsional spring and submerged in an oscillatory flow. The wing system is free to rotate and flap. We first explore the extent to which the flyer can withstand roll perturbations, then study its flapping behavior and performance as a function of spring stiffness. We find an optimal range of spring stiffness that results in large flapping amplitudes, high force generation and good storage of elastic energy. We conclude by conjecturing that insects may select and adjust the muscle spring stiffness to achieve desired movement. These findings may have significant implications on the design principles of wings in micro air-vehicles.

  7. Benchmarking aerodynamic prediction of unsteady rotor aerodynamics of active flaps on wind turbine blades using ranging fidelity tools

    NASA Astrophysics Data System (ADS)

    Barlas, Thanasis; Jost, Eva; Pirrung, Georg; Tsiantas, Theofanis; Riziotis, Vasilis; Navalkar, Sachin T.; Lutz, Thorsten; van Wingerden, Jan-Willem

    2016-09-01

    Simulations of a stiff rotor configuration of the DTU 10MW Reference Wind Turbine are performed in order to assess the impact of prescribed flap motion on the aerodynamic loads on a blade sectional and rotor integral level. Results of the engineering models used by DTU (HAWC2), TUDelft (Bladed) and NTUA (hGAST) are compared to the CFD predictions of USTUTT-IAG (FLOWer). Results show fairly good comparison in terms of axial loading, while alignment of tangential and drag-related forces across the numerical codes needs to be improved, together with unsteady corrections associated with rotor wake dynamics. The use of a new wake model in HAWC2 shows considerable accuracy improvements.

  8. Repair of large full-thickness cartilage defect by activating endogenous peripheral blood stem cells and autologous periosteum flap transplantation combined with patellofemoral realignment.

    PubMed

    Fu, Wei-Li; Ao, Ying-Fang; Ke, Xiao-Yan; Zheng, Zhuo-Zhao; Gong, Xi; Jiang, Dong; Yu, Jia-Kuo

    2014-03-01

    Minimal-invasive procedure and one-step surgery offer autologous mesenchymal stem cells derived from peripheral blood (PB-MSCs) a promising prospective in the field of cartilage regeneration. We report a case of a 19-year-old male athlete of kickboxing with ICRS grade IV chondral lesions at the 60° region of lateral femoral trochlea, which was repaired by activating endogenous PB-MSCs plus autologous periosteum flap transplantation combined with correcting the patellofemoral malalignment. After a 7.5 year follow-up, the result showed that the patient returned to competitive kickboxing. Second-look under arthroscopy showed a smooth surface at 8 months postoperation. The IKDC 2000 subjective score, Lysholm score and Tegner score were 95, 98 and 9 respectively at the final follow up. CT and MRI evaluations showed a significant improvement compared with those of pre-operation.

  9. Use of microbial transglutaminase and nonmeat proteins to improve functional properties of low NaCl, phosphate-free patties made from channel catfish (Ictalurus punctatus) belly flap meat.

    PubMed

    Min, B; Green, B W

    2008-06-01

    This study was aimed at developing value-added low sodium chloride (NaCl), phosphate-free restructured patties using minced channel catfish (Ictalurus punctatus) belly flap meat. The effect of microbial transglutaminase (MTGase) and nonmeat proteins (isolated soy protein, ISP, and whey protein concentrate, WPC; 1.7%, respectively) alone and in combination were evaluated to improve cooking yield and textural properties in patties with reduced NaCl and no phosphate. The concentration effect of MTGase (0.05% to 0.7%) was also studied. The addition of MTGase increased textural properties such as binding strength, hardness, cohesiveness, chewiness, and springiness, but decreased cooking yield of the patties (P < 0.05). Isolated soy protein increased cooking yield (P < 0.05), but did not affect textural properties. Inclusion of WPC did not increase cooking yield or impact textural properties of patties. The combination of MTGase and ISP significantly increased both the cooking yield and textural properties of patties. As the concentration of MTGase increased at constant ISP, the textural properties of cooked patties significantly increased, but cooking yield decreased (P < 0.05). In conclusion, we suggest that the combination of 0.05% to 0.1% of MTGase with 1.7% ISP is optimal for development of a low NaCl, phosphate-free patty using minced catfish belly flap meat.

  10. Genomic and protein expression analysis reveals Flap endonuclease 1 (FEN1) as a key biomarker in breast and ovarian cancer

    PubMed Central

    Abdel-Fatah, Tarek MA; Russell, Roslin; Albarakati, Nada; Maloney, David J; Dorjsuren, Dorjbal; Rueda, Oscar M; Moseley, Paul; Mohan, Vivek; Sun, Hongmao; Abbotts, Rachel; Mukherjee, Abhik; Agarwal, Devika; Illuzzi, Jennifer L.; Jadhav, Ajit; Simeonov, Anton; Ball, Graham; Chan, Stephen; Caldas, Carlos; Ellis, Ian O; Wilson, David M; Madhusudan, Srinivasan

    2015-01-01

    FEN1 has key roles in Okazaki fragment maturation during replication, long patch base excision repair, rescue of stalled replication forks, maintenance of telomere stability and apoptosis. FEN1 may be dysregulated in breast and ovarian cancers and have clinicopathological significance in patients. We comprehensively investigated FEN1 mRNA expression in multiple cohorts of breast cancer [training set (128), test set (249), external validation (1952)]. FEN1 protein expression was evaluated in 568 oestrogen receptor (ER) negative breast cancers, 894 ER positive breast cancers and 156 ovarian epithelial cancers. FEN1 mRNA overexpression was highly significantly associated with high grade (p=4.89 × 10−57), high mitotic index (p=5.25 × 10−28), pleomorphism (p=6.31 × 10−19), ER negative (p=9.02 × 10−35), PR negative (p=9.24 × 10−24), triple negative phenotype (p=6.67 × 10−21), PAM50.Her2 (p=5.19 × 10−13), PAM50.Basal (p=2.7 × 10−41), PAM50.LumB (p=1.56 × 10−26), integrative molecular cluster 1 (intClust.1) (p=7.47 × 10−12), intClust.5 (p=4.05 × 10−12) and intClust. 10 (p=7.59 × 10−38) breast cancers. FEN1 mRNA overexpression is associated with poor breast cancer specific survival in univariate (p= 4.4 × 10−16) and multivariate analysis (p= 9.19 × 10−7). At the protein level, in ER positive tumours, FEN1 overexpression remains significantly linked to high grade, high mitotic index and pleomorphism (ps<0.01). In ER negative tumours, high FEN1 is significantly associated with pleomorphism, tumour type, lymphovascular invasion, triple negative phenotype, EGFR and HER2 expression (ps<0.05). In ER positive as well as in ER negative tumours, FEN1 protein overexpression is associated with poor survival in univariate and multivariate analysis (ps<0.01). In ovarian epithelial cancers, similarly, FEN1 overexpression is associated with high grade, high stage and poor survival (ps<0.05). We conclude that FEN1 is a promising biomarker in breast

  11. Genomic and protein expression analysis reveals flap endonuclease 1 (FEN1) as a key biomarker in breast and ovarian cancer.

    PubMed

    Abdel-Fatah, Tarek M A; Russell, Roslin; Albarakati, Nada; Maloney, David J; Dorjsuren, Dorjbal; Rueda, Oscar M; Moseley, Paul; Mohan, Vivek; Sun, Hongmao; Abbotts, Rachel; Mukherjee, Abhik; Agarwal, Devika; Illuzzi, Jennifer L; Jadhav, Ajit; Simeonov, Anton; Ball, Graham; Chan, Stephen; Caldas, Carlos; Ellis, Ian O; Wilson, David M; Madhusudan, Srinivasan

    2014-10-01

    FEN1 has key roles in Okazaki fragment maturation during replication, long patch base excision repair, rescue of stalled replication forks, maintenance of telomere stability and apoptosis. FEN1 may be dysregulated in breast and ovarian cancers and have clinicopathological significance in patients. We comprehensively investigated FEN1 mRNA expression in multiple cohorts of breast cancer [training set (128), test set (249), external validation (1952)]. FEN1 protein expression was evaluated in 568 oestrogen receptor (ER) negative breast cancers, 894 ER positive breast cancers and 156 ovarian epithelial cancers. FEN1 mRNA overexpression was highly significantly associated with high grade (p = 4.89 × 10(-57)), high mitotic index (p = 5.25 × 10(-28)), pleomorphism (p = 6.31 × 10(-19)), ER negative (p = 9.02 × 10(-35)), PR negative (p = 9.24 × 10(-24)), triple negative phenotype (p = 6.67 × 10(-21)), PAM50.Her2 (p = 5.19 × 10(-13)), PAM50. Basal (p = 2.7 × 10(-41)), PAM50.LumB (p = 1.56 × 10(-26)), integrative molecular cluster 1 (intClust.1) (p = 7.47 × 10(-12)), intClust.5 (p = 4.05 × 10(-12)) and intClust. 10 (p = 7.59 × 10(-38)) breast cancers. FEN1 mRNA overexpression is associated with poor breast cancer specific survival in univariate (p = 4.4 × 10(-16)) and multivariate analysis (p = 9.19 × 10(-7)). At the protein level, in ER positive tumours, FEN1 overexpression remains significantly linked to high grade, high mitotic index and pleomorphism (ps < 0.01). In ER negative tumours, high FEN1 is significantly associated with pleomorphism, tumour type, lymphovascular invasion, triple negative phenotype, EGFR and HER2 expression (ps < 0.05). In ER positive as well as in ER negative tumours, FEN1 protein overexpression is associated with poor survival in univariate and multivariate analysis (ps < 0.01). In ovarian epithelial cancers, similarly, FEN1 overexpression is associated with high grade, high stage and poor survival (ps < 0.05). We conclude that FEN1

  12. Protein-water dynamics in antifreeze protein III activity

    NASA Astrophysics Data System (ADS)

    Xu, Yao; Bäumer, Alexander; Meister, Konrad; Bischak, Connor G.; DeVries, Arthur L.; Leitner, David M.; Havenith, Martina

    2016-03-01

    We combine Terahertz absorption spectroscopy (THz) and molecular dynamics (MD) simulations to investigate the underlying molecular mechanism for the antifreeze activity of one class of antifreeze protein, antifreeze protein type III (AFP-III) with a focus on the collective water hydrogen bond dynamics near the protein. After summarizing our previous work on AFPs, we present a new investigation of the effects of cosolutes on protein antifreeze activity by adding sodium citrate to the protein solution of AFP-III. Our results reveal that for AFP-III, unlike some other AFPs, the addition of the osmolyte sodium citrate does not affect the hydrogen bond dynamics at the protein surface significantly, as indicated by concentration dependent THz measurements. The present data, in combination with our previous THz measurements and molecular simulations, confirm that while long-range solvent perturbation is a necessary condition for the antifreeze activity of AFP-III, the local binding affinity determines the size of the hysteresis.

  13. Blowing Flap Experiment: PIV Measurements

    NASA Technical Reports Server (NTRS)

    Hutcheson, Florence V.; Stead, Daniel J.; Bremmer, David M.

    2004-01-01

    PIV measurements of the flow in the region of a flap side edge are presented for several flap configurations. The test model is a NACA 63(sub 2)-215 Hicks Mod-B main element airfoil with a half-span Fowler flap. Air is blown from small slots located along the flap side edge on either the top, bottom or side surfaces. The test set up is described and flow measurements for a baseline and three blowing flap configurations are presented. The effects that the flap tip jets have on the structure of the flap side edge flow are discussed for each of the flap configurations tested. The results indicate that blowing air from a slot located along the top surface of the flap greatly weakened the top vortex system and pushed it further off the top surface. Blowing from the bottom flap surface kept the strong side vortex further outboard while blowing from the side surface only strengthened the flap vortex system. It is concluded that blowing from the top or bottom surfaces of the flap may lead to a reduction of flap side edge noise.

  14. Endogenous protein phosphorylation and protein kinase activity in winged bean.

    PubMed

    Mukhopadhyay, K; Singh, M

    1997-10-01

    In winged bean (Psophocarpus tetragonolobus) protein kinases (E.C. 2.7.1.37) were found in all tissues studied. There was a significant increase in kinase activity during seed development, with a concomitant enhancement in the phosphorylation of a number of polypeptides; this was reversed in germinating seed cotyledons. Protein phosphorylation was apparently correlated with the increase in the protein content of the developing seed and the growing axis. At least three distinct autophosphorylating proteins could be distinguished in the developing seeds after SDS-PAGE, indicating the presence of different types of protein kinases in winged bean.

  15. Crystal structures of the structure-selective nuclease Mus81-Eme1 bound to flap DNA substrates

    PubMed Central

    Gwon, Gwang Hyeon; Jo, Aera; Baek, Kyuwon; Jin, Kyeong Sik; Fu, Yaoyao; Lee, Jong-Bong; Kim, YoungChang; Cho, Yunje

    2014-01-01

    The Mus81-Eme1 complex is a structure-selective endonuclease with a critical role in the resolution of recombination intermediates during DNA repair after interstrand cross-links, replication fork collapse, or double-strand breaks. To explain the molecular basis of 3′ flap substrate recognition and cleavage mechanism by Mus81-Eme1, we determined crystal structures of human Mus81-Eme1 bound to various flap DNA substrates. Mus81-Eme1 undergoes gross substrate-induced conformational changes that reveal two key features: (i) a hydrophobic wedge of Mus81 that separates pre- and post-nick duplex DNA and (ii) a “5′ end binding pocket” that hosts the 5′ nicked end of post-nick DNA. These features are crucial for comprehensive protein-DNA interaction, sharp bending of the 3′ flap DNA substrate, and incision strand placement at the active site. While Mus81-Eme1 unexpectedly shares several common features with members of the 5′ flap nuclease family, the combined structural, biochemical, and biophysical analyses explain why Mus81-Eme1 preferentially cleaves 3′ flap DNA substrates with 5′ nicked ends. PMID:24733841

  16. Bilobed flap in sole surgery

    SciTech Connect

    Sanchez-Conejo-Mir, J.; Bueno Montes, J.; Moreno Gimenez, J.C.; Camacho-Martinez, F.

    1985-09-01

    The bilobed flap is a simple reconstructive technique principally used to correct substantial defects in the facial region. The authors present their experience with this local flap in the difficult plantar area, with excellent short-term functional results. They describe the special characteristics of the bilobed flap in this zone, and comment on its indications and possible complications.

  17. Identification of intracellular receptor proteins for activated protein kinase C.

    PubMed Central

    Mochly-Rosen, D; Khaner, H; Lopez, J

    1991-01-01

    Protein kinase C (PKC) translocates from the cytosol to the particulate fraction on activation. This activation-induced translocation of PKC is thought to reflect PKC binding to the membrane lipids. However, immunological and biochemical data suggest that PKC may bind to proteins in the cytoskeletal elements in the particulate fraction and in the nuclei. Here we describe evidence for the presence of intracellular receptor proteins that bind activated PKC. Several proteins from the detergent-insoluble material of the particulate fraction bound PKC in the presence of phosphatidylserine and calcium; binding was further increased with the addition of diacylglycerol. Binding of PKC to two of these proteins was concentration-dependent, saturable, and specific, suggesting that these binding proteins are receptors for activated C-kinase, termed here "RACKs." PKC binds to RACKs via a site on PKC distinct from the substrate binding site. We suggest that binding to RACKs may play a role in activation-induced translocation of PKC. Images PMID:1850844

  18. Activity-Based Protein Profiling of Microbes

    SciTech Connect

    Sadler, Natalie C.; Wright, Aaron T.

    2015-02-01

    Activity-Based Protein Profiling (ABPP) in conjunction with multimodal characterization techniques has yielded impactful findings in microbiology, particularly in pathogen, bioenergy, drug discovery, and environmental research. Using small molecule chemical probes that react irreversibly with specific proteins or protein families in complex systems has provided insights in enzyme functions in central metabolic pathways, drug-protein interactions, and regulatory protein redox, for systems ranging from photoautotrophic cyanobacteria to mycobacteria, and combining live cell or cell extract ABPP with proteomics, molecular biology, modeling, and other techniques has greatly expanded our understanding of these systems. New opportunities for application of ABPP to microbial systems include: enhancing protein annotation, characterizing protein activities in myriad environments, and reveal signal transduction and regulatory mechanisms in microbial systems.

  19. Local Flaps of The Hand

    PubMed Central

    Rehim, Shady A.; Chung, Kevin C.

    2014-01-01

    Synopsis A local flap consists of skin and subcutaneous tissue that is harvested from a site nearby a given defect while maintaining its intrinsic blood supply. When a soft tissue defect of the hand is not amenable to primary closure or skin grafting, local skin flaps can be a used as a reliable source of soft tissue replacement that replaces like with like. Flaps are categorized based on their composition, method of transfer, flap design and blood supply, yet flap circulation is considered the most critical factor for the flap survival. This article reviews the classification of local skin flaps of the hand and offers a practical reconstructive approach for several soft tissue defects of the hand and digits. PMID:24731606

  20. Noise Reduction of Aircraft Flap

    NASA Technical Reports Server (NTRS)

    Hutcheson, Florence V. (Inventor); Brooks, Thomas F. (Inventor)

    2009-01-01

    A reduction in noise radiating from a side of a deployed aircraft flap is achieved by locating a slot adjacent the side of the flap, and then forcing air out through the slot with a suitable mechanism. One, two or even three or more slots are possible, where the slot is located at one;or more locations selected from a group of locations comprising a top surface of the flap, a bottom surface of the flap, an intersection of the top and side surface of the flap, an intersection of the bottom and side surfaces of the flap, and a side surface of the flap. In at least one embodiment the slot is substantially rectangular. A device for adjusting a rate of the air forced out through the slot can also be provided.

  1. The Deltopectoral Flap Revisited: The Internal Mammary Artery Perforator Flap.

    PubMed

    Ibrahim, Amir; Atiyeh, Bishara; Karami, Reem; Adelman, David M; Papazian, Nazareth J

    2016-03-01

    Pharyngo-esophageal and tracheostomal defects pose a challenge in head and neck reconstruction whenever microanastomosis is extremely difficult in hostile neck that is previously dissected and irradiated. The deltopectoral (DP) flap was initially described as a pedicled flap for such reconstruction with acceptable postoperative results. A major drawback is still that the DP flap is based on 3 perforator vessels leading to a decreased arc of rotation. The DP flap also left contour deformities in the donor site. The internal mammary artery perforator flap was described as a refinement of the deltopectoral flap. It is a pedicled fasciocutaneous flap based on a single perforator, with comparable and reliable blood supply compared with the DP flap, giving it the benefit of having a wide arc of rotation. It is both thin and pliable, with good skin color match and texture. The donor site can be closed primarily with no esthetic deformity and minimal morbidity. The procedure is relatively simple and does not require microvascular expertise. In this report, the authors describe a patient in whom bilateral internal mammary artery perforator flaps were used for subtotal pharyngo-esophageal reconstruction and neck resurfacing. The flaps healed uneventfully bilaterally with no postoperative complications. PMID:26854779

  2. Hydrogen peroxide activates activator protein-1 and mitogen-activated protein kinases in pancreatic stellate cells.

    PubMed

    Kikuta, Kazuhiro; Masamune, Atsushi; Satoh, Masahiro; Suzuki, Noriaki; Satoh, Kennichi; Shimosegawa, Tooru

    2006-10-01

    Activated pancreatic stellate cells (PSCs) are implicated in the pathogenesis of pancreatic inflammation and fibrosis, where oxidative stress is thought to play a key role. Reactive oxygen species such as hydrogen peroxide (H(2)O(2)) may act as a second messenger to mediate the actions of growth factors and cytokines. But the role of reactive oxygen species in the activation and regulation of cell functions in PSCs remains largely unknown. We here examined the effects of H(2)O(2) on the activation of signal transduction pathways and cell functions in PSCs. PSCs were isolated from the pancreas of male Wistar rats, and used in their culture-activated, myofibroblast-like phenotype unless otherwise stated. Activation of transcription factors was examined by electrophoretic mobility shift assay and luciferase assay. Activation of mitogen-activated protein (MAP) kinases was assessed by Western blotting using anti-phosphospecific antibodies. The effects of H(2)O(2) on proliferation, alpha(1)(I)procollagen gene expression, and monocyte chemoattractant protein-1 production were evaluated. The effect of H(2)O(2) on the transformation of freshly isolated PSCs in culture was also assessed. H(2)O(2) at non-cytotoxic concentrations (up to 100 microM) induced oxidative stress in PSCs. H(2)O(2) activated activator protein-1, but not nuclear factor kappaB. In addition, H(2)O(2) activated three classes of MAP kinases: extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 MAP kinase. H(2)O(2) induced alpha(1)(I)procollagen gene expression but did not induce proliferation or monocyte chemoattractant protein-1 production. H(2)O(2) did not initiate the transformation of freshly isolated PSCs to myofibroblast-like phenotype. Specific activation of these signal transduction pathways and collagen gene expression by H(2)O(2) may play a role in the pathogenesis of pancreatic fibrosis.

  3. Computational Introduction of Catalytic Activity into Proteins.

    PubMed

    Bertolani, Steve J; Carlin, Dylan Alexander; Siegel, Justin B

    2016-01-01

    Recently, there have been several successful cases of introducing catalytic activity into proteins. One method that has been used successfully to achieve this is the theozyme placement and enzyme design algorithms implemented in Rosetta Molecular Modeling Suite. Here, we illustrate how to use this software to recapitulate the placement of catalytic residues and ligand into a protein using a theozyme, protein scaffold, and catalytic constraints as input. PMID:27094294

  4. Human FAN1 promotes strand incision in 5'-flapped DNA complexed with RPA.

    PubMed

    Takahashi, Daisuke; Sato, Koichi; Hirayama, Emiko; Takata, Minoru; Kurumizaka, Hitoshi

    2015-09-01

    Fanconi anaemia (FA) is a human infantile recessive disorder. Seventeen FA causal proteins cooperatively function in the DNA interstrand crosslink (ICL) repair pathway. Dual DNA strand incisions around the crosslink are critical steps in ICL repair. FA-associated nuclease 1 (FAN1) is a DNA structure-specific endonuclease that is considered to be involved in DNA incision at the stalled replication fork. Replication protein A (RPA) rapidly assembles on the single-stranded DNA region of the stalled fork. However, the effect of RPA on the FAN1-mediated DNA incision has not been determined. In this study, we purified human FAN1, as a bacterially expressed recombinant protein. FAN1 exhibited robust endonuclease activity with 5'-flapped DNA, which is formed at the stalled replication fork. We found that FAN1 efficiently promoted DNA incision at the proper site of RPA-coated 5'-flapped DNA. Therefore, FAN1 possesses the ability to promote the ICL repair of 5'-flapped DNA covered by RPA.

  5. Flap Endonuclease 1 Limits Telomere Fragility on the Leading Strand*

    PubMed Central

    Teasley, Daniel C.; Parajuli, Shankar; Nguyen, Mai; Moore, Hayley R.; Alspach, Elise; Lock, Ying Jie; Honaker, Yuchi; Saharia, Abhishek; Piwnica-Worms, Helen; Stewart, Sheila A.

    2015-01-01

    The existence of redundant replication and repair systems that ensure genome stability underscores the importance of faithful DNA replication. Nowhere is this complexity more evident than in challenging DNA templates, including highly repetitive or transcribed sequences. Here, we demonstrate that flap endonuclease 1 (FEN1), a canonical lagging strand DNA replication protein, is required for normal, complete leading strand replication at telomeres. We find that the loss of FEN1 nuclease activity, but not DNA repair activities, results in leading strand-specific telomere fragility. Furthermore, we show that FEN1 depletion-induced telomere fragility is increased by RNA polymerase II inhibition and is rescued by ectopic RNase H1 expression. These data suggest that FEN1 limits leading strand-specific telomere fragility by processing RNA:DNA hybrid/flap intermediates that arise from co-directional collisions occurring between the replisome and RNA polymerase. Our data reveal the first molecular mechanism for leading strand-specific telomere fragility and the first known role for FEN1 in leading strand DNA replication. Because FEN1 mutations have been identified in human cancers, our findings raise the possibility that unresolved RNA:DNA hybrid structures contribute to the genomic instability associated with cancer. PMID:25922071

  6. Skin flaps and grafts - self-care

    MedlinePlus

    ... Free flap - self-care; Skin autografting - self-care; Pressure ulcer skin flap self-care; Burns skin flap self- ... skin infection Surgery for skin cancer Venous ulcers , pressure ulcers , or diabetic ulcers that DO NOT heal After ...

  7. Flexible Flapping Foils

    NASA Astrophysics Data System (ADS)

    Marais, Catherine; Godoy-Diana, Ramiro; Wesfreid, José. Eduardo

    2010-11-01

    Hydrodynamic tunnel experiments with flexible flapping foils of 4:1 span-to-chord aspect ratio are used in the present work to study the effect of foil compliance in the dynamical features of a propulsive wake. The average thrust force produced by the foil is estimated from 2D PIV measurements and the regime transitions in the wake are characterized according to a flapping frequency-amplitude phase diagram as in Godoy-Diana et al. (Phys. Rev. E 77, 016308, 2008). We show that the thrust production regime occurs on a broader region of the parameter space for flexible foils, with propulsive forces up to 3 times greater than for the rigid case. We examine in detail the vortex generation at the trailing edge of the foils, and propose a mechanism to explain how foil deformation leads to an optimization of propulsion.

  8. MK-886, an inhibitor of the 5-lipoxygenase-activating protein, inhibits cyclooxygenase-1 activity and suppresses platelet aggregation.

    PubMed

    Koeberle, Andreas; Siemoneit, Ulf; Northoff, Hinnak; Hofmann, Bettina; Schneider, Gisbert; Werz, Oliver

    2009-04-17

    MK-886, an inhibitor of the 5-lipoxygenase-activating protein (FLAP), potently suppresses leukotriene biosynthesis in intact cells and is frequently used to define a role of the 5-lipoxygenase (EC 1.13.11.34) pathway in cellular or animal models of inflammation, allergy, cancer, and cardiovascular disease. Here we show that MK-886 also interferes with the activities of cyclooxygenases (COX, EC 1.14.99.1). MK-886 inhibited isolated COX-1 (IC(50)=8 microM) and blocked the formation of the COX-1-derived products 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid (12-HHT) and thromboxane B(2) in washed human platelets in response to collagen as well as from exogenous arachidonic acid (IC(50)=13-15 microM). Isolated COX-2 was less affected (IC(50)=58 microM), and in A549 cells, MK-886 (33 microM) failed to suppress COX-2-dependent 6-keto-prostaglandin (PG)F(1alpha) formation. The distinct susceptibility of MK-886 towards COX-1 and -2 is apparent in automated molecular docking studies that indicate a preferred binding of MK-886 to COX-1 into the active site. MK-886 (10 microM) inhibited COX-1-mediated platelet aggregation induced by collagen or arachidonic acid whereas thrombin- or U-46619-induced (COX-independent) aggregation was not affected. Since leukotrienes and prostaglandins share (patho)physiological properties in the development and regulation of carcinogenesis, inflammation, and vascular functions, caution should be used when interpreting data where MK-886 is used as tool to determine the involvement of FLAP and/or the 5-lipoxygenase pathway in respective experimental models.

  9. Engineered Vascularized Muscle Flap.

    PubMed

    Egozi, Dana; Shandalov, Yulia; Freiman, Alina; Rosenfeld, Dekel; Ben-Shimol, David; Levenberg, Shulamit

    2016-01-01

    One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen supply to the cell microenvironment, as passive diffusion is limited to a very thin layer. Although various materials have been described to restore the integrity of full-thickness defects of the abdominal wall, no material has yet proved to be optimal, due to low graft vascularization, tissue rejection, infection, or inadequate mechanical properties. This protocol describes a means of engineering a fully vascularized flap, with a thickness relevant for muscle tissue reconstruction. Cell-embedded poly L-lactic acid/poly lactic-co-glycolic acid constructs are implanted around the mouse femoral artery and vein and maintained in vivo for a period of one or two weeks. The vascularized graft is then transferred as a flap towards a full thickness defect made in the abdomen. This technique replaces the need for autologous tissue sacrifications and may enable the use of in vitro engineered vascularized flaps in many surgical applications. PMID:26779840

  10. The Versatile Modiolus Perforator Flap

    PubMed Central

    Gunnarsson, Gudjon Leifur; Thomsen, Jorn Bo

    2016-01-01

    Background: Perforator flaps are well established, and their usefulness as freestyle island flaps is recognized. The whereabouts of vascular perforators and classification of perforator flaps in the face are a debated subject, despite several anatomical studies showing similar consistency. In our experience using freestyle facial perforator flaps, we have located areas where perforators are consistently found. This study is focused on a particular perforator lateral to the angle of the mouth; the modiolus and the versatile modiolus perforator flap. Methods: A cohort case series of 14 modiolus perforator flap reconstructions in 14 patients and a color Doppler ultrasonography localization of the modiolus perforator in 10 volunteers. Results: All 14 flaps were successfully used to reconstruct the defects involved, and the location of the perforator was at the level of the modiolus as predicted. The color Doppler ultrasonography study detected a sizeable perforator at the level of the modiolus lateral to the angle of the mouth within a radius of 1 cm. This confirms the anatomical findings of previous authors and indicates that the modiolus perforator is a consistent anatomical finding, and flaps based on it can be recommended for several indications from the reconstruction of defects in the perioral area, cheek and nose. Conclusions: The modiolus is a well-described anatomical area containing a sizeable perforator that is consistently present and readily visualized using color Doppler ultrasonography. We have used the modiolus perforator flap successfully for several indications, and it is our first choice for perioral reconstruction. PMID:27257591

  11. Flag flapping in a channel

    NASA Astrophysics Data System (ADS)

    Alben, Silas; Shoele, Kourosh; Mittal, Rajat; Jha, Sourabh; Glezer, Ari

    2015-11-01

    We study the flapping of a flag in an inviscid channel flow. We focus especially on how quantities vary with channel spacing. As the channel walls move inwards towards the flag, heavier flags become more unstable, while light flags' stability is less affected. We use a vortex sheet model to compute large-amplitude flapping, and find that the flag undergoes a series of jumps to higher flapping modes as the channel walls are moved towards the flag. Meanwhile, the drag on the flag and the energy lost to the wake first rise as the walls become closer, then drop sharply as the flag moves to a higher flapping mode.

  12. DNA-based control of protein activity

    PubMed Central

    Engelen, W.; Janssen, B. M. G.

    2016-01-01

    DNA has emerged as a highly versatile construction material for nanometer-sized structures and sophisticated molecular machines and circuits. The successful application of nucleic acid based systems greatly relies on their ability to autonomously sense and act on their environment. In this feature article, the development of DNA-based strategies to dynamically control protein activity via oligonucleotide triggers is discussed. Depending on the desired application, protein activity can be controlled by directly conjugating them to an oligonucleotide handle, or expressing them as a fusion protein with DNA binding motifs. To control proteins without modifying them chemically or genetically, multivalent ligands and aptamers that reversibly inhibit their function provide valuable tools to regulate proteins in a noncovalent manner. The goal of this feature article is to give an overview of strategies developed to control protein activity via oligonucleotide-based triggers, as well as hurdles yet to be taken to obtain fully autonomous systems that interrogate, process and act on their environments by means of DNA-based protein control. PMID:26812623

  13. Increased flexibility decreases antifreeze protein activity

    PubMed Central

    Patel, Shruti N; Graether, Steffen P

    2010-01-01

    Antifreeze proteins protect several cold-blooded organisms from subzero environments by preventing death from freezing. The Type I antifreeze protein (AFP) isoform from Pseudopleuronectes americanus, named HPLC6, is a 37-residue protein that is a single α-helix. Mutational analysis of the protein showed that its alanine-rich face is important for binding to and inhibiting the growth of macromolecular ice. Almost all structural studies of HPLC6 involve the use of chemically synthesized protein as it requires a native N-terminal aspartate and an amidated C-terminus for full activity. Here, we examine the role of C-terminal amide and C-terminal arginine side chain in the activity, structure, and dynamics of nonamidated Arg37 HPLC6, nonamidated HPLC6 Ala37, amidated HPLC6 Ala37, and fully native HPLC6 using a recombinant bacterial system. The thermal hysteresis (TH) activities of the nonamidated mutants are 35% lower compared with amidated proteins, but analysis of the NMR data and circular dichroism spectra shows that they are all still α-helical. Relaxation data from the two nonamidated mutants indicate that the C-terminal residues are considerably more flexible than the rest of the protein because of the loss of the amide group, whereas the amidated Ala37 mutant has a C-terminus that is as rigid as the wild-type protein and has high TH activity. We propose that an increase in flexibility of the AFP causes it to lose activity because its dynamic nature prevents it from binding strongly to the ice surface. PMID:20936690

  14. A Miniature Controllable Flapping Wing Robot

    NASA Astrophysics Data System (ADS)

    Arabagi, Veaceslav Gheorghe

    The agility and miniature size of nature's flapping wing fliers has long baffled researchers, inspiring biological studies, aerodynamic simulations, and attempts to engineer their robotic replicas. Flapping wing flight is characterized by complex reciprocating wing kinematics, transient aerodynamic effects, and very small body lengths. These characteristics render robotic flapping wing aerial vehicles ideal for surveillance and defense applications, search and rescue missions, and environment monitoring, where their ability to hover and high maneuverability is immensely beneficial. One of the many difficulties in creating flapping wing based miniature robotic aerial vehicles lies in generating a proper wing trajectory that would result in sufficient lift forces for hovering and maneuvering. Since design of a flapping wing system is a balance between overall weight and the number of actuated inputs, we take the approach of having minimal controlled inputs, allowing passive behavior wherever possible. Hence, we propose a completely passive wing pitch reversal design that relies on wing inertial dynamics, an elastic energy storage mechanism, and low Reynolds number aerodynamic effects. Theoretical models, compiling previous research on piezoelectric actuators, four-bar transmissions, and aerodynamics effects, are developed and used as basis for a complete numerical simulation. Limitations of the model are discussed in comparison to experimental results obtained from a working prototype of the proposed passive pitch reversal flapping wing mechanism. Given that the mechanism is under-actuated, methods to control lift force generation by actively varying system parameters are proposed, discussed, and tested experimentally. A dual wing aerial platform is developed based on the passive pitch reversal wing concept. Design considerations are presented, favoring controllability and structural rigidity of the final platform. Finite element analysis and experimental

  15. Reduction of Flap Side Edge Noise - the Blowing Flap

    NASA Technical Reports Server (NTRS)

    Hutcheson, Florence V.; Brooks, THomas F.

    2005-01-01

    A technique to reduce the noise radiating from a wing-flap side edge is being developed. As an airplane wing with an extended flap is exposed to a subsonic airflow, air is blown outward through thin rectangular chord-wise slots at various locations along the side edges and side surface of the flap to weaken and push away the vortices that originate in that region of the flap and are responsible for important noise emissions. Air is blown through the slots at up to twice the local flow velocity. The blowing is done using one or multiple slots, where a slot is located along the top, bottom or side surface of the flap along the side edge, or also along the intersection of the bottom (or top) and side surfaces.

  16. Flap-Edge Blowing Experiments

    NASA Technical Reports Server (NTRS)

    Gaeta, R. J.; Englar, R. J.; Ahuja, K. K.

    2003-01-01

    This Appendix documents the salient results from an effort to mitigate the so-called flap-edge noise generated at the split between a flap edge that is deployed and the undeployed flap. Utilizing a Coanda surface installed at the flap edge, steady blowing was used in an attempt to diminish the vortex strength resulting from the uneven lift distribution. The strength of this lifting vortex was augmented by steady blowing over the deployed flap. The test article for this study was the same 2D airfoil used in the steady blowing program reported earlier (also used in pulsed blowing tests, see Appendix G), however its trailing edge geometry was modified. An exact duplicate of the airfoil shape was made out of fiberglass with no flap, and in the clean configuration. It was attached to the existing airfoil to make an airfoil that has half of its flap deployed and half un-deployed. Figure 1 shows a schematic of the planform showing the two areas where steady blowing was introduced. The flap-edge blowing or the auxiliary blowing was in the direction normal to the freestream velocity vector. Slot heights for the blowing chambers were on the order of 0.0 14 inches.

  17. Molecular Dynamics Simulations of Ligand-Induced Flap Conformational Changes in Cathepsin-D-A Comparative Study.

    PubMed

    Arodola, Olayide A; Soliman, Mahmoud E S

    2016-11-01

    The flap region in aspartic proteases is a unique structural feature to this class of enzymes, and found to have a profound impact on protein overall structure, function, and dynamics. Understanding the structure and dynamic behavior of the flap regions is crucial in the design of selective inhibitors against aspartic proteases. Cathepsin-D, an aspartic protease enzyme, has been implicated in a long list of degenerative diseases as well as breast cancer progression. Presented herein, for the first time, is a comprehensive description of the conformational flap dynamics of cathepsin-D using a comparative 50 ns "multiple" molecular dynamics simulations. Diverse collective metrics were proposed to accurately define flap dynamics. These are distance d1 between the flap tips residues (Gly79 and Met301); dihedral angle ϕ; in addition to TriCα angles Gly79-Asp33-Asp223, θ1 , and Gly79-Asp223-Met301, θ2 . The maximum distance attained throughout the simulation was 17.42 and 11.47 Å for apo and bound cathepsin-D, respectively, while the minimum distance observed was 8.75 and 6.32 Å for apo and bound cathepsin-D, respectively. The movement of the flap as well as the twist of the active pocket can properly be explained by measuring the angle, θ1 , between Gly79-Asp33-Met301 and correlating it with the distance Cα of the flap tip residues. The asymmetrical opening of the binding cavity was best described by the large shift of -6.26° to +20.94° in the dihedral angle, ϕ, corresponding to the full opening of the flap at a range of 31-33 ns. A wide-range of post-dynamic analyses was also applied in this report to supplement our findings. We believe that this report would augment current efforts in designing potent structure-based inhibitors against cathepsin-D in the treatment of breast cancer and other degenerative diseases. J. Cell. Biochem. 117: 2643-2657, 2016. © 2016 Wiley Periodicals, Inc. PMID:27038253

  18. Enamel matrix proteins (Emdogain) in combination with coronally advanced flap or subepithelial connective tissue graft in the treatment of shallow gingival recessions.

    PubMed

    Berlucchi, Ignazio; Francetti, Luca; Del Fabbro, Massimo; Testori, Tiziano; Weinstein, Roberto L

    2002-12-01

    This article describes two different surgical techniques of root coverage using Emdogain and shows preliminary results on 26 shallow recessions in 14 patients. For the treatment of 13 recessions, Emdogain was used in combination with a coronally advanced flap (CAF+EMD group). In the other 13 recessions, Emdogain and the flap were used in combination with a subepithelial connective tissue graft (CAF+CTG+EMD group). For the CAF+EMD group, the root coverage at 6 months was 93.97%, with an attachment gain of 3.2 mm; for the CAF+CTG+EMD group, the root coverage was 93.59%, with an attachment gain of 3.4 mm (no statistically significant difference between groups). When complete root coverage was not achieved, the residual recession was 1 mm in four cases and 2 mm in one case. Keratinized gingiva was increased for both groups, but more for the CAF+CTG+EMD group (1.38 mm versus 0.69 mm; statistically significant difference). Clinical attachment level decreased significantly in both groups, from 4.46 to 1.23 mm in the CAF+EMD group, and from 4.62 to 1.23 mm in the CAF+CTG+EMD group. Preliminary results show that Emdogain, in combination with CAF or CAF+CTG for the treatment of Miller Class I or II gingival recessions, displays good clinical results, with percentage of root coverage comparable or superior to other techniques. Further experimental studies on the dynamics of wound healing are needed to prove that EMD is really responsible for improving the percentage of regenerated versus repaired tissues with respect to other techniques. PMID:12516830

  19. Active Nuclear Import of Membrane Proteins Revisited.

    PubMed

    Laba, Justyna K; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker's yeast. PMID:26473931

  20. Active Nuclear Import of Membrane Proteins Revisited

    PubMed Central

    Laba, Justyna K.; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M.

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker’s yeast. PMID:26473931

  1. Animal flight dynamics II. Longitudinal stability in flapping flight.

    PubMed

    Taylor, G K; Thomas, A L R

    2002-02-01

    Stability is essential to flying and is usually assumed to be especially problematic in flapping flight. If so, problems of stability may have presented a particular hurdle to the evolution of flapping flight. In spite of this, the stability of flapping flight has never been properly analysed. Here we use quasi-static and blade element approaches to analyse the stability provided by a flapping wing. By using reduced order approximations to the natural modes of motion, we show that wing beat frequencies are generally high enough compared to the natural frequencies of motion for a quasi-static approach to be valid as a first approximation. Contrary to expectations, we find that there is noting inherently destabilizing about flapping: beating the wings faster simply amplifies any existing stability or instability, and flapping can even enhance stability compared to gliding at the same air speed. This suggests that aerodynamic stability may not have been a particular hurdle in the evolution of flapping flight. Hovering animals, like hovering helicopters, are predicted to possess neutral static stability. Flapping animals, like fixed wing aircraft, are predicted to be stable in forward flight if the mean flight force acts above and/or behind the centre of gravity. In this case, the downstroke will always be stabilizing. The stabilizing contribution may be diminished by an active upstroke with a low advance ratio and more horizontal stroke plane; other forms of the upstroke may make a small positive contribution to stability. An active upstroke could, therefore, be used to lower stability and enhance manoeuvrability. Translatory mechanisms of unsteady lift production are predicted to amplify the stability predicted by a quasi-static analysis. Non-translatory mechanisms will make little or no contribution to stability. This may be one reason why flies, and other animals which rely upon non-translatory aerodynamic mechanisms, often appear inherently unstable.

  2. Protein kinase activators alter glial cholesterol esterification

    SciTech Connect

    Jeng, I.; Dills, C.; Klemm, N.; Wu, C.

    1986-05-01

    Similar to nonneural tissues, the activity of glial acyl-CoA cholesterol acyltransferase is controlled by a phosphorylation and dephosphorylation mechanism. Manipulation of cyclic AMP content did not alter the cellular cholesterol esterification, suggesting that cyclic AMP is not a bioregulator in this case. Therefore, the authors tested the effect of phorbol-12-myristate 13-acetate (PMA) on cellular cholesterol esterification to determine the involvement of protein kinase C. PMA has a potent effect on cellular cholesterol esterification. PMA depresses cholesterol esterification initially, but cells recover from inhibition and the result was higher cholesterol esterification, suggesting dual effects of protein kinase C. Studies of other phorbol analogues and other protein kinase C activators such as merezein indicate the involvement of protein kinase C. Oleoyl-acetyl glycerol duplicates the effect of PMA. This observation is consistent with a diacyl-glycerol-protein kinase-dependent reaction. Calcium ionophore A23187 was ineffective in promoting the effect of PMA. They concluded that a calcium-independent and protein C-dependent pathway regulated glial cholesterol esterification.

  3. Self-propulsion of flapping bodies in viscous fluids: Recent advances and perspectives

    NASA Astrophysics Data System (ADS)

    Wang, Shizhao; He, Guowei; Zhang, Xing

    2016-08-01

    Flapping-powered propulsion is used by many animals to locomote through air or water. Here we review recent experimental and numerical studies on self-propelled mechanical systems powered by a flapping motion. These studies improve our understanding of the mutual interaction between actively flapping bodies and surrounding fluids. The results obtained in these works provide not only new insights into biolocomotion but also useful information for the biomimetic design of artificial flyers and swimmers.

  4. Active Control of Flow Separation on a High-Lift System with Slotted Flap at High Reynolds Number

    NASA Technical Reports Server (NTRS)

    Khodadoust, Abdollah; Washburn, Anthony

    2007-01-01

    The NASA Energy Efficient Transport (EET) airfoil was tested at NASA Langley's Low- Turbulence Pressure Tunnel (LTPT) to assess the effectiveness of distributed Active Flow Control (AFC) concepts on a high-lift system at flight scale Reynolds numbers for a medium-sized transport. The test results indicate presence of strong Reynolds number effects on the high-lift system with the AFC operational, implying the importance of flight-scale testing for implementation of such systems during design of future flight vehicles with AFC. This paper describes the wind tunnel test results obtained at the LTPT for the EET high-lift system for various AFC concepts examined on this airfoil.

  5. Wuho Is a New Member in Maintaining Genome Stability through its Interaction with Flap Endonuclease 1

    PubMed Central

    Cheng, I-Cheng; Chen, Betty Chamay; Shuai, Hung-Hsun; Chien, Fan-Ching; Chen, Peilin; Hsieh, Tao-shih

    2016-01-01

    Replication forks are vulnerable to wayward nuclease activities. We report here our discovery of a new member in guarding genome stability at replication forks. We previously isolated a Drosophila mutation, wuho (wh, no progeny), characterized by a severe fertility defect and affecting expression of a protein (WH) in a family of conserved proteins with multiple WD40 repeats. Knockdown of WH by siRNA in Drosophila, mouse, and human cultured cells results in DNA damage with strand breaks and apoptosis through ATM/Chk2/p53 signaling pathway. Mice with mWh knockout are early embryonic lethal and display DNA damage. We identify that the flap endonuclease 1 (FEN1) is one of the interacting proteins. Fluorescence microscopy showed the localization of WH at the site of nascent DNA synthesis along with other replication proteins, including FEN1 and PCNA. We show that WH is able to modulate FEN1’s endonucleolytic activities depending on the substrate DNA structure. The stimulatory or inhibitory effects of WH on FEN1’s flap versus gap endonuclease activities are consistent with the proposed WH’s functions in protecting the integrity of replication fork. These results suggest that wh is a new member of the guardians of genome stability because it regulates FEN1’s potential DNA cleavage threat near the site of replication. PMID:26751069

  6. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Flap interconnection. 23.701 Section 23.701... Systems § 23.701 Flap interconnection. (a) The main wing flaps and related movable surfaces as a system must— (1) Be synchronized by a mechanical interconnection between the movable flap surfaces that...

  7. Flap Conformations in HIV-1 Protease are Altered by Mutations

    NASA Astrophysics Data System (ADS)

    Fanucci, Gail; Blackburn, Mandy; Veloro, Angelo; Galiano, Luis; Fangu, Ding; Simmerling, Carlos

    2009-03-01

    HIV-1 protease (PR) is an enzyme that is a major drug target in the treatment of AIDS. Although the structure and function of HIV-1 PR have been studied for over 20 years, questions remain regarding the conformations and dynamics of the β-hairpin turns (flaps) that cover the active site cavity. Distance measurements with pulsed EPR spectroscopy of spin labeled constructs of HIV-1 PR have been used to characterize the flap conformations in the apo and inhibitor bound states. From the most probably distances and the breadth of the distance distribution profiles from analysis of the EPR data, insights regarding the flap conformations and flexibility are gained. The EPR results clearly show how drug pressure selected mutations alter the average conformation of the flaps and the degree of opening of the flaps. Molecular dynamics simulations successfully regenerate the experimentally determined distance distribution profiles, and more importantly, provide structural models for full interpretation of the EPR results. By combining experiment and theory to understand the role that altered flap flexibility/conformations play in the mechanism of drug resistance, key insights are gained toward the rational development of new inhibitors of this important enzyme.

  8. Musculocutaneous flaps in head and neck reconstruction.

    PubMed Central

    Leonard, A. G.

    1989-01-01

    The introduction of musculocutaneous flaps to head and neck reconstructive surgery is described. The flaps available are listed, and the most important ones described and illustrated. Both the latissimus dorsi and pectoralis major flaps are felt to have a role in head and neck reconstruction, though they have largely been superseded by microvascular free flaps such as the radial forearm flap. Images fig. 1 fig. 2 fig. 3 fig. 4 fig. 5 fig. 6 fig. 7 fig. 8 PMID:2686511

  9. Radiated noise from an externally blown flap

    NASA Technical Reports Server (NTRS)

    Reddy, N. N.; Yu, J. C.

    1975-01-01

    The far field noise from subsonic jet impingement on a wing-flap with a 45 deg bend was experimentally investigated. The test parameters are jet Mach number and flap length. For long flaps, the primary source mechanisms are found to be turbulent mixing and flow impingement. For short flaps, the interaction of turbulent flow with the flap trailing edge appears to strongly influence the radiated noise.

  10. Flap Edge Noise Reduction Fins

    NASA Technical Reports Server (NTRS)

    Khorrami, Mehdi R. (Inventor); Choudhan, Meelan M. (Inventor)

    2015-01-01

    A flap of the type that is movably connected to an aircraft wing to provide control of an aircraft in flight includes opposite ends, wherein at least a first opposite end includes a plurality of substantially rigid, laterally extending protrusions that are spaced apart to form a plurality of fluidly interconnected passageways. The passageways have openings adjacent to upper and lower sides of the flap, and the passageways include a plurality of bends such that high pressure fluid flows from a high pressure region to a low pressure region to provide a boundary condition that inhibits noise resulting from airflow around the end of the flap.

  11. Structural insights into 5‧ flap DNA unwinding and incision by the human FAN1 dimer

    NASA Astrophysics Data System (ADS)

    Zhao, Qi; Xue, Xiaoyu; Longerich, Simonne; Sung, Patrick; Xiong, Yong

    2014-12-01

    Human FANCD2-associated nuclease 1 (FAN1) is a DNA structure-specific nuclease involved in the processing of DNA interstrand crosslinks (ICLs). FAN1 maintains genomic stability and prevents tissue decline in multiple organs, yet it confers ICL-induced anti-cancer drug resistance in several cancer subtypes. Here we report three crystal structures of human FAN1 in complex with a 5‧ flap DNA substrate, showing that two FAN1 molecules form a head-to-tail dimer to locate the lesion, orient the DNA and unwind a 5‧ flap for subsequent incision. Biochemical experiments further validate our model for FAN1 action, as structure-informed mutations that disrupt protein dimerization, substrate orientation or flap unwinding impair the structure-specific nuclease activity. Our work elucidates essential aspects of FAN1-DNA lesion recognition and a unique mechanism of incision. These structural insights shed light on the cellular mechanisms underlying organ degeneration protection and cancer drug resistance mediated by FAN1.

  12. Active Wnt proteins are secreted on exosomes.

    PubMed

    Gross, Julia Christina; Chaudhary, Varun; Bartscherer, Kerstin; Boutros, Michael

    2012-10-01

    Wnt signalling has important roles during development and in many diseases. As morphogens, hydrophobic Wnt proteins exert their function over a distance to induce patterning and cell differentiation decisions. Recent studies have identified several factors that are required for the secretion of Wnt proteins; however, how Wnts travel in the extracellular space remains a largely unresolved question. Here we show that Wnts are secreted on exosomes both during Drosophila development and in human cells. We demonstrate that exosomes carry Wnts on their surface to induce Wnt signalling activity in target cells. Together with the cargo receptor Evi/WIs, Wnts are transported through endosomal compartments onto exosomes, a process that requires the R-SNARE Ykt6. Our study demonstrates an evolutionarily conserved functional role of extracellular vesicular transport of Wnt proteins.

  13. Total Cellular RNA Modulates Protein Activity.

    PubMed

    Majumder, Subhabrata; DeMott, Christopher M; Reverdatto, Sergey; Burz, David S; Shekhtman, Alexander

    2016-08-16

    RNA constitutes up to 20% of a cell's dry weight, corresponding to ∼20 mg/mL. This high concentration of RNA facilitates low-affinity protein-RNA quinary interactions, which may play an important role in facilitating and regulating biological processes. In the yeast Pichia pastoris, the level of ubiquitin-RNA colocalization increases when cells are grown in the presence of dextrose and methanol instead of methanol as the sole carbon source. Total RNA isolated from cells grown in methanol increases β-galactosidase activity relative to that seen with RNA isolated from cells grown in the presence of dextrose and methanol. Because the total cellular RNA content changes with growth medium, protein-RNA quinary interactions can alter in-cell protein biochemistry and may play an important role in cell adaptation, critical to many physiological and pathological states. PMID:27456029

  14. Electrochemical Activation of Engineered Protein Switches

    PubMed Central

    Choi, Jay H.; Zayats, Maya; Searson, Peter C.; Ostermeier, Marc

    2016-01-01

    Engineered protein switches have a large dynamic range, high specificity for the activating ligand, and a modular architecture, and have been explored for a wide range of applications including biosensors and therapeutics. The ability to externally control switch function is important in extending applications for protein switches. We recently demonstrated that the on/off state could be controlled by the redox state of disulfide bonds introduced into the switches at select locations. Here, we demonstrate that an electrochemical signal can be used as an exogenous input to control switch function via reduction of the engineered disulfide bonds. This study suggests that disulfide-containing protein switch is a potentially useful platform for bioelectronic sensors with remote control of the sensing ability. PMID:26241391

  15. Analysis of mitogen-activated protein kinase activity in yeast.

    PubMed

    Elion, Elaine A; Sahoo, Rupam

    2010-01-01

    Mitogen-activated protein (MAP) kinases play central roles in transmitting extracellular and intracellular information in a wide variety of situations in eukaryotic cells. Their activities are perturbed in a large number of diseases, and their activating kinases are currently therapeutic targets in cancer. MAPKs are highly conserved among all eukaryotes. MAPKs were first cloned from the yeast Saccharomyces cerevisiae. Yeast has five MAPKs and one MAPK-like kinase. The mating MAPK Fus3 is the best characterized yeast MAPK. Members of all subfamilies of human MAPKs can functionally substitute S. cerevisiae MAPKs, providing systems to use genetic approaches to study the functions of either yeast or human MAPKs and to identify functionally relevant amino acid residues that enhance or reduce the effects of therapeutically relevant inhibitors and regulatory proteins. Here, we describe an assay to measure Fus3 activity in immune complexes prepared from S. cerevisiae extracts. The assay conditions are applicable to other MAPKs, as well. PMID:20811996

  16. De Novo Construction of Redox Active Proteins.

    PubMed

    Moser, C C; Sheehan, M M; Ennist, N M; Kodali, G; Bialas, C; Englander, M T; Discher, B M; Dutton, P L

    2016-01-01

    Relatively simple principles can be used to plan and construct de novo proteins that bind redox cofactors and participate in a range of electron-transfer reactions analogous to those seen in natural oxidoreductase proteins. These designed redox proteins are called maquettes. Hydrophobic/hydrophilic binary patterning of heptad repeats of amino acids linked together in a single-chain self-assemble into 4-alpha-helix bundles. These bundles form a robust and adaptable frame for uncovering the default properties of protein embedded cofactors independent of the complexities introduced by generations of natural selection and allow us to better understand what factors can be exploited by man or nature to manipulate the physical chemical properties of these cofactors. Anchoring of redox cofactors such as hemes, light active tetrapyrroles, FeS clusters, and flavins by His and Cys residues allow cofactors to be placed at positions in which electron-tunneling rates between cofactors within or between proteins can be predicted in advance. The modularity of heptad repeat designs facilitates the construction of electron-transfer chains and novel combinations of redox cofactors and new redox cofactor assisted functions. Developing de novo designs that can support cofactor incorporation upon expression in a cell is needed to support a synthetic biology advance that integrates with natural bioenergetic pathways. PMID:27586341

  17. Synaptic Vesicle Proteins and Active Zone Plasticity.

    PubMed

    Kittel, Robert J; Heckmann, Manfred

    2016-01-01

    Neurotransmitter is released from synaptic vesicles at the highly specialized presynaptic active zone (AZ). The complex molecular architecture of AZs mediates the speed, precision and plasticity of synaptic transmission. Importantly, structural and functional properties of AZs vary significantly, even for a given connection. Thus, there appear to be distinct AZ states, which fundamentally influence neuronal communication by controlling the positioning and release of synaptic vesicles. Vice versa, recent evidence has revealed that synaptic vesicle components also modulate organizational states of the AZ. The protein-rich cytomatrix at the active zone (CAZ) provides a structural platform for molecular interactions guiding vesicle exocytosis. Studies in Drosophila have now demonstrated that the vesicle proteins Synaptotagmin-1 (Syt1) and Rab3 also regulate glutamate release by shaping differentiation of the CAZ ultrastructure. We review these unexpected findings and discuss mechanistic interpretations of the reciprocal relationship between synaptic vesicles and AZ states, which has heretofore received little attention.

  18. Synaptic Vesicle Proteins and Active Zone Plasticity

    PubMed Central

    Kittel, Robert J.; Heckmann, Manfred

    2016-01-01

    Neurotransmitter is released from synaptic vesicles at the highly specialized presynaptic active zone (AZ). The complex molecular architecture of AZs mediates the speed, precision and plasticity of synaptic transmission. Importantly, structural and functional properties of AZs vary significantly, even for a given connection. Thus, there appear to be distinct AZ states, which fundamentally influence neuronal communication by controlling the positioning and release of synaptic vesicles. Vice versa, recent evidence has revealed that synaptic vesicle components also modulate organizational states of the AZ. The protein-rich cytomatrix at the active zone (CAZ) provides a structural platform for molecular interactions guiding vesicle exocytosis. Studies in Drosophila have now demonstrated that the vesicle proteins Synaptotagmin-1 (Syt1) and Rab3 also regulate glutamate release by shaping differentiation of the CAZ ultrastructure. We review these unexpected findings and discuss mechanistic interpretations of the reciprocal relationship between synaptic vesicles and AZ states, which has heretofore received little attention. PMID:27148040

  19. Flap monitoring using infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Keller, Alex; Wright, Leigh P.; Elmandjra, Mohamed; Mao, Jian-min

    2006-02-01

    We report results of clinical trials on flap monitoring in 65 plastic surgeries. Hemoglobin oxygen saturation of flap tissue (StO II) was monitored non-invasively by using ODISsey TM tissue oximeter, an infrared spectroscopic device. StO II measurements were conducted both intra-operatively and post-operatively. From the intra-operative measurements, we observed that StO II values dropped when the main blood vessels supplying the flap were clamped in surgery, and that StO II jumped after anastomosis to a value close to its pre-operative value. From post-operative monitoring measurements for the 65 flap cases, each lasted two days or so, we found that the StO II values approach to a level close to the baseline if the surgery was successful, and that the StO II value dropped to a value below 30% if there is a perfusion compromise, such as vascular thrombosis.

  20. Managing Flap Vortices via Separation Control

    NASA Technical Reports Server (NTRS)

    Greenblatt, David

    2006-01-01

    A pilot study was conducted on a flapped semi-span model to investigate the concept and viability of near-wake vortex management by means of boundary layer separation control. Passive control was achieved using a simple fairing and active control was achieved via zero mass-flux blowing slots. Vortex sheet strength, estimated by integrating surface pressures, was used to predict vortex characteristics based on inviscid rollup relations and vortices trailing the flaps were mapped using a seven-hole probe. Separation control was found to have a marked effect on vortex location, strength, tangential velocity, axial velocity and size over a wide range of angles of attack and control conditions. In general, the vortex trends were well predicted by the inviscid rollup relations. Manipulation of the separated flow near the flap edges exerted significant control over either outboard or inboard edge vortices while producing small lift and moment excursions. Unsteady surface pressures indicated that dynamic separation and attachment control can be exploited to perturb vortices at wavelengths shorter than a typical wingspan. In summary, separation control has the potential for application to time-independent or time-dependent wake alleviation schemes, where the latter can be deployed to minimize adverse effects on ride-quality and dynamic structural loading.

  1. Development of Novel Adenosine Monophosphate-Activated Protein Kinase Activators

    PubMed Central

    Guh, Jih-Hwa; Chang, Wei-Ling; Yang, Jian; Lee, Su-Lin; Wei, Shuo; Wang, Dasheng; Kulp, Samuel K.; Chen, Ching-Shih

    2010-01-01

    In light of the unique ability of thiazolidinediones to mediate peroxisome proliferator-activated receptor (PPAR)γ-independent activation of adenosine monophosphate-activated protein kinase (AMPK) and suppression of interleukin (IL)-6 production, we conducted a screening of an in-house, thiazolidinedione-based focused compound library to identify novel agents with these dual pharmacological activities. Cell-based assays pertinent to the activation status of AMPK and mammalian homolog of target of rapamycin (i.e., phosphorylation of AMPK and p70 ribosomal protein S6 kinase, respectively), and IL-6/IL-6 receptor signaling (i.e., IL-6 production and signal transducer and activator of transcription 3 phosphorylation, respectively) in lipopolysaccharide (LPS)-stimulated THP-1 human macrophages were used to screen this compound library, which led to the identification of compound 53 (N-{4-[3-(1-Methylcyclohexylmethyl)-2,4-dioxo-thiazolidin-5-ylidene-methyl]-phenyl}-4-nitro-3-trifluoromethyl-benzenesulfonamide) as the lead agent. Evidence indicates that this drug-induced suppression of LPS-stimulated IL-6 production was attributable to AMPK activation. Furthermore, compound 53-mediated AMPK activation was demonstrated in C-26 colon adenocarcinoma cells, indicating that it is not a cell line-specific event. PMID:20170185

  2. Full scale upper surface blown flap noise

    NASA Technical Reports Server (NTRS)

    Heidelberg, L. J.; Homyak, L.; Jones, W. L.

    1975-01-01

    A highly noise suppressed TF 34 engine was used to investigate the noise of several powered lift configurations involving upper surface blown (USB) flaps. The configuration variables were nozzle type (i.e. slot and circular with deflector), flap chord length, and flap angle. The results of velocity surveys at both the nozzle exit and the flap trailing edge are also presented and used for correlation of the noise data. Configurations using a long flap design were 4 db quieter than a short flap typical of current trends in USB flap design. The lower noise for the long flap is attributed primarily to the greater velocity decay of the jet at the flap trailing edge. The full-scale data revealed substantially more quadrupole noise in the region near the deflected jet than observed in previous sub-scale tests.

  3. Physiological, aerodynamic and geometric constraints of flapping account for bird gaits, and bounding and flap-gliding flight strategies.

    PubMed

    Usherwood, James Richard

    2016-11-01

    Aerodynamically economical flight is steady and level. The high-amplitude flapping and bounding flight style of many small birds departs considerably from any aerodynamic or purely mechanical optimum. Further, many large birds adopt a flap-glide flight style in cruising flight which is not consistent with purely aerodynamic economy. Here, an account is made for such strategies by noting a well-described, general, physiological cost parameter of muscle: the cost of activation. Small birds, with brief downstrokes, experience disproportionately high costs due to muscle activation for power during contraction as opposed to work. Bounding flight may be an adaptation to modulate mean aerodynamic force production in response to (1) physiological pressure to extend the duration of downstroke to reduce power demands during contraction; (2) the prevention of a low-speed downstroke due to the geometric constraints of producing thrust; (3) an aerodynamic cost to flapping with very low lift coefficients. In contrast, flap-gliding birds, which tend to be larger, adopt a strategy that reduces the physiological cost of work due both to activation and contraction efficiency. Flap-gliding allows, despite constraints to modulation of aerodynamic force lever-arm, (1) adoption of moderately large wing-stroke amplitudes to achieve suitable muscle strains, thereby reducing the activation costs for work; (2) reasonably quick downstrokes, enabling muscle contraction at efficient velocities, while being (3) prevented from very slow weight-supporting upstrokes due to the cost of performing 'negative' muscle work.

  4. Physiological, aerodynamic and geometric constraints of flapping account for bird gaits, and bounding and flap-gliding flight strategies.

    PubMed

    Usherwood, James Richard

    2016-11-01

    Aerodynamically economical flight is steady and level. The high-amplitude flapping and bounding flight style of many small birds departs considerably from any aerodynamic or purely mechanical optimum. Further, many large birds adopt a flap-glide flight style in cruising flight which is not consistent with purely aerodynamic economy. Here, an account is made for such strategies by noting a well-described, general, physiological cost parameter of muscle: the cost of activation. Small birds, with brief downstrokes, experience disproportionately high costs due to muscle activation for power during contraction as opposed to work. Bounding flight may be an adaptation to modulate mean aerodynamic force production in response to (1) physiological pressure to extend the duration of downstroke to reduce power demands during contraction; (2) the prevention of a low-speed downstroke due to the geometric constraints of producing thrust; (3) an aerodynamic cost to flapping with very low lift coefficients. In contrast, flap-gliding birds, which tend to be larger, adopt a strategy that reduces the physiological cost of work due both to activation and contraction efficiency. Flap-gliding allows, despite constraints to modulation of aerodynamic force lever-arm, (1) adoption of moderately large wing-stroke amplitudes to achieve suitable muscle strains, thereby reducing the activation costs for work; (2) reasonably quick downstrokes, enabling muscle contraction at efficient velocities, while being (3) prevented from very slow weight-supporting upstrokes due to the cost of performing 'negative' muscle work. PMID:27418386

  5. Comparison of Metalloproteinase Protein and Activity Profiling

    PubMed Central

    Giricz, Orsi; Lauer, Janelle L.; Fields, Gregg B.

    2010-01-01

    Proteolytic enzymes play fundamental roles in many biological processes. Members of the matrix metalloproteinase (MMP) family have been shown to take part in processes crucial in disease progression. The present study used the ExcelArray Human MMP/TIMP Array to quantify MMP and tissue inhibitor of metalloproteinase (TIMP) production in the lysates and media of 14 cancer and one normal cell line. The overall patterns were very similar in terms of which MMPs and TIMPs were secreted in the media versus associated with the cells in the individual samples. However, more MMP was found in the media, both in amount and in variety. TIMP-1 was produced in all cell lines. MMP activity assays with three different FRET substrates were then utilized to determine if protein production correlated with function for the WM-266-4 and BJ cell lines. Metalloproteinase activity was observed for both cell lines with a general MMP substrate (Knight SSP), consistent with protein production data. However, although both cell lines promoted the hydrolysis of a more selective MMP substrate (NFF-3), metalloproteinase activity was only confirmed in the BJ cell line. The use of inhibitors to confirm metalloproteinase activities pointed to the strengths and weaknesses of in situ FRET substrate assays. PMID:20920458

  6. Crowding Activates Heat Shock Protein 90.

    PubMed

    Halpin, Jackson C; Huang, Bin; Sun, Ming; Street, Timothy O

    2016-03-18

    Hsp90 is a dimeric ATP-dependent chaperone involved in the folding, maturation, and activation of diverse target proteins. Extensive in vitro structural analysis has led to a working model of Hsp90's ATP-driven conformational cycle. An implicit assumption is that dilute experimental conditions do not significantly perturb Hsp90 structure and function. However, Hsp90 undergoes a dramatic open/closed conformational change, which raises the possibility that this assumption may not be valid for this chaperone. Indeed, here we show that the ATPase activity of Hsp90 is highly sensitive to molecular crowding, whereas the ATPase activities of Hsp60 and Hsp70 chaperones are insensitive to crowding conditions. Polymer crowders activate Hsp90 in a non-saturable manner, with increasing efficacy at increasing concentration. Crowders exhibit a non-linear relationship between their radius of gyration and the extent to which they activate Hsp90. This experimental relationship can be qualitatively recapitulated with simple structure-based volume calculations comparing open/closed configurations of Hsp90. Thermodynamic analysis indicates that crowding activation of Hsp90 is entropically driven, which is consistent with a model in which excluded volume provides a driving force that favors the closed active state of Hsp90. Multiple Hsp90 homologs are activated by crowders, with the endoplasmic reticulum-specific Hsp90, Grp94, exhibiting the highest sensitivity. Finally, we find that crowding activation works by a different mechanism than co-chaperone activation and that these mechanisms are independent. We hypothesize that Hsp90 has a higher intrinsic activity in the cell than in vitro. PMID:26797120

  7. The Role of Muscle Flaps for Salvage of Failed Perforator Free Flaps

    PubMed Central

    2015-01-01

    Background: Despite the most heroic efforts, sometimes free flaps fail. Perforator free flaps are not invincible and can suffer the same fate. The real challenge is how to decide what is the next best choice for achieving the desired outcome. Methods: Over the past decade, 298 free perforator flaps were used in our institution. Total failure occurred in 16 patients, and partial failure requiring a second free flap occurred in an additional 6 patients for a true success rate of 93%. All failures had some form of secondary vascularized tissue transfer, which included the use of muscle flaps in 9 (41%) different patients. Results: Initial flap salvage after a failed perforator free flap was attempted with 12 perforator and 5 muscle free flaps as well as 1 perforator and 2 muscle local flaps. These were not all successful, with loss of 3 muscle free flaps and 3 perforator flaps. Tertiary free flap coverage was successful in 3 cases using 2 muscle flaps and 1 perforator free flap. Local fasciocutaneous flaps or primary wound closure was used in the remaining individuals. Conclusions: Microsurgical tissue transfers can be the most rewarding and at the same time the most challenging reconstructive endeavor. Persistence in achieving the desired outcome can require multiple steps. Perforator flaps are an important asset to obtain this goal. However, muscle flaps can still be a useful alternative, and the message is that they should not be overlooked as sometimes a viable option. PMID:26893989

  8. Polycarboxylates Enhance Beetle Antifreeze Protein Activity

    PubMed Central

    Amornwittawat, Natapol; Wang, Sen; Duman, John G.; Wen, Xin

    2008-01-01

    Summary Antifreeze proteins (AFPs) lower the noncolligative freezing point of water in the presence of ice below the ice melting point. The temperature difference between the melting point and the noncolligative freezing point is termed thermal hysteresis (TH). The magnitude of the TH depends on the specific activity and the concentration of AFP, and the concentration of enhancers in the solution. Known enhancers are certain low molecular mass molecules and proteins. Here, we investigated a series of polycarboxylates that enhance the TH activity of an AFP from the beetle Dendroides canadensis (DAFP) using differential scanning calorimetry (DSC). Triethylenetetramine-N,N,N′,N″,N‴,N‴-hexaacetate, the most efficient enhancer identified in this work, can increase the TH of DAFP by nearly 1.5 fold over than that of the published best enhancer, citrate. The Zn2+ coordinated carboxylate results in loss of the enhancement ability of the carboxylate on antifreeze activity. There is not an additional increase in TH when a weaker enhancer is added to a stronger enhancer solution. These observations suggest that the more carboxylate groups per enhancer molecule the better the efficiency of the enhancer and that the freedom of motion of these molecules is necessary for them to serve as enhancers for AFP. The hydroxyl groups in the enhancer molecules can also positively affect their TH enhancement efficiency, though not as strongly as carboxylate groups. Mechanisms are discussed. PMID:18620083

  9. PIV Measurements on a Blowing Flap

    NASA Technical Reports Server (NTRS)

    Hutcheson, Florence V.; Stead, Daniel J.

    2004-01-01

    PIV measurements of the flow in the region of a flap side edge are presented for several blowing flap configurations. The test model is a NACA 63(sub 2)-215 Hicks Mod-B main-element airfoil with a half-span Fowler flap. Air is blown from small slots located along the flap side edge on either the top, bottom or side surfaces. The test set up is described and flow measurements for a baseline and three blowing flap configurations are presented. The effects that the flap tip jets have on the structure of the flap side edge flow are discussed for each of the flap configurations tested. The results indicate that blowing air from a slot located along the top surface of the flap greatly weakened the top vortex system and pushed it further off the top surface. Blowing from the bottom flap surface kept the strong side vortex further outboard while blowing from the side surface only strengthened the vortex system or accelerated the merging of the side vortex to the flap top surface. It is concluded that blowing from the top or bottom surfaces of the flap may lead to a reduction of flap side edge noise.

  10. Experimental Study of Wake / Flap Interaction Noise and the Reduction of Flap Side Edge Noise

    NASA Technical Reports Server (NTRS)

    Hutcheson, Florence V.; Stead, Daniel J.; Plassman, Gerald E.

    2016-01-01

    The effects of the interaction of a wake with a half-span flap on radiated noise are examined. The incident wake is generated by bars of various widths and lengths or by a simplified landing gear model. Single microphone and phased array measurements are used to isolate the effects of the wake interaction on the noise radiating from the flap side edge and flap cove regions. The effects on noise of the wake generator's geometry and relative placement with respect to the flap are assessed. Placement of the wake generators upstream of the flap side edge is shown to lead to the reduction of flap side edge noise by introducing a velocity deficit and likely altering the instabilities in the flap side edge vortex system. Significant reduction in flap side edge noise is achieved with a bar positioned directly upstream of the flap side edge. The noise reduction benefit is seen to improve with increased bar width, length and proximity to the flap edge. Positioning of the landing gear model upstream of the flap side edge also leads to decreased flap side edge noise. In addition, flap cove noise levels are significantly lower than when the landing gear is positioned upstream of the flap mid-span. The impact of the local flow velocity on the noise radiating directly from the landing gear is discussed. The effects of the landing gear side-braces on flap side edge, flap cove and landing gear noise are shown.

  11. [Protein kinase C activation induces platelet apoptosis].

    PubMed

    Zhao, Li-Li; Chen, Meng-Xing; Zhang, Ming-Yi; Dai, Ke-Sheng

    2013-10-01

    Platelet apoptosis elucidated by either physical or chemical compound or platelet storage occurs wildly, which might play important roles in controlling the numbers and functions of circulated platelets, or in the development of some platelet-related diseases. However, up to now, a little is known about the regulatory mechanisms of platelet apoptosis. Protein kinase C (PKC) is highly expressed in platelets and plays central roles in regulating platelet functions. Although there is evidence indicating that PKC is involved in the regulation of apoptosis of nucleated cells, it is still unclear whether PKC plays a role in platelet apoptosis. The aim of this study was to investigate the role of PKC in platelet apoptosis. The effects of PKC on mitochondrial membrane potential (ΔΨm), phosphatidylserine (PS) exposure, and caspase-3 activation of platelets were analyzed by flow cytometry and Western blot. The results showed that the ΔΨm depolarization in platelets was induced by PKC activator in time-dependent manner, and the caspase-3 activation in platelets was induced by PKC in concentration-dependent manner. However, the platelets incubated with PKC inhibitor did not results in ΔΨm depolarization and PS exposure. It is concluded that the PKC activation induces platelet apoptosis through influencing the mitochondrial functions and activating caspase 3. The finds suggest a novel mechanism for PKC in regulating platelet numbers and functions, which has important pathophysiological implications for thrombosis and hemostasis.

  12. Beneficial Effects of Aminoguanidine on Skin Flap Survival in Diabetic Rats

    PubMed Central

    Ozturk, Ayse; Fırat, Cemal; Parlakpınar, Hakan; Bay-Karabulut, Aysun; Kirimlioglu, Hale; Gurlek, Ali

    2012-01-01

    Random flaps in DM patients have poor reliability for wound coverage, and flap loss remains a complex challenge. The protective effects of aminoguanidine (AG) administration on the survival of dorsal random flaps and oxidative stress were studied in diabetic rats. Two months after the onset of DM, dorsal McFarlane flaps were raised. Forty rats were divided into four groups: (1) control, (2) AG, (3) DM, and (4) DM + AG groups. Flap viability, determined with the planimetric method, and free-radical measurements were investigated. In addition, HbA1c and blood glucose levels, body weight measurements, and histopathological examinations were evaluated. The mean flap necrotic areas (%) in Groups I to IV were 50.9 ± 13.0, 32.9 ± 12.5, 65.2 ± 11.5, and 43.5 ± 14.7, respectively. The malondialdehyde (MDA) and nitric oxide (NO) levels were higher in the DM group than in the nondiabetic group, while the reduced glutathione (GSH) levels and superoxide dismutase (SOD) activity were reduced as a result of flap injury. In the diabetic and nondiabetic groups, AG administration significantly reduced the MDA and NO levels and significantly increased GSH content and SOD enzyme activity. We concluded that AG plays an important role in preventing random pattern flap necrosis. PMID:23304118

  13. Heat dissipation guides activation in signaling proteins

    PubMed Central

    Weber, Jeffrey K.; Shukla, Diwakar; Pande, Vijay S.

    2015-01-01

    Life is fundamentally a nonequilibrium phenomenon. At the expense of dissipated energy, living things perform irreversible processes that allow them to propagate and reproduce. Within cells, evolution has designed nanoscale machines to do meaningful work with energy harnessed from a continuous flux of heat and particles. As dictated by the Second Law of Thermodynamics and its fluctuation theorem corollaries, irreversibility in nonequilibrium processes can be quantified in terms of how much entropy such dynamics produce. In this work, we seek to address a fundamental question linking biology and nonequilibrium physics: can the evolved dissipative pathways that facilitate biomolecular function be identified by their extent of entropy production in general relaxation processes? We here synthesize massive molecular dynamics simulations, Markov state models (MSMs), and nonequilibrium statistical mechanical theory to probe dissipation in two key classes of signaling proteins: kinases and G-protein–coupled receptors (GPCRs). Applying machinery from large deviation theory, we use MSMs constructed from protein simulations to generate dynamics conforming to positive levels of entropy production. We note the emergence of an array of peaks in the dynamical response (transient analogs of phase transitions) that draw the proteins between distinct levels of dissipation, and we see that the binding of ATP and agonist molecules modifies the observed dissipative landscapes. Overall, we find that dissipation is tightly coupled to activation in these signaling systems: dominant entropy-producing trajectories become localized near important barriers along known biological activation pathways. We go on to classify an array of equilibrium and nonequilibrium molecular switches that harmonize to promote functional dynamics. PMID:26240354

  14. Phospholipases as GTPase activity accelerating proteins (GAPs) in plants.

    PubMed

    Pandey, Sona

    2016-05-01

    GTPase activity accelerating proteins (GAPs) are key regulators of the G-protein signaling cycle. By facilitating effective hydrolysis of the GTP bound on Gα proteins, GAPs control the timing and amplitude of the signaling cycle and ascertain the availability of the inactive heterotrimer for the next round of activation. Until very recently, the studies of GAPs in plants were focused exclusively on the regulator of G-protein signaling (RGS) protein. We now show that phospholipase Dα1 (PLDα1) is also a bona fide GAP in plants and together with the RGS protein controls the level of activeprotein. PMID:27124090

  15. Development of Potent Adenosine Monophosphate Activated Protein Kinase (AMPK) Activators.

    PubMed

    Dokla, Eman M E; Fang, Chun-Sheng; Lai, Po-Ting; Kulp, Samuel K; Serya, Rabah A T; Ismail, Nasser S M; Abouzid, Khaled A M; Chen, Ching-Shih

    2015-11-01

    Previously, we reported the identification of a thiazolidinedione-based adenosine monophosphate activated protein kinase (AMPK) activator, compound 1 (N-[4-({3-[(1-methylcyclohexyl)methyl]-2,4-dioxothiazolidin-5-ylidene}methyl)phenyl]-4-nitro-3-(trifluoromethyl)benzenesulfonamide), which provided a proof of concept to delineate the intricate role of AMPK in regulating oncogenic signaling pathways associated with cell proliferation and epithelial-mesenchymal transition (EMT) in cancer cells. In this study, we used 1 as a scaffold to conduct lead optimization, which generated a series of derivatives. Analysis of the antiproliferative and AMPK-activating activities of individual derivatives revealed a distinct structure-activity relationship and identified 59 (N-(3-nitrophenyl)-N'-{4-[(3-{[3,5-bis(trifluoromethyl)phenyl]methyl}-2,4-dioxothiazolidin-5-ylidene)methyl]phenyl}urea) as the optimal agent. Relative to 1, compound 59 exhibits multifold higher potency in upregulating AMPK phosphorylation in various cell lines irrespective of their liver kinase B1 (LKB1) functional status, accompanied by parallel changes in the phosphorylation/expression levels of p70S6K, Akt, Foxo3a, and EMT-associated markers. Consistent with its predicted activity against tumors with activated Akt status, orally administered 59 was efficacious in suppressing the growth of phosphatase and tensin homologue (PTEN)-null PC-3 xenograft tumors in nude mice. Together, these findings suggest that 59 has clinical value in therapeutic strategies for PTEN-negative cancer and warrants continued investigation in this regard.

  16. Receptor activity-modifying proteins; multifunctional G protein-coupled receptor accessory proteins.

    PubMed

    Hay, Debbie L; Walker, Christopher S; Gingell, Joseph J; Ladds, Graham; Reynolds, Christopher A; Poyner, David R

    2016-04-15

    Receptor activity-modifying proteins (RAMPs) are single pass membrane proteins initially identified by their ability to determine the pharmacology of the calcitonin receptor-like receptor (CLR), a family B G protein-coupled receptor (GPCR). It is now known that RAMPs can interact with a much wider range of GPCRs. This review considers recent developments on the structure of the complexes formed between the extracellular domains (ECDs) of CLR and RAMP1 or RAMP2 as these provide insights as to how the RAMPs direct ligand binding. The range of RAMP interactions is also considered; RAMPs can interact with numerous family B GPCRs as well as examples of family A and family C GPCRs. They influence receptor expression at the cell surface, trafficking, ligand binding and G protein coupling. The GPCR-RAMP interface offers opportunities for drug targeting, illustrated by examples of drugs developed for migraine. PMID:27068971

  17. New model of flap-gliding flight.

    PubMed

    Sachs, Gottfried

    2015-07-21

    A new modelling approach is presented for describing flap-gliding flight in birds and the associated mechanical energy cost of travelling. The new approach is based on the difference in the drag characteristics between flapping and non-flapping due to the drag increase caused by flapping. Thus, the possibility of a gliding flight phase, as it exists in flap-gliding flight, yields a performance advantage resulting from the decrease in the drag when compared with continuous flapping flight. Introducing an appropriate non-dimensionalization for the mathematical relations describing flap-gliding flight, results and findings of generally valid nature are derived. It is shown that there is an energy saving of flap-gliding flight in the entire speed range compared to continuous flapping flight. The energy saving reaches the highest level in the lower speed region. The travelling speed of flap-gliding flight is composed of the weighted average of the differing speeds in the flapping and gliding phases. Furthermore, the maximum range performance achievable with flap-gliding flight and the associated optimal travelling speed are determined.

  18. Pyrrolopyridine inhibitors of mitogen-activated protein kinase-activated protein kinase 2 (MK-2).

    PubMed

    Anderson, David R; Meyers, Marvin J; Vernier, William F; Mahoney, Matthew W; Kurumbail, Ravi G; Caspers, Nicole; Poda, Gennadiy I; Schindler, John F; Reitz, David B; Mourey, Robert J

    2007-05-31

    A new class of potent kinase inhibitors selective for mitogen-activated protein kinase-activated protein kinase 2 (MAPKAP-K2 or MK-2) for the treatment of rheumatoid arthritis has been prepared and evaluated. These inhibitors have IC50 values as low as 10 nM against the target and have good selectivity profiles against a number of kinases including CDK2, ERK, JNK, and p38. These MK-2 inhibitors have been shown to suppress TNFalpha production in U397 cells and to be efficacious in an acute inflammation model. The structure-activity relationships of this series, the selectivity for MK-2 and their activity in both in vitro and in vivo models are discussed. The observed selectivity is discussed with the aid of an MK-2/inhibitor crystal structure.

  19. Benefit of HSP90α intervention on ischemia-reperfusion injury of venous blood-congested flaps

    PubMed Central

    HU, XIAO-YING; CHEN, ZHEN-YU; ZHANG, BIN; LENG, XIANG-FENG; FAN, XIAO-JIAN; LIU, TAO

    2016-01-01

    In order to decrease the incidence of flap necrosis after reconstructive surgeries, new approaches are required. In the present study, a model of venous congested flaps in rats was established to test the heat shock protein (HSP) 90α, ‘F-5’, protein as an intervention therapy to alleviate ischemia-reperfusion injury. A recombinant plasmid pET15b-F-5 carrying the HSP90α gene was constructed and the induced protein was purified from bacterial cell cultures. The rats in the study were divided into three different intervention groups: group A rats were treated with normal saline prior to flap establishment, group B rats were treated with HSP90α, ‘F-5’, protein prior to flap establishment, and group C rats were treated with the same ‘F-5’ protein after the surgical procedure. Additionally, the reperfusion time-points, ischemia for 6 or 8 h (5 rats each), were established in each group. After set periods of time, the flaps were observed for skin appearance, blood flow, survival rate and histological changes including neovascularization and re-epithelialization. The results showed that the flaps in the rats pre-treated with ‘F-5’ protein performed better than the flaps of rats in the other two groups: the blood flow was higher, flap survival rate was increased, inflammatory cell infiltration was decreased and angiogenesis increased, and new skin structure was better completed by the end of the experiment. The parameters examind were improved for all the groups when the ischemia time was 6 h instead of 8 h. In conclusion, HSP90α intervention prior to flap establishment was shown to be beneficial in the model of ischemia-reperfusion injury in venous-congested flaps. PMID:27347036

  20. Efficient flapping flight of pterosaurs

    NASA Astrophysics Data System (ADS)

    Strang, Karl Axel

    In the late eighteenth century, humans discovered the first pterosaur fossil remains and have been fascinated by their existence ever since. Pterosaurs exploited their membrane wings in a sophisticated manner for flight control and propulsion, and were likely the most efficient and effective flyers ever to inhabit our planet. The flapping gait is a complex combination of motions that sustains and propels an animal in the air. Because pterosaurs were so large with wingspans up to eleven meters, if they could have sustained flapping flight, they would have had to achieve high propulsive efficiencies. Identifying the wing motions that contribute the most to propulsive efficiency is key to understanding pterosaur flight, and therefore to shedding light on flapping flight in general and the design of efficient ornithopters. This study is based on published results for a very well-preserved specimen of Coloborhynchus robustus, for which the joints are well-known and thoroughly described in the literature. Simplifying assumptions are made to estimate the characteristics that can not be inferred directly from the fossil remains. For a given animal, maximizing efficiency is equivalent to minimizing power at a given thrust and speed. We therefore aim at finding the flapping gait, that is the joint motions, that minimize the required flapping power. The power is computed from the aerodynamic forces created during a given wing motion. We develop an unsteady three-dimensional code based on the vortex-lattice method, which correlates well with published results for unsteady motions of rectangular wings. In the aerodynamic model, the rigid pterosaur wing is defined by the position of the bones. In the aeroelastic model, we add the flexibility of the bones and of the wing membrane. The nonlinear structural behavior of the membrane is reduced to a linear modal decomposition, assuming small deflections about the reference wing geometry. The reference wing geometry is computed for

  1. Schooling of flapping wings: Simulations

    NASA Astrophysics Data System (ADS)

    Masoud, Hassan; Becker, Alexander; Ristroph, Leif; Shelley, Michael

    2014-11-01

    We examine the locomotion of an infinite array of wings that heave vertically with a prescribed sinusoidal motion and are free to translate in the horizontal direction. To do this, we simulate the motion of a freely translating flapping airfoil in a domain with periodic horizontal boundary conditions. These simulations indicate that the wings can ``take advantage'' of their collectively generated wake flows. In agreement with our experiments in a rotational geometry, we find ranges of flapping frequency over which there are multiple stable states of locomotion, with one of these swimming states having both higher speeds and efficiencies than an isolated flapping and locomoting wing. A simple mathematical model, which emphasizes the importance of history dependence in vortical flows, explains this multi-stability. These results may be important to understanding the role of hydrodynamic interactions in fish schooling and bird flocking.

  2. Deep digital burns treated with 2 abdominal flaps: case report.

    PubMed

    Wu, Choulang; Zhou, Lichun; Zhu, Lili; Zheng, Jinman

    2013-11-01

    We present the case of a 22-year-old man with third- to fourth-degree flame burns to all fingers of the right hand. After removing the necrotic tissues and amputating the carbonized distal segments of each finger, we covered the injured thumb with a paraumbilical tubular flap and covered the other fingers with an abdominal wall marsupial flap. Fifty days after injury, all the wounds healed, and the remaining fingers were salvaged. Four months after injury, the grip strength and the first web span of the right hand was 23.6 kg and 53°, and the patient could fulfill almost all the activities of daily living.

  3. Development of a Wind Turbine Test Rig and Rotor for Trailing Edge Flap Investigation: Static Flap Angles Case

    NASA Astrophysics Data System (ADS)

    Abdelrahman, Ahmed; Johnson, David A.

    2014-06-01

    One of the strategies used to improve performance and increase the life-span of wind turbines is active flow control. It involves the modification of the aerodynamic characteristics of a wind turbine blade by means of moveable aerodynamic control surfaces. Trailing edge flaps are relatively small moveable control surfaces placed at the trailing edge of a blade's airfoil that modify the lift of a blade or airfoil section. An instrumented wind turbine test rig and rotor were specifically developed to enable a wide-range of experiments to investigate the potential of trailing edge flaps as an active control technique. A modular blade based on the S833 airfoil was designed to allow accurate instrumentation and customizable settings. The blade is 1.7 meters long, had a constant 178mm chord and a 6° pitch. The modular aerodynamic parts were 3D printed using plastic PC-ABS material. The blade design point was within the range of wind velocities in the available large test facility. The wind facility is a large open jet wind tunnel with a maximum velocity of 11m/s in the test area. The capability of the developed system was demonstrated through an initial study of the effect of stationary trailing edge flaps on blade load and performance. The investigation focused on measuring the changes in flapwise bending moment and power production for different trailing edge flap spanwise locations and deflection angles. The relationship between the load reduction and deflection angle was linear as expected from theory and the highest reduction was caused by the flap furthest from the rotor center. Overall, the experimental setup proved to be effective in measuring small changes in flapwise bending moment within the wind turbine blade and will provide insight when (active) flap control is targeted.

  4. Etanercept protects myocutaneous flaps from ischaemia reperfusion injury: An experimental study in a rat tram flap model.

    PubMed

    Ersoy, Burak; Çevik, Özge; Çilingir, Özlem Tuğçe

    2016-08-01

    Background Being an inevitable component of free tissue transfer, ischemia-reperfusion injury tends to contribute to flap failure. TNF-α is an important proinflammatory cytokine and a prominent mediator of the ischemia-reperfusion injury. Etanercept, a soluble TNF-α binding protein, has shown anti-inflammatory and anti-apoptotic effects in animal models of renal and myocardial ischemia-reperfusion injury. We have designed an experimental study to investigate the effect of etanercept on myocutaneous ischemia-reperfusion injury on transverse rectus abdominis myocutaneous flap model in rats. Methods Twenty-four male Sprague-Dawley rats were divided into 3 groups: In group 1 (sham), the TRAM flap was raised and sutured back without further intervention. In group 2 (control), the flap was raised and the ischemia-reperfusion protocol was followed. In group 3, etanercept (10 mg/kg, i.v.) was administered 10 minutes before reperfusion. At the end of the reperfusion period, biochemical and histolopathological evaluations were performed on serum and tissue samples. Results In the etanercept group the IMA and 8-OHdG levels (p = 0.005 and p = 0.004, respectively) were found significantly lower, and the GSH and SOD levels (p = 0.01 and p < 0.001, respectively) significantly higher in comparison to the control group. The histopathological analysis has revealed a lower degree of hyalinization, degenerated muscle fibers and nuclear change in the etanercept group compared to the control group. Conclusion The results of our experimental study indicate that etanercept offers protection against ischemia-reperfusion injury in skeletal muscle tissue, enhancing the TRAM flap viability. The ability of etanercept to induce ischemic tolerance suggests that it may be applicable in free-flap surgery. PMID:26950289

  5. Arabinogalactan proteins: focus on carbohydrate active enzymes

    PubMed Central

    Knoch, Eva; Dilokpimol, Adiphol; Geshi, Naomi

    2014-01-01

    Arabinogalactan proteins (AGPs) are a highly diverse class of cell surface proteoglycans that are commonly found in most plant species. AGPs play important roles in many cellular processes during plant development, such as reproduction, cell proliferation, pattern formation and growth, and in plant-microbe interaction. However, little is known about the molecular mechanisms of their function. Numerous studies using monoclonal antibodies that recognize different AGP glycan epitopes have shown the appearance of a slightly altered AGP glycan in a specific stage of development in plant cells. Therefore, it is anticipated that the biosynthesis and degradation of AGP glycan is tightly regulated during development. Until recently, however, little was known about the enzymes involved in the metabolism of AGP glycans. In this review, we summarize recent discoveries of carbohydrate active enzymes (CAZy; http://www.cazy.org/) involved in the biosynthesis and degradation of AGP glycans, and we discuss the biological role of these enzymes in plant development. PMID:24966860

  6. Observation of microtubule-based motor protein activity.

    PubMed

    Sloboda, Roger D

    2015-02-01

    It is possible to detect the presence of motor proteins that have the ability to translocate particles along microtubules. The two procedures described here were developed to detect microtubule-dependent motor protein activity in cell lysates or of purified proteins. In the first procedure, latex beads bound to the putative motor protein are assayed for their ability to translocate along microtubules in an ATP-dependent fashion. If motor protein activity is present, it will bind to the beads and translocate them unidirectionally along the microtubules. In the second procedure, motor proteins induce microtubule gliding over a glass coverslip surface that is coated with active motor protein. Because the mass of a microtubule is negligible compared to that of a coverslip or slide, the microtubule glides over the glass surface when the surface is coated with active motor protein. Also included here are descriptions of assays designed to determine the directionality of movement of microtubule-based motor proteins. PMID:25646501

  7. Proline Scanning Mutagenesis Reveals a Role for the Flap Endonuclease-1 Helical Cap in Substrate Unpairing*

    PubMed Central

    Patel, Nikesh; Exell, Jack C.; Jardine, Emma; Ombler, Ben; Finger, L. David; Ciani, Barbara; Grasby, Jane A.

    2013-01-01

    The prototypical 5′-nuclease, flap endonuclease-1 (FEN1), catalyzes the essential removal of single-stranded flaps during DNA replication and repair. FEN1 hydrolyzes a specific phosphodiester bond one nucleotide into double-stranded DNA. This specificity arises from double nucleotide unpairing that places the scissile phosphate diester on active site divalent metal ions. Also related to FEN1 specificity is the helical arch, through which 5′-flaps, but not continuous DNAs, can thread. The arch contains basic residues (Lys-93 and Arg-100 in human FEN1 (hFEN1)) that are conserved by all 5′-nucleases and a cap region only present in enzymes that process DNAs with 5′ termini. Proline mutations (L97P, L111P, L130P) were introduced into the hFEN1 helical arch. Each mutation was severely detrimental to reaction. However, all proteins were at least as stable as wild-type (WT) hFEN1 and bound substrate with comparable affinity. Moreover, all mutants produced complexes with 5′-biotinylated substrate that, when captured with streptavidin, were resistant to challenge with competitor DNA. Removal of both conserved basic residues (K93A/R100A) was no more detrimental to reaction than the single mutation R100A, but much less severe than L97P. The ability of protein-Ca2+ to rearrange 2-aminopurine-containing substrates was monitored by low energy CD. Although L97P and K93A/R100A retained the ability to unpair substrates, the cap mutants L111P and L130P did not. Taken together, these data challenge current assumptions related to 5′-nuclease family mechanism. Conserved basic amino acids are not required for double nucleotide unpairing and appear to act cooperatively, whereas the helical cap plays an unexpected role in hFEN1-substrate rearrangement. PMID:24126913

  8. Natural Hirudin Increases Rat Flap Viability by Anti-Inflammation via PARs/p38/NF-κB Pathway

    PubMed Central

    Peng, Liu; Pan, Xinyuan; Yin, Guoqian

    2015-01-01

    The present study aimed to evaluate the effect of natural hirudin on rat random skin flap viability and to determine the mechanism. Forty-eight rats were randomly divided into 2 groups. After the dorsal skin flap operation (3 cm × 10 cm in size), subcutaneous injections of 6 ATU hirudin were administered to group H (n = 24) every 12 h, while group C (n = 24) received an equal volume of 0.9% normal saline. Six rats from each group were euthanized 1, 2, 4, and 7 days after the operation. A full skin sample was collected from these rats to measure the p38-mitogen-activated protein kinase (p38-MAPK), phospho-p38- (Pp38-) MAPK, nuclear factor-κB (NF-κB) p65, phosphor-NF-κB (pNF-κB) p65, tumour necrosis factor- (TNF-) α, interleukin- (IL-) 6, and intercellular adhesion molecule- (ICAM-) 1 levels via western blot (WB) assays. The results showed that flap viability was significantly higher in the hirudin-treated group, which showed a reduced inflammatory response compared with the control group. The Pp38/p38, pNF-κB p65/NF-κB p65, TNF-α, IL-6, and ICAM-1 levels in the hirudin-treated group were lower than those in the control group. The results demonstrated that hirudin could improve random skin flap viability and suggested that this effect maybe occurs by blocking the thrombin/proteinase-activated receptors (PARs)/p38/NF-κB signalling pathway, thus decreasing the inflammatory response. PMID:26770977

  9. Sternocleidomastoid Muscle Flap after Parotidectomy

    PubMed Central

    Nofal, Ahmad Abdel-Fattah; Mohamed, Morsi

    2015-01-01

    Introduction Most patients after either superficial or total parotidectomy develop facial deformity and Frey syndrome, which leads to a significant degree of patient dissatisfaction. Objective Assess the functional outcome and esthetic results of the superiorly based sternocleidomastoid muscle (SCM) flap after superficial or total parotidectomy. Methods A prospective cohort study for 11 patients subjected to parotidectomy using a partial-thickness superiorly based SCM flap. The functional outcome (Frey syndrome, facial nerve involvement, and ear lobule sensation) and the esthetic results were evaluated subjectively and objectively. Results Facial nerve palsy occurred in 5 cases (45%), and all of them recovered completely within 6 months. The Minor starch iodine test was positive in 3 patients (27%), although only 1 (9%) subjectively complained of gustatory sweating. The designed visual analog score completed by the patients themselves ranged from 0 to 3 with a mean of 1.55 ± 0.93; the scores from the blinded evaluators ranged from 1 to 3 with a mean 1.64 ± 0.67. Conclusion The partial-thickness superiorly based SCM flap offers a reasonable cosmetic option for reconstruction following either superficial or total parotidectomy by improving the facial deformity. The flap also lowers the incidence of Frey syndrome objectively and subjectively with no reported hazard of the spinal accessory nerve. PMID:26491478

  10. Fibrin-embedded administration of VEGF plasmid enhances skin flap survival.

    PubMed

    Michlits, Wolfgang; Mittermayr, Rainer; Schäfer, Romana; Redl, Heinz; Aharinejad, Seyedhossein

    2007-01-01

    The aim of the present study was to experimentally evaluate whether topical fibrin-mediated administration of a vascular endothelial growth factor (VEGF)-A plasmid to the wound bed can protect skin flaps from necrosis. A plasmid expression vector containing the VEGF-A cDNA was constructed. The plasmid was then administered to the wound bed of rat abdominal skin flaps in a fibrin sealant. The percentage of viable, ischemic and necrotic tissue was assessed postoperatively as a baseline and after 3 and 7 days using digital surface area morphometry. Laser Doppler imaging of the flaps and VEGF-A Western blot analysis of flap tissue were performed to assess angiogenesis and VEGF-A tissue levels. Flaps treated with VEGF plasmids in the presence of uptake enhancing Lipofectamine transfection reagent increased flap survival 7 days postoperatively significantly associated with markedly elevated tissue perfusion and enhanced tissue VEGF-A protein expression. Our results indicate that topical fibrin-mediated administration of a VEGF-A plasmid may serve as an alternative to previous strategies in treating ischemic skin flaps. The suggested therapeutic approach is easily applicable and inexpensive in preparation. Thus, this protocol may also enhance wound healing in posttrauma skin lacerations or in skin grafts.

  11. A new agent for flap survival - Hippophae rhamnoides L. (sea buckthorn): An experimental study in rats.

    PubMed

    Emsen, Ilteris Murat

    2005-01-01

    Hippophae rhamnoides L. (sea buckthorn) is a member of the Elaeagnaceae family, and is a temperate bush native to Europe and Asia. The antioxidant activity of H rhamnoides L. has been shown in vitro cell culture and animal studies. Different fractions of H rhamnoides L. fruits inhibit 2,2-azobis-(2,4 dimethylvaleronitrile) and ascorbate iron-induced lipid peroxidations in vitro. H rhamnoides L., as well as vitamin E, decrease the malondialdehyde content in hyperlipidemic rabbit serum-cultured smooth muscle cells. The aim of the present study was to investigate, in a rat model, the potential effect of H rhamnoides L. on survival of random pattern skin flaps. For this purpose, 30 Wistar Albino rats were used, and a McFarlane-type caudally based skin flap was created on the dorsum of the rat (2.5 cm × 8 cm). Rats were divided into three groups: one control (group A) and two treatment groups (groups B and C). H rhamnoides L. was administered orally to the experimental groups: group B received a single 15 mg/kg dose per day and group C received 15 mg/kg twice per day. The areas and lengths of flap necrosis were measured in each group. The extent of necrotic flap areas were evaluated as length and area of total flap area, and differences were studied by Student's t tests. The areas and lengths of necrosis of skin flaps decreased depending on H rhamnoides L., but viability of the flaps treated with 15 mg/kg/day was not significantly different from the control group. The rats receiving H rhamnoides L. 15 mg/kg twice per day had the highest flap survival rate (P<0.001). In conclusion, H rhamnoides L. may have a dose-dependent effect to increase flap survival in random skin flaps. PMID:24227931

  12. The possibility for use of venous flaps in plastic surgery

    NASA Astrophysics Data System (ADS)

    Baytinger, V. F.; Kurochkina, O. S.; Selianinov, K. V.; Baytinger, A. V.; Dzyuman, A. N.

    2015-11-01

    The use of venous flaps is controversial. The mechanism of perfusion of venous flaps is still not fully understood. The research was conducted on 56 white rats. In our experimental work we studied two different models of venous flaps: pedicled venous flap (PVF) and pedicled arterialized venous flap (PAVF). Our results showed that postoperative congestion was present in all flaps. However 66.7% of all pedicled venous flaps and 100% of all pedicled arterialized venous flaps eventually survived. Histological examination revealed that postoperatively the blood flow in the skin of the pedicled arterialized venous flap became «re-reversed» again; there were no differences between mechanism of survival of venous flaps and other flaps. On the 7-14th day in the skin of all flaps were processes of neoangiogenesis and proliferation. Hence the best scenario for the clinical use of venous flaps unfolds when both revascularization and skin coverage are required.

  13. The possibility for use of venous flaps in plastic surgery

    SciTech Connect

    Baytinger, V. F. Kurochkina, O. S. Selianinov, K. V.; Baytinger, A. V.; Dzyuman, A. N.

    2015-11-17

    The use of venous flaps is controversial. The mechanism of perfusion of venous flaps is still not fully understood. The research was conducted on 56 white rats. In our experimental work we studied two different models of venous flaps: pedicled venous flap (PVF) and pedicled arterialized venous flap (PAVF). Our results showed that postoperative congestion was present in all flaps. However 66.7% of all pedicled venous flaps and 100% of all pedicled arterialized venous flaps eventually survived. Histological examination revealed that postoperatively the blood flow in the skin of the pedicled arterialized venous flap became «re-reversed» again; there were no differences between mechanism of survival of venous flaps and other flaps. On the 7-14th day in the skin of all flaps were processes of neoangiogenesis and proliferation. Hence the best scenario for the clinical use of venous flaps unfolds when both revascularization and skin coverage are required.

  14. Activating AMP-activated protein kinase (AMPK) slows renal cystogenesis.

    PubMed

    Takiar, Vinita; Nishio, Saori; Seo-Mayer, Patricia; King, J Darwin; Li, Hui; Zhang, Li; Karihaloo, Anil; Hallows, Kenneth R; Somlo, Stefan; Caplan, Michael J

    2011-02-01

    Renal cyst development and expansion in autosomal dominant polycystic kidney disease (ADPKD) involves both fluid secretion and abnormal proliferation of cyst-lining epithelial cells. The chloride channel of the cystic fibrosis transmembrane conductance regulator (CFTR) participates in secretion of cyst fluid, and the mammalian target of rapamycin (mTOR) pathway may drive proliferation of cyst epithelial cells. CFTR and mTOR are both negatively regulated by AMP-activated protein kinase (AMPK). Metformin, a drug in wide clinical use, is a pharmacological activator of AMPK. We find that metformin stimulates AMPK, resulting in inhibition of both CFTR and the mTOR pathways. Metformin induces significant arrest of cystic growth in both in vitro and ex vivo models of renal cystogenesis. In addition, metformin administration produces a significant decrease in the cystic index in two mouse models of ADPKD. Our results suggest a possible role for AMPK activation in slowing renal cystogenesis as well as the potential for therapeutic application of metformin in the context of ADPKD. PMID:21262823

  15. Comprehensive Evaluation of Risk Factors and Management of Impending Flap Loss in 2138 Breast Free Flaps.

    PubMed

    Chang, Edward I; Chang, Eric I; Soto-Miranda, Miguel A; Zhang, Hong; Nosrati, Naveed; Crosby, Melissa A; Reece, Gregory P; Robb, Geoffrey L; Chang, David W

    2016-01-01

    Loss of a breast free flap is a relatively rare but catastrophic occurrence. Our study aims to identify risk factors for flap loss and to assess whether different salvage techniques affect flap salvage. We performed a retrospective review of all breast free flaps performed at a single institution from 2000 to 2010. Overall, 2138 flaps were performed in 1608 patients (unilateral, 1120 and bilateral, 488) with 44 flap losses (2.1%). Age, body mass index, smoking, radiation, chemotherapy, and surgeon experience did not affect flap loss. Abdominal flaps based on a single perforator were at significantly higher risk for flap loss compared with flaps based on multiple perforators (P = 0.0007). Subgroup analysis of the subset of 166 compromised free flaps (flaps requiring a return to the operating room, an intraoperative anastomotic revision, or loss/partial loss of a free flap) demonstrated deep inferior epigastric perforator, and other flaps (superficial inferior epigastric artery and superior gluteal artery perforator) were significantly associated with flap loss [odds ratio (OR) 5.20; P = 0.03 and OR 6.91; P = 0.0004, respectively] compared with transverse rectus abdominis myocutaneous and muscle-sparing transverse rectus abdominis myocutaneous flaps. Although an intraoperative complication was not associated with a flap loss, the need for a reoperation was strongly predictive (P < 0.0001). Flap salvage was the highest within the first 24 hours (83.7%) and significantly less between days 1 and 3 (38.6%; P < 0.0001) and beyond 4 days (29.4%; P < 0.0001). Longer ischemia time was significantly associated with flap loss (P = 0.04). Salvage techniques (aspirin, heparinzation, thrombectomy, and thrombolytic) had no impact on flap salvage rates. Heparinization and thrombolytics were associated with higher loss rates (OR 3.40; P = 0.003 and OR 10.36; P < 0.0001, respectively). Free flap loss following breast reconstruction is multifactorial with higher losses in superficial

  16. Forward flight of swallowtail butterfly with simple flapping motion.

    PubMed

    Tanaka, Hiroto; Shimoyama, Isao

    2010-06-01

    Unlike other flying insects, the wing motion of swallowtail butterflies is basically limited to flapping because their fore wings partly overlap their hind wings, structurally restricting the feathering needed for active control of aerodynamic force. Hence, it can be hypothesized that the flight of swallowtail butterflies is realized with simple flapping, requiring little feedback control of the feathering angle. To verify this hypothesis, we fabricated an artificial butterfly mimicking the wing motion and wing shape of a swallowtail butterfly and analyzed its flights using images taken with a high-speed video camera. The results demonstrated that stable forward flight could be realized without active feathering or feedback control of the wing motion. During the flights, the artificial butterfly's body moved up and down passively in synchronization with the flapping, and the artificial butterfly followed an undulating flight trajectory like an actual swallowtail butterfly. Without feedback control of the wing motion, the body movement is directly affected by change of aerodynamic force due to the wing deformation; the degree of deformation was determined by the wing venation. Unlike a veinless wing, a mimic wing with veins generated a much higher lift coefficient during the flapping flight than in a steady flow due to the large body motion.

  17. Biologically active LIL proteins built with minimal chemical diversity

    PubMed Central

    Heim, Erin N.; Marston, Jez L.; Federman, Ross S.; Edwards, Anne P. B.; Karabadzhak, Alexander G.; Petti, Lisa M.; Engelman, Donald M.; DiMaio, Daniel

    2015-01-01

    We have constructed 26-amino acid transmembrane proteins that specifically transform cells but consist of only two different amino acids. Most proteins are long polymers of amino acids with 20 or more chemically distinct side-chains. The artificial transmembrane proteins reported here are the simplest known proteins with specific biological activity, consisting solely of an initiating methionine followed by specific sequences of leucines and isoleucines, two hydrophobic amino acids that differ only by the position of a methyl group. We designate these proteins containing leucine (L) and isoleucine (I) as LIL proteins. These proteins functionally interact with the transmembrane domain of the platelet-derived growth factor β-receptor and specifically activate the receptor to transform cells. Complete mutagenesis of these proteins identified individual amino acids required for activity, and a protein consisting solely of leucines, except for a single isoleucine at a particular position, transformed cells. These surprisingly simple proteins define the minimal chemical diversity sufficient to construct proteins with specific biological activity and change our view of what can constitute an active protein in a cellular context. PMID:26261320

  18. G protein activation by G protein coupled receptors: ternary complex formation or catalyzed reaction?

    PubMed

    Roberts, David J; Waelbroeck, Magali

    2004-09-01

    G protein coupled receptors catalyze the GDP/GTP exchange on G proteins, thereby activating them. The ternary complex model, designed to describe agonist binding in the absence of GTP, is often extended to G protein activation. This is logically unsatisfactory as the ternary complex does not accumulate when G proteins are activated by GTP. Extended models taking into account nucleotide binding exist, but fail to explain catalytic G protein activation. This review puts forward an enzymatic model of G protein activation and compares its predictions with the ternary complex model and with observed receptor phenomenon. This alternative model does not merely provide a new set of formulae but leads to a new philosophical outlook and more readily accommodates experimental observations. The ternary complex model implies that, HRG being responsible for efficient G protein activation, it should be as stable as possible. In contrast, the enzyme model suggests that although a limited stabilization of HRG facilitates GDP release, HRG should not be "too stable" as this might trap the G protein in an inactive state and actually hinder G protein activation. The two models also differ completely in the definition of the receptor "active state": the ternary complex model implies that the active state corresponds to a single active receptor conformation (HRG); in contrast, the catalytic model predicts that the active receptor state is mobile, switching smoothly through various conformations with high and low affinities for agonists (HR, HRG, HRGGDP, HRGGTP, etc.).

  19. The binding landscape of plasmepsin V and the implications for flap dynamics.

    PubMed

    L, McGillewie; Soliman, Mahmoud E

    2016-04-26

    Plasmepsin V belongs to the plasmepsin family of aspartic proteases. PlmV is unique compared to other plasmepsins, as this membrane bound aspartic protease resides in the endoplasmic reticulum and is responsible for the cleavage of PEXEL tagged proteins destined for export outside of the host red blood cell. Plasmepsin V is highly conserved throughout the Plasmodium species, and is essential to the survival of the parasite. Recently, two potent inhibitors of Plmv have been identified, WEHI-916 and WEHI-842. Of these inhibitors, WEHI-842 has a higher binding affinity for P. vivax PlmV, and a crystal structure of PlmV in complex with WEHI-842 has recently been resolved (). The structure of PlmV is unique compared to other plasmepsins; it is stabilised internally by seven disulfide bonds, a NAP1 insert/fold is associated with the movement of the flap covering the active site and a highly conserved helix-turn-helix is situated towards the C-terminus. Flap motion and dynamics play an important role in enzyme selectivity and function. To better understand the impact of ligand binding on the flap dynamics, molecular dynamic simulations and post-dynamic analysis were employed in the present study on PlmV in complex with WEHI-842. Previously defined parameters, which accurately accounted for the opening and closing of the active site, were used to assess the conformational changes induced in the absence and presence of WEHI-842. From the simulations it can be seen that inhibitor binding significantly reduces the flexibility and mobility of not only the flap and flexible loop but areas outside of the active site. Ligand binding leads to the formation of a more stable compact structure. This being said, there is a possibility of reducing the flexibility even further with potentially more lethal effects on the plasmodium parasite. We believe that results presented herein would assist researchers in the discovery of potent PlmV inhibitors as potential antimalarial therapies

  20. Receptor activity modifying protein-3 mediates the protumorigenic activity of lysyl oxidase-like protein-2.

    PubMed

    Brekhman, Vera; Lugassie, Jennie; Zaffryar-Eilot, Shelly; Sabo, Edmond; Kessler, Ofra; Smith, Victoria; Golding, Hana; Neufeld, Gera

    2011-01-01

    Lysyl oxidase-like protein-2 (LOXL2) induces epithelial to mesenchymal transition and promotes invasiveness. To understand the mechanisms involved, we examined the effect of LOXL2 overexpression in MCF-7 cells on gene expression. We found that LOXL2 up-regulated the expression of receptor activity modifying protein-3 (RAMP3). Expression of RAMP3 in MDA-MB-231 cells in which LOXL2 expression was inhibited restored vimentin expression, invasiveness, and tumor development. Inhibition of RAMP3 expression in MDA-MB-231 cells mimicked the effects produced by inhibition of LOXL2 expression and was accompanied by inhibition of p38 phosphorylation. LOXL2 overexpression in these cells did not restore invasiveness, suggesting that RAMP3 functions downstream to LOXL2. LOXL2 and RAMP3 are strongly coexpressed in human colon, breast, and gastric carcinomas but not in normal colon or gastric epithelial cells. RAMP3 associates with several G-protein-coupled receptors forming receptors for peptides, such as adrenomedullin and amylin. We hypothesized that RAMP3 could function as a transducer of autocrine signals induced by such peptides. However, the proinvasive effects of RAMP3 could not be abrogated following inhibition of the expression or activity of these peptides. Our experiments suggest that the protumorigenic effects of LOXL2 are partially mediated by RAMP3 and that RAMP3 inhibitors may function as antitumorigenic agents. -

  1. Modeling Protein Folding and Applying It to a Relevant Activity

    ERIC Educational Resources Information Center

    Nelson, Allan; Goetze, Jim

    2004-01-01

    The different levels of protein structure that can be easily understood by creating a model that simulates protein folding, which can then be evaluated by applying it to a relevant activity, is presented. The materials required and the procedure for constructing a protein folding model are mentioned.

  2. Freestyle Local Perforator Flaps for Facial Reconstruction.

    PubMed

    Lee, Jun Yong; Kim, Ji Min; Kwon, Ho; Jung, Sung-No; Shim, Hyung Sup; Kim, Sang Wha

    2015-01-01

    For the successful reconstruction of facial defects, various perforator flaps have been used in single-stage surgery, where tissues are moved to adjacent defect sites. Our group successfully performed perforator flap surgery on 17 patients with small to moderate facial defects that affected the functional and aesthetic features of their faces. Of four complicated cases, three developed venous congestion, which resolved in the subacute postoperative period, and one patient with partial necrosis underwent minor revision. We reviewed the literature on freestyle perforator flaps for facial defect reconstruction and focused on English articles published in the last five years. With the advance of knowledge regarding the vascular anatomy of pedicled perforator flaps in the face, we found that some perforator flaps can improve functional and aesthetic reconstruction for the facial defects. We suggest that freestyle facial perforator flaps can serve as alternative, safe, and versatile treatment modalities for covering small to moderate facial defects. PMID:26236734

  3. Davis flap: the glory still present

    PubMed Central

    El-Sabbagh, Ahmed Hassan

    2016-01-01

    Background: Upper third defects of the ear are too large to be closed primarily without distorting the auricle. Full thickness defects can be reconstructed with local flaps. In this article, Davis flap was used to fill the upper third defects of the ear with some modifications. Patients and methods: Eight patients underwent reconstruction of full thickness auricular defects with Davis flaps from July 2012 to December 2014. The posterior surface of the flap and the raw area of conchal area were covered by full thickness graft taken from posterior surface of ear. Results: All flaps survived. No congestion was noted. The donor sites and skin grafts healed uneventfully. Conclusion: Davis flap is a simple and reproducible tool for reconstruction of upper third of ear. PMID:27274439

  4. Dissecting the active site of a photoreceptor protein

    NASA Astrophysics Data System (ADS)

    Hoff, Wouter; Hara, Miwa; Ren, Jie; Moghadam, Farzaneh; Xie, Aihua; Kumauchi, Masato

    While enzymes are quite large molecules, functionally important chemical events are often limited to a small region of the protein: the active site. The physical and chemical properties of residues at such active sites are often strongly altered compared to the same groups dissolved in water. Understanding such effects is important for unraveling the mechanisms underlying protein function and for protein engineering, but has proven challenging. Here we report on our ongoing efforts on using photoactive yellow protein (PYP), a bacterial photoreceptor, as a model system for such effects. We will report on the following questions: How many residues affect active site properties? Are these residues in direct physical contact with the active site? Can functionally important residues be recognized in the crystal structure of a protein? What structural resolution is needed to understand active sites? What spectroscopic techniques are most informative? Which weak interactions dominate active site properties?

  5. Determination of Protein Carbonylation and Proteasome Activity in Seeds.

    PubMed

    Xia, Qiong; El-Maarouf-Bouteau, Hayat; Bailly, Christophe; Meimoun, Patrice

    2016-01-01

    Reactive oxygen species (ROS) have been shown to be toxic but also function as signaling molecules in a process called redox signaling. In seeds, ROS are produced at different developmental stages including dormancy release and germination. Main targets of oxidation events by ROS in cell are lipids, nucleic acids, and proteins. Protein oxidation has various effects on their function, stability, location, and degradation. Carbonylation represents an irreversible and unrepairable modification that can lead to protein degradation through the action of the 20S proteasome. Here, we present techniques which allow the quantification of protein carbonyls in complex protein samples after derivatization by 2,4-dinitrophenylhydrazine (DNPH) and the determination proteasome activity by an activity-based protein profiling (ABPP) using the probe MV151. These techniques, routinely easy to handle, allow the rapid assessment of protein carbonyls and proteasome activity in seeds in various physiological conditions where ROS may act as signaling or toxic elements. PMID:27424756

  6. Activated protein C anticoagulant system dysfunction and thrombophilia in Asia.

    PubMed

    Hamasaki, Naotaka; Kuma, Hiroyuki; Tsuda, Hiroko

    2013-01-01

    Thrombophilia that is common among Caucasians is caused by genetic polymorphisms of coagulation factor V Leiden (R506Q) and prothrombin G20210A. Unlike that in Caucasians, thrombophilia that is common in the Japanese and Chinese involve dysfunction of the activated protein C (APC) anticoagulant system caused by abnormal protein S and protein C molecules. Approximately 50% of Japanese and Chinese individuals who develop venous thrombosis have reduced activities of protein S. The abnormal sites causing the protein S molecule abnormalities are distributed throughout the protein S gene, PROS1. One of the most common abnormalities is protein S Tokushima (K155E), which accounts for about 30% of the protein S molecule abnormalities in the Japanese. Whether APC dysfunction occurs in other Asian countries is an important aspect of mapping thrombophilia among Asians. International surveys using an accurate assay system are needed to determine this.

  7. Global Analysis of Protein Activities Using Proteome Chips

    NASA Astrophysics Data System (ADS)

    Zhu, Heng; Bilgin, Metin; Bangham, Rhonda; Hall, David; Casamayor, Antonio; Bertone, Paul; Lan, Ning; Jansen, Ronald; Bidlingmaier, Scott; Houfek, Thomas; Mitchell, Tom; Miller, Perry; Dean, Ralph A.; Gerstein, Mark; Snyder, Michael

    2001-09-01

    To facilitate studies of the yeast proteome, we cloned 5800 open reading frames and overexpressed and purified their corresponding proteins. The proteins were printed onto slides at high spatial density to form a yeast proteome microarray and screened for their ability to interact with proteins and phospholipids. We identified many new calmodulin- and phospholipid-interacting proteins; a common potential binding motif was identified for many of the calmodulin-binding proteins. Thus, microarrays of an entire eukaryotic proteome can be prepared and screened for diverse biochemical activities. The microarrays can also be used to screen protein-drug interactions and to detect posttranslational modifications.

  8. Optimal propulsive flapping in Stokes flows.

    PubMed

    Was, Loïc; Lauga, Eric

    2014-03-01

    Swimming fish and flying insects use the flapping of fins and wings to generate thrust. In contrast, microscopic organisms typically deform their appendages in a wavelike fashion. Since a flapping motion with two degrees of freedom is able, in theory, to produce net forces from a time-periodic actuation at all Reynolds numbers, we compute in this paper the optimal flapping kinematics of a rigid spheroid in a Stokes flow. The hydrodynamics for the force generation and energetics of the flapping motion is solved exactly. We then compute analytically the gradient of a flapping efficiency in the space of all flapping gaits and employ it to derive numerically the optimal flapping kinematics as a function of the shape of the flapper and the amplitude of the motion. The kinematics of optimal flapping are observed to depend weakly on the flapper shape and are very similar to the figure-eight motion observed in the motion of insect wings. Our results suggest that flapping could be a exploited experimentally as a propulsion mechanism valid across the whole range of Reynolds numbers.

  9. Refining the intrinsic chimera flap: a review.

    PubMed

    Agarwal, Jayant P; Agarwal, Shailesh; Adler, Neta; Gottlieb, Lawrence J

    2009-10-01

    Reconstruction of complex tissue deficiencies in which each missing component is in a different spatial relationship to each other can be particularly challenging, especially in patients with limited recipient vessels. The chimera flap design is uniquely suited to reconstruct these deformities. Chimera flaps have been previously defined in many ways with 2 main categories: prefabricated or intrinsic. Herein we attempt to clarify the definition of a true intrinsic chimeric flap and provide examples of how these constructs provide a method for reconstruction of complex defects. The versatility of the intrinsic chimera flap and its procurement from 7 different vascular systems is described. A clarification of the definition of a true intrinsic chimera flap is described. In addition, construction of flaps from the lateral femoral circumflex, deep circumflex iliac, inferior gluteal, peroneal, subscapular, thoracodorsal, and radial arterial systems is described to showcase the versatility of these chimera flaps. A true intrinsic chimera flap must consist of more than a single tissue type. Each of the tissue components receives its blood flow from separate vascular branches or perforators that are connected to a single vascular source. These vascular branches must be of appropriate length to allow for insetting with 3-dimensional spatial freedom. There are a multitude of sites from which true intrinsic chimera flaps may be harvested.

  10. Optimal propulsive flapping in Stokes flows.

    PubMed

    Was, Loïc; Lauga, Eric

    2014-03-01

    Swimming fish and flying insects use the flapping of fins and wings to generate thrust. In contrast, microscopic organisms typically deform their appendages in a wavelike fashion. Since a flapping motion with two degrees of freedom is able, in theory, to produce net forces from a time-periodic actuation at all Reynolds numbers, we compute in this paper the optimal flapping kinematics of a rigid spheroid in a Stokes flow. The hydrodynamics for the force generation and energetics of the flapping motion is solved exactly. We then compute analytically the gradient of a flapping efficiency in the space of all flapping gaits and employ it to derive numerically the optimal flapping kinematics as a function of the shape of the flapper and the amplitude of the motion. The kinematics of optimal flapping are observed to depend weakly on the flapper shape and are very similar to the figure-eight motion observed in the motion of insect wings. Our results suggest that flapping could be a exploited experimentally as a propulsion mechanism valid across the whole range of Reynolds numbers. PMID:24343130

  11. Pressure Available for Cooling with Cowling Flaps

    NASA Technical Reports Server (NTRS)

    Stickle, George W; Naiman, Irven; Crigler, John L

    1941-01-01

    Report presents the results of a full-scale investigation conducted in the NACA 20-foot tunnel to determine the pressure difference available for cooling with cowling flaps. The flaps were applied to an exit slot of smooth contour at 0 degree flap angle. Flap angles of 0 degree, 15 degrees, and 30 degrees were tested. Two propellers were used; propeller c which has conventional round blade shanks and propeller f which has airfoil sections extending closer to the hub. The pressure available for cooling is shown to be a direct function of the thrust disk-loading coefficient of the propeller.

  12. Basic Perforator Flap Hemodynamic Mathematical Model

    PubMed Central

    Tao, Youlun; Ding, Maochao; Wang, Aiguo; Zhuang, Yuehong; Chang, Shi-Min; Mei, Jin; Hallock, Geoffrey G.

    2016-01-01

    Background: A mathematical model to help explain the hemodynamic characteristics of perforator flaps based on blood flow resistance systems within the flap will serve as a theoretical guide for the future study and clinical applications of these flaps. Methods: There are 3 major blood flow resistance network systems of a perforator flap. These were defined as the blood flow resistance of an anastomosis between artery and artery of adjacent perforasomes, between artery and vein within a perforasome, and then between vein and vein corresponding to the outflow of that perforasome. From this, a calculation could be made of the number of such blood flow resistance network systems that must be crossed for all perforasomes within a perforator flap to predict whether that arrangement would be viable. Results: The summation of blood flow resistance networks from each perforasome in a given perforator flap could predict which portions would likely survive. This mathematical model shows how this is directly dependent on the location of the vascular pedicle to the flap and whether supercharging or superdrainage maneuvers have been added. These configurations will give an estimate of the hemodynamic characteristics for the given flap design. Conclusions: This basic mathematical model can (1) conveniently determine the degree of difficulty for each perforasome within a perforator flap to survive; (2) semiquantitatively allow the calculation of basic hemodynamic parameters; and (3) allow the assessment of the pros and cons expected for each pattern of perforasomes encountered clinically based on predictable hemodynamic observations. PMID:27579238

  13. Arterialized Venous Bone Flaps: An Experimental Investigation

    PubMed Central

    Borumandi, Farzad; Higgins, James P.; Buerger, Heinz; Vasilyeva, Anna; Benlidayi, Memmet Emre; Sencar, Leman; Gaggl, Alexander

    2016-01-01

    In arterialized venous flaps (AVFs) the venous network is used to revascularize the flap. While the feasibility of AVFs in soft tissues has been reported there is no study on osseous AVFs. In this study we aim to assess the flap survival of osseous AVFs in a pig model. Medial femoral condyle flaps were elevated in 18 pigs. Three groups were created: AVF (n = 6), conventional arterial flap (cAF, n = 6) and bone graft (BG, n = 6). The AVFs were created by anastomosis of genicular artery with one vena comitans while leaving one efferent vein for drainage. After 6 months the specimens were harvested. The histology and histomorphometry of of the bone in cAF and AVF was significantly superior to bone grafts with a higher bone volume in AVFs (p = 0.01). This study demonstrates that osseous free flaps may be supported and survive using the technique of arterialization of the venous network. The concept of AVFs in osseous flaps may be feasible for revascularization of free flaps with an inadequate artery but well developed veins. Further experimental and clinical studies are needed to assess the feasibility of clinical use of arterialized venous bone flaps. PMID:27558705

  14. Unraveling HIV protease flaps dynamics by Constant pH Molecular Dynamics simulations.

    PubMed

    Soares, Rosemberg O; Torres, Pedro H M; da Silva, Manuela L; Pascutti, Pedro G

    2016-08-01

    The active site of HIV protease (HIV-PR) is covered by two flaps. These flaps are known to be essential for the catalytic activity of the HIV-PR, but their exact conformations at the different stages of the enzymatic pathway remain subject to debate. Understanding the correct functional dynamics of the flaps might aid the development of new HIV-PR inhibitors. It is known that, the HIV-PR catalytic efficiency is pH-dependent, likely due to the influence of processes such as charge transfer and protonation/deprotonation of ionizable residues. Several Molecular Dynamics (MD) simulations have reported information about the HIV-PR flaps. However, in MD simulations the protonation of a residue is fixed and thus it is not possible to study the correlation between conformation and protonation state. To address this shortcoming, this work attempts to capture, through Constant pH Molecular Dynamics (CpHMD), the conformations of the apo, substrate-bound and inhibitor-bound HIV-PR, which differ drastically in their flap arrangements. The results show that the HIV-PR flaps conformations are defined by the protonation of the catalytic residues Asp25/Asp25' and that these residues are sensitive to pH changes. This study suggests that the catalytic aspartates can modulate the opening of the active site and substrate binding. PMID:27291071

  15. [The super extended sub-mental flap or combo sub-mental flap].

    PubMed

    Martin, D

    2014-08-01

    The author presents a technical variation of the sub-mental flap including in a conventional pedicled flap both sub-mental axes and their anastomoses on the midline. The assessment of the first flaps raised according to this method confirms the improvement of the distal blood supply. It allows the possibility to harvest "super extended" flaps reaching the contralateral auricular lobula. This variation can be considered as an axial flap which only the tip, located beyond the mandibular angle, is at random. The evolution of the sub-mental flap from its original description to this variation called "combo sub-mental flap" is then presented. Its reliability and the technical simplification it provides will have to be assessed in the future. PMID:24840945

  16. Protein C activity in dogs envenomed by Vipera palaestinae.

    PubMed

    Hadar, Gil; Kelmer, Efrat; Segev, Gilad; Bruchim, Yaron; Aroch, Itamar

    2014-09-01

    Vipera palaestinae is responsible for most envenomations in humans and domestic animal in Israel. Its venom has pro- and anticoagulant properties. Protein C is a major natural anticoagulant, preventing excess clotting and thrombosis. This study investigated protein C activity and its prognostic value, as well as several other hemostatic analytes in dogs (Canis familiaris) accidently envenomed by V. palaestinae. Protein C activity was compared between envenomed dogs and 33 healthy control dogs. Mean protein C was lower in dogs envenomed by V. palaestinae compared to controls (12.9% vs. 22.9%, respectively; P < 0.01). It was positively correlated with antithrombin activity (r = 0.3, P = 0.04), but not with other hemostatic analytes. The overall mortality rate was 13%, and at presentation no significant protein C activity difference was noted between survivors and non-survivors. A receiver operator characteristics analysis of protein C activity as a predictor of mortality had an area under the curve of 0.7 (95% confidence interval 0.52-0.87). A protein C cutoff point of 8% corresponded to sensitivity and specificity of 70% and 57%, respectively. Dogs diagnosed with consumptive coagulopathy (14%) tended to have lower protein C activity compared to others; however, their mortality did differ from that of other dogs. This is the first study assessing protein C activity in V. palaestinae victims. Decreased protein C activity in such dogs may play a role in formation of thrombosis and hemostatic derangement as well as inflammation in V. palaestinae envenomations.

  17. 14 CFR 25.457 - Wing flaps.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Wing flaps. 25.457 Section 25.457 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Control Surface and System Loads § 25.457 Wing flaps....

  18. 14 CFR 25.457 - Wing flaps.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Wing flaps. 25.457 Section 25.457 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Control Surface and System Loads § 25.457 Wing flaps....

  19. 14 CFR 25.457 - Wing flaps.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Wing flaps. 25.457 Section 25.457 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Control Surface and System Loads § 25.457 Wing flaps....

  20. 14 CFR 25.457 - Wing flaps.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Wing flaps. 25.457 Section 25.457 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Control Surface and System Loads § 25.457 Wing flaps....

  1. 14 CFR 25.457 - Wing flaps.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Wing flaps. 25.457 Section 25.457 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Control Surface and System Loads § 25.457 Wing flaps....

  2. Anthocyanidins inhibit activator protein 1 activity and cell transformation: structure-activity relationship and molecular mechanisms.

    PubMed

    Hou, De-Xing; Kai, Keiko; Li, Jian-Jian; Lin, Shigang; Terahara, Norihiko; Wakamatsu, Mika; Fujii, Makoto; Young, Mattew R; Colburn, Nancy

    2004-01-01

    Anthocyanins are the chemical components that give the intense color to many fruits and vegetables, such as blueberries, red cabbages and purple sweet potatoes. Extensive studies have indicated that anthocyanins have strong antioxidant activities. To investigate the mechanism of anthocyanidins as an anticancer food source, six kinds of anthocyanidins representing the aglycons of most anthocyanins, were used to examine their effects on tumor promotion in mouse JB6 cells, a validated model for screening cancer chemopreventive agents and elucidating the molecular mechanisms. Of the six anthocyanins tested, only those with an ortho-dihydroxyphenyl structure on the B-ring suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell transformation and activator protein-1 transactivation, suggesting that the ortho-dihydroxyphenyl may contribute to the inhibitory action. Delphinidin, but not peonidin, blocked the phosphorylation of protein kinases in the extracellular signal-regulated protein kinase (ERK) pathway at early times and the c-Jun N-terminal kinase (JNK) signaling pathway at later times. p38 kinase was not inhibited by delphinidin. Furthermore, two mitogen-activated protein kinase (MAPK) specific inhibitors (SP600125 for JNK and UO126 for ERK) could specifically block the activation of JNK and ERK and cell transformation. Those results demonstrate that anthocyanidins contribute to the inhibition of tumorigenesis by blocking activation of the MAPK pathway. These findings provide the first molecular basis for the anticarcinogenic action of anthocyanidins. PMID:14514663

  3. Anthocyanidins inhibit activator protein 1 activity and cell transformation: structure-activity relationship and molecular mechanisms.

    PubMed

    Hou, De-Xing; Kai, Keiko; Li, Jian-Jian; Lin, Shigang; Terahara, Norihiko; Wakamatsu, Mika; Fujii, Makoto; Young, Mattew R; Colburn, Nancy

    2004-01-01

    Anthocyanins are the chemical components that give the intense color to many fruits and vegetables, such as blueberries, red cabbages and purple sweet potatoes. Extensive studies have indicated that anthocyanins have strong antioxidant activities. To investigate the mechanism of anthocyanidins as an anticancer food source, six kinds of anthocyanidins representing the aglycons of most anthocyanins, were used to examine their effects on tumor promotion in mouse JB6 cells, a validated model for screening cancer chemopreventive agents and elucidating the molecular mechanisms. Of the six anthocyanins tested, only those with an ortho-dihydroxyphenyl structure on the B-ring suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell transformation and activator protein-1 transactivation, suggesting that the ortho-dihydroxyphenyl may contribute to the inhibitory action. Delphinidin, but not peonidin, blocked the phosphorylation of protein kinases in the extracellular signal-regulated protein kinase (ERK) pathway at early times and the c-Jun N-terminal kinase (JNK) signaling pathway at later times. p38 kinase was not inhibited by delphinidin. Furthermore, two mitogen-activated protein kinase (MAPK) specific inhibitors (SP600125 for JNK and UO126 for ERK) could specifically block the activation of JNK and ERK and cell transformation. Those results demonstrate that anthocyanidins contribute to the inhibition of tumorigenesis by blocking activation of the MAPK pathway. These findings provide the first molecular basis for the anticarcinogenic action of anthocyanidins.

  4. Breadboard activities for advanced protein crystal growth

    NASA Technical Reports Server (NTRS)

    Rosenberger, Franz; Banish, Michael

    1993-01-01

    The proposed work entails the design, assembly, testing, and delivery of a turn-key system for the semi-automated determination of protein solubilities as a function of temperature. The system will utilize optical scintillation as a means of detecting and monitoring nucleation and crystallite growth during temperature lowering (or raising, with retrograde solubility systems). The deliverables of this contract are: (1) turn-key scintillation system for the semi-automatic determination of protein solubilities as a function of temperature, (2) instructions and software package for the operation of the scintillation system, and (3) one semi-annual and one final report including the test results obtained for ovostatin with the above scintillation system.

  5. Visualizing active membrane protein complexes by electron cryotomography

    PubMed Central

    Gold, Vicki A.M.; Ieva, Raffaele; Walter, Andreas; Pfanner, Nikolaus; van der Laan, Martin; Kühlbrandt, Werner

    2014-01-01

    Unravelling the structural organization of membrane protein machines in their active state and native lipid environment is a major challenge in modern cell biology research. Here we develop the STAMP (Specifically TArgeted Membrane nanoParticle) technique as a strategy to localize protein complexes in situ by electron cryotomography (cryo-ET). STAMP selects active membrane protein complexes and marks them with quantum dots. Taking advantage of new electron detector technology that is currently revolutionizing cryotomography in terms of achievable resolution, this approach enables us to visualize the three-dimensional distribution and organization of protein import sites in mitochondria. We show that import sites cluster together in the vicinity of crista membranes, and we reveal unique details of the mitochondrial protein import machinery in action. STAMP can be used as a tool for site-specific labelling of a multitude of membrane proteins by cryo-ET in the future. PMID:24942077

  6. Piezoelectrically actuated insect scale flapping wing

    NASA Astrophysics Data System (ADS)

    Mukherjee, Sujoy; Ganguli, Ranjan

    2010-04-01

    An energy method is used in order to derive the non-linear equations of motion of a smart flapping wing. Flapping wing is actuated from the root by a PZT unimorph in the piezofan configuration. Dynamic characteristics of the wing, having the same size as dragonfly Aeshna Multicolor, are analyzed using numerical simulations. It is shown that flapping angle variations of the smart flapping wing are similar to the actual dragonfly wing for a specific feasible voltage. An unsteady aerodynamic model based on modified strip theory is used to obtain the aerodynamic forces. It is found that the smart wing generates sufficient lift to support its own weight and carry a small payload. It is therefore a potential candidate for flapping wing of micro air vehicles.

  7. Scrubbing noise of externally blown flaps

    NASA Technical Reports Server (NTRS)

    Fink, M. R.

    1975-01-01

    An experimental study was conducted to examine the aeroacoustic mechanism that produces externally blown flap (EBF) scrubbing noise, i.e. a surface-radiated noise which is generally strongest normal to UTW deflected flaps. Scrubbing noise was not radiated from portions of the surface adjacent to strong, locally coherent turbulent eddies. Instead, scrubbing noise seemed to come from weak loading fluctuations that were coherent along the scrubbed span. These loading fluctuations probably were induced by the convected large-scale vortex structure of the attached exhaust jet. Deflecting a UTW flap would reduce the distance between the vortex trajectory and the flap surface, increasing the resulting dipole noise and rotating its directivity. In contrast, deflecting a USB flap would increase this distance, so that observable scrubbing noise would be radiated only from the undeflected forward portion of the wing.

  8. Facial artery flaps in facial oncoplastic reconstruction.

    PubMed

    Fabrizio, Tommaso

    2013-10-01

    The face is one of the common sites for cutaneous cancer localization. It is well known that the face is the localization of more than 50% of skin cancers. Nowadays, the principles of modern "oncoplasty" recommend the complete excision of the cancer and the reconstruction with respect to cosmetic features of the face in terms of good color, good softness, and good texture of the flaps, utilized in cancer repair. The oncological and cosmetic results of facial reconstruction are strictly linked and the modern plastic and reconstructive surgeon must respect both oncological and cosmetic aspects. For that reason the best solution in facial cancer repair is the utilization of locoregional flaps based on the tributary vessels of the facial artery. In consideration of the dimension of recipient area to repair, the retroangular flap (RAF) or the submental flap could be used. This article is voted to illustrate a very large and long-term casuistry dedicated to these flaps.

  9. Antioxidant activities of protein hydrolysates obtained from the housefly larvae.

    PubMed

    Zhang, Huan; Wang, Pan; Zhang, Ai-Jun; Li, Xuan; Zhang, Ji-Hong; Qin, Qi-Lian; Wu, Yi-Jun

    2016-09-01

    The housefly is an important resource insect and the housefly larvae are ideal source of food additives. The housefly larvae protein hydrolysates were obtained by enzymatic hydrolysis by alcalase and neutral proteinase. Their antioxidant activities were investigated, including the superoxide and hydroxyl radicalscavenging activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, reducing power and metal chelating activity. The antioxidant activities of both hydrolysates increased with their increasing concentrations. The alcalase hydrolysate (AH) showed higher scavenging activities against hydroxyl radical and superoxide anion radical at low concentrations and higher metal-chelating activity than the neutral proteinase hydrolysate (NPH). The NPH exhibited higher scavenging activity against DPPH free radical and higher reducing power than the AH. Both hydrolysates showed more than 50% superoxide anion radical-scavenging activity at 10 μg/mL. These results indicate that both housefly larvae protein hydrolysates display high antioxidant activities and they could serve as potential natural antioxidant food additives. PMID:27630047

  10. Modeling of flap endonuclease interactions with DNA substrate.

    PubMed

    Allawi, Hatim T; Kaiser, Michael W; Onufriev, Alexey V; Ma, Wu-Po; Brogaard, Andrew E; Case, David A; Neri, Bruce P; Lyamichev, Victor I

    2003-05-01

    Structure-specific 5' nucleases play an important role in DNA replication and repair uniquely recognizing an overlap flap DNA substrate and processing it into a DNA nick. However, in the absence of a high-resolution structure of the enzyme/DNA complex, the mechanism underlying this recognition and substrate specificity, which is key to the enzyme's function, remains unclear. Here, we propose a three-dimensional model of the structure-specific 5' flap endonuclease from Pyrococcus furiosus in its complex with DNA. The model is based on the known X-ray structure of the enzyme and a variety of biochemical and molecular dynamics (MD) data utilized in the form of distance restraints between the enzyme and the DNA. Contacts between the 5' flap endonuclease and the sugar-phosphate backbone of the overlap flap substrate were identified using enzyme activity assays on substrates with methylphosphonate or 2'-O-methyl substitutions. The enzyme footprint extends two to four base-pairs upstream and eight to nine base-pairs downstream of the cleavage site, thus covering 10-13 base-pairs of duplex DNA. The footprint data are consistent with a model in which the substrate is bound in the DNA-binding groove such that the downstream duplex interacts with the helix-hairpin-helix motif of the enzyme. MD simulations to identify the substrate orientation in this model are consistent with the results of the enzyme activity assays on the methylphosphonate and 2'-O-methyl-modified substrates. To further refine the model, 5' flap endonuclease variants with alanine point substitutions at amino acid residues expected to contact phosphates in the substrate and one deletion mutant were tested in enzyme activity assays on the methylphosphonate-modified substrates. Changes in the enzyme footprint observed for two point mutants, R64A and R94A, and for the deletion mutant in the enzyme's beta(A)/beta(B) region, were interpreted as being the result of specific interactions in the enzyme/DNA complex

  11. Flow visualization of vortices locked by spanwise blowing over wings featuring a unique leading and trailing-edge flap system

    NASA Technical Reports Server (NTRS)

    Erickson, G. E.; Campbell, J. F.

    1975-01-01

    Flow visualization studies were conducted to qualitatively determine the effects of active generation and augmentation of vortex flow over wings by blowing a discrete jet in a spanwise direction in the channel formed by extension of upper surface leading- and trailing-edge flaps. Spanwise blowing from a reflection plane over a rectangular wing was found to generate and lock a dual corotating vortex system within the channel and, at sufficient blowing rates, cause the separated flow off the upper end of the leading-edge flap to reattach to the trailing-edge flap. Test parameters included wing angle of attack, jet momentum coefficient, leading- and trailing-edge flap deflection angle, and jet location above the wing surface. Effects due to removal of the leading- and trailing-edge flap were also investigated.

  12. Double strand binding – single strand incision mechanism for human flap endonuclease: implications for the superfamily

    PubMed Central

    Tsutakawa, Susan E.; Tainer, John A.

    2012-01-01

    Detailed structural, mutational, and biochemical analyses of human FEN1/DNA complexes have revealed the mechanism for recognition of 5′ flaps formed during lagging strand replication and DNA repair. FEN1 processes 5′ flaps through a previously unknown, but structurally elegant double-stranded (ds) recognition/single stranded (ss) incision mechanism that both selects for 5′ flaps and selects against ss DNA or RNA, intact dsDNA, and 3′ flaps. Two major DNA binding interfaces, including a K+ bridge between the DNA and the H2TH motif, are spaced one helical turn apart and together select for substrates with dsDNA. A conserved helical gateway and a helical cap protects the two-metal active site and selects for ss flaps with free termini. Structures of substrate and product reveal an unusual step between binding substrate and incision that involves a double base unpairing with incision occurring in the resulting unpaired DNA or RNA. Ordering of the active site requires a disorder-to-order transition induced by binding of an unpaired 3′ flap, which ensures that the product is ligatable. Comparison with FEN superfamily members, including XPG, EXO1, and GEN1, identifies superfamily motifs such as the helical gateway that select for ss-dsDNA junctions and provides key biological insights into nuclease specificity and regulation. PMID:22244820

  13. Local Injection of Deferoxamine Improves Neovascularization in Ischemic Diabetic Random Flap by Increasing HIF-1α and VEGF Expression

    PubMed Central

    Zhang, Yun; Xiong, Zhuyou; Li, Guangzao; Cui, Lei

    2014-01-01

    Background Although the systemic administration of deferoxamine (DFO) is protective in experimental models of normal ischemic flap and diabetic wound, its effect on diabetic flap ischemia using a local injection remains unknown. Objective To explore the feasibility of local injection of DFO to improve the survival of ischemic random skin flaps in streptozotocin (STZ)-induced diabetic mice. Methods Ischemic random skin flaps were made in 125 mice. Animals were divided into the DFO-treated (n = 20), PBS-treated (n = 16) and untreated (n = 16) groups. Surviving area, vessel density, and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) were evaluated on the seventh day after local injection. Results The viability of DFO-treated flap was significantly enhanced, with increased regional blood perfusion and capillary density compared with those in the two control groups. Fluorescence-activated cell sorting (FACS) analysis demonstrated a marked increase in systemic Flk-1+/CD11b− endothelial progenitor cells (EPCs) in DFO-treated mice. Furthermore, the expression of VEGF and HIF-1α was increased not only in diabetic flap tissue, but also in dermal fibroblasts cultured under hyperglycemic and hypoxic conditions. Conclusions Local injection of DFO could exert preventive effects against skin flap necrosis in STZ-induced diabetic mice by elevating the expression of HIF-1α and VEGF, increased EPC mobilization, which all contributed to promote ischemic diabetic flap survival. PMID:24963878

  14. Controlled Activation of Protein Rotational Dynamics Using Smart Hydrogel Tethering

    SciTech Connect

    Beech, Brenda M.; Xiong, Yijia; Boschek, Curt B.; Baird, Cheryl L.; Bigelow, Diana J.; Mcateer, Kathleen; Squier, Thomas C.

    2014-09-05

    Stimulus-responsive hydrogel materials that stabilize and control protein dynamics have the potential to enable a range of applications to take advantage of the inherent specificity and catalytic efficiencies of proteins. Here we describe the modular construction of a hydrogel using an engineered calmodulin (CaM) within a polyethylene glycol (PEG) matrix that involves the reversible tethering of proteins through an engineered CaM-binding sequence. For these measurements, maltose binding protein (MBP) was isotopically labeled with [13C] and [15N], permitting dynamic structural measurements using TROSY-HSQC NMR spectroscopy. Upon initial formation of hydrogels protein dynamics are suppressed, with concomitant increases in protein stability. Relaxation of the hydrogel matrix following transient heating results in the activation of protein dynamics and restoration of substrate-induced large-amplitude domain motions necessary for substrate binding.

  15. Synthesis, SAR, and series evolution of novel oxadiazole-containing 5-lipoxygenase activating protein inhibitors: discovery of 2-[4-(3-{(r)-1-[4-(2-amino-pyrimidin-5-yl)-phenyl]-1-cyclopropyl-ethyl}-[1,2,4]oxadiazol-5-yl)-pyrazol-1-yl]-N,N-dimethyl-acetamide (BI 665915).

    PubMed

    Takahashi, Hidenori; Riether, Doris; Bartolozzi, Alessandra; Bosanac, Todd; Berger, Valentina; Binetti, Ralph; Broadwater, John; Chen, Zhidong; Crux, Rebecca; De Lombaert, Stéphane; Dave, Rajvee; Dines, Jonathon A; Fadra-Khan, Tazmeen; Flegg, Adam; Garrigou, Michael; Hao, Ming-Hong; Huber, John; Hutzler, J Matthew; Kerr, Steven; Kotey, Adrian; Liu, Weimin; Lo, Ho Yin; Loke, Pui Leng; Mahaney, Paige E; Morwick, Tina M; Napier, Spencer; Olague, Alan; Pack, Edward; Padyana, Anil K; Thomson, David S; Tye, Heather; Wu, Lifen; Zindell, Renee M; Abeywardane, Asitha; Simpson, Thomas

    2015-02-26

    The synthesis, structure-activity relationship (SAR), and evolution of a novel series of oxadiazole-containing 5-lipoxygenase-activating protein (FLAP) inhibitors are described. The use of structure-guided drug design techniques provided compounds that demonstrated excellent FLAP binding potency (IC50 < 10 nM) and potent inhibition of LTB4 synthesis in human whole blood (IC50 < 100 nM). Optimization of binding and functional potencies, as well as physicochemical properties resulted in the identification of compound 69 (BI 665915) that demonstrated an excellent cross-species drug metabolism and pharmacokinetics (DMPK) profile and was predicted to have low human clearance. In addition, 69 was predicted to have a low risk for potential drug-drug interactions due to its cytochrome P450 3A4 profile. In a murine ex vivo whole blood study, 69 demonstrated a linear dose-exposure relationship and a dose-dependent inhibition of LTB4 production. PMID:25671290

  16. Hox proteins: sculpting body parts by activating localized cell death.

    PubMed

    Alonso, Claudio R

    2002-11-19

    Hox proteins shape animal structures by eliciting different developmental programs along the anteroposterior body axis. A recent study reveals that the Drosophila Hox protein Deformed directly activates the cell-death-promoting gene reaper to maintain the boundaries between distinct head segments.

  17. Modeling the SHG activities of diverse protein crystals

    SciTech Connect

    Haupert, Levi M.; DeWalt, Emma L.; Simpson, Garth J.

    2012-11-01

    The origins of the diversity in the SHG signal from protein crystals are investigated and potential protein-crystal coverage by SHG microscopy is assessed. A symmetry-additive ab initio model for second-harmonic generation (SHG) activity of protein crystals was applied to assess the likely protein-crystal coverage of SHG microscopy. Calculations were performed for 250 proteins in nine point-group symmetries: a total of 2250 crystals. The model suggests that the crystal symmetry and the limit of detection of the instrument are expected to be the strongest predictors of coverage of the factors considered, which also included secondary-structural content and protein size. Much of the diversity in SHG activity is expected to arise primarily from the variability in the intrinsic protein response as well as the orientation within the crystal lattice. Two or more orders-of-magnitude variation in intensity are expected even within protein crystals of the same symmetry. SHG measurements of tetragonal lysozyme crystals confirmed detection, from which a protein coverage of ∼84% was estimated based on the proportion of proteins calculated to produce SHG responses greater than that of tetragonal lysozyme. Good agreement was observed between the measured and calculated ratios of the SHG intensity from lysozyme in tetragonal and monoclinic lattices.

  18. Hemagglutinating activity of proteins from Parkia speciosa seeds.

    PubMed

    Chankhamjon, Kanokwan; Petsom, Amorn; Sawasdipuksa, Narumon; Sangvanich, Polkit

    2010-01-01

    Proteins from Parkia speciosa Hassk. (Fabaceae) seeds were extracted and stepwise precipitated using ammonium sulfate. Proteins precipitated with 25% ammonium sulfate were separated by affinity chromatography on Affi-Gel Blue gel followed by protein liquid chromatography on Superdex 200. The protein Gj, which was identified as a protein similar to putative aristolochene synthase, 3'-partial from Oryza sativa L. (Poaceae), had hemagglutinating activity of 0.39 mug/muL. Moreover, fraction C2 from the proteins precipitated with 60% ammonium sulfate, separated by lectin-specific adsorption chromatography using Con A Sepharose, had hemagglutinating activity of 1.17 mug/muL. Using gel electrophoresis, two proteins C2a and C2b were separated, having molecular weights of 45 kDa and 23 kDa, respectively. From protein identification, C2a was found to be similar to the hypothetical protein B1342F01.11 from Oryza sativa, and C2b was similar to the hypothetical protein At1g51560 from Arabidopsis thaliana (L.) Heynh. (Brassicaceae). PMID:20645760

  19. Flapping flexible fish

    NASA Astrophysics Data System (ADS)

    Root, Robert G.; Courtland, Hayden-William; Shepherd, William; Long, John H.

    In order to analyze and model the body kinematics used by fish in a wide range of swimming behaviors, we developed a technique to separate the periodic whole-body motions that characterize steady swimming from the secular motions that characterize changes in whole-body shape. We applied this harmonic analysis technique to the study of the forward and backward swimming of lamprey. We found that in order to vary the unsteadiness of swimming, lamprey superimpose periodic and secular components of their body motion, modulate the patterns and magnitudes of those components, and change shape. These kinematic results suggest the following hydromechanical hypothesis: steady swimming is a maneuver that requires active suppression of secular body reconfigurations.

  20. Differential regulation of protein subdomain activity with caged bivalent ligands.

    PubMed

    Mayer, Günter; Müller, Jens; Mack, Timo; Freitag, Daniel F; Höver, Thomas; Pötzsch, Bernd; Heckel, Alexander

    2009-03-01

    Subtle change: Spatiotemporal modulation of individual protein subdomains with light as the trigger signal becomes possible by using bivalent aptamers and introducing photolabile "caging groups" to switch individual aptamer modules ON or OFF differentially. To the best of our knowledge, this is the first study to show that it is possible to modulate individual domain activity in aptamers, and thus also domain activity in proteins, with light.

  1. Cloning of three novel neuronal Cdk5 activator binding proteins.

    PubMed

    Ching, Y P; Qi, Z; Wang, J H

    2000-01-25

    Neuronal Cdc2-like kinase (Nclk) is involved in the regulation of neuronal differentiation and neuro-cytoskeleton dynamics. The active kinase consists of a catalytic subunit, Cdk5, and a 25 kDa activator protein (p25nck5a) derived from a 35 kDa neuronal-specific protein (p35nck5a). As an extension of our previous study (Qi, Z., Tang, D., Zhu, X., Fujita, D.J., Wang, J.H., 1998. Association of neurofilament proteins with neuronal Cdk5 activator. J. Biol. Chem. 270, 2329-2335), which showed that neurofilament is one of the p35nck5a-associated proteins, we now report the isolation of three other novel p35nck5a-associated proteins using the yeast two-hybrid screen. The full-length forms of these three novel proteins, designated C42, C48 and C53, have a molecular mass of 66, 24, and 57 kDa, respectively. Northern analysis indicates that these novel proteins are widely expressed in human tissues, including the heart, brain, skeletal muscle, placenta, lung, liver, kidney and pancreas. The bacterially expressed glutathione S-transferase (GST)-fusion forms of these three proteins were able to co-precipitate p35nck5a complexed with Cdk5 from insect cell lysate. Among these three proteins, only C48 and C53 can be phosphorylated by Nclk, suggesting that they may be the substrates of Nclk. Sequence homology searches have suggested that the C48 protein is marginally related to restin protein, whereas the C42 protein has homologues of unknown function in Caenorhabditis elegans and Arabidopsis thaliana. PMID:10721722

  2. Activation of an Endoribonuclease by Non-intein Protein Splicing.

    PubMed

    Campbell, Stephen J; Stern, David B

    2016-07-29

    The Chlamydomonas reinhardtii chloroplast-localized poly(A)-binding protein RB47 is predicted to contain a non-conserved linker (NCL) sequence flanked by highly conserved N- and C-terminal sequences, based on the corresponding cDNA. RB47 was purified from chloroplasts in association with an endoribonuclease activity; however, protein sequencing failed to detect the NCL. Furthermore, while recombinant RB47 including the NCL did not display endoribonuclease activity in vitro, versions lacking the NCL displayed strong activity. Both full-length and shorter forms of RB47 could be detected in chloroplasts, with conversion to the shorter form occurring in chloroplasts isolated from cells grown in the light. This conversion could be replicated in vitro in chloroplast extracts in a light-dependent manner, where epitope tags and protein sequencing showed that the NCL was excised from a full-length recombinant substrate, together with splicing of the flanking sequences. The requirement for endogenous factors and light differentiates this protein splicing from autocatalytic inteins, and may allow the chloroplast to regulate the activation of RB47 endoribonuclease activity. We speculate that this protein splicing activity arose to post-translationally repair proteins that had been inactivated by deleterious insertions or extensions. PMID:27311716

  3. Hydrodynamic schooling of flapping swimmers

    DOE PAGES

    Becker, Alexander D.; Masoud, Hassan; Newbolt, Joel W.; Shelley, Michael; Ristroph, Leif

    2015-10-06

    Fish schools and bird flocks are fascinating examples of collective behaviours in which many individuals generate and interact with complex flows. Motivated by animal groups on the move, here we explore how the locomotion of many bodies emerges from their flow-mediated interactions. Through experiments and simulations of arrays of flapping wings that propel within a collective wake, we discover distinct modes characterized by the group swimming speed and the spatial phase shift between trajectories of neighbouring wings. For identical flapping motions, slow and fast modes coexist and correspond to constructive and destructive wing–wake interactions. Simulations show that swimming in amore » group can enhance speed and save power, and we capture the key phenomena in a mathematical model based on memory or the storage and recollection of information in the flow field. Lastly, these results also show that fluid dynamic interactions alone are sufficient to generate coherent collective locomotion, and thus might suggest new ways to characterize the role of flows in animal groups.« less

  4. Hydrodynamic schooling of flapping swimmers

    SciTech Connect

    Becker, Alexander D.; Masoud, Hassan; Newbolt, Joel W.; Shelley, Michael; Ristroph, Leif

    2015-10-06

    Fish schools and bird flocks are fascinating examples of collective behaviours in which many individuals generate and interact with complex flows. Motivated by animal groups on the move, here we explore how the locomotion of many bodies emerges from their flow-mediated interactions. Through experiments and simulations of arrays of flapping wings that propel within a collective wake, we discover distinct modes characterized by the group swimming speed and the spatial phase shift between trajectories of neighbouring wings. For identical flapping motions, slow and fast modes coexist and correspond to constructive and destructive wing–wake interactions. Simulations show that swimming in a group can enhance speed and save power, and we capture the key phenomena in a mathematical model based on memory or the storage and recollection of information in the flow field. Lastly, these results also show that fluid dynamic interactions alone are sufficient to generate coherent collective locomotion, and thus might suggest new ways to characterize the role of flows in animal groups.

  5. Hydrodynamic schooling of flapping swimmers

    PubMed Central

    Becker, Alexander D.; Masoud, Hassan; Newbolt, Joel W.; Shelley, Michael; Ristroph, Leif

    2015-01-01

    Fish schools and bird flocks are fascinating examples of collective behaviours in which many individuals generate and interact with complex flows. Motivated by animal groups on the move, here we explore how the locomotion of many bodies emerges from their flow-mediated interactions. Through experiments and simulations of arrays of flapping wings that propel within a collective wake, we discover distinct modes characterized by the group swimming speed and the spatial phase shift between trajectories of neighbouring wings. For identical flapping motions, slow and fast modes coexist and correspond to constructive and destructive wing–wake interactions. Simulations show that swimming in a group can enhance speed and save power, and we capture the key phenomena in a mathematical model based on memory or the storage and recollection of information in the flow field. These results also show that fluid dynamic interactions alone are sufficient to generate coherent collective locomotion, and thus might suggest new ways to characterize the role of flows in animal groups. PMID:26439509

  6. Hydrodynamic schooling of flapping swimmers

    NASA Astrophysics Data System (ADS)

    Becker, Alexander D.; Masoud, Hassan; Newbolt, Joel W.; Shelley, Michael; Ristroph, Leif

    2015-10-01

    Fish schools and bird flocks are fascinating examples of collective behaviours in which many individuals generate and interact with complex flows. Motivated by animal groups on the move, here we explore how the locomotion of many bodies emerges from their flow-mediated interactions. Through experiments and simulations of arrays of flapping wings that propel within a collective wake, we discover distinct modes characterized by the group swimming speed and the spatial phase shift between trajectories of neighbouring wings. For identical flapping motions, slow and fast modes coexist and correspond to constructive and destructive wing-wake interactions. Simulations show that swimming in a group can enhance speed and save power, and we capture the key phenomena in a mathematical model based on memory or the storage and recollection of information in the flow field. These results also show that fluid dynamic interactions alone are sufficient to generate coherent collective locomotion, and thus might suggest new ways to characterize the role of flows in animal groups.

  7. Tongue reconstruction with minimal donor site morbidity using a deep inferior epigastric perforator (DIEP) free flap in a 6-year-old girl.

    PubMed

    Yano, Tomoyuki; Okazaki, Mutsumi; Kawaguchi, Runa; Suesada, Nobuko; Tanaka, Kentaro; Kishimoto, Seiji

    2013-09-01

    Tongue reconstruction was performed using a deep inferior epigastric perforator (DIEP) free flap in a 6-year-old girl with undifferentiated sarcoma of the tongue. After hemi-glossectomy with upper neck dissection, a 3-lobed DIEP free flap was used for the reconstruction. Donor site was closed primarily with suturing umbilicus in proper position. No flap loss, leakage, or infection occurred. Postoperatively, the patient was able to consume a normal diet without difficulty or aspiration and displayed good speech function. No donor site morbidity, e.g., herniation or bulging, was observed, and the patient was able to perform their normal daily activities. DIEP flaps provide a pliable skin paddle, an adequate amount of adipose tissue, and reduced donor site morbidity, even in children. We did not have any difficulty harvesting the DIEP flap or with the microvascular anastomosis. We consider DIEP free flaps to be the ideal option for pediatric tongue reconstruction.

  8. 4-Anilino-6-phenyl-quinoline inhibitors of mitogen activated protein kinase-activated protein kinase 2 (MK2).

    PubMed

    Olsson, Henric; Sjö, Peter; Ersoy, Oguz; Kristoffersson, Anna; Larsson, Joakim; Nordén, Bo

    2010-08-15

    A class of inhibitors of mitogen activated protein kinase-activated kinase 2 (MK2) was discovered via high-throughput screening. This compound class demonstrates activity against the enzyme with sub-microM IC(50) values, and suppresses LPS-induced TNFalpha levels in THP-1 cells. MK2 inhibition kinetic measurements indicated mixed binding approaching non-ATP competitive inhibition.

  9. Comparison of the effects of troxerutin and heparinoid on flap necrosis.

    PubMed

    Celik, Alper; Ersoy, Omer F; Ozkan, Namik; Kayaoglu, Huseyin A; Ozugurlu, Fikret; Cakir, Ebru A; Lordlar, Nese; Omeroglu, Suna

    2010-05-01

    We aimed to assess the effects of local troxerutin and heparinoid (HEP) treatments in a model of flap necrosis. Three groups of Wistar albino rats, each comprising 10 animals were used. A cranially based 6x3-cm full-thickness random-pattern skin flap was raised and sutured to the same area in each model. The control group was treated daily with normal saline, the second with topical HEP and the third with topical troxerutin. The amount of flap necrosis was measured in all groups by the end of the seventh day. Flap tissues were excised for histological analysis and evaluation of the expression of vascular endothelial growth factor (VEGF) levels. Assessment of the blood levels of nitric oxide was also performed in each animal by cardiac puncture. The mean area of flap necrosis was 110.6mm(2) in the control, 39.44 mm(2) in the troxerutin and 47.11 mm(2) in the heparinoid-treated rats. The treatment arms exhibited significant reduction in areas of flap necrosis, compared with the control group (p<0.001), but it was similar among treatment groups (p=0.60). The rates of fibroblast proliferation were decreased in control group as compared to HEP and troxerutin arms (p<0.001). The mean level of collagen density, collagen organisation, granulation tissue and demarcation were similar in all rats. Measurement of VEGF expression did not show any significant difference between the groups (p=0.30). Nitric oxide levels were significantly higher in control rats, as compared to treatment groups (p<0.0001), but were similar in treatment arms (p=0.45). Our results suggest that troxerutin and HEP effectively reduce the flap necrosis and improve flap survival. The observed effects might be due to their anti-oedematogenic, radical-scavenging, antioxidant effects and supportive activities on capillary permeability and transudation.

  10. Phosphorylation of platelet actin-binding protein during platelet activation

    SciTech Connect

    Carroll, R.C.; Gerrard, J.M.

    1982-03-01

    In this study we have followed the 32P-labeling of actin-binding protein as a function of platelet activation. Utilizing polyacrylamide-sodium dodecyl sulfate gel electrophoresis to resolve total platelet protein samples, we found 2 to 3-fold labeling increases in actin-binding protein 30 to 60 sec after thrombin stimulation. Somewhat larger increases were observed for 40,000 and 20,000 apparent molecular weight peptides. The actin-binding protein was identified on the gels by coelectrophoresis with purified actin-binding protein, its presence in cytoskeletal cores prepared by detergent extraction of activated 32P-labeled platelets, and by direct immunoprecipitation with antibodies against guinea pig vas deferens filamin (actin-binding protein). In addition, these cytoskeletal cores indicated that the 32P-labeled actin-binding protein was closely associated with the activated platelet's cytoskeleton. Following the 32P-labeling of actin-binding protein over an 8-min time course revealed that in aggregating platelet samples rapid dephosphorylation to almost initial levels occurred between 3 and 5 min. A similar curve was obtained for the 20,000 apparent molecular weight peptide. However, rapid dephosphorylation was not observed if platelet aggregation was prevented by chelating external calcium or by using thrombasthenic platelets lacking the aggregation response. Thus, cell-cell contact would seem to be crucial in initiating the rapid dephosphorylation response.

  11. Aerodynamic characteristics of a wing with Fowler flaps including flap loads, downwash, and calculated effect on take-off

    NASA Technical Reports Server (NTRS)

    Platt, Robert C

    1936-01-01

    This report presents the results of wind tunnel tests of a wing in combination with each of three sizes of Fowler flap. The purpose of the investigation was to determine the aerodynamic characteristics as affected by flap chord and position, the air loads on the flaps, and the effect of flaps on the downwash.

  12. Detecting protein complexes from active protein interaction networks constructed with dynamic gene expression profiles

    PubMed Central

    2013-01-01

    Background Protein interaction networks (PINs) are known to be useful to detect protein complexes. However, most available PINs are static, which cannot reflect the dynamic changes in real networks. At present, some researchers have tried to construct dynamic networks by incorporating time-course (dynamic) gene expression data with PINs. However, the inevitable background noise exists in the gene expression array, which could degrade the quality of dynamic networkds. Therefore, it is needed to filter out contaminated gene expression data before further data integration and analysis. Results Firstly, we adopt a dynamic model-based method to filter noisy data from dynamic expression profiles. Then a new method is proposed for identifying active proteins from dynamic gene expression profiles. An active protein at a time point is defined as the protein the expression level of whose corresponding gene at that time point is higher than a threshold determined by a standard variance involved threshold function. Furthermore, a noise-filtered active protein interaction network (NF-APIN) is constructed. To demonstrate the efficiency of our method, we detect protein complexes from the NF-APIN, compared with those from other dynamic PINs. Conclusion A dynamic model based method can effectively filter out noises in dynamic gene expression data. Our method to compute a threshold for determining the active time points of noise-filtered genes can make the dynamic construction more accuracy and provide a high quality framework for network analysis, such as protein complex prediction. PMID:24565281

  13. Structural mechanism of G protein activation by G protein-coupled receptor.

    PubMed

    Duc, Nguyen Minh; Kim, Hee Ryung; Chung, Ka Young

    2015-09-15

    G protein-coupled receptors (GPCRs) are a family of membrane receptors that regulate physiology and pathology of various organs. Consequently, about 40% of drugs in the market targets GPCRs. Heterotrimeric G proteins are composed of α, β, and γ subunits, and act as the key downstream signaling molecules of GPCRs. The structural mechanism of G protein activation by GPCRs has been of a great interest, and a number of biochemical and biophysical studies have been performed since the late 80's. These studies investigated the interface between GPCR and G proteins and the structural mechanism of GPCR-induced G protein activation. Recently, arrestins are also reported to be important molecular switches in GPCR-mediated signal transduction, and the physiological output of arrestin-mediated signal transduction is different from that of G protein-mediated signal transduction. Understanding the structural mechanism of the activation of G proteins and arrestins would provide fundamental information for the downstream signaling-selective GPCR-targeting drug development. This review will discuss the structural mechanism of GPCR-induced G protein activation by comparing previous biochemical and biophysical studies.

  14. Protein expression, characterization and activity comparisons of wild type and mutant DUSP5 proteins

    SciTech Connect

    Nayak, Jaladhi; Gastonguay, Adam J.; Talipov, Marat R.; Vakeel, Padmanabhan; Span, Elise A.; Kalous, Kelsey S.; Kutty, Raman G.; Jensen, Davin R.; Pokkuluri, Phani Raj; Sem, Daniel S.; Rathore, Rajendra; Ramchandran, Ramani

    2014-12-18

    Background: The mitogen-activated protein kinases (MAPKs) pathway is critical for cellular signaling, and proteins such as phosphatases that regulate this pathway are important for normal tissue development. Based on our previous work on dual specificity phosphatase-5 (DUSP5), and its role in embryonic vascular development and disease, we hypothesized that mutations in DUSP5 will affect its function. Results: In this study, we tested this hypothesis by generating full-length glutathione-S-transferase-tagged DUSP5 and serine 147 proline mutant (S147P) proteins from bacteria. Light scattering analysis, circular dichroism, enzymatic assays and molecular modeling approaches have been performed to extensively characterize the protein form and function. We demonstrate that both proteins are active and, interestingly, the S147P protein is hypoactive as compared to the DUSP5 WT protein in two distinct biochemical substrate assays. Furthermore, due to the novel positioning of the S147P mutation, we utilize computational modeling to reconstruct full-length DUSP5 and S147P to predict a possible mechanism for the reduced activity of S147P. Conclusion: Taken together, this is the first evidence of the generation and characterization of an active, full-length, mutant DUSP5 protein which will facilitate future structure-function and drug development-based studies.

  15. Protein expression, characterization and activity comparisons of wild type and mutant DUSP5 proteins

    DOE PAGES

    Nayak, Jaladhi; Gastonguay, Adam J.; Talipov, Marat R.; Vakeel, Padmanabhan; Span, Elise A.; Kalous, Kelsey S.; Kutty, Raman G.; Jensen, Davin R.; Pokkuluri, Phani Raj; Sem, Daniel S.; et al

    2014-12-18

    Background: The mitogen-activated protein kinases (MAPKs) pathway is critical for cellular signaling, and proteins such as phosphatases that regulate this pathway are important for normal tissue development. Based on our previous work on dual specificity phosphatase-5 (DUSP5), and its role in embryonic vascular development and disease, we hypothesized that mutations in DUSP5 will affect its function. Results: In this study, we tested this hypothesis by generating full-length glutathione-S-transferase-tagged DUSP5 and serine 147 proline mutant (S147P) proteins from bacteria. Light scattering analysis, circular dichroism, enzymatic assays and molecular modeling approaches have been performed to extensively characterize the protein form and function.more » We demonstrate that both proteins are active and, interestingly, the S147P protein is hypoactive as compared to the DUSP5 WT protein in two distinct biochemical substrate assays. Furthermore, due to the novel positioning of the S147P mutation, we utilize computational modeling to reconstruct full-length DUSP5 and S147P to predict a possible mechanism for the reduced activity of S147P. Conclusion: Taken together, this is the first evidence of the generation and characterization of an active, full-length, mutant DUSP5 protein which will facilitate future structure-function and drug development-based studies.« less

  16. TRAM flap breast reconstruction after radiation treatment.

    PubMed Central

    Williams, J K; Bostwick, J; Bried, J T; Mackay, G; Landry, J; Benton, J

    1995-01-01

    OBJECTIVE: Patients with and without radiation treatment before their breast reconstruction were compared to study the relationship of radiation to flap-related complications. SUMMARY BACKGROUND DATA: The transverse rectus abdominis muscle (TRAM) flap for breast reconstruction involves a a vascular pedicle and recipient bed, both included in the radiated field of patients undergoing adjunctive therapy. Detailed reviews of flap-related complications in this subgroup of patients have been limited. METHODS: One hundred eight patients with radiation treatment who subsequently underwent a TRAM flap breast reconstruction were compared with 572 patients with no radiation treatment before similar reconstruction. Flap-related complications, radiation dosage, time, fields, relationships between risk factors, and complications were studied. RESULTS: Overall complication rates were comparable between the two groups. Only fat necrosis (> 10% of total reconstruction) was found to be statistically significant (17.6% vs. 10.1%, p = 0.03228). No difference was found for fat necrosis in unipedicled vs. bipedicled flaps controlled for radiation (17.7% vs. 17.4%). Obesity and radiation therapy were associated with fat necrosis and major infection in a logistic regression. Significant abdominal scarring was also associated with major infection (p = 0.0044). CONCLUSIONS: In this, the largest reported series, radiation therapy was associated with increased fat necrosis and major infection. The use of the TRAM flap was not found to be prohibitive in radiated patients and should still be the first choice in this subgroup of patients. Images Figure 1. Figure 2. PMID:7794079

  17. Preputial flaps to correct buried penis.

    PubMed

    Chu, Chih-Chun; Chen, Yi-Hsin; Diau, Guan-Yeu; Loh, Ih-Wei; Chen, Ke-Chi

    2007-11-01

    The authors developed a preputial skin flap technique to correct the buried penis which was simple and practical. This simple procedure can be applied to most boys with buried penis. In the last 3 years, we have seen 12 boys with buried penis and have been treated by using preputial flaps. The mean age is about 5.1 (from 3 to 12). By making a longitudinal incision on the ventral side of penis, the tightness of the foreskin is released and leave a diamond-shaped skin defect. It allows the penile shaft to extend out. A circumferential incision is made about 5 mm proximal to the coronal sulcus. Pedicled preputial flaps are obtained leaving optimal penile skin on the dorsal side. The preputial skin flaps are rotated onto the ventral side and tailored to cover the defect. All patients are followed for at least 3 months. Edema and swelling on the flaps are common, but improves with time. None of our patients need a second operation. The preputial flaps technique is a simple technique which allows surgeons to deal with most cases of buried penis by tailoring the flaps providing good cosmetic and functional results.

  18. Notum deacylates Wnt proteins to suppress signalling activity.

    PubMed

    Kakugawa, Satoshi; Langton, Paul F; Zebisch, Matthias; Howell, Steven A; Chang, Tao-Hsin; Liu, Yan; Feizi, Ten; Bineva, Ganka; O'Reilly, Nicola; Snijders, Ambrosius P; Jones, E Yvonne; Vincent, Jean-Paul

    2015-03-12

    Signalling by Wnt proteins is finely balanced to ensure normal development and tissue homeostasis while avoiding diseases such as cancer. This is achieved in part by Notum, a highly conserved secreted feedback antagonist. Notum has been thought to act as a phospholipase, shedding glypicans and associated Wnt proteins from the cell surface. However, this view fails to explain specificity, as glypicans bind many extracellular ligands. Here we provide genetic evidence in Drosophila that Notum requires glypicans to suppress Wnt signalling, but does not cleave their glycophosphatidylinositol anchor. Structural analyses reveal glycosaminoglycan binding sites on Notum, which probably help Notum to co-localize with Wnt proteins. They also identify, at the active site of human and Drosophila Notum, a large hydrophobic pocket that accommodates palmitoleate. Kinetic and mass spectrometric analyses of human proteins show that Notum is a carboxylesterase that removes an essential palmitoleate moiety from Wnt proteins and thus constitutes the first known extracellular protein deacylase. PMID:25731175

  19. Critical Mach Numbers of Thin Airfoil Sections with Plain Flaps

    NASA Technical Reports Server (NTRS)

    Pardee, Otway O'm.; Heaslet, Max A.

    1946-01-01

    Critical Mach number as function of lift coefficient is determined for certain moderately thick NACA low-drag airfoils. Results, given graphically, included calculations on same airfoil sections with plain flaps for small flap deflections. Curves indicate optimum critical conditions for airfoils with flaps in such form that they can be compared with corresponding results for zero flap deflections. Plain flaps increase life-coefficient range for which critical Mach number is in region of high values characteristic of low-drag airfoils.

  20. Magnetostrictively actuated control flaps for vibration reduction in helicopter rotors

    SciTech Connect

    Millott, T.; Friedmann, P.P.

    1994-12-31

    High vibration levels can impose constraints on helicopter operations and hinder passenger acceptance. Vibration reduction using blade root pitch control introduces a significant power penalty and may adversely affect the airworthiness of the flight control system. Comparable levels of vibration reduction can be achieved using considerably less power through an actively controlled trailing edge flap mounted on the blade. Such a device would have no effect on helicopter airworthiness since it is controlled by a loop separate from the primary flight control system which utilizes the swashplate. Control flap actuation using the magnetostrictive material Terfenol-D is studied in this paper by designing a minimum weight actuator, subject to a set of actuation and stress constraints. The resulting device is capable of producing vibration reduction in excess of 90% at cruise conditions.

  1. Ontogeny of aerial righting and wing flapping in juvenile birds.

    PubMed

    Evangelista, Dennis; Cam, Sharlene; Huynh, Tony; Krivitskiy, Igor; Dudley, Robert

    2014-08-01

    Mechanisms of aerial righting in juvenile chukar partridge (Alectoris chukar) were studied from hatching to 14 days-post-hatching (dph). Asymmetric movements of the wings were used from 1 to 8 dph to effect progressively more successful righting behaviour via body roll. Following 8 dph, wing motions transitioned to bilaterally symmetric flapping that yielded aerial righting via nose-down pitch, along with substantial increases in vertical force production during descent. Ontogenetically, the use of such wing motions to effect aerial righting precedes both symmetric flapping and a previously documented behaviour in chukar (i.e. wing-assisted incline running) hypothesized to be relevant to incipient flight evolution in birds. These findings highlight the importance of asymmetric wing activation and controlled aerial manoeuvres during bird development and are potentially relevant to understanding the origins of avian flight. PMID:25165451

  2. Regulatory crosstalk by protein kinases on CFTR trafficking and activity

    NASA Astrophysics Data System (ADS)

    Farinha, Carlos Miguel; Swiatecka-Urban, Agnieszka; Brautigan, David; Jordan, Peter

    2016-01-01

    Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e. channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease.

  3. Regulatory Crosstalk by Protein Kinases on CFTR Trafficking and Activity.

    PubMed

    Farinha, Carlos M; Swiatecka-Urban, Agnieszka; Brautigan, David L; Jordan, Peter

    2016-01-01

    Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e., channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease.

  4. Regulatory Crosstalk by Protein Kinases on CFTR Trafficking and Activity

    PubMed Central

    Farinha, Carlos M.; Swiatecka-Urban, Agnieszka; Brautigan, David L.; Jordan, Peter

    2016-01-01

    Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e., channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease. PMID:26835446

  5. Pedicled Supraclavicular Artery Island Flap Versus Free Radial Forearm Flap for Tongue Reconstruction Following Hemiglossectomy.

    PubMed

    Zhang, Senlin; Chen, Wei; Cao, Gang; Dong, Zhen

    2015-09-01

    This study investigated the tongue function and donor-site morbidity of patients with malignant tumors who had undergone immediate flap reconstruction surgery. Twenty-seven patients who had undergone immediate reconstruction after hemiglossectomy were observed. Twelve patients were reconstructed using the pedicled supraclavicular artery island flap (PSAIF) and 15 patients using the free radial forearm flap (FRFF). Flap survival, speech and swallowing function, and donor-site morbidity at the 6-month follow-up were evaluated. All the flaps were successfully transferred. No obvious complications were found in either the transferred flaps or donor regions. Age, sex, defect extent, speech and swallowing function were comparable between the 2 groups. Donor-site complications were less frequent with PSAIF reconstruction than FRFF reconstruction. The PSAIF is reliable and well suited for hemiglossectomy defect. It has few significant complications, and allows preservation of oral function.

  6. On the generation of side-edge flap noise. [part span trailing edge flaps

    NASA Technical Reports Server (NTRS)

    Howe, M. S.

    1981-01-01

    A theory is proposed for estimating the noise generated at the side edges of part span trailing edge flaps in terms of pressure fluctuations measured just in-board of the side edge of the upper surface of the flap. Asymptotic formulae are developed in the opposite extremes of Lorentz contracted acoustic wavelength large/small compared with the chord of the flap. Interpolation between these limiting results enables the field shape and its dependence on subsonic forward flight speed to be predicted over the whole frequency range. It is shown that the mean width of the side edge gap between the flap and the undeflected portion of the airfoil has a significant influence on the intensity of the radiated sound. It is estimated that the noise generated at a single side edge of a full scale part span flap can exceed that produced along the whole of the trailing edge of the flap by 3 dB or more.

  7. A review of propeller flaps for distal lower extremity soft tissue reconstruction: Is flap loss too high?

    PubMed

    Nelson, Jonas A; Fischer, John P; Brazio, Philip S; Kovach, Stephen J; Rosson, Gedge D; Rad, Ariel N

    2013-10-01

    Soft tissue coverage in the distal lower extremity remains a significant challenge. While free flaps are often utilized for larger defects, local perforator-based propeller flaps may be ideal for smaller wounds requiring coverage. Propeller flaps can provide excellent form and function for both traumatic and atraumatic defects with minimal donor site morbidity but can have concerning rates of flap loss. We reviewed the literature, identifying 21 studies presenting 310 propeller flaps for distal lower extremity reconstruction. Total flap necrosis was noted in 5.5% of flaps, with partial necrosis in 11.6%. While these flaps do enable transfer of local, healthy tissue to the defect site without the need for a microsurgical anastomosis, this rate of flap loss is concerning and appropriate patient selection is crucial. This review provides a brief history and overview of the clinical application and research into distal lower extremity perforator propeller flaps to place this technique into a clinical context.

  8. Cellular reprogramming through mitogen-activated protein kinases

    PubMed Central

    Lee, Justin; Eschen-Lippold, Lennart; Lassowskat, Ines; Böttcher, Christoph; Scheel, Dierk

    2015-01-01

    Mitogen-activated protein kinase (MAPK) cascades are conserved eukaryote signaling modules where MAPKs, as the final kinases in the cascade, phosphorylate protein substrates to regulate cellular processes. While some progress in the identification of MAPK substrates has been made in plants, the knowledge on the spectrum of substrates and their mechanistic action is still fragmentary. In this focused review, we discuss the biological implications of the data in our original paper (Sustained mitogen-activated protein kinase activation reprograms defense metabolism and phosphoprotein profile in Arabidopsis thaliana; Frontiers in Plant Science 5: 554) in the context of related research. In our work, we mimicked in vivo activation of two stress-activated MAPKs, MPK3 and MPK6, through transgenic manipulation of Arabidopsis thaliana and used phosphoproteomics analysis to identify potential novel MAPK substrates. Here, we plotted the identified putative MAPK substrates (and downstream phosphoproteins) as a global protein clustering network. Based on a highly stringent selection confidence level, the core networks highlighted a MAPK-induced cellular reprogramming at multiple levels of gene and protein expression—including transcriptional, post-transcriptional, translational, post-translational (such as protein modification, folding, and degradation) steps, and also protein re-compartmentalization. Additionally, the increase in putative substrates/phosphoproteins of energy metabolism and various secondary metabolite biosynthesis pathways coincides with the observed accumulation of defense antimicrobial substances as detected by metabolome analysis. Furthermore, detection of protein networks in phospholipid or redox elements suggests activation of downstream signaling events. Taken in context with other studies, MAPKs are key regulators that reprogram cellular events to orchestrate defense signaling in eukaryotes. PMID:26579181

  9. The plane problem of the flapping wing

    NASA Technical Reports Server (NTRS)

    Birnbaum, Walter

    1954-01-01

    In connection with an earlier report on the lifting vortex sheet which forms the basis of the following investigations this will show how the methods developed there are also suitable for dealing with the air forces for a wing with a circulation variable with time. The theory of a propulsive wing flapping up and down periodically in the manner of a bird's wing is developed. This study shows how the lift and its moment result as a function of the flapping motion, what thrust is attainable, and how high is the degree of efficiency of this flapping propulsion unit if the air friction is disregarded.

  10. Flap-augmented shrouds for aerogenerators

    NASA Technical Reports Server (NTRS)

    Seginer, A.

    1976-01-01

    Axisymmetrical shrouds for windmills are augmented by ring-shaped 'flaps' and their performance is studied experimentally. The concept of the shroud as an annular 'wing' is justified, leading to the conclusion that high-lift techniques should be used in shroud design, and that high-lift devices, such as flaps, would increase the power output of the windmill. It is shown experimentally that the ideal power output of a flap-augmented shrouded turbine can be more than 4 times the power of unshrouded turbines of the same diameter.

  11. Dynamic response of a piezoelectric flapping wing

    NASA Astrophysics Data System (ADS)

    Kumar, Alok; Khandwekar, Gaurang; Venkatesh, S.; Mahapatra, D. R.; Dutta, S.

    2015-03-01

    Piezo-composite membranes have advantages over motorized flapping where frequencies are high and certain coupling between bending and twisting is useful to generate lift and forward flight. We draw examples of fruit fly and bumble bee. Wings with Piezo ceramic PZT coating are realized. The passive mechanical response of the wing is characterized experimentally and validated using finite element simulation. Piezoelectric actuation with uniform electrode coating is characterized and optimal frequencies for flapping are identified. The experimental data are used in an empirical model and advanced ratio for a flapping insect like condition for various angular orientations is estimated.

  12. Oxidative Stress Impairs the Stimulatory Effect of S100 Proteins on Protein Phosphatase 5 Activity.

    PubMed

    Yamaguchi, Fuminori; Tsuchiya, Mitsumasa; Shimamoto, Seiko; Fujimoto, Tomohito; Tokumitsu, Hiroshi; Tokuda, Masaaki; Kobayashi, Ryoji

    2016-01-01

    Oxidative stress is the consequence of an imbalance between the production of harmful reactive oxygen species and the cellular antioxidant system for neutralization, and it activates multiple intracellular signaling pathways, including apoptosis signal-regulating kinase 1 (ASK1). Protein phosphatase 5 (PP5) is a serine/threonine phosphatase involved in oxidative stress responses. Previously, we reported that S100 proteins activate PP5 in a calcium-dependent manner. S100 proteins belong to a family of small EF-hand calcium-binding proteins involved in many processes such as cell proliferation, differentiation, apoptosis, and inflammation. Therefore, we investigated the effects of oxidative stress on S100 proteins, their interaction with PP5, and PP5 enzyme activity. Recombinant S100A2 was easily air-oxidized or Cu-oxidized, and oxidized S100A2 formed cross-linked dimers and higher molecular-mass complexes. The binding of oxidized S100A2 to PP5 was reduced, resulting in decreased PP5 activation in vitro. Oxidation also impaired S100A1, S100A6, S100B, and S100P to activate PP5, although the low dose of oxidized S100 proteins still activated PP5. Hydrogen peroxide (H2O2) induced S100A2 oxidation in human keratinocytes (HaCaT) and human hepatocellular carcinoma (Huh-7) cells. Furthermore, H2O2 reduced the binding of S100A2 to PP5 and decreased PP5 activation in HaCaT and Huh-7 cells. Importantly, even the low dose of S100A2 achieved by knocking down increased dephosphorylation of ASK1 and reduced caspase 3/7 activity in Huh-7 cells treated with H2O2. These results indicate that oxidative stress impairs the ability of S100 proteins to bind and activate PP5, which in turn modulates the ASK1-mediated signaling cascades involved in apoptosis. PMID:27600583

  13. A conserved patch of hydrophobic amino acids modulates Myb activity by mediating protein-protein interactions.

    PubMed

    Dukare, Sandeep; Klempnauer, Karl-Heinz

    2016-07-01

    The transcription factor c-Myb plays a key role in the control of proliferation and differentiation in hematopoietic progenitor cells and has been implicated in the development of leukemia and certain non-hematopoietic tumors. c-Myb activity is highly dependent on the interaction with the coactivator p300 which is mediated by the transactivation domain of c-Myb and the KIX domain of p300. We have previously observed that conservative valine-to-isoleucine amino acid substitutions in a conserved stretch of hydrophobic amino acids have a profound effect on Myb activity. Here, we have explored the function of the hydrophobic region as a mediator of protein-protein interactions. We show that the hydrophobic region facilitates Myb self-interaction and binding of the histone acetyl transferase Tip60, a previously identified Myb interacting protein. We show that these interactions are affected by the valine-to-isoleucine amino acid substitutions and suppress Myb activity by interfering with the interaction of Myb and the KIX domain of p300. Taken together, our work identifies the hydrophobic region in the Myb transactivation domain as a binding site for homo- and heteromeric protein interactions and leads to a picture of the c-Myb transactivation domain as a composite protein binding region that facilitates interdependent protein-protein interactions of Myb with regulatory proteins.

  14. Organization, structure and activity of proteins in monolayers.

    PubMed

    Boucher, Julie; Trudel, Eric; Méthot, Mario; Desmeules, Philippe; Salesse, Christian

    2007-08-01

    Many different processes take place at the cell membrane interface. Indeed, for instance, ligands bind membrane proteins which in turn activate peripheral membrane proteins, some of which are enzymes whose action is also located at the membrane interface. Native cell membranes are difficult to use to gain information on the activity of individual proteins at the membrane interface because of the large number of different proteins involved in membranous processes. Model membrane systems, such as monolayers at the air-water interface, have thus been extensively used during the last 50 years to reconstitute proteins and to gain information on their organization, structure and activity in membranes. In the present paper, we review the recent work we have performed with membrane and peripheral proteins as well as enzymes in monolayers at the air-water interface. We show that the structure and orientation of gramicidin has been determined by combining different methods. Furthermore, we demonstrate that the secondary structure of rhodopsin and bacteriorhodopsin is indistinguishable from that in native membranes when appropriate conditions are used. We also show that the kinetics and extent of monolayer binding of myristoylated recoverin is much faster than that of the nonmyristoylated form and that this binding is highly favored by the presence polyunsaturated phospholipids. Moreover, we show that the use of fragments of RPE65 allow determine which region of this protein is most likely involved in membrane binding. Monomolecular films were also used to further understand the hydrolysis of organized phospholipids by phospholipases A2 and C.

  15. Anthelmintic activity of Leucaena leucocephala protein extracts on Haemonchus contortus.

    PubMed

    Soares, Alexandra Martins dos Santos; de Araújo, Sandra Alves; Lopes, Suzana Gomes; Costa Junior, Livio Martins

    2015-01-01

    The objective of this study was to evaluate the effects of protein extracts obtained from the plant Leucaena leucocephala on the nematode parasite Haemonchus contortus. The seeds, shell and cotyledon of L. leucocephala were separated and their proteins extracted using a sodium phosphate buffer, and named as TE (total seed extract), SE (shell extract) and CE (cotyledon extract). Soluble protein content, protease, protease inhibitory and chitinase activity assays were performed. Exsheathment inhibition of H. contortus larvae were performed at concentrations of 0.6 mg mL-1, and egg hatch assays were conducted at protein concentrations of 0.8, 0.4, 0.2, 0.1 and 0.05 mg mL-1. The effective concentration for 50% hatching inhibition (EC50) was estimated by probit. Different proportions of soluble proteins, protease and chitinase were found in TE and CE. Protease inhibitory activity was detected in all extracts. The EC50 of the CE and TE extracts were 0.48 and 0.33 mg mL-1, respectively. No ovicidal effects on H. contortus were detected in SE extracts, and none of the protein extracts demonstrated larvicidal effects on H. contortus. We therefore conclude that protein extracts of L. leucocephala had a detrimental effect on nematode eggs, which can be correlated with the high protease and chitinase activity of these extracts. PMID:26689178

  16. Organization, Structure and Activity of Proteins in Monolayers

    SciTech Connect

    Boucher,J.; Trudel, E.; Methot, M.; Desmeules, P.; Salesse, C.

    2007-01-01

    Many different processes take place at the cell membrane interface. Indeed, for instance, ligands bind membrane proteins which in turn activate peripheral membrane proteins, some of which are enzymes whose action is also located at the membrane interface. Native cell membranes are difficult to use to gain information on the activity of individual proteins at the membrane interface because of the large number of different proteins involved in membranous processes. Model membrane systems, such as monolayers at the air-water interface, have thus been extensively used during the last 50 years to reconstitute proteins and to gain information on their organization, structure and activity in membranes. In the present paper, we review the recent work we have performed with membrane and peripheral proteins as well as enzymes in monolayers at the air-water interface. We show that the structure and orientation of gramicidin has been determined by combining different methods. Furthermore, we demonstrate that the secondary structure of rhodopsin and bacteriorhodopsin is indistinguishable from that in native membranes when appropriate conditions are used. We also show that the kinetics and extent of monolayer binding of myristoylated recoverin is much faster than that of the nonmyristoylated form and that this binding is highly favored by the presence polyunsaturated phospholipids. Moreover, we show that the use of fragments of RPE65 allow determine which region of this protein is most likely involved in membrane binding. Monomolecular films were also used to further understand the hydrolysis of organized phospholipids by phospholipases A2 and C.

  17. Biologically active protein fragments containing specific binding regions of serum albumin or related proteins

    NASA Technical Reports Server (NTRS)

    Carter, Daniel C. (Inventor)

    1998-01-01

    In accordance with the present invention, biologically active protein fragments can be constructed which contain only those specific portions of the serum albumin family of proteins such as regions known as subdomains IIA and IIIA which are primarily responsible for the binding properties of the serum albumins. The artificial serums that can be prepared from these biologically active protein fragments are advantageous in that they can be produced much more easily than serums containing the whole albumin, yet still retain all or most of the original binding potential of the full albumin proteins. In addition, since the protein fragment serums of the present invention can be made from non-natural sources using conventional recombinant DNA techniques, they are far safer than serums containing natural albumin because they do not carry the potentially harmful viruses and other contaminants that will be found in the natural substances.

  18. Propeller Flap Reconstruction in Post Oncological Thigh Defect: "The Move in Flap".

    PubMed

    Nambi, G I; Salunke, Abhijeet Ashok

    2015-06-01

    Reconstruction of soft tissue defects of the limb after tumor resection is challenging question for oncosurgeons. The management differs from reconstruction of post traumatic defects due to the complexity of the primary surgery and subsequent radiation. The conventional propeller flap is based on a perforator which is located close to the defect; but in present case the perforator was located far away from the defect. So we describe it as "Move in flap" as the flap rotated a large volume of soft tissue lying between the defect and the perforator. We present a case of post oncological thigh defect with reconstruction using a propeller flap based on distal anteromedial perforator.

  19. DNA-dependent protein phosphorylation activity in Xenopus is coupled to a Ku-like protein.

    PubMed

    Kanungo, J; Cameron, R S; Takeda, Y; Hardin, J A

    1997-10-01

    DNA-dependent protein kinase (DNA-PK) is a nuclear enzyme and functions as a serine/threonine kinase that has been well characterized in both the human and the mouse. The regulatory subunit of DNA-PK is the Ku autoantigen. To demonstrate that a Ku-like protein is present in Xenopus oocytes, we used immunoprecipitation analysis with a monoclonal antibody raised against human Ku antigen and autoimmune serum containing anti-Ku antibodies. Metabolic labeling studies indicate that the Ku-like protein is synthesized mainly in late vitellogenic oocytes. By using a specific peptide substrate for DNA-PK, we demonstrate the activity of a DNA-dependent protein kinase in oocyte extracts. The kinase activity requires the Ku-like protein, since extracts depleted of Ku protein by immunoadsorption with human anti-Ku antibodies fail to demonstrate the DNA-dependent phosphorylation activity. The increased enzyme activity in vitellogenic oocytes may be correlated to the increased levels of Ku protein observed in these oocytes compared to the pre- and early vitellogenic oocytes.

  20. Factor H-related proteins determine complement-activating surfaces.

    PubMed

    Józsi, Mihály; Tortajada, Agustin; Uzonyi, Barbara; Goicoechea de Jorge, Elena; Rodríguez de Córdoba, Santiago

    2015-06-01

    Complement factor H-related proteins (FHRs) are strongly associated with different diseases involving complement dysregulation, which suggests a major role for these proteins regulating complement activation. Because FHRs are evolutionarily and structurally related to complement inhibitor factor H (FH), the initial assumption was that the FHRs are also negative complement regulators. Whereas weak complement inhibiting activities were originally reported for these molecules, recent developments indicate that FHRs may enhance complement activation, with important implications for the role of these proteins in health and disease. We review these findings here, and propose that FHRs represent a complex set of surface recognition molecules that, by competing with FH, provide improved discrimination of self and non-self surfaces and play a central role in determining appropriate activation of the complement pathway.

  1. Energy transfer at the active sites of heme proteins

    SciTech Connect

    Dlott, D.D.; Hill, J.R.

    1995-12-31

    Experiments using a picosecond pump-probe apparatus at the Picosecond Free-electron Laser Center at Stanford University, were performed to investigate the relaxation of carbon monoxide bound to the active sites of heme proteins. The significance of these experiments is two-fold: (1) they provide detailed information about molecular dynamics occurring at the active sites of proteins; and (2) they provide insight into the nature of vibrational relaxation processes in condensed matter. Molecular engineering is used to construct various molecular systems which are studied with the FEL. We have studied native proteins, mainly myoglobin obtained from different species, mutant proteins produced by genetic engineering using recombinant DNA techniques, and a variety of model systems which mimic the structures of the active sites of native proteins, which are produced using molecular synthesis. Use of these different systems permits us to investigate how specific molecular structural changes affect dynamical processes occurring at the active sites. This research provides insight into the problems of how different species needs are fulfilled by heme proteins which have greatly different functionality, which is induced by rather small structural changes.

  2. Utilizing avidity to improve antifreeze protein activity: a type III antifreeze protein trimer exhibits increased thermal hysteresis activity.

    PubMed

    Can, Özge; Holland, Nolan B

    2013-12-01

    Antifreeze proteins (AFPs) are ice growth inhibitors that allow the survival of several species living at temperatures colder than the freezing point of their bodily fluids. AFP activity is commonly defined in terms of thermal hysteresis, which is the difference observed for the solution freezing and melting temperatures. Increasing the thermal hysteresis activity of these proteins, particularly at low concentrations, is of great interest because of their wide range of potential applications. In this study, we have designed and expressed one-, two-, and three-domain antifreeze proteins to improve thermal hysteresis activity through increased binding avidity. The three-domain type III AFP yielded significantly greater activity than the one- and two-domain proteins, reaching a thermal hysteresis of >1.6 °C at a concentration of <1 mM. To elucidate the basis of this increase, the data were fit to a multidomain protein adsorption model based on the classical Langmuir isotherm. Fits of the data to the modified isotherms yield values for the equilibrium binding constants for the adsorption of AFP to ice and indicate that protein surface coverage is proportional to thermal hysteresis activity.

  3. Effects of diammonium glycyrrhizinate on random skin flap survival in rats: An experimental study

    PubMed Central

    Lv, Qing-Bo; Gao, Xiang; Lin, Ding-Sheng; Chen, Yun; Cao, Bin; Zhou, Kai-Liang

    2016-01-01

    Partial necrosis of skin flaps continues to restrict the survival of local skin flaps following plastic and reconstructive surgeries. The aim of the present study was to investigate the effects of diammonium glycyrrhizinate (DG), a salt of glycyrrhetinic acid that has been widely used in the therapy of chronic hepatitis and human immunodeficiency virus infection, on random skin flap survival in rats. McFarlane flaps were established in 60 male Sprague-Dawley rats randomly divided into three groups. Group I served as the control group and was injected with saline (10 mg/kg) once per day. Group II and group III were the experimental groups, and were injected with 10 mg/kg DG once and twice per day, respectively. On day 7, the survival area of the flap was measured. Tissue samples were stained with hematoxylin and eosin and immunohistochemically evaluated. Tissue edema, neutrophil density, superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were evaluated. The mean survival areas of the flaps of group II were significantly larger when compared with those of group I (P<0.05), and the rats of group III exhibited significantly higher survival areas than group II (P<0.05). Histologic and immunohistochemical evaluation showed that microvessel development and the expression level of vascular endothelial growth factor were higher in the two experimental groups than in the control group. Furthermore, SOD activity was significantly increased (P<0.05), while the neutrophil density and MDA level were significantly reduced (P<0.05) in group II when compared with group I. Significant differences between group II and group III with regard to SOD activity and MDA level were also observed (P<0.05). Thus, DG may have a dose-dependent effect on promoting the survival of random skin flaps. PMID:27588181

  4. Gc protein (vitamin D-binding protein): Gc genotyping and GcMAF precursor activity.

    PubMed

    Nagasawa, Hideko; Uto, Yoshihiro; Sasaki, Hideyuki; Okamura, Natsuko; Murakami, Aya; Kubo, Shinichi; Kirk, Kenneth L; Hori, Hitoshi

    2005-01-01

    The Gc protein (human group-specific component (Gc), a vitamin D-binding protein or Gc globulin), has important physiological functions that include involvement in vitamin D transport and storage, scavenging of extracellular G-actin, enhancement of the chemotactic activity of C5a for neutrophils in inflammation and macrophage activation (mediated by a GalNAc-modified Gc protein (GcMAF)). In this review, the structure and function of the Gc protein is focused on especially with regard to Gc genotyping and GcMAF precursor activity. A discussion of the research strategy "GcMAF as a target for drug discovery" is included, based on our own research. PMID:16302727

  5. Fluorogen-Activating-Proteins as Universal Affinity Biosensors for Immunodetection

    PubMed Central

    Gallo, Eugenio; Vasilev, Kalin V.; Jarvik, Jonathan

    2014-01-01

    Fluorogen-activating-proteins (FAPs) are a novel platform of fluorescence biosensors utilized for protein discovery. The technology currently demands molecular manipulation methods that limit its application and adaptability. Here, we highlight an alternative approach based on universal affinity reagents for protein detection. The affinity reagents were engineered as bi-partite fusion proteins, where the specificity moiety is derived from IgG-binding proteinsProtein-A or Protein-G – and the signaling element is a FAP. In this manner, primary antibodies provide the antigenic selectivity against a desired protein in biological samples, while FAP affinity reagents target the constant region (Fc) of antibodies and provide the biosensor component of detection. Fluorescence results using various techniques indicate minimal background and high target specificity for exogenous and endogenous proteins in mammalian cells. Additionally, FAP-based affinity reagents provide enhanced properties of detection previously absent using conventional affinity systems. Distinct features explored in this report include: (1) unfixed signal wavelengths (excitation and emission) determined by the particular fluorogen chosen, (2) real-time user controlled fluorescence on-set and off-set, (3) signal wavelength substitution while performing live analysis, and (4) enhanced resistance to photobleaching. PMID:24122476

  6. Counteracting Protein Kinase Activity in the Heart: The Multiple Roles of Protein Phosphatases

    PubMed Central

    Weber, Silvio; Meyer-Roxlau, Stefanie; Wagner, Michael; Dobrev, Dobromir; El-Armouche, Ali

    2015-01-01

    Decades of cardiovascular research have shown that variable and flexible levels of protein phosphorylation are necessary to maintain cardiac function. A delicate balance between phosphorylated and dephosphorylated states of proteins is guaranteed by a complex interplay of protein kinases (PKs) and phosphatases. Serine/threonine phosphatases, in particular members of the protein phosphatase (PP) family govern dephosphorylation of the majority of these cardiac proteins. Recent findings have however shown that PPs do not only dephosphorylate previously phosphorylated proteins as a passive control mechanism but are capable to actively control PK activity via different direct and indirect signaling pathways. These control mechanisms can take place on (epi-)genetic, (post-)transcriptional, and (post-)translational levels. In addition PPs themselves are targets of a plethora of proteinaceous interaction partner regulating their endogenous activity, thus adding another level of complexity and feedback control toward this system. Finally, novel approaches are underway to achieve spatiotemporal pharmacologic control of PPs which in turn can be used to fine-tune misleaded PK activity in heart disease. Taken together, this review comprehensively summarizes the major aspects of PP-mediated PK regulation and discusses the subsequent consequences of deregulated PP activity for cardiovascular diseases in depth. PMID:26617522

  7. Peptides and proteins with antimicrobial activity.

    PubMed

    Coutinho, Henrique Douglas Melo; Lôbo, Katiuscia Menezes; Bezerra, Denise Aline Casimiro; Lôbo, Inalzuir

    2008-01-01

    The increase of microbial resistance to antibiotics has led to a continuing search for newer and more effective drugs. Antimicrobial peptides are generally found in animals, plants, and microorganisms and are of great interest to medicine, pharmacology, and the food industry. These peptides are capable of inhibiting pathogenic microorganisms. They can attack parasites, while causing little or no harm to the host cells. The defensins are peptides found in granules in the polymorphonuclear neutrophils (PMNs) and are responsible for the defense of the organism. Several animal defensins, like dermaseptin, antileukoprotease, protegrin, and others, have had their activities and efficacy tested and been shown to be effective against bacteria, fungi, and protists; there are also specific defensins from invertebrates, e.g., drosomycin and heliomicin; from plants, e.g., the types A and B; and the bacteriocins, e.g., acrocin, marcescin, etc. The aim of the present work was to compile a comprehensive bibliographic review of the diverse potentially antimicrobial peptides in an effort to systematize the current knowledge on these substances as a contribution for further researches. The currently available bibliography does not give a holistic approach on this subject. The present work intends to show that the mechanism of defense represented by defensins is promising from the perspective of its application in the treatment of infectious diseases in human, animals and plants.

  8. Route Flap Damping Made Usable

    NASA Astrophysics Data System (ADS)

    Pelsser, Cristel; Maennel, Olaf; Mohapatra, Pradosh; Bush, Randy; Patel, Keyur

    The Border Gateway Protocol (BGP), the de facto inter-domain routing protocol of the Internet, is known to be noisy. The protocol has two main mechanisms to ameliorate this, MinRouteAdvertisementInterval (MRAI), and Route Flap Damping (RFD). MRAI deals with very short bursts on the order of a few to 30 seconds. RFD deals with longer bursts, minutes to hours. Unfortunately, RFD was found to severely penalize sites for being well-connected because topological richness amplifies the number of update messages exchanged. So most operators have disabled it. Through measurement, this paper explores the avenue of absolutely minimal change to code, and shows that a few RFD algorithmic constants and limits can be trivially modified, with the result being damping a non-trivial amount of long term churn without penalizing well-behaved prefixes' normal convergence process.

  9. Heated Proteins are Still Active in a Functionalized Nanoporous Support

    SciTech Connect

    Chen, Baowei; Qi, Wen N.; Li, Xiaolin; Lei, Chenghong; Liu, Jun

    2013-07-08

    We report that even under the heated condition, the conformation and activity of a protein can be hoarded in a functionalized nanoporous support via non-covalent interaction, although the hoarded protein was not exhibiting the full protein activity, the protein released subsequently still maintained its native conformation and activity. Glucose oxidase (GOX) was spontaneously and largely entrapped in aminopropyl-functionalized mesoporous silica (NH2-FMS) at 20 oC via a dominant electrostatic interaction. Although FMS-GOX displayed 45% activity of the free enzyme in solution, the GOX released from FMS exhibited its 100% activity prior to the entrapment. Surprisingly, the released GOX from FMS still maintained 89% of its initial activity prior to the entrapment after FMS-GOX was incubated at 60 oC for 1 h prior to release, while the free GOX in solution lost nearly all activity under the same incubation. Intrinsic fluorescence emission of GOX and native electrophoresis demonstrated that the heating resulted in significant conformational changes and oligomeric structures of the free GOX, but FMS efficiently maintained the thermal stability of GOX therein and resisted the thermal denaturation and oligomeric aggregation.

  10. Staphylokinase as a Plasminogen Activator Component in Recombinant Fusion Proteins

    PubMed Central

    Szarka, S. J.; Sihota, E. G.; Habibi, H. R.; Wong, S.-L.

    1999-01-01

    The plasminogen activator staphylokinase (SAK) is a promising thrombolytic agent for treatment of myocardial infarction. It can specifically stimulate the thrombolysis of both erythrocyte-rich and platelet-rich clots. However, SAK lacks fibrin-binding and thrombin inhibitor activities, two functions which would supplement and potentially improve its thrombolytic potency. Creating a recombinant fusion protein is one approach for combining protein domains with complementary functions. To evaluate SAK for use in a translational fusion protein, both N- and C-terminal fusions to SAK were constructed by using hirudin as a fusion partner. Recombinant fusion proteins were secreted from Bacillus subtilis and purified from culture supernatants. The rate of plasminogen activation by SAK was not altered by the presence of an additional N- or C-terminal protein sequence. However, cleavage at N-terminal lysines within SAK rendered the N-terminal fusion unstable in the presence of plasmin. The results of site-directed mutagenesis of lysine 10 and lysine 11 in SAK suggested that a plasmin-resistant variant cannot be created without interfering with the plasmin processing necessary for activation of SAK. Although putative plasmin cleavage sites are located at the C-terminal end of SAK at lysine 135 and lysine 136, these sites were resistant to plasmin cleavage in vitro. Therefore, C-terminal fusions represent stable configurations for developing improved thrombolytic agents based on SAK as the plasminogen activator component. PMID:9925575

  11. Ca2+ activates human homologous recombination protein Rad51 by modulating its ATPase activity

    PubMed Central

    Bugreev, Dmitry V.; Mazin, Alexander V.

    2004-01-01

    Human Rad51 (hRad51) protein plays a key role in homologous recombination and DNA repair. hRad51 protein forms a helical filament on single-stranded DNA (ssDNA), which performs the basic steps of homologous recombination: a search for homologous double-stranded DNA (dsDNA) and DNA strand exchange. hRad51 protein possesses DNA-dependent ATPase activity; however, the role of this activity has not been understood. Our current results show that Ca2+ greatly stimulates DNA strand exchange activity of hRad51 protein. We found that Ca2+ exerts its stimulatory effect by modulating the ATPase activity of hRad51 protein. Our data demonstrate that, in the presence of Mg2+, the hRad51-ATP-ssDNA filament is quickly converted to an inactive hRad51-ADP-ssDNA form, due to relatively rapid ATP hydrolysis and slow dissociation of ADP. Ca2+ maintains the active hRad51-ATP-ssDNA filament by reducing the ATP hydrolysis rate. These findings demonstrate a crucial role of the ATPase activity in regulation of DNA strand exchange activity of hRad51 protein. This mechanism of Rad51 protein regulation by modulating its ATPase activity is evolutionarily recent; we found no such mechanism for yeast Rad51 (yRad51) protein. PMID:15226506

  12. Phytochrome activation of two nuclear genes requires cytoplasmic protein synthesis.

    PubMed Central

    Lam, E; Green, P J; Wong, M; Chua, N H

    1989-01-01

    We have investigated the effects of protein synthesis inhibitors on light-induced expression of two plant nuclear genes, Cab and rbcS, in wheat, pea and transgenic tobacco. Light activation of these two genes is very sensitive to cycloheximide, an inhibitor of cytoplasmic protein synthesis but not to chloramphenicol, an inhibitor of organellar protein synthesis. Studies with chimeric gene constructs in transgenic tobacco seedlings show that cycloheximide exerts its effect at the transcriptional level. As a control, we show that the expression of the cauliflower mosaic virus (CaMV) 35S promoter is enhanced by cycloheximide treatment, irrespective of the coding sequence used. Escape-time analyses with green wheat seedlings show that the cycloheximide block for Cab gene expression is after the primary signal transduction step linked to phytochrome photoconversion. Our results suggest that phytochrome activation of Cab and rbcS is mediated by a labile protein factor(s) synthesized on cytoplasmic ribosomes. Images PMID:2583082

  13. Mucocele formation under pedicled nasoseptal flap.

    PubMed

    Vaezeafshar, Reza; Hwang, Peter H; Harsh, Griffith; Turner, Justin H

    2012-01-01

    The pedicled nasoseptal flap has become an indispensible tool for the reconstruction of skull base defects. This flap is easily harvested, provides a large surface area of vascularized tissue, and has few reported complications. We describe the case of a 60-year-old man who underwent endoscopic, endonasal transsphenoidal surgery with septal flap reconstruction who developed a sphenoid sinus mucocele postoperatively. We also have reviewed the literature for similar findings and discuss this complication in the setting of pituitary surgery and endoscopic skull base repair. Although likely a rare occurrence, mucocele formation after septal flap reconstruction should be recognized and monitored with postoperative nasal endoscopy and radiologic imaging. Reoperation or mucocele drainage may be necessary if symptomatic or in cases of rapid enlargement.

  14. Postirradiation flap infection about the oral cavity

    SciTech Connect

    Cabbabe, E.B.; Herbold, D.R.; Sunwoo, Y.C.; Baroudi, I.F.

    1983-06-01

    Postirradiation alteration of oral flora is well documented in the literature. Infection as a complication leading to partial or complete loss of a flap used to reconstruct a defect in the oral cavity is a worrisome outcome. We describe how a flap that was judged clinically to be viable became overwhelmingly infected with the Klebsiella oxytoca, an oral cavity pathogen encountered in this patient following irradiation. Local and systemic changes led to detachment of the flap. This complication may be explained, in view of the absence of venous congestion or arterial ischemia both clinically and pathologically, by the proven contamination of the flap by the Klebsiella pathogen. Local factors resulted in lower resistance and subsequent overwhelming infection. Discussion of the case, review of pertinent literature, and proposed solutions are presented.

  15. A dynamical system for interacting flapping swimmers

    NASA Astrophysics Data System (ADS)

    Oza, Anand; Ramananarivo, Sophie; Ristroph, Leif; Shelley, Michael

    2015-11-01

    We present the results of a theoretical investigation into the dynamics of interacting flapping swimmers. Our study is motivated by the recent experiments of Becker et al., who studied a one-dimensional array of self-propelled flapping wings that swim within each other's wakes in a water tank. They discovered that the system adopts certain ``schooling modes'' characterized by specific spatial phase relationships between swimmers. To rationalize these phenomena, we develop a discrete dynamical system in which the swimmers are modeled as heaving airfoils that shed point vortices during each flapping cycle. We then apply our model to recent experiments in the Applied Math Lab, in which two tandem flapping airfoils are free to choose both their speed and relative positions. We expect that our model may be used to understand how schooling behavior is influenced by hydrodynamics in more general contexts. Thanks to the NSF for its support.

  16. Management of Vortices Trailing Flapped Wings via Separation Control

    NASA Technical Reports Server (NTRS)

    Greenblatt, David

    2005-01-01

    A pilot study was conducted on a flapped semi-span model to investigate the concept and viability of near-wake vortex management via separation control. Passive control was achieved by means of a simple fairing and active control was achieved via zero mass-flux blowing slots. Vortex sheet strength, estimated by integrating surface pressure ports, was used to predict vortex characteristics by means of inviscid rollup relations. Furthermore, vortices trailing the flaps were mapped using a seven-hole probe. Separation control was found to have a marked effect on vortex location, strength, tangential velocity, axial velocity and size over a wide range of angles of attack and control conditions. In general, the vortex trends were well predicted by the inviscid rollup relations. Manipulation of the separated flow near the flap edges exerted significant control over both outboard and inboard edge vortices while producing negligible lift excursions. Dynamic separation and attachment control was found to be an effective means for dynamically perturbing the vortex from arbitrarily long wavelengths down to wavelengths less than a typical wingspan. In summary, separation control has the potential for application to time-independent or time-dependent wake alleviation schemes, where the latter can be deployed to minimize adverse effects on ride-quality and dynamic structural loading.

  17. Endocytosis of Seven-Transmembrane RGS Protein Activates G- protein Coupled Signaling in Arabidopsis

    PubMed Central

    Urano, Daisuke; Phan, Nguyen; Jones, Janice C.; Yang, Jing; Huang, Jirong; Grigston, Jeffrey; Taylor, J. Philip; Jones, Alan M.

    2012-01-01

    Signal transduction typically begins by ligand-dependent activation of a concomitant partner which is otherwise in its resting state. However, in cases where signal activation is constitutive by default, the mechanism of regulation is unknown. The Arabidopsis thaliana heterotrimeric Gα protein self-activates without accessory proteins, and is kept in its resting state by the negative regulator, AtRGS1 (Regulator of G protein Signaling 1), which is the prototype of a seven transmembrane receptor fused with an RGS domain. Endocytosis of AtRGS1 by ligand-dependent endocytosis physically uncouples the GTPase accelerating activity of AtRGS1 from the Gα protein, permitting sustained activation. Phosphorylation of AtRGS1 by AtWNK8 kinase causes AtRGS1 endocytosis, required both for G protein-mediated sugar signaling and cell proliferation. In animals, receptor endocytosis results in signal desensitization, whereas in plants, endocytosis results in signal activation. These findings reveal how different organisms rearrange a regulatory system to result in opposite outcomes using similar phosphorylation-dependent endocytosis. PMID:22940907

  18. Beyond the zone: protein needs of active individuals.

    PubMed

    Lemon, P W

    2000-10-01

    There has been debate among athletes and nutritionists regarding dietary protein needs for centuries. Although contrary to traditional belief, recent scientific information collected on physically active individuals tends to indicate that regular exercise increases daily protein requirements; however, the precise details remain to be worked out. Based on laboratory measures, daily protein requirements are increased by perhaps as much as 100% vs. recommendations for sedentary individuals (1.6-1.8 vs. 0.8 g/kg). Yet even these intakes are much less than those reported by most athletes. This may mean that actual requirements are below what is needed to optimize athletic performance, and so the debate continues. Numerous interacting factors including energy intake, carbohydrate availability, exercise intensity, duration and type, dietary protein quality, training history, gender, age, timing of nutrient intake and the like make this topic extremely complex. Many questions remain to be resolved. At the present time, substantial data indicate that the current recommended protein intake should be adjusted upward for those who are physically active, especially in populations whose needs are elevated for other reasons, e.g., growing individuals, dieters, vegetarians, individuals with muscle disease-induced weakness and the elderly. For these latter groups, specific supplementation may be appropriate, but for most North Americans who consume a varied diet, including complete protein foods (meat, eggs, fish and dairy products), and sufficient energy the increased protein needs induced by a regular exercise program can be met in one's diet.

  19. Interpreting laser Doppler recordings from free flaps.

    PubMed

    Svensson, H; Holmberg, J; Svedman, P

    1993-01-01

    Although the transfer of free flaps is nowadays accomplished with an increasing degree of safety, thrombosis of the microvascular anastomoses is still a problem. In order to avoid delay in re-operating, various methods for objective blood flow monitoring have been tried, among them Laser Doppler Flowmetry (LDF). When one reviews the literature, it is apparent that opinions differ about whether or not LDF is a reliable technique for this purpose. To focus on the need to interpret continuous recordings, this paper reports our findings in six latissimus dorsi free flaps chosen from our series of LDF monitoring procedures. One uneventful flap, no. 1, had an immediate postoperative LDF value of 4.5 perfusion units (PU). LDF values improved during the recovery period and the graphic recording showed fluctuations due to normal physiological variations of the blood flow in the flap. Another uneventful flap, no. 4, showed the same pattern, though at an appreciably lower level, 2 PU, on average. Flap no. 2 had an acceptably high value of 3.5 PU despite suffering a venous thrombosis. However, the LDF recording showed no fluctuations and the value declined gradually. Another flap, no. 3, showed fluctuations and blood flow was normal although the value decreased to 2.5 PU. In flap no. 5, any value between 2 and 3.5 PU could be obtained merely by adjusting the position of the probe in the holder. In no. 6, the LDF value suddenly dropped, accompanied by a decrease in the total amount of backscattered light, indicating venous obstruction which was confirmed at re-operation.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. Extending the extended V-Y flap.

    PubMed

    Prowse, Phoebe; Morton, Jonathan

    2012-06-01

    This case report demonstrates a modification of the so-called 'Extended V-Y Flap' used to simultaneously reconstruct a defect involving the upper lip, floor of nose and alar rim following tumour excision. We hope that this case serves as a reminder of the versatility of the V-Y flap in the nasolabial region, and its considerable capacity for augmentation. PMID:22018837

  1. The origin of the temporalis muscle flap.

    PubMed

    Speculand, B

    1992-12-01

    This paper explores the origin of the temporalis muscle flap which has been attributed to Golovine, a Moscow ophthalmic surgeon, who described a forehead skin transposition flap. Small temporalis muscle transpositions were used in surgery for TMJ ankylosis, but the use of the majority of the muscle for reconstruction of facial defects was first described by Sir Harold Gillies during the 1914-18 war.

  2. Aerodynamic effects of flexibility in flapping wings.

    PubMed

    Zhao, Liang; Huang, Qingfeng; Deng, Xinyan; Sane, Sanjay P

    2010-03-01

    Recent work on the aerodynamics of flapping flight reveals fundamental differences in the mechanisms of aerodynamic force generation between fixed and flapping wings. When fixed wings translate at high angles of attack, they periodically generate and shed leading and trailing edge vortices as reflected in their fluctuating aerodynamic force traces and associated flow visualization. In contrast, wings flapping at high angles of attack generate stable leading edge vorticity, which persists throughout the duration of the stroke and enhances mean aerodynamic forces. Here, we show that aerodynamic forces can be controlled by altering the trailing edge flexibility of a flapping wing. We used a dynamically scaled mechanical model of flapping flight (Re approximately 2000) to measure the aerodynamic forces on flapping wings of variable flexural stiffness (EI). For low to medium angles of attack, as flexibility of the wing increases, its ability to generate aerodynamic forces decreases monotonically but its lift-to-drag ratios remain approximately constant. The instantaneous force traces reveal no major differences in the underlying modes of force generation for flexible and rigid wings, but the magnitude of force, the angle of net force vector and centre of pressure all vary systematically with wing flexibility. Even a rudimentary framework of wing veins is sufficient to restore the ability of flexible wings to generate forces at near-rigid values. Thus, the magnitude of force generation can be controlled by modulating the trailing edge flexibility and thereby controlling the magnitude of the leading edge vorticity. To characterize this, we have generated a detailed database of aerodynamic forces as a function of several variables including material properties, kinematics, aerodynamic forces and centre of pressure, which can also be used to help validate computational models of aeroelastic flapping wings. These experiments will also be useful for wing design for small

  3. Aerodynamic effects of flexibility in flapping wings

    PubMed Central

    Zhao, Liang; Huang, Qingfeng; Deng, Xinyan; Sane, Sanjay P.

    2010-01-01

    Recent work on the aerodynamics of flapping flight reveals fundamental differences in the mechanisms of aerodynamic force generation between fixed and flapping wings. When fixed wings translate at high angles of attack, they periodically generate and shed leading and trailing edge vortices as reflected in their fluctuating aerodynamic force traces and associated flow visualization. In contrast, wings flapping at high angles of attack generate stable leading edge vorticity, which persists throughout the duration of the stroke and enhances mean aerodynamic forces. Here, we show that aerodynamic forces can be controlled by altering the trailing edge flexibility of a flapping wing. We used a dynamically scaled mechanical model of flapping flight (Re ≈ 2000) to measure the aerodynamic forces on flapping wings of variable flexural stiffness (EI). For low to medium angles of attack, as flexibility of the wing increases, its ability to generate aerodynamic forces decreases monotonically but its lift-to-drag ratios remain approximately constant. The instantaneous force traces reveal no major differences in the underlying modes of force generation for flexible and rigid wings, but the magnitude of force, the angle of net force vector and centre of pressure all vary systematically with wing flexibility. Even a rudimentary framework of wing veins is sufficient to restore the ability of flexible wings to generate forces at near-rigid values. Thus, the magnitude of force generation can be controlled by modulating the trailing edge flexibility and thereby controlling the magnitude of the leading edge vorticity. To characterize this, we have generated a detailed database of aerodynamic forces as a function of several variables including material properties, kinematics, aerodynamic forces and centre of pressure, which can also be used to help validate computational models of aeroelastic flapping wings. These experiments will also be useful for wing design for small robotic

  4. A theoretical approach to spot active regions in antimicrobial proteins

    PubMed Central

    2009-01-01

    Background Much effort goes into identifying new antimicrobial compounds able to evade the increasing resistance of microorganisms to antibiotics. One strategy relies on antimicrobial peptides, either derived from fragments released by proteolytic cleavage of proteins or designed from known antimicrobial protein regions. Results To identify these antimicrobial determinants, we developed a theoretical approach that predicts antimicrobial proteins from their amino acid sequence in addition to determining their antimicrobial regions. A bactericidal propensity index has been calculated for each amino acid, using the experimental data reported from a high-throughput screening assay as reference. Scanning profiles were performed for protein sequences and potentially active stretches were identified by the best selected threshold parameters. The method was corroborated against positive and negative datasets. This successful approach means that we can spot active sequences previously reported in the literature from experimental data for most of the antimicrobial proteins examined. Conclusion The method presented can correctly identify antimicrobial proteins with an accuracy of 85% and a sensitivity of 90%. The method can also predict their key active regions, making this a tool for the design of new antimicrobial drugs. PMID:19906288

  5. Angiogenic activity of an Onchocerca volvulus Ancylostoma secreted protein homologue.

    PubMed

    Higazi, Tarig B; Pearlman, Eric; Whikehart, David R; Unnasch, Thomas R

    2003-06-01

    Angiogenesis is an important step in the development of ocular onchocercaisis. In previous studies, it has been demonstrated that Onchocerca volvulus homologues of the Ancylostoma secreted protein family have pronounced angiogenic activity. The overall goal of the current study was to determine if this angiogenic effect is exerted through a direct or indirect mechanism. These studies focused on one member of this family, OvASP-2, as this protein is expressed in microfilaria, the stage of the parasite that causes ocular onchocercaisis. Clones encoding truncated and full length open reading frames were expressed as fusion proteins with Escherichia coli maltose binding protein (MBP), and angiogenic activity was compared in vitro and in vivo with MBP alone. Truncated constructs expressing only the first 105 amino acids of OvASP-2 were as active as the full length protein in inducing new blood vessel formation. The full length fusion protein did not stimulate proliferation or production of vascular endothelial growth factor in vascular endothelial cells in vitro, indicating that OvASP-2 does not directly stimulate angiogenesis. Sequence analysis demonstrated that the gene encoding OvASP-2 contained five introns. Sequence comparisons of the genomic loci from West African blinding and non-blinding strains of O. volvulus revealed that some polymorphism existed among the various isolates tested. However, none of these polymorphisms could be used to differentiate the parasite strains, suggesting that qualitative variation in OvASP-2 could not explain the difference in ocular pathogenic potential of the two parasite strains.

  6. Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi.

    PubMed Central

    Ulloa, R M; Mesri, E; Esteva, M; Torres, H N; Téllez-Iñón, M T

    1988-01-01

    A cyclic AMP-dependent protein kinase activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns, giving two peaks of kinase activity, which were eluted at 0.15 M- and 0.32 M-NaCl respectively. The second activity peak was stimulated by nanomolar concentrations of cyclic AMP. In addition, a cyclic AMP-binding protein co-eluted with the second kinase activity peak. Cyclic AMP-dependent protein kinase activity was further purified by gel filtration, affinity chromatography on histone-agarose and cyclic AMP-agarose, as well as by chromatography on CM-Sephadex. The enzyme ('holoenzyme') could be partially dissociated into two different components: 'catalytic' and 'regulatory'. The 'regulatory' component had specific binding for cyclic AMP, and it inhibited phosphotransferase activity of the homologous 'catalytic component' or of the 'catalytic subunit' from bovine heart. Cyclic AMP reversed these inhibitions. A 'holoenzyme preparation' was phosphorylated in the absence of exogenous phosphate acceptor and analysed by polyacrylamide-gel electrophoresis. A 56 kDa band was phosphorylated. The same preparation was analysed by Western blotting, by using polyclonal antibodies to the regulatory subunits of protein kinases type I or II. Both antibodies reacted with the 56 kDa band. Images Fig. 7. Fig. 8. PMID:2848508

  7. Hydrodynamic collective effects of active proteins in biological membranes.

    PubMed

    Koyano, Yuki; Kitahata, Hiroyuki; Mikhailov, Alexander S

    2016-08-01

    Lipid bilayers forming biological membranes are known to behave as viscous two-dimensional fluids on submicrometer scales; usually they contain a large number of active protein inclusions. Recently, it was shown [A. S. Mikhailov and R. Kapral, Proc. Natl. Acad. Sci. USA 112, E3639 (2015)PNASA60027-842410.1073/pnas.1506825112] that such active proteins should induce nonthermal fluctuating lipid flows leading to diffusion enhancement and chemotaxislike drift for passive inclusions in biomembranes. Here, a detailed analytical and numerical investigation of such effects is performed. The attention is focused on the situations when proteins are concentrated within lipid rafts. We demonstrate that passive particles tend to become attracted by active rafts and are accumulated inside them. PMID:27627343

  8. MAPK-Activated Protein Kinases (MKs): Novel Insights and Challenges.

    PubMed

    Gaestel, Matthias

    2015-01-01

    Downstream of MAPKs, such as classical/atypical ERKs and p38 MAPKs, but not of JNKs, signaling is often mediated by protein kinases which are phosphorylated and activated by MAPKs and, therefore, designated MAPK-activated protein kinases (MAPKAPKs). Recently, novel insights into the specificity of the assembly of MAPK/MAPKAPK hetero-dimeric protein kinase signaling complexes have been gained. In addition, new functional aspects of MKs have been described and established functions have been challenged. This short review will summarize recent developments including the linear motif (LM) in MKs, the ERK-independent activation of RSK, the RSK-independent effects of some RSK-inhibitors and the challenged role of MK5/PRAK in tumor suppression. PMID:26779481

  9. Sterol carrier protein2-like activity in rat intestine.

    PubMed

    Kharroubi, A; Wadsworth, J A; Chanderbhan, R; Wiesenfeld, P; Noland, B; Scallen, T; Vahouny, G V; Gallo, L L

    1988-03-01

    A sterol carrier protein2 (SCP2)-like activity has been demonstrated in rat intestinal mucosal homogenates and in isolated intestinal cells from both crypt and villus zones. The results indicate the presence of a protein with similar molecular weight and antigenicity to that of authentic SCP2 purified from rat liver cytosol. Like liver SCP2, mucosal cytosol stimulates pregnenolone production in rat adrenal mitochondria and acyl coenzyme A:cholesterol acyltransferase activity of liver and mucosal microsomes. The distribution of SCP2-like activity as determined by radioimmunoassay indicates high levels in mitochondria and cytosol and relatively lower levels in microsomes and in brush-border membranes. The widespread distribution of SCP2-like protein in the intestine is consistent with potential transfer functions in all phases of cholesterol processing. PMID:3379341

  10. Hydrodynamic collective effects of active proteins in biological membranes

    NASA Astrophysics Data System (ADS)

    Koyano, Yuki; Kitahata, Hiroyuki; Mikhailov, Alexander S.

    2016-08-01

    Lipid bilayers forming biological membranes are known to behave as viscous two-dimensional fluids on submicrometer scales; usually they contain a large number of active protein inclusions. Recently, it was shown [A. S. Mikhailov and R. Kapral, Proc. Natl. Acad. Sci. USA 112, E3639 (2015), 10.1073/pnas.1506825112] that such active proteins should induce nonthermal fluctuating lipid flows leading to diffusion enhancement and chemotaxislike drift for passive inclusions in biomembranes. Here, a detailed analytical and numerical investigation of such effects is performed. The attention is focused on the situations when proteins are concentrated within lipid rafts. We demonstrate that passive particles tend to become attracted by active rafts and are accumulated inside them.

  11. Investigation of Energy Harvesting Using Flapping Foils

    NASA Astrophysics Data System (ADS)

    Mivehchi, Amin; Persichetti, Amanda; Dunham, Brandon; Dahl, Jason M.

    2013-11-01

    When harvesting kinetic energy using a flapping foil, the separation of coherent structures in the wake is crucial for determining forces on the body. Applications for utilizing energy harvesting with a flapping foil include powering of local, low power equipment and recharging AUV batteries that use flapping foils for propulsion and maneuvering. In each of these cases, it is critical to accurately predict the physical behavior and location of vortices in relation to the motion of the body in order to maximize energy output. A two-dimensional open source boundary data immersion method (LilyPad) is used for simulating the flapping motion of a foil for energy harvesting in a current. Forced motion of the flapping body indicates theoretical efficiencies for energy harvesting near 43 percent under specific flapping conditions. A simple control scheme based on pressure sensing on the surface of the foil is developed to control pitch of the foil while energy harvesting occurs in the heave direction. The control scheme is tested through real time numerical simulation. Comparisons are made with physical laboratory experiments, demonstrating high efficiencies in energy harvesting.

  12. Signal peptides are allosteric activators of the protein translocase

    PubMed Central

    Gouridis, Giorgos; Karamanou, Spyridoula; Gelis, Ioannis; Kalodimos, Charalampos G.; Economou, Anastassios

    2010-01-01

    Extra-cytoplasmic polypeptides are usually synthesized as “preproteins” carrying aminoterminal, cleavable signal peptides1 and secreted across membranes by translocases. The main bacterial translocase comprises the SecYEG protein-conducting channel and the peripheral ATPase motor SecA2,3. Most proteins destined for the periplasm and beyond are exported post-translationally by SecA2,3. Preprotein targeting to SecA is thought to involve signal peptides4 and chaperones like SecB5,6. Here we reveal that signal peptides have a novel role beyond targeting: they are essential allosteric activators of the translocase. Upon docking on their binding groove on SecA, signal peptides act in trans to drive three successive states: first, “triggering” that drives the translocase to a lower activation energy state; then “trapping” that engages non-native preprotein mature domains docked with high affinity on the secretion apparatus and, finally, “secretion” during which trapped mature domains undergo multiple turnovers of translocation in segments7. A significant contribution by mature domains renders signal peptides less critical in bacterial secretory protein targeting than currently assumed. Rather, it is their function as allosteric activators of the translocase that renders signal peptides essential for protein secretion. A role for signal peptides and targeting sequences as allosteric activators may be universal in protein translocases. PMID:19924216

  13. The dog-ear flap as an alternative for breast reconstruction in patients who have already undergone a DIEAP flap.

    PubMed

    Colebunders, Britt; Depypere, Bernard; Van Landuyt, Koenraad

    2016-05-01

    Breast reconstruction in patients who have previously undergone deep inferior epigastric artery perforator flap (DIEAP) reconstruction or abdominoplasty is often challenging. Depending on patients' body habitus, several second-choice flaps have been described such as the transverse upper gracilis (TUG) flap, profundus femoris artery perforator (PFAP) flap, superior gluteal artery perforator (SGAP) flap, and lumbar artery perforator (LAP) flap. Patients who have undergone a DIEAP flap reconstruction or abdominoplasty occasionally present with dog ears on both sides of the abdominal scar. The adipose tissue and skin of these dog ears are supplied by perforators of the deep circumflex iliac artery (DCIA). The DCIA flap was first described in 1979 by Taylor. We introduce this abdominal "dog-ear" flap for autologous breast reconstruction. PMID:26951847

  14. Complement activation and protein adsorption by carbon nanotubes.

    PubMed

    Salvador-Morales, Carolina; Flahaut, Emmanuel; Sim, Edith; Sloan, Jeremy; Green, Malcolm L H; Sim, Robert B

    2006-02-01

    As a first step to validate the use of carbon nanotubes as novel vaccine or drug delivery devices, their interaction with a part of the human immune system, complement, has been explored. Haemolytic assays were conducted to investigate the activation of the human serum complement system via the classical and alternative pathways. Western blot and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) techniques were used to elucidate the mechanism of activation of complement via the classical pathway, and to analyse the interaction of complement and other plasma proteins with carbon nanotubes. We report for the first time that carbon nanotubes activate human complement via both classical and alternative pathways. We conclude that complement activation by nanotubes is consistent with reported adjuvant effects, and might also in various circumstances promote damaging effects of excessive complement activation, such as inflammation and granuloma formation. C1q binds directly to carbon nanotubes. Protein binding to carbon nanotubes is highly selective, since out of the many different proteins in plasma, very few bind to the carbon nanotubes. Fibrinogen and apolipoproteins (AI, AIV and CIII) were the proteins that bound to carbon nanotubes in greatest quantity.

  15. L-Alanylglutamine inhibits signaling proteins that activate protein degradation, but does not affect proteins that activate protein synthesis after an acute resistance exercise.

    PubMed

    Wang, Wanyi; Choi, Ran Hee; Solares, Geoffrey J; Tseng, Hung-Min; Ding, Zhenping; Kim, Kyoungrae; Ivy, John L

    2015-07-01

    Sustamine™ (SUS) is a dipeptide composed of alanine and glutamine (AlaGln). Glutamine has been suggested to increase muscle protein accretion; however, the underlying molecular mechanisms of glutamine on muscle protein metabolism following resistance exercise have not been fully addressed. In the present study, 2-month-old rats climbed a ladder 10 times with a weight equal to 75 % of their body mass attached at the tail. Rats were then orally administered one of four solutions: placebo (PLA-glycine = 0.52 g/kg), whey protein (WP = 0.4 g/kg), low dose of SUS (LSUS = 0.1 g/kg), or high dose of SUS (HSUS = 0.5 g/kg). An additional group of sedentary (SED) rats was intubated with glycine (0.52 g/kg) at the same time as the ladder-climbing rats. Blood samples were collected immediately after exercise and at either 20 or 40 min after recovery. The flexor hallucis longus (FHL), a muscle used for climbing, was excised at 20 or 40 min post exercise and analyzed for proteins regulating protein synthesis and degradation. All supplements elevated the phosphorylation of FOXO3A above SED at 20 min post exercise, but only the SUS supplements significantly reduced the phosphorylation of AMPK and NF-kB p65. SUS supplements had no effect on mTOR signaling, but WP supplementation yielded a greater phosphorylation of mTOR, p70S6k, and rpS6 compared with PLA at 20 min post exercise. However, by 40 min post exercise, phosphorylation of mTOR and rpS6 in PLA had risen to levels not different than WP. These results suggest that SUS blocks the activation of intracellular signals for MPB, whereas WP accelerates mRNA translation.

  16. Methods to distinguish various types of protein phosphatase activity

    SciTech Connect

    Brautigan, D.L.; Shriner, C.L.

    1988-01-01

    To distinguish the action of protein Tyr(P) and protein Ser(P)/Thr(P) phosphatases on /sup 32/P-labeled phosphoproteins in subcellular fractions different inhibitors and activators are utilized. Comparison of the effects of added compounds provides a convenient, indirect method to characterize dephosphorylation reactions. Protein Tyr(P) phosphatases are specifically inhibited by micromolar Zn2+ or vanadate, and show maximal activity in the presence of EDTA. The other class of cellular phosphatases, specific for protein Ser(P) and Thr(P) residues, are inhibited by fluoride and EDTA. In this class of enzymes two major functional types can be distinguished: those sensitive to inhibition by the heat-stable protein inhibitor-2 and not stimulated by polycations, and those not sensitive to inhibition and stimulated by polycations. Preparation of /sup 32/P-labeled Tyr(P) and Ser(P) phosphoproteins also is presented for the direct measurement of phosphatase activities in preparations by the release of acid-soluble (/sup 32/P)phosphate.

  17. Actions of Rho family small G proteins and p21-activated protein kinases on mitogen-activated protein kinase family members.

    PubMed Central

    Frost, J A; Xu, S; Hutchison, M R; Marcus, S; Cobb, M H

    1996-01-01

    The mitogen-activated protein (MAP) kinases are a family of serine/threonine kinases that are regulated by distinct extracellular stimuli. The currently known members include extracellular signal-regulated protein kinase 1 (ERK1), ERK2, the c-Jun N-terminal kinase/stress-activated protein kinases (JNK/SAPKs), and p38 MAP kinases. We find that overexpression of the Ste20-related enzymes p21-activated kinase 1 (PAK1) and PAK2 in 293 cells is sufficient to activate JNK/SAPK and to a lesser extent p38 MAP kinase but not ERK2. Rat MAP/ERK kinase kinase 1 can stimulate the activity of each of these MAP kinases. Although neither activated Rac nor the PAKs stimulate ERK2 activity, overexpression of either dominant negative Rac2 or the N-terminal regulatory domain of PAK1 inhibits Ras-mediated activation of ERK2, suggesting a permissive role for Rac in the control of the ERK pathway. Furthermore, constitutively active Rac2, Cdc42hs, and RhoA synergize with an activated form of Raf to increase ERK2 activity. These findings reveal a previously unrecognized connection between Rho family small G proteins and the ERK pathway. PMID:8668187

  18. Comparing semilunar coronally positioned flap to standard coronally positioned flap using periodontal clinical parameters.

    PubMed

    Nassar, Carlos Augusto; da Silva, Wilson Aparecido Dias; Tonet, Karine; Secundes, Mayron Barros; Nassar, Patricia Oehlmeyer

    2014-01-01

    This study compared the effectiveness of 2 surgical root coverage techniques--semilunar coronally positioned flap and coronally advanced flap--using the clinical parameters of periodontal tissues from patients with Miller Class I gingival recession. Twenty patients (20-50 years of age) were selected. Basic periodontal treatment was performed, and plaque index, gingival index, probing depth, clinical attachment level, and height of the attached gingiva were determined. Each patient was placed into 1 of 2 groups: Group 1 patients underwent the semilunar coronally positioned flap technique, and Group 2 patients underwent the coronally advanced flap technique. Patients were assessed for 180 days. Both groups showed significant reduction of plaque and gingival indices and an improvement in clinical attachment levels and probing depth. However, results showed the standard coronally positioned flap technique was deemed more effective due to significant clinical attachment level gains. PMID:24598495

  19. AMP-activated protein kinase and metabolic control

    PubMed Central

    Viollet, Benoit; Andreelli, Fabrizio

    2011-01-01

    AMP-activated protein kinase (AMPK), a phylogenetically conserved serine/threonine protein kinase, is a major regulator of cellular and whole-body energy homeostasis that coordinates metabolic pathways in order to balance nutrient supply with energy demand. It is now recognized that pharmacological activation of AMPK improves blood glucose homeostasis, lipid profile and blood pressure in insulin-resistant rodents. Indeed, AMPK activation mimics the beneficial effects of physical activity or those of calorie restriction by acting on multiple cellular targets. In addition it is now demonstrated that AMPK is one of the probable (albeit indirect) targets of major antidiabetic drugs including, the biguanides (metformin) and thiazolidinediones, as well as of insulin sensitizing adipokines (e.g., adiponectin). Taken together, such findings highlight the logic underlying the concept of targeting the AMPK pathway for the treatment of metabolic syndrome and type 2 diabetes. PMID:21484577

  20. Detergent activation of the binding protein in the folate radioassay

    SciTech Connect

    Hansen, S.I.; Holm, J.; Lyngbye, J.

    1982-01-01

    A minor cow's whey protein associated with ..beta..-lactoglobulin is used as binding protein in the competitive radioassay for serum and erythrocyte folate. Seeking to optimize the assay, we tested the performance of binder solutions of increasing purity. The folate binding protein was isolated from cow's whey by means of CM-Sepharose CL-6B cation-exchange chromatography, and further purified on a methotrexate-AH-Sepharose 4B affinity matrix. In contrast to ..beta..-lactoglobulin, the purified protein did not bind folate unless the detergents cetyltrimethylammonium (10 mmol/Ll) or Triton X-100 (1 g/L) were present. Such detergent activation was not needed in the presence of serum. There seems to be a striking analogy between these phenomena and the well-known reactivation of certain purified membrane-derived enzymes by surfactants (lipids/detergents).

  1. Controlling Protein Activity and Degradation Using Blue Light.

    PubMed

    Lutz, Anne P; Renicke, Christian; Taxis, Christof

    2016-01-01

    Regulation of protein stability is a fundamental process in eukaryotic cells and pivotal to, e.g., cell cycle progression, faithful chromosome segregation, or protein quality control. Synthetic regulation of protein stability requires conditional degradation sequences (degrons) that induce a stability switch upon a specific signal. Fusion to a selected target protein permits to influence virtually every process in a cell. Light as signal is advantageous due to its precise applicability in time, space, quality, and quantity. Light control of protein stability was achieved by fusing the LOV2 photoreceptor domain of Arabidopsis thaliana phototropin1 with a synthetic degron (cODC1) derived from the carboxy-terminal degron of ornithine decarboxylase to obtain the photosensitive degron (psd) module. The psd module can be attached to the carboxy terminus of target proteins that are localized to the cytosol or nucleus to obtain light control over their stability. Blue light induces structural changes in the LOV2 domain, which in turn lead to activation of the degron and thus proteasomal degradation of the whole fusion protein. Variants of the psd module with diverse characteristics are useful to fine-tune the stability of a selected target at permissive (darkness) and restrictive conditions (blue light). PMID:26965116

  2. Altered Activation of Protein Kinase PKR and Enhanced Apoptosis in Dystonia Cells Carrying a Mutation in PKR Activator Protein PACT*

    PubMed Central

    Vaughn, Lauren S; Bragg, D. Cristopher; Sharma, Nutan; Camargos, Sarah; Cardoso, Francisco; Patel, Rekha C

    2015-01-01

    PACT is a stress-modulated activator of the interferon-induced double-stranded RNA-activated protein kinase (PKR). Stress-induced phosphorylation of PACT is essential for PACT's association with PKR leading to PKR activation. PKR activation leads to phosphorylation of translation initiation factor eIF2α inhibition of protein synthesis and apoptosis. A recessively inherited form of early-onset dystonia DYT16 has been recently identified to arise due to a homozygous missense mutation P222L in PACT. To examine if the mutant P222L protein alters the stress-response pathway, we examined the ability of mutant P222L to interact with and activate PKR. Our results indicate that the substitution mutant P222L activates PKR more robustly and for longer duration albeit with slower kinetics in response to the endoplasmic reticulum stress. In addition, the affinity of PACT-PACT and PACT-PKR interactions is enhanced in dystonia patient lymphoblasts, thereby leading to intensified PKR activation and enhanced cellular death. P222L mutation also changes the affinity of PACT-TRBP interaction after cellular stress, thereby offering a mechanism for the delayed PKR activation in response to stress. Our results demonstrate the impact of a dystonia-causing substitution mutation on stress-induced cellular apoptosis. PMID:26231208

  3. Steroidogenic Acute Regulatory Protein Overexpression Correlates with Protein Kinase A Activation in Adrenocortical Adenoma

    PubMed Central

    Xie, Jing; Su, Tingwei; Jiang, Lei; Jiang, Yiran; Cao, Yanan; Liu, Jianmin; Ning, Guang; Wang, Weiqing

    2016-01-01

    The association of pathological features of cortisol-producing adrenocortical adenomas (ACAs) with somatic driver mutations and their molecular classification remain unclear. In this study, we explored the association between steroidogenic acute regulatory protein (StAR) expression and the driver mutations activating cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling to identify the pathological markers of ACAs. Immunohistochemical staining for StAR and mutations in the protein kinase cAMP-activated catalytic subunit alpha (PRKACA), protein kinase cAMP-dependent type I regulatory subunit alpha (PRKAR1A) and guanine nucleotide binding protein, alpha stimulating (GNAS) genes were examined in 97 ACAs. The association of StAR expression with the clinical and mutational features of the ACAs was analyzed. ACAs with mutations in PRKACA, GNAS, and PRKAR1A showed strong immunopositive staining for StAR. The concordance between high StAR expression and mutations activating cAMP/PKA signaling in the ACAs was 99.0%. ACAs with high expression of StAR had significantly smaller tumor volume (P < 0.001) and higher urinary cortisol per tumor volume (P = 0.032) than those with low expression of StAR. Our findings suggest that immunohistochemical staining for StAR is a reliable pathological approach for the diagnosis and classification of ACAs with cAMP/PKA signaling-activating mutations. PMID:27606678

  4. Steroidogenic Acute Regulatory Protein Overexpression Correlates with Protein Kinase A Activation in Adrenocortical Adenoma.

    PubMed

    Zhou, Weiwei; Wu, Luming; Xie, Jing; Su, Tingwei; Jiang, Lei; Jiang, Yiran; Cao, Yanan; Liu, Jianmin; Ning, Guang; Wang, Weiqing

    2016-01-01

    The association of pathological features of cortisol-producing adrenocortical adenomas (ACAs) with somatic driver mutations and their molecular classification remain unclear. In this study, we explored the association between steroidogenic acute regulatory protein (StAR) expression and the driver mutations activating cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling to identify the pathological markers of ACAs. Immunohistochemical staining for StAR and mutations in the protein kinase cAMP-activated catalytic subunit alpha (PRKACA), protein kinase cAMP-dependent type I regulatory subunit alpha (PRKAR1A) and guanine nucleotide binding protein, alpha stimulating (GNAS) genes were examined in 97 ACAs. The association of StAR expression with the clinical and mutational features of the ACAs was analyzed. ACAs with mutations in PRKACA, GNAS, and PRKAR1A showed strong immunopositive staining for StAR. The concordance between high StAR expression and mutations activating cAMP/PKA signaling in the ACAs was 99.0%. ACAs with high expression of StAR had significantly smaller tumor volume (P < 0.001) and higher urinary cortisol per tumor volume (P = 0.032) than those with low expression of StAR. Our findings suggest that immunohistochemical staining for StAR is a reliable pathological approach for the diagnosis and classification of ACAs with cAMP/PKA signaling-activating mutations.

  5. Steroidogenic Acute Regulatory Protein Overexpression Correlates with Protein Kinase A Activation in Adrenocortical Adenoma.

    PubMed

    Zhou, Weiwei; Wu, Luming; Xie, Jing; Su, Tingwei; Jiang, Lei; Jiang, Yiran; Cao, Yanan; Liu, Jianmin; Ning, Guang; Wang, Weiqing

    2016-01-01

    The association of pathological features of cortisol-producing adrenocortical adenomas (ACAs) with somatic driver mutations and their molecular classification remain unclear. In this study, we explored the association between steroidogenic acute regulatory protein (StAR) expression and the driver mutations activating cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling to identify the pathological markers of ACAs. Immunohistochemical staining for StAR and mutations in the protein kinase cAMP-activated catalytic subunit alpha (PRKACA), protein kinase cAMP-dependent type I regulatory subunit alpha (PRKAR1A) and guanine nucleotide binding protein, alpha stimulating (GNAS) genes were examined in 97 ACAs. The association of StAR expression with the clinical and mutational features of the ACAs was analyzed. ACAs with mutations in PRKACA, GNAS, and PRKAR1A showed strong immunopositive staining for StAR. The concordance between high StAR expression and mutations activating cAMP/PKA signaling in the ACAs was 99.0%. ACAs with high expression of StAR had significantly smaller tumor volume (P < 0.001) and higher urinary cortisol per tumor volume (P = 0.032) than those with low expression of StAR. Our findings suggest that immunohistochemical staining for StAR is a reliable pathological approach for the diagnosis and classification of ACAs with cAMP/PKA signaling-activating mutations. PMID:27606678

  6. Blade-Mounted Flap Control for BVI Noise Reduction Proof-of-Concept Test

    NASA Technical Reports Server (NTRS)

    Dawson, Seth; Hassan, Ahmed; Straub, Friedrich; Tadghighi, Hormoz

    1995-01-01

    This report describes a wind tunnel test of the McDonnell Douglas Helicopter Systems (MDHS) Active Flap Model Rotor at the NASA Langley 14- by 22-Foot Subsonic Tunnel. The test demonstrated that BVI noise reductions and vibration reductions were possible with the use of an active flap. Aerodynamic results supported the acoustic data trends, showing a reduction in the strength of the tip vortex with the deflection of the flap. Acoustic results showed that the flap deployment, depending on the peak deflection angle and azimuthal shift in its deployment schedule, can produce BVI noise reductions as much as 6 dB on the advancing and retreating sides. The noise reduction was accompanied by an increase in low frequency harmonic noise and high frequency broadband noise. A brief assessment of the effect of the flap on vibration showed that significant reductions were possible. The greatest vibration reductions (as much as 76%) were found in the four per rev pitching moment at the hub. Performance improvement cam results were inconclusive, as the improvements were predicted to be smaller than the resolution of the rotor balance.

  7. A bio-inspired study on tidal energy extraction with flexible flapping wings.

    PubMed

    Liu, Wendi; Xiao, Qing; Cheng, Fai

    2013-09-01

    Previous research on the flexible structure of flapping wings has shown an improved propulsion performance in comparison to rigid wings. However, not much is known about this function in terms of power efficiency modification for flapping wing energy devices. In order to study the role of the flexible wing deformation in the hydrodynamics of flapping wing energy devices, we computationally model the two-dimensional flexible single and twin flapping wings in operation under the energy extraction conditions with a large Reynolds number of 106. The flexible motion for the present study is predetermined based on a priori structural result which is different from a passive flexibility solution. Four different models are investigated with additional potential local distortions near the leading and trailing edges. Our simulation results show that the flexible structure of a wing is beneficial to enhance power efficiency by increasing the peaks of lift force over a flapping cycle, and tuning the phase shift between force and velocity to a favourable trend. Moreover, the impact of wing flexibility on efficiency is more profound at a low nominal effective angle of attack (AoA). At a typical flapping frequency f * = 0.15 and nominal effective AoA of 10°, a flexible integrated wing generates 7.68% higher efficiency than a rigid wing. An even higher increase, around six times that of a rigid wing, is achievable if the nominal effective AoA is reduced to zero degrees at feathering condition. This is very attractive for a semi-actuated flapping energy system, where energy input is needed to activate the pitching motion. The results from our dual-wing study found that a parallel twin-wing device can produce more power compared to a single wing due to the strong flow interaction between the two wings.

  8. Installing hydrolytic activity into a completely de novo protein framework

    NASA Astrophysics Data System (ADS)

    Burton, Antony J.; Thomson, Andrew R.; Dawson, William M.; Brady, R. Leo; Woolfson, Derek N.

    2016-09-01

    The design of enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here we install hydrolytic activity predictably into a completely de novo and thermostable α-helical barrel, which comprises seven helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine-histidine-glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies that match the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such a predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis.

  9. Installing hydrolytic activity into a completely de novo protein framework

    NASA Astrophysics Data System (ADS)

    Burton, Antony J.; Thomson, Andrew R.; Dawson, William M.; Brady, R. Leo; Woolfson, Derek N.

    2016-09-01

    The design of enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here we install hydrolytic activity predictably into a completely de novo and thermostable α-helical barrel, which comprises seven helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine–histidine–glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies that match the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such a predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis.

  10. Installing hydrolytic activity into a completely de novo protein framework.

    PubMed

    Burton, Antony J; Thomson, Andrew R; Dawson, William M; Brady, R Leo; Woolfson, Derek N

    2016-09-01

    The design of enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here we install hydrolytic activity predictably into a completely de novo and thermostable α-helical barrel, which comprises seven helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine-histidine-glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies that match the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such a predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis. PMID:27554410

  11. Aerodynamic flight performance in flap-gliding birds and bats.

    PubMed

    Muijres, Florian T; Henningsson, Per; Stuiver, Melanie; Hedenström, Anders

    2012-08-01

    Many birds use a flight mode called undulating or flap-gliding flight, where they alternate between flapping and gliding phases, while only a few bats make use of such a flight mode. Among birds, flap-gliding is commonly used by medium to large species, where it is regarded to have a lower energetic cost than continuously flapping flight. Here, we introduce a novel model for estimating the energetic flight economy of flap-gliding animals, by determining the lift-to-drag ratio for flap-gliding based on empirical lift-to-drag ratio estimates for continuous flapping flight and for continuous gliding flight, respectively. We apply the model to flight performance data of the common swift (Apus apus) and of the lesser long-nosed bat (Leptonycteris yerbabuenae). The common swift is a typical flap-glider while-to the best of our knowledge-the lesser long-nosed bat does not use flap-gliding. The results show that, according to the model, the flap-gliding common swift saves up to 15% energy compared to a continuous flapping swift, and that this is primarily due to the exceptionally high lift-to-drag ratio in gliding flight relative to that in flapping flight for common swifts. The lesser long-nosed bat, on the other hand, seems not to be able to reduce energetic costs by flap-gliding. The difference in relative costs of flap-gliding flight between the common swift and the lesser long-nosed bat can be explained by differences in morphology, flight style and wake dynamics. The model presented here proves to be a valuable tool for estimating energetic flight economy in flap-gliding animals. The results show that flap-gliding flight that is naturally used by common swifts is indeed the most economic one of the two flight modes, while this is not the case for the non-flap-gliding lesser long-nosed bat.

  12. Extended thoracodorsal artery perforator flap for breast reconstruction.

    PubMed

    Angrigiani, Claudio; Rancati, Alberto; Escudero, Ezequiel; Artero, Guillermo

    2015-12-01

    A total of 45 patients underwent partial or total autologous breast reconstruction after skin-sparing mastectomy, skin-reducing mastectomy, and quadrantectomy using a thoracodorsal artery perforator (TDAP) flap. The detailed surgical technique with its variations is explained in this report. The propeller, flip-over, conventional perforator, and muscle-sparing flaps have been described and evaluated. The flaps were partially or completely de-epithelialized. The conventional TDAP can be enlarged or "extended" as the traditional latissimus dorsi musculocutaneous (LD-MC) flap by incorporating the superior and inferior fat compartments. It can be referred to as the "extended TDAP flap". This technique augments the flap volume. In addition, this flap can serve as a scaffold for lipofilling to obtain autologous breast reconstruction in medium to large cases. There were two complete failures due to technical errors during flap elevation. Distal partial tissue suffering was observed in four flaps. These flaps were longer than usual; they reached the midline of the back. It is advisable to discard the distal medial quarter of the flap when it is designed up to the midline to avoid steatonecrosis or fibrosis. A retrospective analysis of the 39 flaps that survived completely revealed a satisfactory result in 82% of the cases. The main disadvantage of this procedure is the final scar. The TDAP flap is a reliable and safe method for partial or total breast autologous reconstruction. PMID:26645006

  13. Protein kinase A activity and Hedgehog signaling pathway.

    PubMed

    Kotani, Tomoya

    2012-01-01

    Protein kinase A (PKA) is a well-known kinase that plays fundamental roles in a variety of biological processes. In Hedgehog-responsive cells, PKA plays key roles in proliferation and fate specification by modulating the transduction of Hedgehog signaling. In the absence of Hedgehog, a basal level of PKA activity represses the transcription of Hedgehog target genes. The main substrates of PKA in this process are the Ci/Gli family of bipotential transcription factors, which activate and repress Hedgehog target gene expression. PKA phosphorylates Ci/Gli, promoting the production of the repressor forms of Ci/Gli and thus repressing Hedgehog target gene expression. In contrast, the activation of Hedgehog signaling in response to Hedgehog increases the active forms of Ci/Gli, resulting in Hedgehog target gene expression. Because both decreased and increased levels of PKA activity cause abnormal cell proliferation and alter cell fate specification, the basal level of PKA activity in Hedgehog-responsive cells should be precisely regulated. However, the mechanism by which PKA activity is regulated remains obscure and appears to vary between cell types, tissues, and organisms. To date, two mechanisms have been proposed. One is a classical mechanism in which PKA activity is regulated by a small second messenger, cAMP; the other is a novel mechanism in which PKA activity is regulated by a protein, Misty somites. PMID:22391308

  14. Monitoring Brain Activity with Protein Voltage and Calcium Sensors

    PubMed Central

    Storace, Douglas A.; Braubach, Oliver R.; Jin, Lei; Cohen, Lawrence B.; Sung, Uhna

    2015-01-01

    Understanding the roles of different cell types in the behaviors generated by neural circuits requires protein indicators that report neural activity with high spatio-temporal resolution. Genetically encoded fluorescent protein (FP) voltage sensors, which optically report the electrical activity in distinct cell populations, are, in principle, ideal candidates. Here we demonstrate that the FP voltage sensor ArcLight reports odor-evoked electrical activity in the in vivo mammalian olfactory bulb in single trials using both wide-field and 2-photon imaging. ArcLight resolved fast odorant-responses in individual glomeruli, and distributed odorant responses across a population of glomeruli. Comparisons between ArcLight and the protein calcium sensors GCaMP3 and GCaMP6f revealed that ArcLight had faster temporal kinetics that more clearly distinguished activity elicited by individual odorant inspirations. In contrast, the signals from both GCaMPs were a saturating integral of activity that returned relatively slowly to the baseline. ArcLight enables optical electrophysiology of mammalian neuronal population activity in vivo. PMID:25970202

  15. Monitoring brain activity with protein voltage and calcium sensors.

    PubMed

    Storace, Douglas A; Braubach, Oliver R; Jin, Lei; Cohen, Lawrence B; Sung, Uhna

    2015-05-13

    Understanding the roles of different cell types in the behaviors generated by neural circuits requires protein indicators that report neural activity with high spatio-temporal resolution. Genetically encoded fluorescent protein (FP) voltage sensors, which optically report the electrical activity in distinct cell populations, are, in principle, ideal candidates. Here we demonstrate that the FP voltage sensor ArcLight reports odor-evoked electrical activity in the in vivo mammalian olfactory bulb in single trials using both wide-field and 2-photon imaging. ArcLight resolved fast odorant-responses in individual glomeruli, and distributed odorant responses across a population of glomeruli. Comparisons between ArcLight and the protein calcium sensors GCaMP3 and GCaMP6f revealed that ArcLight had faster temporal kinetics that more clearly distinguished activity elicited by individual odorant inspirations. In contrast, the signals from both GCaMPs were a saturating integral of activity that returned relatively slowly to the baseline. ArcLight enables optical electrophysiology of mammalian neuronal population activity in vivo.

  16. Protein synthesis inhibitors reveal differential regulation of mitogen-activated protein kinase and stress-activated protein kinase pathways that converge on Elk-1.

    PubMed Central

    Zinck, R; Cahill, M A; Kracht, M; Sachsenmaier, C; Hipskind, R A; Nordheim, A

    1995-01-01

    Inhibitors of protein synthesis, such as anisomycin and cycloheximide, lead to superinduction of immediate-early genes. We demonstrate that these two drugs activate intracellular signaling pathways involving both the mitogen-activated protein kinase (MAPK) and stress-activated protein kinase (SAPK) cascades. The activation of either pathway correlates with phosphorylation of the c-fos regulatory transcription factor Elk-1. In HeLa cells, anisomycin stabilizes c-fos mRNA when protein synthesis is inhibited to only 50%. Under these conditions, anisomycin, in contrast to cycloheximide, rapidly induces kinase activation and efficient Elk-1 phosphorylation. However, full inhibition of translation by either drug leads to prolonged activation of SAPK activity, while MAPK induction is transient. This correlates with prolonged Elk-1 phosphorylation and c-fos transcription. Elk-1 induction and c-fos activation are also observed in KB cells, in which anisomycin strongly induces SAPKs but not MAPKs. Purified p54 SAPK alpha efficiently phosphorylates the Elk-1 C-terminal domain in vitro and comigrates with anisomycin-activated kinases in in-gel kinase assays. Thus, Elk-1 provides a potential convergence point for the MAPK and SAPK signaling pathways. The activation of signal cascades and control of transcription factor function therefore represent prominent processes in immediate-early gene superinduction. PMID:7651411

  17. Immersion freezing of ice nucleating active protein complexes

    NASA Astrophysics Data System (ADS)

    Hartmann, S.; Augustin, S.; Clauss, T.; Voigtländer, J.; Niedermeier, D.; Wex, H.; Stratmann, F.

    2012-08-01

    Biological particles, e.g. bacteria and their Ice Nucleating Active (INA) protein complexes, might play an important role for the ice formation in atmospheric mixed-phase clouds. Therefore, the immersion freezing behavior of INA protein complexes generated from a SnomaxTM solution/suspension was investigated as function of temperature in a range of -5 °C to -38 °C at the Leipzig Aerosol Cloud Interaction Simulator (LACIS). The immersion freezing of droplets containing small numbers of INA protein complexes occurs in a temperature range of -7 °C and -10 °C. The experiments performed in the lower temperature range, where all droplets freeze which contain at least one INA protein complex, are used to determine the average number of INA protein complexes present, assuming that the INA protein complexes are Poisson distributed over the droplet ensemble. Knowing the average number of INA protein complexes, the heterogeneous ice nucleation rate and rate coefficient of a single INA protein complex is determined by using the newly-developed CHESS model (stoCHastic model of idEntical poiSSon distributed ice nuclei). Therefore, we assume the ice nucleation process to be of stochastic nature, and a parameterization of the INA protein complex's nucleation rate. Analyzing the results of immersion freezing experiments from literature (SnomaxTM and Pseudomonas syringae bacteria), to results gained in this study, demonstrates that first, a similar temperature dependence of the heterogeneous ice nucleation rate for a single INA protein complex was found in all experiments, second, the shift of the ice fraction curves to higher temperatures can be explained consistently by a higher average number of INA protein complexes being present in the droplet ensemble, and finally the heterogeneous ice nucleation rate of one single INA protein complex might be also applicable for intact Pseudomonas syringae bacteria cells. The results obtained in this study allow a new perspective on the

  18. Activation of immobilized, biotinylated choleragen AI protein by a 19-kilodalton guanine nucleotide-binding protein.

    PubMed

    Noda, M; Tsai, S C; Adamik, R; Bobak, D A; Moss, J; Vaughan, M

    1989-09-19

    Cholera toxin catalyzes the ADP-ribosylation that results in activation of the stimulatory guanine nucleotide-binding protein of the adenylyl cyclase system, known as Gs. The toxin also ADP-ribosylates other proteins and simple guanidino compounds and auto-ADP-ribosylates its AI protein (CTA1). All of the ADP-ribosyltransferase activities of CTAI are enhanced by 19-21-kDa guanine nucleotide-binding proteins known as ADP-ribosylation factors, or ARFs. CTAI contains a single cysteine located near the carboxy terminus. CTAI was immobilized through this cysteine by reaction with iodoacetyl-N-biotinyl-hexylenediamine and binding of the resulting biotinylated protein to avidin-agarose. Immobilized CTAI catalyzed the ARF-stimulated ADP-ribosylation of agmatine. The reaction was enhanced by detergents and phospholipid, but the fold stimulation by purified sARF-II from bovine brain was considerably less than that observed with free CTA. ADP-ribosylation of Gsa by immobilized CTAI, which was somewhat enhanced by sARF-II, was much less than predicted on the basis of the NAD:agmatine ADP-ribosyltransferase activity. Immobilized CTAI catalyzed its own auto-ADP-ribosylation as well as the ADP-ribosylation of the immobilized avidin and CTA2, with relatively little stimulation by sARF-II. ADP-ribosylation of CTA2 by free CTAI is minimal. These observations are consistent with the conclusion that the cysteine near the carboxy terminus of the toxin is not critical for ADP-ribosyltransferase activity or for its regulation by sARF-II. Biotinylation and immobilization of the toxin through this cysteine may, however, limit accessibility to Gsa or SARF-II, or perhaps otherwise reduce interaction with these proteins whether as substrates or activator.

  19. Reassessing the Potential Activities of Plant CGI-58 Protein

    PubMed Central

    Khatib, Abdallah; Arhab, Yani; Bentebibel, Assia; Abousalham, Abdelkarim; Noiriel, Alexandre

    2016-01-01

    Comparative Gene Identification-58 (CGI-58) is a widespread protein found in animals and plants. This protein has been shown to participate in lipolysis in mice and humans by activating Adipose triglyceride lipase (ATGL), the initial enzyme responsible for the triacylglycerol (TAG) catabolism cascade. Human mutation of CGI-58 is the cause of Chanarin-Dorfman syndrome, an orphan disease characterized by a systemic accumulation of TAG which engenders tissue disorders. The CGI-58 protein has also been shown to participate in neutral lipid metabolism in plants and, in this case, a mutation again provokes TAG accumulation. Although its roles as an ATGL coactivator and in lipid metabolism are quite clear, the catalytic activity of CGI-58 is still in question. The acyltransferase activities of CGI-58 have been speculated about, reported or even dismissed and experimental evidence that CGI-58 expressed in E. coli possesses an unambiguous catalytic activity is still lacking. To address this problem, we developed a new set of plasmids and site-directed mutants to elucidate the in vivo effects of CGI-58 expression on lipid metabolism in E. coli. By analyzing the lipid composition in selected E. coli strains expressing CGI-58 proteins, and by reinvestigating enzymatic tests with adequate controls, we show here that recombinant plant CGI-58 has none of the proposed activities previously described. Recombinant plant and mouse CGI-58 both lack acyltransferase activity towards either lysophosphatidylglycerol or lysophosphatidic acid to form phosphatidylglycerol or phosphatidic acid and recombinant plant CGI-58 does not catalyze TAG or phospholipid hydrolysis. However, expression of recombinant plant CGI-58, but not mouse CGI-58, led to a decrease in phosphatidylglycerol in all strains of E. coli tested, and a mutation of the putative catalytic residues restored a wild-type phenotype. The potential activities of plant CGI-58 are subsequently discussed. PMID:26745266

  20. Reassessing the Potential Activities of Plant CGI-58 Protein.

    PubMed

    Khatib, Abdallah; Arhab, Yani; Bentebibel, Assia; Abousalham, Abdelkarim; Noiriel, Alexandre

    2016-01-01

    Comparative Gene Identification-58 (CGI-58) is a widespread protein found in animals and plants. This protein has been shown to participate in lipolysis in mice and humans by activating Adipose triglyceride lipase (ATGL), the initial enzyme responsible for the triacylglycerol (TAG) catabolism cascade. Human mutation of CGI-58 is the cause of Chanarin-Dorfman syndrome, an orphan disease characterized by a systemic accumulation of TAG which engenders tissue disorders. The CGI-58 protein has also been shown to participate in neutral lipid metabolism in plants and, in this case, a mutation again provokes TAG accumulation. Although its roles as an ATGL coactivator and in lipid metabolism are quite clear, the catalytic activity of CGI-58 is still in question. The acyltransferase activities of CGI-58 have been speculated about, reported or even dismissed and experimental evidence that CGI-58 expressed in E. coli possesses an unambiguous catalytic activity is still lacking. To address this problem, we developed a new set of plasmids and site-directed mutants to elucidate the in vivo effects of CGI-58 expression on lipid metabolism in E. coli. By analyzing the lipid composition in selected E. coli strains expressing CGI-58 proteins, and by reinvestigating enzymatic tests with adequate controls, we show here that recombinant plant CGI-58 has none of the proposed activities previously described. Recombinant plant and mouse CGI-58 both lack acyltransferase activity towards either lysophosphatidylglycerol or lysophosphatidic acid to form phosphatidylglycerol or phosphatidic acid and recombinant plant CGI-58 does not catalyze TAG or phospholipid hydrolysis. However, expression of recombinant plant CGI-58, but not mouse CGI-58, led to a decrease in phosphatidylglycerol in all strains of E. coli tested, and a mutation of the putative catalytic residues restored a wild-type phenotype. The potential activities of plant CGI-58 are subsequently discussed. PMID:26745266

  1. Reassessing the Potential Activities of Plant CGI-58 Protein.

    PubMed

    Khatib, Abdallah; Arhab, Yani; Bentebibel, Assia; Abousalham, Abdelkarim; Noiriel, Alexandre

    2016-01-01

    Comparative Gene Identification-58 (CGI-58) is a widespread protein found in animals and plants. This protein has been shown to participate in lipolysis in mice and humans by activating Adipose triglyceride lipase (ATGL), the initial enzyme responsible for the triacylglycerol (TAG) catabolism cascade. Human mutation of CGI-58 is the cause of Chanarin-Dorfman syndrome, an orphan disease characterized by a systemic accumulation of TAG which engenders tissue disorders. The CGI-58 protein has also been shown to participate in neutral lipid metabolism in plants and, in this case, a mutation again provokes TAG accumulation. Although its roles as an ATGL coactivator and in lipid metabolism are quite clear, the catalytic activity of CGI-58 is still in question. The acyltransferase activities of CGI-58 have been speculated about, reported or even dismissed and experimental evidence that CGI-58 expressed in E. coli possesses an unambiguous catalytic activity is still lacking. To address this problem, we developed a new set of plasmids and site-directed mutants to elucidate the in vivo effects of CGI-58 expression on lipid metabolism in E. coli. By analyzing the lipid composition in selected E. coli strains expressing CGI-58 proteins, and by reinvestigating enzymatic tests with adequate controls, we show here that recombinant plant CGI-58 has none of the proposed activities previously described. Recombinant plant and mouse CGI-58 both lack acyltransferase activity towards either lysophosphatidylglycerol or lysophosphatidic acid to form phosphatidylglycerol or phosphatidic acid and recombinant plant CGI-58 does not catalyze TAG or phospholipid hydrolysis. However, expression of recombinant plant CGI-58, but not mouse CGI-58, led to a decrease in phosphatidylglycerol in all strains of E. coli tested, and a mutation of the putative catalytic residues restored a wild-type phenotype. The potential activities of plant CGI-58 are subsequently discussed.

  2. Rat C-reactive protein activates the autologous complement system.

    PubMed

    Diaz Padilla, Niubel; Bleeker, Wim K; Lubbers, Yvonne; Rigter, Gemma M M; Van Mierlo, Gerard J; Daha, Mohamed R; Hack, C Erik

    2003-08-01

    Activation of complement is a biological function of human C-reactive protein (hCRP), whereas rat CRP (rCRP) has been claimed to be unable to activate complement. As important biological functions of proteins are probably conserved among species, we re-evaluated, using various ligands, the capability of rCRP to activate complement. The activation of complement by hCRP and rCRP was investigated in solid- and fluid-phase systems. In the solid-phase system, purified CRP was fixed to enzyme-linked immunosorbent assay (ELISA) plates and incubated with human or rat recalcified plasma. Dose-dependent binding of human and rat C3 and C4 was observed to human and rat CRP, respectively. In the fluid-phase system, recalcified rat plasma, which contains about 500 mg/l of CRP, or human plasma supplemented with hCRP, were incubated with lyso-phosphatidylcholine. A dose-dependent activation of complement was observed upon incubation with this ligand, as reflected by the generation of activated C4 as well as of CRP-complement complexes. This activation was, in both cases, inhibited by preincubation of plasma with p-aminophosphorylcholine, a specific inhibitor of the interaction of CRP with its ligands, or by chelation of calcium ions. We conclude that rat CRP, similarly to human CRP, can activate autologous complement. These results support the notion that opsonization of ligands with complement is an important biological function of CRP.

  3. Auxin efflux by PIN-FORMED proteins is activated by two different protein kinases, D6 PROTEIN KINASE and PINOID.

    PubMed

    Zourelidou, Melina; Absmanner, Birgit; Weller, Benjamin; Barbosa, Inês C R; Willige, Björn C; Fastner, Astrid; Streit, Verena; Port, Sarah A; Colcombet, Jean; de la Fuente van Bentem, Sergio; Hirt, Heribert; Kuster, Bernhard; Schulze, Waltraud X; Hammes, Ulrich Z; Schwechheimer, Claus

    2014-06-19

    The development and morphology of vascular plants is critically determined by synthesis and proper distribution of the phytohormone auxin. The directed cell-to-cell distribution of auxin is achieved through a system of auxin influx and efflux transporters. PIN-FORMED (PIN) proteins are proposed auxin efflux transporters, and auxin fluxes can seemingly be predicted based on the--in many cells--asymmetric plasma membrane distribution of PINs. Here, we show in a heterologous Xenopus oocyte system as well as in Arabidopsis thaliana inflorescence stems that PIN-mediated auxin transport is directly activated by D6 PROTEIN KINASE (D6PK) and PINOID (PID)/WAG kinases of the Arabidopsis AGCVIII kinase family. At the same time, we reveal that D6PKs and PID have differential phosphosite preferences. Our study suggests that PIN activation by protein kinases is a crucial component of auxin transport control that must be taken into account to understand auxin distribution within the plant.

  4. Pivotal Role of Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 in Inflammatory Pulmonary Diseases

    PubMed Central

    Qian, Feng; Deng, Jing; Wang, Gang; Ye, Richard D.; Christman, John W.

    2016-01-01

    Mitogen-activated protein kinase (MAPK)-activated protein kinase (MK2) is exclusively regulated by p38 MAPK in vivo. Upon activation of p38 MAPK, MK2 binds with p38 MAPK, leading to phosphorylation of TTP, Hsp27, Akt and Cdc25 that are involved in regulation of various essential cellular functions. In this review, we discuss current knowledge about molecular mechanisms of MK2 in regulation of TNF-α production, NADPH oxidase activation, neutrophil migration, and DNA-damage-induced cell cycle arrest which are involved in the molecular pathogenesis of acute lung injury, pulmonary fibrosis, and non-small-cell lung cancer. Collectively current and emerging new information indicate that developing MK2 inhibitors and blocking MK2-mediated signal pathways is a potential therapeutic strategy for treatment of inflammatory and fibrotic lung diseases and lung cancer. PMID:26119506

  5. RNF4-Dependent Oncogene Activation by Protein Stabilization.

    PubMed

    Thomas, Jane J; Abed, Mona; Heuberger, Julian; Novak, Rostislav; Zohar, Yaniv; Beltran Lopez, Angela P; Trausch-Azar, Julie S; Ilagan, Ma Xenia G; Benhamou, David; Dittmar, Gunnar; Kopan, Raphael; Birchmeier, Walter; Schwartz, Alan L; Orian, Amir

    2016-09-20

    Ubiquitylation regulates signaling pathways critical for cancer development and, in many cases, targets proteins for degradation. Here, we report that ubiquitylation by RNF4 stabilizes otherwise short-lived oncogenic transcription factors, including β-catenin, Myc, c-Jun, and the Notch intracellular-domain (N-ICD) protein. RNF4 enhances the transcriptional activity of these factors, as well as Wnt- and Notch-dependent gene expression. While RNF4 is a SUMO-targeted ubiquitin ligase, protein stabilization requires the substrate's phosphorylation, rather than SUMOylation, and binding to RNF4's arginine-rich motif domain. Stabilization also involves generation of unusual polyubiquitin chains and docking of RNF4 to chromatin. Biologically, RNF4 enhances the tumor phenotype and is essential for cancer cell survival. High levels of RNF4 mRNA correlate with poor survival of a subgroup of breast cancer patients, and RNF4 protein levels are elevated in 30% of human colon adenocarcinomas. Thus, RNF4-dependent ubiquitylation translates transient phosphorylation signal(s) into long-term protein stabilization, resulting in enhanced oncoprotein activation. PMID:27653698

  6. A Novel Method for Assessing the Chaperone Activity of Proteins

    PubMed Central

    Hristozova, Nevena; Tompa, Peter; Kovacs, Denes

    2016-01-01

    Protein chaperones are molecular machines which function both during homeostasis and stress conditions in all living organisms. Depending on their specific function, molecular chaperones are involved in a plethora of cellular processes by playing key roles in nascent protein chain folding, transport and quality control. Among stress protein families–molecules expressed during adverse conditions, infection, and diseases–chaperones are highly abundant. Their molecular functions range from stabilizing stress-susceptible molecules and membranes to assisting the refolding of stress-damaged proteins, thereby acting as protective barriers against cellular damage. Here we propose a novel technique to test and measure the capability for protective activity of known and putative chaperones in a semi-high throughput manner on a plate reader. The current state of the art does not allow the in vitro measurements of chaperone activity in a highly parallel manner with high accuracy or high reproducibility, thus we believe that the method we report will be of significant benefit in this direction. The use of this method may lead to a considerable increase in the number of experimentally verified proteins with such functions, and may also allow the dissection of their molecular mechanism for a better understanding of their function. PMID:27564234

  7. Redox Control of Protein Arginine Methyltransferase 1 (PRMT1) Activity*

    PubMed Central

    Morales, Yalemi; Nitzel, Damon V.; Price, Owen M.; Gui, Shanying; Li, Jun; Qu, Jun; Hevel, Joan M.

    2015-01-01

    Elevated levels of asymmetric dimethylarginine (ADMA) correlate with risk factors for cardiovascular disease. ADMA is generated by the catabolism of proteins methylated on arginine residues by protein arginine methyltransferases (PRMTs) and is degraded by dimethylarginine dimethylaminohydrolase. Reports have shown that dimethylarginine dimethylaminohydrolase activity is down-regulated and PRMT1 protein expression is up-regulated under oxidative stress conditions, leading many to conclude that ADMA accumulation occurs via increased synthesis by PRMTs and decreased degradation. However, we now report that the methyltransferase activity of PRMT1, the major PRMT isoform in humans, is impaired under oxidative conditions. Oxidized PRMT1 displays decreased activity, which can be rescued by reduction. This oxidation event involves one or more cysteine residues that become oxidized to sulfenic acid (-SOH). We demonstrate a hydrogen peroxide concentration-dependent inhibition of PRMT1 activity that is readily reversed under physiological H2O2 concentrations. Our results challenge the unilateral view that increased PRMT1 expression necessarily results in increased ADMA synthesis and demonstrate that enzymatic activity can be regulated in a redox-sensitive manner. PMID:25911106

  8. Mitogen-activated protein kinase and abscisic acid signal transduction.

    PubMed

    Heimovaara-Dijkstra, S; Testerink, C; Wang, M

    2000-01-01

    The phytohormone abscisic acid (ABA) is a classical plant hormone, responsible for regulation of abscission, diverse aspects of plant and seed development, stress responses and germination. It was found that ABA signal transduction in plants can involve the activity of type 2C-phosphatases (PP2C), calcium, potassium, pH and a transient activation of MAP kinase. The ABA signal transduction cascades have been shown to be tissue-specific, the transient activation of MAP kinase has until now only been found in barley aleurone cells. However, type 2C phosphatases are involved in the induction of most ABA responses, as shown by the PP2C-deficient abi-mutants. These phosphatases show high homology with phosphatases that regulate MAP kinase activity in yeast. In addition, the role of farnesyl transferase as a negative regulator of ABA responses also indicates towards involvement of MAP kinase in ABA signal transduction. Farnesyl transferase is known to regulate Ras proteins, Ras proteins in turn are known to regulate MAP kinase activation. Interestingly, Ras-like proteins were detected in barley aleurone cells. Further establishment of the involvement of MAP kinase in ABA signal transduction and its role therein, still awaits more study.

  9. Protein kinase C controls activation of the DNA integrity checkpoint

    PubMed Central

    Soriano-Carot, María; Quilis, Inma; Bañó, M. Carmen; Igual, J. Carlos

    2014-01-01

    The protein kinase C (PKC) superfamily plays key regulatory roles in numerous cellular processes. Saccharomyces cerevisiae contains a single PKC, Pkc1, whose main function is cell wall integrity maintenance. In this work, we connect the Pkc1 protein to the maintenance of genome integrity in response to genotoxic stresses. Pkc1 and its kinase activity are necessary for the phosphorylation of checkpoint kinase Rad53, histone H2A and Xrs2 protein after deoxyribonucleic acid (DNA) damage, indicating that Pkc1 is required for activation of checkpoint kinases Mec1 and Tel1. Furthermore, Pkc1 electrophoretic mobility is delayed after inducing DNA damage, which reflects that Pkc1 is post-translationally modified. This modification is a phosphorylation event mediated by Tel1. The expression of different mammalian PKC isoforms at the endogenous level in yeast pkc1 mutant cells revealed that PKCδ is able to activate the DNA integrity checkpoint. Finally, downregulation of PKCδ activity in HeLa cells caused a defective activation of checkpoint kinase Chk2 when DNA damage was induced. Our results indicate that the control of the DNA integrity checkpoint by PKC is a mechanism conserved from yeast to humans. PMID:24792164

  10. Activation of autophagy by unfolded proteins during endoplasmic reticulum stress.

    PubMed

    Yang, Xiaochen; Srivastava, Renu; Howell, Stephen H; Bassham, Diane C

    2016-01-01

    Endoplasmic reticulum stress is defined as the accumulation of unfolded proteins in the endoplasmic reticulum, and is caused by conditions such as heat or agents that cause endoplasmic reticulum stress, including tunicamycin and dithiothreitol. Autophagy, a major pathway for degradation of macromolecules in the vacuole, is activated by these stress agents in a manner dependent on inositol-requiring enzyme 1b (IRE1b), and delivers endoplasmic reticulum fragments to the vacuole for degradation. In this study, we examined the mechanism for activation of autophagy during endoplasmic reticulum stress in Arabidopsis thaliana. The chemical chaperones sodium 4-phenylbutyrate and tauroursodeoxycholic acid were found to reduce tunicamycin- or dithiothreitol-induced autophagy, but not autophagy caused by unrelated stresses. Similarly, over-expression of BINDING IMMUNOGLOBULIN PROTEIN (BIP), encoding a heat shock protein 70 (HSP70) molecular chaperone, reduced autophagy. Autophagy activated by heat stress was also found to be partially dependent on IRE1b and to be inhibited by sodium 4-phenylbutyrate, suggesting that heat-induced autophagy is due to accumulation of unfolded proteins in the endoplasmic reticulum. Expression in Arabidopsis of the misfolded protein mimics zeolin or a mutated form of carboxypeptidase Y (CPY*) also induced autophagy in an IRE1b-dependent manner. Moreover, zeolin and CPY* partially co-localized with the autophagic body marker GFP-ATG8e, indicating delivery to the vacuole by autophagy. We conclude that accumulation of unfolded proteins in the endoplasmic reticulum is a trigger for autophagy under conditions that cause endoplasmic reticulum stress. PMID:26616142

  11. Flow field of flexible flapping wings

    NASA Astrophysics Data System (ADS)

    Sallstrom, Erik

    The agility and maneuverability of natural fliers would be desirable to incorporate into engineered micro air vehicles (MAVs). However, there is still much for engineers to learn about flapping flight in order to understand how such vehicles can be built for efficient flying. The goal of this study is to develop a methodology for capturing high quality flow field data around flexible flapping wings in a hover environment and to interpret it to gain a better understanding of how aerodynamic forces are generated. The flow field data was captured using particle image velocimetry (PIV) and required that measurements be taken around a repeatable flapping motion to obtain phase-averaged data that could be studied throughout the flapping cycle. Therefore, the study includes the development of flapping devices with a simple repeatable single degree of freedom flapping motion. The acquired flow field data has been examined qualitatively and quantitatively to investigate the mechanisms behind force production in hovering flight and to relate it to observations in previous research. Specifically, the flow fields have been investigated around a rigid wing and several carbon fiber reinforced flexible membrane wings. Throughout the whole study the wings were actuated with either a sinusoidal or a semi-linear flapping motion. The semi-linear flapping motion holds the commanded angular velocity nearly constant through half of each half-stroke while the sinusoidal motion is always either accelerating or decelerating. The flow fields were investigated by examining vorticity and vortex structures, using the Q criterion as the definition for the latter, in two and three dimensions. The measurements were combined with wing deflection measurements to demonstrate some of the key links in how the fluid-structure interactions generated aerodynamic forces. The flow fields were also used to calculate the forces generated by the flapping wings using momentum balance methods which yielded

  12. Current activities of the Yersinia effector protein YopM.

    PubMed

    Höfling, Sabrina; Grabowski, Benjamin; Norkowski, Stefanie; Schmidt, M Alexander; Rüter, Christian

    2015-05-01

    Yersinia outer protein M (YopM) belongs to the group of Yop effector proteins, which are highly conserved among pathogenic Yersinia species. During infection, the effectors are delivered into the host cell cytoplasm via the type 3 secretion system to subvert the host immune response and support the survival of Yersinia. In contrast to the other Yop effectors, YopM does not possess a known enzymatic activity and its molecular mechanism(s) of action remain(s) poorly understood. However, YopM was shown to promote colonization and dissemination of Yersinia, thus being crucial for the pathogen's virulence in vivo. Moreover, YopM interacts with several host cell proteins and might utilize them to execute its anti-inflammatory activities. The results obtained so far indicate that YopM is a multifunctional protein that counteracts the host immune defense by multiple activities, which are at least partially independent of each other. Finally, its functions seem to be also influenced by differences between the specific YopM isoforms expressed by Yersinia subspecies. In this review, we focus on the global as well as more specific contribution of YopM to virulence of Yersinia during infection and point out the various extra- and intracellular molecular functions of YopM. In addition, the novel cell-penetrating ability of recombinant YopM and its potential applications as a self-delivering immunomodulatory therapeutic will be discussed.

  13. Pharmacological activities in thermal proteins: relationships in molecular evolution

    NASA Technical Reports Server (NTRS)

    Fox, S. W.; Hefti, F.; Hartikka, J.; Junard, E.; Przybylski, A. T.; Vaughan, G.

    1987-01-01

    The model of protobiological events that has been presented in these pages has increasing relevance to pharmacological research. The thermal proteins that function as key substances in the proteinoid theory have recently been found to prolong the survival of rat forebrain neurons in culture and to stimulate the growth of neurites. A search for such activity in thermal proteins added to cultures of modern neurons was suggested by the fact that some of the microspheres assembled from proteinoids rich in hydrophobic amino acids themselves generate fibrous outgrowths.

  14. Pharmacological activities in thermal proteins: relationships in molecular evolution.

    PubMed

    Fox, S W; Hefti, F; Hartikka, J; Junard, E; Przybylski, A T; Vaughan, G

    1987-01-01

    The model of protobiological events that has been presented in these pages has increasing relevance to pharmacological research. The thermal proteins that function as key substances in the proteinoid theory have recently been found to prolong the survival of rat forebrain neurons in culture and to stimulate the growth of neurites. A search for such activity in thermal proteins added to cultures of modern neurons was suggested by the fact that some of the microspheres assembled from proteinoids rich in hydrophobic amino acids themselves generate fibrous outgrowths.

  15. Deltopectoral Flap in the Era of Microsurgery

    PubMed Central

    Chan, R. C. L.; Chan, J. Y. W.

    2014-01-01

    Background. Our study aimed to review the role of deltopectoral (DP) flap as a reconstructive option for defects in the head and neck region in the microvascular era. Methods. All patients who received DP flap reconstruction surgery at the Department of Surgery, Queen Mary Hospital, between 1999 and 2011 were recruited. Demographic data, indications for surgery, defect for reconstruction, and surgical outcomes were analyzed. Results. Fifty-four patients were included. All but two patients were operated for reconstruction after tumour resection. The remaining two patients were operated for necrotizing fasciitis and osteoradionecrosis. The majority of DP flaps were used to cover neck skin defect (63.0%). Other reconstructed defects included posterior pharyngeal wall (22.2%), facial skin defect (11.1%), and tracheal wall (3.7%). All donor sites were covered with partial thickness skin graft. Two patients developed partial flap necrosis at the tip and were managed conservatively. The overall flap survival rate was 96.3%. Conclusions. Albeit the technical advancements in microvascular surgery, DP still possesses multiple advantages (technical simplicity, reliable axial blood supply, large size, thinness, and pliability) which allows it to remain as a useful, reliable, and versatile surgical option for head and neck reconstruction. PMID:25374953

  16. Protein Kinase Cδ Mediates Neurogenic but Not Mitogenic Activation of Mitogen-Activated Protein Kinase in Neuronal Cells

    PubMed Central

    Corbit, Kevin C.; Foster, David A.; Rosner, Marsha Rich

    1999-01-01

    In several neuronal cell systems, fibroblast-derived growth factor (FGF) and nerve growth factor (NGF) act as neurogenic agents, whereas epidermal growth factor (EGF) acts as a mitogen. The mechanisms responsible for these different cellular fates are unclear. We report here that although FGF, NGF, and EGF all activate mitogen-activated protein (MAP) kinase (extracellular signal-related kinase [ERK]) in rat hippocampal (H19-7) and pheochromocytoma (PC12) cells, the activation of ERK by the neurogenic agents FGF and NGF is dependent upon protein kinase Cδ (PKCδ), whereas ERK activation in response to the mitogenic EGF is independent of PKCδ. Antisense PKCδ oligonucleotides or the PKCδ-specific inhibitor rottlerin inhibited FGF- and NGF-induced, but not EGF-induced, ERK activation. In contrast, EGF-induced ERK activation was inhibited by the phosphatidylinositol-3-kinase inhibitor wortmannin, which had no effect upon FGF-induced ERK activation. Rottlerin also inhibited the activation of MAP kinase kinase (MEK) in response to activated Raf, but had no effect upon c-Raf activity or ERK activation by activated MEK. These results indicate that PKCδ functions either downstream from or in parallel with c-Raf, but upstream of MEK. Inhibition of PKCδ also blocked neurite outgrowth induced by FGF and NGF in PC12 cells and by activated Raf in H19-7 cells, indicating a role for PKCδ in the neurogenic effects of FGF, NGF, and Raf. Interestingly, the PKCδ requirement is apparently cell type specific, since FGF-induced ERK activation was independent of PKCδ in NIH 3T3 murine fibroblasts, in which FGF is a mitogen. These data demonstrate that PKCδ contributes to growth factor specificity and response in neuronal cells and may also promote cell-type-specific differences in growth factor signaling. PMID:10330161

  17. Novel condensation products having high activity to insolubilize proteins and protein-insolubilized products

    SciTech Connect

    Krasnobajew, V.; Boeniger, R.

    1980-01-01

    According to the invention a substantially more active product with respect to the fixing or insolubilization pf proteins, including enzymes, is obtained when 1,3 phenylenediamine is condensed with glutardialdehyde. One application of the process is the enzymatic hydrolysis of lactose in milk products by lactase.

  18. Pharmacokinetics of activated protein C in guinea pigs

    SciTech Connect

    Berger, H. Jr.; Kirstein, C.G.; Orthner, C.L. )

    1991-05-15

    Protein C is a vitamin K-dependent zymogen of the serine protease, activated protein C (APC), an important regulatory enzyme in hemostasis. In view of the potential of human APC as an anticoagulant and profibrinolytic agent, the pharmacokinetics and tissue distribution of APC were studied in guinea pigs. The plasma elimination of a trace dose of {sup 125}I-APC was biphasic following an initial rapid elimination of approximately 15% of the injected dose within 1 to 2 minutes. This rapid removal of {sup 125}I-APC from the circulation was found to be a result of an association with the liver regardless of the route of injection. Essentially identical results were obtained with active site-blocked forms of APC generated with either diisopropylfluorophosphate or D-phenylalanyl-L-prolyl-L-arginine chloromethyl ketone, which indicates that the active site was not essential for the liver association. Accumulation of all three forms of APC in the liver peaked at 30 minutes and then declined as increasing amounts of degraded radiolabeled material appeared in the gastrointestinal tract and urine. Removal of the gamma-carboxyglutamic acid (gla) domain of diisopropylphosphoryl-APC resulted in a 50% reduction in the association with liver and an accumulation in the kidneys. Protein C and protein S were cleared from the circulation at rates approximately one-half and one-fourth, respectively, that of APC. Both in vitro and in vivo, APC was found to form complexes with protease inhibitors present in guinea pig plasma. Complex formation resulted in a more rapid disappearance of the enzymatic activity of APC than elimination of the protein moiety. These findings indicate two distinct mechanisms for the elimination of APC. One mechanism involves reaction with plasma protease inhibitors and subsequent elimination by specific hepatic receptors. (Abstract Truncated)

  19. 14 CFR 25.1511 - Flap extended speed.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Flap extended speed. 25.1511 Section 25... Limitations § 25.1511 Flap extended speed. The established flap extended speed V FE must be established so that it does not exceed the design flap speed V F chosen under §§ 25.335(e) and 25.345, for...

  20. 14 CFR 25.1511 - Flap extended speed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Flap extended speed. 25.1511 Section 25... Limitations § 25.1511 Flap extended speed. The established flap extended speed V FE must be established so that it does not exceed the design flap speed V F chosen under §§ 25.335(e) and 25.345, for...

  1. 14 CFR 25.1511 - Flap extended speed.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Flap extended speed. 25.1511 Section 25... Limitations § 25.1511 Flap extended speed. The established flap extended speed V FE must be established so that it does not exceed the design flap speed V F chosen under §§ 25.335(e) and 25.345, for...

  2. 14 CFR 25.1511 - Flap extended speed.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Flap extended speed. 25.1511 Section 25... Limitations § 25.1511 Flap extended speed. The established flap extended speed V FE must be established so that it does not exceed the design flap speed V F chosen under §§ 25.335(e) and 25.345, for...

  3. 14 CFR 25.1511 - Flap extended speed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Flap extended speed. 25.1511 Section 25... Limitations § 25.1511 Flap extended speed. The established flap extended speed V FE must be established so that it does not exceed the design flap speed V F chosen under §§ 25.335(e) and 25.345, for...

  4. Acute Deep Hand Burns Covered by a Pocket Flap-Graft

    PubMed Central

    Pradier, Jean-Philippe; Oberlin, Christophe; Bey, Eric

    2007-01-01

    Objective: We evaluated the long-term outcome of the “pocket flap-graft” technique, used to cover acute deep burns of the dorsum of the hand, and analyzed surgical alternatives. Methods: This was a 6-year, retrospective study of 8 patients with extensive burns and 1 patient with a single burn (11 hands in all) treated by defatted abdominal wall pockets. We studied the medical records of the patients, and conducted a follow-up examination. Results: All hands had fourth-degree thermal burns caused by flames, with exposure of tendons, bones, and joints, and poor functional prognosis. One third of patients had multiple injuries. Burns affected an average of 36% of the hand surface, and mean coverage was 92.8 cm2. One patient died. The 8 others were seen at 30-month follow-up: the skin quality of the flap was found to be good in 55% of the cases, the score on the Vancouver Scar Scale was 2.4, the Kapandji score was 4.5, and total active motion was 37% of that of a normal hand. Hand function was limited in only 2 cases, 8 patients were able to drive, and 3 patients had gone back to work. Conclusion: The pocket flap-graft allows preservation of hand function following severe burns, when local or free flaps are impossible to perform. Debulking of the flap at the time of elevation limits the need for secondary procedures. PMID:17268577

  5. Functions of AMP-activated protein kinase in adipose tissue

    PubMed Central

    Daval, Marie; Foufelle, Fabienne; Ferré, Pascal

    2006-01-01

    AMP-activated protein kinase (AMPK) is involved in cellular energy homeostasis. Its functions have been extensively studied in muscles and liver. AMPK stimulates pathways which increase energy production (glucose transport, fatty acid oxidation) and switches off pathways which consume energy (lipogenesis, protein synthesis, gluconeogenesis). This has led to the concept that AMPK has an interesting pharmaceutical potential in situations of insulin resistance and it is indeed the target of existing drugs and hormones which improve insulin sensitivity. Adipose tissue is a key player in energy metabolism through the release of substrates and hormones involved in metabolism and insulin sensitivity. Activation of AMPK in adipose tissue can be achieved through situations such as fasting and exercise. Leptin and adiponectin as well as hypoglycaemic drugs are activators of adipose tissue AMPK. This activation probably involves changes in the AMP/ATP ratio and the upstream kinase LKB1. When activated, AMPK limits fatty acid efflux from adipocytes and favours local fatty acid oxidation. Since fatty acids have a key role in insulin resistance, especially in muscles, activating AMPK in adipose tissue might be found to be beneficial in insulin-resistant states, particularly as AMPK activation also reduces cytokine secretion in adipocytes. PMID:16709632

  6. Activation of G Proteins by Guanine Nucleotide Exchange Factors Relies on GTPase Activity.

    PubMed

    Stanley, Rob J; Thomas, Geraint M H

    2016-01-01

    G proteins are an important family of signalling molecules controlled by guanine nucleotide exchange and GTPase activity in what is commonly called an 'activation/inactivation cycle'. The molecular mechanism by which guanine nucleotide exchange factors (GEFs) catalyse the activation of monomeric G proteins is well-established, however the complete reversibility of this mechanism is often overlooked. Here, we use a theoretical approach to prove that GEFs are unable to positively control G protein systems at steady-state in the absence of GTPase activity. Instead, positive regulation of G proteins must be seen as a product of the competition between guanine nucleotide exchange and GTPase activity--emphasising a central role for GTPase activity beyond merely signal termination. We conclude that a more accurate description of the regulation of G proteins via these processes is as a 'balance/imbalance' mechanism. This result has implications for the understanding of intracellular signalling processes, and for experimental strategies that rely on modulating G protein systems. PMID:26986850

  7. Calculation of the longitudinal aerodynamic characteristics of wing-flap configurations with externally blown flaps

    NASA Technical Reports Server (NTRS)

    Mendenhall, M. R.; Spangler, S. B.; Nielsen, J. N.; Goodwin, F. K.

    1976-01-01

    A theoretical investigation was carried out to extend and improve an existing method for predicting the longitudinal characteristics of wing flap configurations with externally blown flaps (EBF). Two potential flow models were incorporated into the prediction method: a wing and flap lifting-surface model and a turbofan engine wake model. The wing-flap model uses a vortex-lattice approach to represent the wing and flaps. The jet wake model consists of a series of closely spaced vortex rings normal to a centerline which may have vertical and lateral curvature to conform to the local flow field beneath the wing and flaps. Comparisons of measured and predicted pressure distributions, span load distributions on each lifting surface, and total lift and pitching moment coefficients on swept and unswept EBF configurations are included. A wide range of thrust coefficients and flap deflection angles is considered at angles of attack up to the onset of stall. Results indicate that overall lift and pitching-moment coefficients are predicted reasonably well over the entire range. The predicted detailed load distributions are qualitatively correct and show the peaked loads at the jet impingement points, but the widths and heights of the load peaks are not consistently predicted.

  8. Regulation of taurine transporter activity in LLC-PK1 cells: role of protein synthesis and protein kinase C activation.

    PubMed

    Jones, D P; Miller, L A; Dowling, C; Chesney, R W

    1991-11-01

    Taurine transporter activity increases after exposure of cultured renal epithelial cells to taurine-free medium for 24 h and decreases after incubation in high (500 microM) taurine. This adaptive response mimics that observed in rat kidney after manipulation of dietary taurine. In order to elucidate potential mechanisms involved in the regulation of beta-amino acid transporter activity, the role of RNA transcription, protein synthesis, and protein import (trafficking), as well as protein kinase C activation, on the control of taurine transport was examined in the continuous proximally derived LLC-PK1 renal cell line. Inhibition of RNA transcription with actinomycin D did not alter the up-regulatory and down-regulatory adaptive responses. Inhibition of protein synthesis with cycloheximide prevented the increased taurine transport in response to taurine-free medium as well as the decrease in taurine transport after exposure to high taurine. Colchicine prevented the response to taurine-free medium but had no effect on the response to high-taurine medium. Exposure of confluent cell monolayers to the active phorbol esters, phorbol 12-myristate 13-acetate and phorbol 12,13 dibutyrate, resulted in a reduction in taurine uptake. The effect was seen within minutes of exposure but was not observed in the presence of the inactive phorbol 4-alpha. This inhibitory action was blocked by staurosporin, an inhibitor of protein kinase C (PKC). Treatment of cells with the diacylglycerol kinase inhibitor R59022, which results in increased intracellular diacylglycerol, a natural stimulant of PKC, also inhibited taurine uptake, providing further evidence for a specific effect of PKC activation.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. L-Alanylglutamine inhibits signaling proteins that activate protein degradation, but does not affect proteins that activate protein synthesis after an acute resistance exercise.

    PubMed

    Wang, Wanyi; Choi, Ran Hee; Solares, Geoffrey J; Tseng, Hung-Min; Ding, Zhenping; Kim, Kyoungrae; Ivy, John L

    2015-07-01

    Sustamine™ (SUS) is a dipeptide composed of alanine and glutamine (AlaGln). Glutamine has been suggested to increase muscle protein accretion; however, the underlying molecular mechanisms of glutamine on muscle protein metabolism following resistance exercise have not been fully addressed. In the present study, 2-month-old rats climbed a ladder 10 times with a weight equal to 75 % of their body mass attached at the tail. Rats were then orally administered one of four solutions: placebo (PLA-glycine = 0.52 g/kg), whey protein (WP = 0.4 g/kg), low dose of SUS (LSUS = 0.1 g/kg), or high dose of SUS (HSUS = 0.5 g/kg). An additional group of sedentary (SED) rats was intubated with glycine (0.52 g/kg) at the same time as the ladder-climbing rats. Blood samples were collected immediately after exercise and at either 20 or 40 min after recovery. The flexor hallucis longus (FHL), a muscle used for climbing, was excised at 20 or 40 min post exercise and analyzed for proteins regulating protein synthesis and degradation. All supplements elevated the phosphorylation of FOXO3A above SED at 20 min post exercise, but only the SUS supplements significantly reduced the phosphorylation of AMPK and NF-kB p65. SUS supplements had no effect on mTOR signaling, but WP supplementation yielded a greater phosphorylation of mTOR, p70S6k, and rpS6 compared with PLA at 20 min post exercise. However, by 40 min post exercise, phosphorylation of mTOR and rpS6 in PLA had risen to levels not different than WP. These results suggest that SUS blocks the activation of intracellular signals for MPB, whereas WP accelerates mRNA translation. PMID:25837301

  10. Reconstruction of eyelids with Washio flap in anophthalmia.

    PubMed

    Tvrdek, M; Kozák, J

    2014-01-01

    The authors present a case report of a patient with anophthalmia in whom retroauriculo-temporal flap (Washio flap) was used for reconstruction of eyelids. This flap, which is mostly used for reconstructions of nasal defects, was not used in this way according to available literature.

  11. Flap Thickness in Eyes with Ectasia after LASIK

    PubMed Central

    Randleman, J. Bradley; Hebson, Carolyn B.; Larson, Paul M.

    2015-01-01

    Purpose To measure central flap thickness in eyes with ectasia after LASIK and to compare these values with estimated anticipated flap thickness based on average published values for each device used for flap creation. Setting Emory Vision at Emory University, Atlanta GA, USA Methods Confocal microscopic analysis using the Confoscan 3 (Nidek technologies) to measure central flap thickness in eyes with ectasia after LASIK, and evaluation of pre-LASIK records, including basic patient demographics, preoperative corneal topographies, estimated anticipated flap thickness based on published average thickness values, and residual stromal bed thickness (RSB) calculations using both measured and estimated flap thicknesses. Results Fifty eyes from 29 patients were evaluated. Average measured flap thickness was (138 ± 26 μ, range 90 to 220 μ). There were no significant differences between measured and estimated flap thicknesses (138 vs. 135 μ, p = 0.5) or RSB (329 vs. 332 μ, p = 0.7), nor were there any differences in flap thickness between eyes with normal or abnormal corneal topographies. Only one eye had a measured flap resulting in unintended RSB < 250 microns; this occurred in an eye with abnormal topography. Conclusions Measured central flap thickness was not thicker than estimated for the vast majority of eyes developing ectasia after LASIK. Thus, excessively thick flaps do not appear to be a major contributing factor to the pathogenesis of ectasia after LASIK. PMID:22424807

  12. The distally based forearm island flap in hand reconstruction.

    PubMed

    Jeng, S F; Wei, F C

    1998-08-01

    The distally based forearm island flap is vascularized by the perforators of the distal radial artery. The skin flap is along the axis of the radial artery, and the pivot point of its subcutaneous pedicle is about 2 to 4 cm above the radial styloid process. We have treated 12 patients with 12 flaps for soft-tissue defects of the hand. Of these recipient sites, seven were in dorsal hands, two were in thumbs, two were in forearms, and one was in the palmar area. The donor-tissue variants included eight skin flaps, two adipofascial flaps, and two sensate flaps. The sizes of the flaps ranged from 6 x 4 cm to 14 x 6 cm. The donor site wound could be closed primarily in five patients. Two sensate flaps, innervated by the lateral antebrachial cutaneous nerve, could provide sensation for thumb reconstruction. The advantage of this flap is its constant and reliable blood supply without sacrifice of the main radial artery. The elevation of the flap is simple and rapid. There is the potential that this flap can be used as an innervated flap, and there is no need of microsurgical technique.

  13. Sural Versus Perforator Flaps for Distal Medial Leg Wounds.

    PubMed

    Schannen, Andrew P; Truchan, Lisa; Goshima, Kaoru; Bentley, Roger; DeSilva, Gregory L

    2015-12-01

    Soft tissue coverage of distal medial ankle wounds is a challenging problem in orthopedic surgery because of the limited local tissues and prominent instrumentation. Traditionally, these wounds required free tissue transfer to achieve suitable coverage and subsequent bony union. To better respect the reconstructive ladder and to avoid the inherent difficulty of free flap coverage, rotational flaps have been used to cover these wounds. Both sural fasciocutaneous flaps and rotational fasciocutaneous perforator (propeller) flaps have been described for distal medial soft tissue coverage. The authors performed a retrospective chart review of patients who underwent distal medial leg coverage with the use of either sural flaps or rotational fasciocutaneous perforator flaps. The authors identified 14 patients by Current Procedural Terminology code who met the study criteria. The average age and degree of medical comorbidities were comparable in the 2 groups. The authors reviewed their medical records to evaluate fracture healing, flap size, complications, and return to normal shoe wear. All 7 sural flaps healed without incident, with underlying fracture healing. Of the 7 perforator flaps, 6 healed without incident, with underlying fracture healing. One perforator-based flap was complicated by superficial tip necrosis and went on to heal with local wound care. All patients returned to normal shoe wear. Both sural artery rotational flaps and posterior tibial artery-based rotational flaps are viable options for coverage of the distal medial leg. Coverage can be achieved reliably without microsurgery, anticoagulation, or monitoring in the intensive care unit. PMID:26652325

  14. 14 CFR 23.1511 - Flap extended speed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Flap extended speed. 23.1511 Section 23.1511 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Information § 23.1511 Flap extended speed. (a) The flap extended speed V FE must be established so that it...

  15. 14 CFR 23.1511 - Flap extended speed.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Flap extended speed. 23.1511 Section 23.1511 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Information § 23.1511 Flap extended speed. (a) The flap extended speed V FE must be established so that it...

  16. 14 CFR 23.1511 - Flap extended speed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Flap extended speed. 23.1511 Section 23.1511 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Information § 23.1511 Flap extended speed. (a) The flap extended speed V FE must be established so that it...

  17. 14 CFR 23.1511 - Flap extended speed.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Flap extended speed. 23.1511 Section 23.1511 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Information § 23.1511 Flap extended speed. (a) The flap extended speed V FE must be established so that it...

  18. 14 CFR 23.1511 - Flap extended speed.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Flap extended speed. 23.1511 Section 23.1511 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Information § 23.1511 Flap extended speed. (a) The flap extended speed V FE must be established so that it...

  19. Posttraumatic eyebrow reconstruction with hair-bearing temporoparietal fascia flap.

    PubMed

    Denadai, Rafael; Raposo-Amaral, Cassio Eduardo; Marques, Frederico Figueiredo; Raposo-Amaral, Cesar Augusto

    2015-01-01

    The temporoparietal fascia flap has been extensively used in craniofacial reconstructions. However, its use for eyebrow reconstruction has been sporadically reported. We describe a successfully repaired hair-bearing temporoparietal fascia flap after traumatic avulsion of eyebrow. Temporoparietal fascia flap is a versatile tool and should be considered as a therapeutic option by all plastic surgeons.

  20. 14 CFR 23.699 - Wing flap position indicator.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Wing flap position indicator. 23.699 Section 23.699 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION... Construction Control Systems § 23.699 Wing flap position indicator. There must be a wing flap...

  1. 14 CFR 23.699 - Wing flap position indicator.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Wing flap position indicator. 23.699 Section 23.699 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION... Construction Control Systems § 23.699 Wing flap position indicator. There must be a wing flap...

  2. 14 CFR 23.699 - Wing flap position indicator.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Wing flap position indicator. 23.699 Section 23.699 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION... Construction Control Systems § 23.699 Wing flap position indicator. There must be a wing flap...

  3. 14 CFR 23.699 - Wing flap position indicator.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Wing flap position indicator. 23.699 Section 23.699 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION... Construction Control Systems § 23.699 Wing flap position indicator. There must be a wing flap...

  4. 14 CFR 23.699 - Wing flap position indicator.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Wing flap position indicator. 23.699 Section 23.699 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION... Construction Control Systems § 23.699 Wing flap position indicator. There must be a wing flap...

  5. Methods of measuring Protein Disulfide Isomerase activity: a critical overview

    NASA Astrophysics Data System (ADS)

    Watanabe, Monica; Laurindo, Francisco; Fernandes, Denise

    2014-09-01

    Protein disulfide isomerase is an essential redox chaperone from the endoplasmic reticulum (ER) and is responsible for correct disulfide bond formation in nascent proteins. PDI is also found in other cellular locations in the cell, particularly the cell surface. Overall, PDI contributes to ER and global cell redox homeostasis and signaling. The knowledge about PDI structure and function progressed substantially based on in vitro studies using recombinant PDI and chimeric proteins. In these experimental scenarios, PDI reductase and chaperone activities are readily approachable. In contrast, assays to measure PDI isomerase activity, the hallmark of PDI family, are more complex. Assessment of PDI roles in cells and tissues mainly relies on gain- or loss-of-function studies. However, there is limited information regarding correlation of experimental readouts with the distinct types of PDI activities. In this mini-review, we evaluate the main methods described for measuring the different kinds of PDI activity: thiol reductase, thiol oxidase, thiol isomerase and chaperone. We emphasize the need to use appropriate controls and the role of critical interferents (e.g., detergent, presence of reducing agents). We also discuss the translation of results from in vitro studies with purified recombinant PDI to cellular and tissue samples, with critical comments on the interpretation of results.

  6. Analysis of antifreeze protein activity using colorimetric gold nanosensors

    NASA Astrophysics Data System (ADS)

    Jing, Xu; Choi, Ho-seok; Park, Ji-In; Kim, Young-Pil

    2015-07-01

    High activity and long stability of antifreeze proteins (AFPs), also known as ice-binding proteins (IBPs), are necessary for exerting their physiological functions in biotechnology and cryomedicine. Here we report a simple analysis of antifreeze protein activity and stability based on self-assembly of gold nanoparticles (AuNPs) via freezing and thawing cycles. While the mercaptosuccinic acid-capped AuNP (MSA-AuNP) was easily self-assembled after a freezing/thawing cycle, due to the mechanical attack of ice crystal on the MSA-AuNP surface, the presence of AFP impeded the self-assembly of MSA-AuNP via the interaction of AFP with ice crystals via freezing and thawing cycles, which led to a strong color in the MSA-AuNP solution. As a result, the aggregation parameter (E520/E650) of MSA-AuNP showed the rapid detection of both activity and stability of AFPs. We suggest that our newly developed method is very suitable for measuring antifreeze activity and stability in a simple and rapid manner with reliable quantification.

  7. Treatment of Ischial Pressure Sores with Both Profunda Femoris Artery Perforator Flaps and Muscle Flaps

    PubMed Central

    Kim, Chae Min; Yun, In Sik; Lee, Dong Won; Lew, Dae Hyun; Rah, Dong Kyun

    2014-01-01

    Background Reconstruction of ischial pressure sore defects is challenging due to extensive bursas and high recurrence rates. In this study, we simultaneously applied a muscle flap that covered the exposed ischium and large bursa with sufficient muscular volume and a profunda femoris artery perforator fasciocutaneous flap for the management of ischial pressure sores. Methods We retrospectively analyzed data from 14 patients (16 ischial sores) whose ischial defects had been reconstructed using both a profunda femoris artery perforator flap and a muscle flap between January 2006 and February 2014. We compared patient characteristics, operative procedure, and clinical course. Results All flaps survived the entire follow-up period. Seven patients (50%) had a history of surgery at the site of the ischial pressure sore. The mean age of the patients included was 52.8 years (range, 18-85 years). The mean follow-up period was 27.9 months (range, 3-57 months). In two patients, a biceps femoris muscle flap was used, while a gracilis muscle flap was used in the remaining patients. In four cases (25%), wound dehiscence occurred, but healed without further complication after resuturing. Additionally, congestion occurred in one case (6%), but resolved with conservative treatment. Among 16 cases, there was only one (6%) recurrence at 34 months. Conclusions The combination of a profunda femoris artery perforator fasciocutaneous flap and muscle flap for the treatment of ischial pressure sores provided pliability, adequate bulkiness and few long-term complications. Therefore, this may be used as an alternative treatment method for ischial pressure sores. PMID:25075362

  8. Antibacterial activity of a lectin-like Burkholderia cenocepacia protein.

    PubMed

    Ghequire, Maarten G K; De Canck, Evelien; Wattiau, Pierre; Van Winge, Iris; Loris, Remy; Coenye, Tom; De Mot, René

    2013-08-01

    Bacteriocins of the LlpA family have previously been characterized in the γ-proteobacteria Pseudomonas and Xanthomonas. These proteins are composed of two MMBL (monocot mannose-binding lectin) domains, a module predominantly and abundantly found in lectins from monocot plants. Genes encoding four different types of LlpA-like proteins were identified in genomes from strains belonging to the Burkholderia cepacia complex (Bcc) and the Burkholderia pseudomallei group. A selected recombinant LlpA-like protein from the human isolate Burkholderia cenocepacia AU1054 displayed narrow-spectrum genus-specific antibacterial activity, thus representing the first functionally characterized bacteriocin within this β-proteobacterial genus. Strain-specific killing was confined to other members of the Bcc, with mostly Burkholderia ambifaria strains being susceptible. In addition to killing planktonic cells, this bacteriocin also acted as an antibiofilm agent.

  9. Efficient expression and purification of biologically active human cystatin proteins.

    PubMed

    Chauhan, Sakshi; Tomar, Raghuvir S

    2016-02-01

    Cystatins are reversible cysteine protease inhibitor proteins. They are known to play important roles in controlling cathepsins, neurodegenerative disease, and in immune system regulation. Production of recombinant cystatin proteins is important for biochemical and function characterization. In this study, we cloned and expressed human stefin A, stefin B and cystatin C in Escherichia coli. Human stefin A, stefin B and cystatin C were purified from soluble fraction. For cystatin C, we used various chaperone plasmids to make cystatin C soluble, as it is reported to localize in inclusion bodies. Trigger factor, GroES-GroEL, DnaK-DnaJ-GrpE chaperones lead to the presence of cystatin C in the soluble fraction. Immobilized metal affinity chromatography, glutathione sepharose and anion exchange chromatography techniques were employed for efficient purification of these proteins. Their biological activities were tested by inhibition assays against cathepsin L and H3 protease.

  10. Antibacterial activity of a lectin-like Burkholderia cenocepacia protein.

    PubMed

    Ghequire, Maarten G K; De Canck, Evelien; Wattiau, Pierre; Van Winge, Iris; Loris, Remy; Coenye, Tom; De Mot, René

    2013-08-01

    Bacteriocins of the LlpA family have previously been characterized in the γ-proteobacteria Pseudomonas and Xanthomonas. These proteins are composed of two MMBL (monocot mannose-binding lectin) domains, a module predominantly and abundantly found in lectins from monocot plants. Genes encoding four different types of LlpA-like proteins were identified in genomes from strains belonging to the Burkholderia cepacia complex (Bcc) and the Burkholderia pseudomallei group. A selected recombinant LlpA-like protein from the human isolate Burkholderia cenocepacia AU1054 displayed narrow-spectrum genus-specific antibacterial activity, thus representing the first functionally characterized bacteriocin within this β-proteobacterial genus. Strain-specific killing was confined to other members of the Bcc, with mostly Burkholderia ambifaria strains being susceptible. In addition to killing planktonic cells, this bacteriocin also acted as an antibiofilm agent. PMID:23737242

  11. Flap effectiveness appraisal for winged re-entry vehicles

    NASA Astrophysics Data System (ADS)

    de Rosa, Donato; Pezzella, Giuseppe; Donelli, Raffaele S.; Viviani, Antonio

    2016-05-01

    The interactions between shock waves and boundary layer are commonplace in hypersonic aerodynamics. They represent a very challenging design issue for hypersonic vehicle. A typical example of shock wave boundary layer interaction is the flowfield past aerodynamic surfaces during control. As a consequence, such flow interaction phenomena influence both vehicle aerodynamics and aerothermodynamics. In this framework, the present research effort describes the numerical activity performed to simulate the flowfield past a deflected flap in hypersonic flowfield conditions for a winged re-entry vehicle.

  12. Novel mechanisms for activated protein C cytoprotective activities involving noncanonical activation of protease-activated receptor 3.

    PubMed

    Burnier, Laurent; Mosnier, Laurent O

    2013-08-01

    The direct cytoprotective activities of activated protein C (APC) on cells convey therapeutic, relevant, beneficial effects in injury and disease models in vivo and require the endothelial protein C receptor (EPCR) and protease activated receptor 1 (PAR1). Thrombin also activates PAR1, but its effects on cells contrast APC's cytoprotective effects. To gain insights into mechanisms for these contrasting cellular effects, protease activated receptor 3 (PAR3) activation by APC and thrombin was studied. APC cleaved PAR3 on transfected and endothelial cells in the presence of EPCR. Remarkably, APC cleaved a synthetic PAR3 N-terminal peptide at Arg41, whereas thrombin cleaved at Lys38. On cells, APC failed to cleave R41Q-PAR3, whereas K38Q-PAR3 was still cleaved by APC but not by thrombin. PAR3 tethered-ligand peptides beginning at amino acid 42, but not those beginning at amino acid 39, conveyed endothelial barrier-protective effects. In vivo, the APC-derived PAR3 tethered-ligand peptide, but not the thrombin-derived PAR3 peptide, blunted vascular endothelial growth factor (VEGF)-induced vascular permeability. These data indicate that PAR3 cleavage by APC at Arg41 can initiate distinctive APC-like cytoprotective effects. These novel insights help explain the differentiation of APC's cytoprotective versus thrombin's proinflammatory effects on cells and suggest a unique contributory role for PAR3 in the complex mechanisms underlying APC cytoprotective effects. PMID:23788139

  13. Protein-tyrosine phosphatase activity of CD45 is activated by sequential phosphorylation by two kinases.

    PubMed Central

    Stover, D R; Walsh, K A

    1994-01-01

    We describe a potential regulatory mechanism for the transmembrane protein-tyrosine phosphatase CD45. Phosphorylation on both tyrosine and serine residues in vitro results in an activation of CD45 specifically toward one artificial substrate but not another. The activation of these kinases appears to be order dependent, as it is enhanced when phosphorylation of tyrosine precedes that of serine but phosphorylation in the reverse order yields no activation. Any of four protein-tyrosine kinases tested, in combination with the protein-serine/threonine kinase, casein kinase II, was capable of mediating this activation in vitro. The time course of phosphorylation of CD45 in response to T-cell activation is consistent with the possibility that this regulatory mechanism is utilized in vivo. Images PMID:7518565

  14. Numerical linearized MHD model of flapping oscillations

    NASA Astrophysics Data System (ADS)

    Korovinskiy, D. B.; Ivanov, I. B.; Semenov, V. S.; Erkaev, N. V.; Kiehas, S. A.

    2016-06-01

    Kink-like magnetotail flapping oscillations in a Harris-like current sheet with earthward growing normal magnetic field component Bz are studied by means of time-dependent 2D linearized MHD numerical simulations. The dispersion relation and two-dimensional eigenfunctions are obtained. The results are compared with analytical estimates of the double-gradient model, which are found to be reliable for configurations with small Bz up to values ˜ 0.05 of the lobe magnetic field. Coupled with previous results, present simulations confirm that the earthward/tailward growth direction of the Bz component acts as a switch between stable/unstable regimes of the flapping mode, while the mode dispersion curve is the same in both cases. It is confirmed that flapping oscillations may be triggered by a simple Gaussian initial perturbation of the Vz velocity.

  15. Interleukin 2 activates extracellular signal-regulated protein kinase 2

    PubMed Central

    1993-01-01

    Interleukin 2 (IL-2) stimulated activation of the 42-kD extracellular signal-regulated kinase 2 (Erk2) in murine IL-3-dependent cells, expressing either high or intermediate affinity IL-2 receptors. Activation was both rapid, occurring within 5 min of IL-2 addition, and prolonged, remaining elevated for 30 min. Activation of Erk2 appeared to be necessary for IL-2 stimulation of proliferation, as deletion of a region of the cytoplasmic domain of the IL-2 receptor beta chain, essential for IL-2 stimulation of proliferation, abolished Erk2 activation by IL-2. Furthermore, cells that had been deprived of cytokine for 24 h were then refractory to IL-2 stimulation of both Erk2 activity and proliferation. However, elevation of Erk2 activity was not sufficient to stimulate proliferation, as protein kinase C activation stimulated Erk2 activity but not DNA synthesis. Also, cells exposed to IL-2 in the presence of rapamycin showed full Erk2 activation but not DNA synthesis. These data suggest that IL-2 must stimulate both Erk2 activity and a further pathway(s) to trigger cell proliferation. PMID:8376945

  16. Novel Biosensor of Membrane Protein Proximity Based on Fluorogen Activated Proteins.

    PubMed

    Vasilev, Kalin V; Gallo, Eugenio; Shank, Nathaniel; Jarvik, Jonathan W

    2016-01-01

    We describe a novel biosensor system for reporting proximity between cell surface proteins in live cultured cells. The biosensor takes advantage of recently developed fluorogen-activating proteins (FAPs) that display fluorescence only when bound to otherwise-nonfluorescent fluorogen molecules. To demonstrate feasibility for the approach, two recombinant rapamycin-binding proteins were expressed as single-pass plasma membrane proteins in HeLa cells; one of the proteins (scAvd- FRB) carried an extracellular avidin tag; the other (HL1-TO1-FKBP) carried an extracellular FAP. Cells were incubated with a membrane-impermeable bivalent ligand (biotin-PEG2000-DIR) consisting of biotin joined to a dimethyl-indole red (DIR) fluorogen by a polyethylene glycol linker, thus tethering the fluorogen to the scAvd-FRB fusion protein. Addition of rapamycin, which promotes FKBP-FRB dimerization and thereby brings the FAP in close proximity to the tethered fluorogen, led to a significant increase in DIR fluorescence. We call the new proximity assay TEFLA, for tethered fluorogen assay. PMID:27055753

  17. Strategies for the recovery of active proteins through refolding of bacterial inclusion body proteins

    PubMed Central

    Vallejo, Luis Felipe; Rinas, Ursula

    2004-01-01

    Recent advances in generating active proteins through refolding of bacterial inclusion body proteins are summarized in conjunction with a short overview on inclusion body isolation and solubilization procedures. In particular, the pros and cons of well-established robust refolding techniques such as direct dilution as well as less common ones such as diafiltration or chromatographic processes including size exclusion chromatography, matrix- or affinity-based techniques and hydrophobic interaction chromatography are discussed. Moreover, the effect of physical variables (temperature and pressure) as well as the presence of buffer additives on the refolding process is elucidated. In particular, the impact of protein stabilizing or destabilizing low- and high-molecular weight additives as well as micellar and liposomal systems on protein refolding is illustrated. Also, techniques mimicking the principles encountered during in vivo folding such as processes based on natural and artificial chaperones and propeptide-assisted protein refolding are presented. Moreover, the special requirements for the generation of disulfide bonded proteins and the specific problems and solutions, which arise during process integration are discussed. Finally, the different strategies are examined regarding their applicability for large-scale production processes or high-throughput screening procedures. PMID:15345063

  18. Genipin Suppresses NLRP3 Inflammasome Activation Through Uncoupling Protein-2

    PubMed Central

    Rajanbabu, Venugopal; Galam, Lakshmi; Fukumoto, Jutaro; Enciso, Juan; Tadikonda, Pratima; Lane, Troy N.; Bandyopadhyay, Sayantani; Parthasarathy, Prasanna Tamarapu; Cho, Young; Cho, Seong Ho; Lee, Yong Chul; Lockey, Richard F.; Kolliputi, Narasaiah

    2015-01-01

    Incomplete clearance of apoptotic cells and reactive oxygen species (ROS) release are known to trigger inflammasome activation causing severe inflammation in acute lung injury and various metabolic and autoimmune diseases. Moreover, it has been reported that apoptotic cell clearance and ROS-mediated apoptosis critically depend on mitochondrial uncoupling protein-2 (UCP2). However, the relationship between UCP2 and inflammasome activation has not been studied. This report investigates the role of UCP2 in the expression and activation of NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome in human macrophages. We found that UCP2 overexpression significantly enhanced the expression levels of NLRP3. The NLRP3 expression levels were significantly suppressed in THP1 cells treated with genipin, a UCP2 inhibitor, compared to controls. In addition, genipin altered adenosine triphosphate (ATP)- and hydrogen peroxide (H2O2)-mediated interleukin-1 beta (IL-1β) secretion and significantly suppressed caspase-1 activity in inflammasome-activated human macrophages. Taken together, our results suggest that genipin modulates NLRP3 inflammasome activation and ATP- or H2O2-mediated IL-1β release. PMID:26123077

  19. Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity[S

    PubMed Central

    Anheuser, Susi; Breiden, Bernadette; Schwarzmann, Günter; Sandhoff, Konrad

    2015-01-01

    Ganglioside GM2 is the major lysosomal storage compound of Tay-Sachs disease. It also accumulates in Niemann-Pick disease types A and B with primary storage of SM and with cholesterol in type C. Reconstitution of GM2 catabolism with β-hexosaminidase A and GM2 activator protein (GM2AP) at uncharged liposomal surfaces carrying GM2 as substrate generated only a physiologically irrelevant catabolic rate, even at pH 4.2. However, incorporation of anionic phospholipids into the GM2 carrying liposomes stimulated GM2 hydrolysis more than 10-fold, while the incorporation of plasma membrane stabilizing lipids (SM and cholesterol) generated a strong inhibition of GM2 hydrolysis, even in the presence of anionic phospholipids. Mobilization of membrane lipids by GM2AP was also inhibited in the presence of cholesterol or SM, as revealed by surface plasmon resonance studies. These lipids also reduced the interliposomal transfer rate of 2-NBD-GM1 by GM2AP, as observed in assays using Förster resonance energy transfer. Our data raise major concerns about the usage of recombinant His-tagged GM2AP compared with untagged protein. The former binds more strongly to anionic GM2-carrying liposomal surfaces, increases GM2 hydrolysis, and accelerates intermembrane transfer of 2-NBD-GM1, but does not mobilize membrane lipids. PMID:26175473

  20. Localization microscopy using noncovalent fluorogen activation by genetically encoded fluorogen activating proteins

    PubMed Central

    Maji, Suvrajit; Huang, Fang; Szent-Gyorgyi, Chris; Lidke, Diane S.; Lidke, Keith A.; Bruchez, Marcel P.

    2014-01-01

    The noncovalent equilibrium activation of a fluorogenic malachite green dye and its cognate fluorogen activating protein has been exploited to produce a sparse labeling distribution of densely tagged genetically encoded proteins, enabling single molecule detection and superresolution imaging in fixed and living cells. These sparse labeling conditions are achieved by control of the dye concentration in the milieu, and do not require any photoswitching or photoactivation. The labeling is achieved using physiological buffers and cellular media, and does not require additives or switching buffer to obtain superresolution images. We evaluate superresolution properties and images obtained from a selected fluorogen activating protein clone fused to actin, and show that the photon counts per object fall between those typically reported for fluorescent proteins and switching dye-pairs, resulting in 10-30 nm localization precision per object. This labeling strategy complements existing approaches, and may simplify multicolor labeling of cellular structures. PMID:24194371

  1. Mitogen-activated protein kinase cascades in Vitis vinifera

    PubMed Central

    Çakır, Birsen; Kılıçkaya, Ozan

    2015-01-01

    Protein phosphorylation is one of the most important mechanisms to control cellular functions in response to external and endogenous signals. Mitogen-activated protein kinases (MAPK) are universal signaling molecules in eukaryotes that mediate the intracellular transmission of extracellular signals resulting in the induction of appropriate cellular responses. MAPK cascades are composed of four protein kinase modules: MAPKKK kinases (MAPKKKKs), MAPKK kinases (MAPKKKs), MAPK kinases (MAPKKs), and MAPKs. In plants, MAPKs are activated in response to abiotic stresses, wounding, and hormones, and during plant pathogen interactions and cell division. In this report, we performed a complete inventory of MAPK cascades genes in Vitis vinifera, the whole genome of which has been sequenced. By comparison with MAPK, MAPK kinases, MAPK kinase kinases and MAPK kinase kinase kinase kinase members of Arabidopsis thaliana, we revealed the existence of 14 MAPKs, 5 MAPKKs, 62 MAPKKKs, and 7 MAPKKKKs in Vitis vinifera. We identified orthologs of V. vinifera putative MAPKs in different species, and ESTs corresponding to members of MAPK cascades in various tissues. This work represents the first complete inventory of MAPK cascades in V. vinifera and could help elucidate the biological and physiological functions of these proteins in V. vinifera. PMID:26257761

  2. Interaction of receptor-activity-modifying protein1 with tubulin.

    PubMed

    Kunz, Thomas H; Mueller-Steiner, Sarah; Schwerdtfeger, Kerstin; Kleinert, Peter; Troxler, Heinz; Kelm, Jens M; Ittner, Lars M; Fischer, Jan A; Born, Walter

    2007-08-01

    Receptor-activity-modifying protein (RAMP) 1 is an accessory protein of the G protein-coupled calcitonin receptor-like receptor (CLR). The CLR/RAMP1 heterodimer defines a receptor for the potent vasodilatory calcitonin gene-related peptide. A wider tissue distribution of RAMP1, as compared to that of the CLR, is consistent with additional biological functions. Here, glutathione S-transferase (GST) pull-down, coimmunoprecipitation and yeast two-hybrid experiments identified beta-tubulin as a novel RAMP1-interacting protein. GST pull-down experiments indicated interactions between the N- and C-terminal domains of RAMP1 and beta-tubulin. Yeast two-hybrid experiments confirmed the interaction between the N-terminal region of RAMP1 and beta-tubulin. Interestingly, alpha-tubulin was co-extracted with beta-tubulin in pull-down experiments and immunoprecipitation of RAMP1 coprecipitated alpha- and beta-tubulin. Confocal microscopy indicated colocalization of RAMP1 and tubulin predominantly in axon-like processes of neuronal differentiated human SH-SY5Y neuroblastoma cells. In conclusion, the findings point to biological roles of RAMP1 beyond its established interaction with G protein-coupled receptors. PMID:17493758

  3. Antibacterial activity of a lectin-like Burkholderia cenocepacia protein

    PubMed Central

    Ghequire, Maarten G K; Canck, Evelien; Wattiau, Pierre; Winge, Iris; Loris, Remy; Coenye, Tom; Mot, René

    2013-01-01

    Abstract Bacteriocins of the LlpA family have previously been characterized in the γ-proteobacteria Pseudomonas and Xanthomonas. These proteins are composed of two MMBL (monocot mannose-binding lectin) domains, a module predominantly and abundantly found in lectins from monocot plants. Genes encoding four different types of LlpA-like proteins were identified in genomes from strains belonging to the Burkholderia cepacia complex (Bcc) and the Burkholderia pseudomallei group. A selected recombinant LlpA-like protein from the human isolate Burkholderia cenocepacia AU1054 displayed narrow-spectrum genus-specific antibacterial activity, thus representing the first functionally characterized bacteriocin within this β-proteobacterial genus. Strain-specific killing was confined to other members of the Bcc, with mostly Burkholderia ambifaria strains being susceptible. In addition to killing planktonic cells, this bacteriocin also acted as an antibiofilm agent. Bacteriocins mediate highly selective antagonism among closely related bacteria but such antimicrobial proteins have not yet been reported in Burkholderia. We identified a lectin-like protein of the LlpA family in a Burkholderia cenocepacia human isolate that strain-specifically and selectively kills planktonic and biofilm cells of other Burkholderia cepacia complex members. PMID:23737242

  4. 50 CFR Figure 16 to Part 223 - Escape Opening and Flap Dimensions for the Double Cover Flap TED

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 9 2011-10-01 2011-10-01 false Escape Opening and Flap Dimensions for the Double Cover Flap TED 16 Figure 16 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE... AND ANADROMOUS SPECIES Pt. 223, Fig. 16 Figure 16 to Part 223—Escape Opening and Flap Dimensions...

  5. 50 CFR Figure 16 to Part 223 - Escape Opening and Flap Dimensions for the Double Cover Flap TED

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 7 2010-10-01 2010-10-01 false Escape Opening and Flap Dimensions for the Double Cover Flap TED 16 Figure 16 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE... AND ANADROMOUS SPECIES Pt. 223, Fig. 16 Figure 16 to Part 223—Escape Opening and Flap Dimensions...

  6. 50 CFR Figure 16 to Part 223 - Escape Opening and Flap Dimensions for the Double Cover Flap TED

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 10 2014-10-01 2014-10-01 false Escape Opening and Flap Dimensions for the Double Cover Flap TED 16 Figure 16 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES... for the Double Cover Flap TED ER02JN04.003...

  7. 50 CFR Figure 16 to Part 223 - Escape Opening and Flap Dimensions for the Double Cover Flap TED

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 10 2013-10-01 2013-10-01 false Escape Opening and Flap Dimensions for the Double Cover Flap TED 16 Figure 16 to Part 223 Wildlife and Fisheries NATIONAL MARINE FISHERIES... for the Double Cover Flap TED ER02JN04.003...

  8. Detection of expansin proteins and activity during tomato fruit ontogeny.

    PubMed

    Rose, J K; Cosgrove, D J; Albersheim, P; Darvill, A G; Bennett, A B

    2000-08-01

    Expansins are plant proteins that have the capacity to induce extension in isolated cell walls and are thought to mediate pH-dependent cell expansion. J.K.C. Rose, H.H. Lee, and A.B. Bennett ([1997] Proc Natl Acad Sci USA 94: 5955-5960) reported the identification of an expansin gene (LeExp1) that is specifically expressed in ripening tomato (Lycopersicon esculentum) fruit where cell wall disassembly, but not cell expansion, is prominent. Expansin expression during fruit ontogeny was examined using antibodies raised to recombinant LeExp1 or a cell elongation-related expansin from cucumber (CsExp1). The LeExp1 antiserum detected expansins in extracts from ripe, but not preripe tomato fruit, in agreement with the pattern of LeExp1 mRNA accumulation. In contrast, antibodies to CsExp1 cross-reacted with expansins in early fruit development and the onset of ripening, but not at a later ripening stage. These data suggest that ripening-related and expansion-related expansin proteins have distinct antigenic epitopes despite overall high sequence identity. Expansin proteins were detected in a range of fruit species and showed considerable variation in abundance; however, appreciable levels of expansin were not present in fruit of the rin or Nr tomato mutants that exhibit delayed and reduced softening. LeExp1 protein accumulation was ethylene-regulated and matched the previously described expression of mRNA, suggesting that expression is not regulated at the level of translation. We report the first detection of expansin activity in several stages of fruit development and while characteristic creep activity was detected in young and developing tomato fruit and in ripe pear, avocado, and pepper, creep activity in ripe tomato showed qualitative differences, suggesting both hydrolytic and expansin activities.

  9. Airplane wing leading edge variable camber flap

    NASA Technical Reports Server (NTRS)

    Cole, J. B.

    1980-01-01

    The invention and design of an aerodynamic high lift device which provided a solution to an aircraft performance problem are described. The performance problem of converting a high speed cruise airfoil into a low speed aerodynamic shape that would provide landing and take-off characteristics superior to those available with contemporary high lift devices are addressed. The need for an improved wing leading edge device that would complement the high lift performance of a triple slotted trailing edge flap is examined. The mechanical and structural aspects of the variable camber flap are discussed and the aerodynamic performance aspects only as they relate to the invention and design of the device are presented.

  10. Flap Edge Aeroacoustic Measurements and Predictions

    NASA Technical Reports Server (NTRS)

    Brooks, Thomas F.; Humphreys, William M., Jr.

    2000-01-01

    An aeroacoustic model test has been conducted to investigate the mechanisms of sound generation on high-lift wing configurations. This paper presents an analysis of flap side-edge noise, which is often the most dominant source. A model of a main element wing section with a half-span flap was tested at low speeds of up to a Mach number of 0.17, corresponding to a wing chord Reynolds number of approximately 1.7 million. Results are presented for flat (or blunt), flanged, and round flap-edge geometries, with and without boundary-layer tripping, deployed at both moderate and high flap angles. The acoustic database is obtained from a Small Aperture Directional Array (SADA) of microphones, which was constructed to electronically steer to different regions of the model and to obtain farfield noise spectra and directivity from these regions. The basic flap-edge aerodynamics is established by static surface pressure data, as well as by Computational Fluid Dynamics (CFD) calculations and simplified edge flow analyses. Distributions of unsteady pressure sensors over the flap allow the noise source regions to be defined and quantified via cross-spectral diagnostics using the SADA output. It is found that shear layer instability and related pressure scatter is the primary noise mechanism. For the flat edge flap, two noise prediction methods based on unsteady-surface-pressure measurements are evaluated and compared to measured noise. One is a new causality spectral approach developed here. The other is a new application of an edge-noise scatter prediction method. The good comparisons for both approaches suggest that much of the physics is captured by the prediction models. Areas of disagreement appear to reveal when the assumed edge noise mechanism does not fully define, the noise production. For the different edge conditions, extensive spectra and directivity are presented. Significantly, for each edge configuration, the spectra for different flow speeds, flap angles, and

  11. Forehead flap in maxillofacial surgery: Our experiences

    PubMed Central

    Ahmed, Syed S.; Ghassemi, Alireza; Rehman, Sajjad A.; Ansari, Md. Kaleem

    2015-01-01

    The forehead flap is a commonly used technique to reconstruct the deep and large nasal defects. It can be conveniently performed under local or general anesthesia and provides a very good color and texture matching to the nasal skin, which makes it a suitable graft harvesting site for nasal reconstruction. It has only single disadvantage disadvantagethat it is a two-stage procedure and “finishing” surgeries are needed occasionally for best surgical and cosmetic outcome. In this paper, we describe three different applications of forehead flaps. PMID:26668457

  12. C-reactive protein activates complement in infarcted human myocardium.

    PubMed

    Nijmeijer, Remco; Lagrand, Wim K; Lubbers, Yvonne T P; Visser, Cees A; Meijer, Chris J L M; Niessen, Hans W M; Hack, C Erik

    2003-07-01

    Circulating levels of C-reactive protein (CRP) constitute a cardiovascular risk marker. Immunohistochemical studies have revealed co-localization of CRP and activated complement in human infarcted myocardium suggesting CRP to enhance inflammation in ischemic myocardium by inducing local complement activation. The aim was to establish whether CRP activates complement in infarcted human myocardium and to assess the relationship between this activation and the duration of infarction. Myocardial tissue samples from 56 patients that had died from acute myocardial infarction were evaluated. Specimens were taken from infarcted as well as noninfarcted sites of the heart. CRP-mediated complement activation was assessed by immunohistochemistry and by measuring levels of complement, CRP, and CRP-complement complexes, specific markers for CRP-mediated activation, in homogenates of the heart. Infarctions of 12 hours to 5 days had significantly more extensive depositions of complement and CRP and contained significantly more CRP, activated complement, and CRP-complement complexes than infarctions that were less than 12 hours old. Levels of CRP complexes correlated significantly with CRP and complement concentrations in the infarctions, as well as with the extent of complement and CRP depositions as measured via immunohistochemistry. Specific activation products of CRP-mediated activation of complement are increased in infarcts of more than 12 hours in duration and correlate with the extent of complement depositions. Hence, CRP seems to enhance local inflammatory reactions ensuing in human myocardial infarcts of more than 12 hours duration.

  13. An aeroelastic instability provides a possible basis for the transition from gliding to flapping flight.

    PubMed

    Curet, Oscar M; Swartz, Sharon M; Breuer, Kenneth S

    2013-03-01

    The morphology, kinematics and stiffness properties of lifting surfaces play a key role in the aerodynamic performance of vertebrate flight. These surfaces, as a result of their flexible nature, may move both actively, owing to muscle contraction, and passively, in reaction to fluid forces. However, the nature and implications of this fluid-structure interaction are not well understood. Here, we study passive flight (flight with no active wing actuation) and explore a physical mechanism that leads to the emergence of a natural flapping motion. We model a vertebrate wing with a compliant shoulder and the ability to camber with an idealized physical model consisting of a cantilevered flat plate with a hinged trailing flap. We find that at low wind speed the wing is stationary, but at a critical speed the wing spontaneously flaps. The lift coefficient is significantly enhanced once the wing starts to oscillate, although this increase in lift generation is accompanied by an increase in drag. Flow visualization suggests that a strong leading edge vortex attached to the wing during downstroke is the primary mechanism responsible for the enhanced lift. The flapping instability we observe suggests a possible scenario for an evolutionary transition from gliding to powered flapping flight in animals that possess compliant wings capable of passive camber. Although the flapping state is accompanied by a lower lift-to-drag ratio, the increased lifting capability it confers might have enabled increased body mass, improved foraging performance and/or flight at lower speeds, any of which might have been selectively advantageous. PMID:23303221

  14. An aeroelastic instability provides a possible basis for the transition from gliding to flapping flight

    PubMed Central

    Curet, Oscar M.; Swartz, Sharon M.; Breuer, Kenneth S.

    2013-01-01

    The morphology, kinematics and stiffness properties of lifting surfaces play a key role in the aerodynamic performance of vertebrate flight. These surfaces, as a result of their flexible nature, may move both actively, owing to muscle contraction, and passively, in reaction to fluid forces. However, the nature and implications of this fluid–structure interaction are not well understood. Here, we study passive flight (flight with no active wing actuation) and explore a physical mechanism that leads to the emergence of a natural flapping motion. We model a vertebrate wing with a compliant shoulder and the ability to camber with an idealized physical model consisting of a cantilevered flat plate with a hinged trailing flap. We find that at low wind speed the wing is stationary, but at a critical speed the wing spontaneously flaps. The lift coefficient is significantly enhanced once the wing starts to oscillate, although this increase in lift generation is accompanied by an increase in drag. Flow visualization suggests that a strong leading edge vortex attached to the wing during downstroke is the primary mechanism responsible for the enhanced lift. The flapping instability we observe suggests a possible scenario for an evolutionary transition from gliding to powered flapping flight in animals that possess compliant wings capable of passive camber. Although the flapping state is accompanied by a lower lift-to-drag ratio, the increased lifting capability it confers might have enabled increased body mass, improved foraging performance and/or flight at lower speeds, any of which might have been selectively advantageous. PMID:23303221

  15. Negative regulation of lymphocyte activation by the adaptor protein LAX.

    PubMed

    Zhu, Minghua; Granillo, Olivia; Wen, Renren; Yang, Kaiyong; Dai, Xuezhi; Wang, Demin; Zhang, Weiguo

    2005-05-01

    The membrane-associated adaptor protein LAX is a linker for activation of T cells (LAT)-like molecule that is expressed in lymphoid tissues. Upon stimulation of T or B cells, it is phosphorylated and interacts with Grb2 and the p85 subunit of PI3K. LAX, however, is not capable of replacing LAT in the TCR signaling pathway. In this study we report that upon T or B cell activation, the LAX protein was up-regulated dramatically. Although disruption of the LAX gene by homologous recombination had no major impact on lymphocyte development, it caused a significant reduction in CD23 expression on mature B cells. Interestingly, naive LAX(-/-) mice had spontaneous germinal center formation. Compared with normal T and B cells, LAX(-/-) T and B cells were hyperresponsive and had enhanced calcium flux, protein tyrosine phosphorylation, MAPK and Akt activation, and cell survival upon engagement of the T or B AgRs. Our data demonstrate that LAX functions as a negative regulator in lymphocyte signaling.

  16. T cell-activating protein on murine lymphocytes.

    PubMed

    Yeh, E T; Reiser, H; Benacerraf, B; Rock, K L

    1986-12-01

    A functional T cell surface molecule, T cell-activating protein (TAP), has been identified on murine lymphocytes. TAP is a protein with an apparent molecular mass of 10-12 kilodaltons (kDa) nonreduced, 16-17 kDa reduced. Cross-linking of TAP can result in profound activation of T lymphocytes to produce lymphokines and to enter the cell cycle. Furthermore, antibody binding to TAP can modulate antigen-driven T cell stimulation. Current data suggest that TAP is physically distinct from the T cell receptor complex. On unstimulated lymphocytes, TAP is expressed on T cells and defines heterogeneity within the major T cell subsets. Its profile of expression is rapidly altered on cell activation. Ontologically, it is first detected in the thymus, where it is found on both the most immature and the most mature cell subsets, and it is functional on both. Together, these TAP+ cells constitute a small fraction of thymocytes. TAP expression, however, defines the immunocompetent compartment of the thymus, and thus could be involved in functional maturation. Finally, the gene controlling TAP expression has been mapped, and is tightly linked to a locus of hematopoietic antigens (Ly-6). TAP is molecularly distinct from these antigens. Furthermore, all of these proteins show a markedly distinct developmental regulation in their cell surface expression.

  17. Protein inhibitor of activated STAT3 inhibits adipogenic gene expression

    SciTech Connect

    Deng Jianbei; Hua Kunjie; Caveney, Erica J.; Takahashi, Nobuyuki; Harp, Joyce B. . E-mail: jharp@unc.edu

    2006-01-20

    Protein inhibitor of activated STAT3 (PIAS3), a cytokine-induced repressor of signal transducer and activator of transcription 3 (STAT3) and a modulator of a broad array of nuclear proteins, is expressed in white adipose tissue, but its role in adipogenesis is not known. Here, we determined that PIAS3 was constitutively expressed in 3T3-L1 cells at all stages of adipogenesis. However, it translocated from the nucleus to the cytoplasm 4 days after induction of differentiation by isobutylmethylxanthine, dexamethasone, and insulin (MDI). In ob/ob mice, PIAS3 expression was increased in white adipose tissue depots compared to lean mice and was found in the cytoplasm of adipocytes. Overexpression of PIAS3 in differentiating preadipocytes, which localized primarily to the nucleus, inhibited mRNA level gene expression of adipogenic transcription factors C/EBP{alpha} and PPAR{gamma}, as well as their downstream target genes aP2 and adiponectin. PIAS3 also inhibited C/EBP{alpha} promoter activation mediated specifically by insulin, but not dexamethasone or isobutylmethylxanthine. Taken together, these data suggest that PIAS3 may play an inhibitory role in adipogenesis by modulating insulin-activated transcriptional activation events. Increased PIAS3 expression in adipose tissue may play a role in the metabolic disturbances of obesity.

  18. AMP-activated Protein Kinase Is Activated as a Consequence of Lipolysis in the Adipocyte

    Technology Transfer Automated Retrieval System (TEKTRAN)

    AMP-activated protein kinase (AMPK) is activated in adipocytes during exercise and other states in which lipolysis is stimulated. However, the mechanism(s) responsible for this effect and its physiological relevance are unclear. To examine these questions, 3T3-L1 adipocytes were treated with agents...

  19. Pokeweed Antiviral Protein, a Ribosome Inactivating Protein: Activity, Inhibition and Prospects

    PubMed Central

    Domashevskiy, Artem V.; Goss, Dixie J.

    2015-01-01

    Viruses employ an array of elaborate strategies to overcome plant defense mechanisms and must adapt to the requirements of the host translational systems. Pokeweed antiviral protein (PAP) from Phytolacca americana is a ribosome inactivating protein (RIP) and is an RNA N-glycosidase that removes specific purine residues from the sarcin/ricin (S/R) loop of large rRNA, arresting protein synthesis at the translocation step. PAP is thought to play an important role in the plant’s defense mechanism against foreign pathogens. This review focuses on the structure, function, and the relationship of PAP to other RIPs, discusses molecular aspects of PAP antiviral activity, the novel inhibition of this plant toxin by a virus counteraction—a peptide linked to the viral genome (VPg), and possible applications of RIP-conjugated immunotoxins in cancer therapeutics. PMID:25635465

  20. Pokeweed antiviral protein, a ribosome inactivating protein: activity, inhibition and prospects.

    PubMed

    Domashevskiy, Artem V; Goss, Dixie J

    2015-01-28

    Viruses employ an array of elaborate strategies to overcome plant defense mechanisms and must adapt to the requirements of the host translational systems. Pokeweed antiviral protein (PAP) from Phytolacca americana is a ribosome inactivating protein (RIP) and is an RNA N-glycosidase that removes specific purine residues from the sarcin/ricin (S/R) loop of large rRNA, arresting protein synthesis at the translocation step. PAP is thought to play an important role in the plant's defense mechanism against foreign pathogens. This review focuses on the structure, function, and the relationship of PAP to other RIPs, discusses molecular aspects of PAP antiviral activity, the novel inhibition of this plant toxin by a virus counteraction-a peptide linked to the viral genome (VPg), and possible applications of RIP-conjugated immunotoxins in cancer therapeutics.

  1. Crystal Structure of the Protein Kinase Domain of Yeast AMP-Activated Protein Kinase Snf1

    SciTech Connect

    Rudolph,M.; Amodeo, G.; Bai, Y.; Tong, L.

    2005-01-01

    AMP-activated protein kinase (AMPK) is a master metabolic regulator, and is an important target for drug development against diabetes, obesity, and other diseases. AMPK is a hetero-trimeric enzyme, with a catalytic ({alpha}) subunit, and two regulatory ({beta} and {gamma}) subunits. Here we report the crystal structure at 2.2 Angstrom resolution of the protein kinase domain (KD) of the catalytic subunit of yeast AMPK (commonly known as SNF1). The Snf1-KD structure shares strong similarity to other protein kinases, with a small N-terminal lobe and a large C-terminal lobe. Two negative surface patches in the structure may be important for the recognition of the substrates of this kinase.

  2. [Virucidal activity of disinfectants. Influence of the serum protein upon the virucidal activity of disinfectants].

    PubMed

    Noda, M; Matsuda, S; Kobayashi, M

    2000-08-01

    Five disinfectants were tested for virucidal activity on three DNA viruses and three RNA viruses in the presence or absence of serum protein. Disinfectants of the aldehyde and halogen groups had a virucidal activity on human herpes virus, bovine rhabdo virus, human immunodeficiency virus, human adeno virus, porcine parvo virus, and polio virus. Disinfectants of the invert and amphoteric soap groups, and biganide group had a destructive effect on RNA and DNA viruses possessing an envelope. The presence of serum protein exerted great influence upon the virucidal activity of disinfectants of the invert and amphoteric soap groups. PMID:11019515

  3. [Aseptic bone flap osteonecrosis following cranioplasty after decompressive cranietomy].

    PubMed

    Smoll, Nicolas R; Stienen, Martin N; Schaller, Karl; Gautschi, Oliver P

    2013-06-19

    This case report discusses a case of aseptic osteonecrosis in a cranioplasty bone flap after decompressive craniectomy, which is a known, but rare complication after autologous cranioplasty. We suggest that the pathophysiology of cranial bone flap necrosis may have a similar pathophysiology to free flap necrosis/failure. The key suggested problem causing the osteonecrosis is vessel thrombosis within the smaller vessels of the bone flap due to the prothrombotic effects of the factors released during drilling of the bone flap. Suspicious local findings like wound dehiscence or fluid leakage should lead to a head computed tomography in order to discuss a prophylactic artificial second cranioplasty if necessary. PMID:23773942

  4. Rapamycin induces mitogen-activated protein (MAP) kinase phosphatase-1 (MKP-1) expression through activation of protein kinase B and mitogen-activated protein kinase kinase pathways.

    PubMed

    Rastogi, Ruchi; Jiang, Zhongliang; Ahmad, Nisar; Rosati, Rita; Liu, Yusen; Beuret, Laurent; Monks, Robert; Charron, Jean; Birnbaum, Morris J; Samavati, Lobelia

    2013-11-22

    Mitogen-activated protein kinase phosphatase-1 (MKP-1), also known as dual specificity phosphatase-1 (DUSP-1), plays a crucial role in the deactivation of MAPKs. Several drugs with immune-suppressive properties modulate MKP-1 expression as part of their mechanism of action. We investigated the effect of mTOR inhibition through rapamycin and a dual mTOR inhibitor (AZD2014) on MKP-1 expression. Low dose rapamycin led to a rapid activation of both AKT and ERK pathways with a subsequent increase in MKP-1 expression. Rapamycin treatment led to phosphorylation of CREB, transcription factor 1 (ATF1), and ATF2, three transcription factors that bind to the cyclic AMP-responsive elements on the Mkp-1 promoter. Inhibition of either the MEK/ERK or the AKT pathway attenuated rapamycin-mediated MKP-1 induction. AZD2014 did not activate AKT but activated the ERK pathway, leading to a moderate MKP-1 induction. Using bone marrow-derived macrophages (BMDMs) derived from wild-type (WT) mice or mice deficient in AKT1 and AKT2 isoforms or BMDM from targeted deficiency in MEK1 and MEK2, we show that rapamycin treatment led to an increased MKP1 expression in BMDM from WT but failed to do so in BMDMs lacking the AKT1 isoform or MEK1 and MEK2. Importantly, rapamycin pretreatment inhibited LPS-mediated p38 activation and decreased nitric oxide and IL-6 production. Our work provides a conceptual framework for the observed immune modulatory effect of mTOR inhibition.

  5. Fast calcium sensor proteins for monitoring neural activity

    PubMed Central

    Badura, Aleksandra; Sun, Xiaonan Richard; Giovannucci, Andrea; Lynch, Laura A.; Wang, Samuel S.-H.

    2014-01-01

    Abstract. A major goal of the BRAIN Initiative is the development of technologies to monitor neuronal network activity during active information processing. Toward this goal, genetically encoded calcium indicator proteins have become widely used for reporting activity in preparations ranging from invertebrates to awake mammals. However, slow response times, the narrow sensitivity range of Ca2+ and in some cases, poor signal-to-noise ratio still limit their usefulness. Here, we review recent improvements in the field of neural activity-sensitive probe design with a focus on the GCaMP family of calcium indicator proteins. In this context, we present our newly developed Fast-GCaMPs, which have up to 4-fold accelerated off-responses compared with the next-fastest GCaMP, GCaMP6f. Fast-GCaMPs were designed by destabilizing the association of the hydrophobic pocket of calcium-bound calmodulin with the RS20 binding domain, an intramolecular interaction that protects the green fluorescent protein chromophore. Fast-GCaMP6f-RS06 and Fast-GCaMP6f-RS09 have rapid off-responses in stopped-flow fluorimetry, in neocortical brain slices, and in the intact cerebellum in vivo. Fast-GCaMP6f variants should be useful for tracking action potentials closely spaced in time, and for following neural activity in fast-changing compartments, such as axons and dendrites. Finally, we discuss strategies that may allow tracking of a wider range of neuronal firing rates and improve spike detection. PMID:25558464

  6. Positive feedback of protein kinase C proteolytic activation during apoptosis.

    PubMed Central

    Leverrier, Sabrina; Vallentin, Alice; Joubert, Dominique

    2002-01-01

    In contrast with protein kinase Calpha (PKCalpha) and PKCepsilon, which are better known for promoting cell survival, PKCdelta is known for its pro-apoptotic function, which is exerted mainly through a caspase-3-dependent proteolytic activation pathway. In the present study, we used the rat GH3B6 pituitary adenoma cell line to show that PKCalpha and PKCepsilon are activated and relocalized together with PKCdelta when apoptosis is induced by a genotoxic stress. Proteolytic activation is a crucial step used by the three isoforms since: (1) the catalytic domains of the PKCalpha, PKCepsilon or PKCdelta isoforms (CDalpha, CDepsilon and CDdelta respectively) accumulated, and this accumulation was dependent on the activity of both calpain and caspase; and (2) transient expression of CDalpha, CDepsilon or CDdelta sufficed to induce apoptosis. However, following this initial step of proteolytic activation, the pathways diverge; cytochrome c release and caspase-3 activation are induced by CDepsilon and CDdelta, but not by CDalpha. Another interesting finding of the present study is the proteolysis of PKCdelta induced by CDepsilon expression that revealed the existence of a cross-talk between PKC isoforms during apoptosis. Hence the PKC family may participate in the apoptotic process of pituitary adenoma cells at two levels: downstream of caspase and calpain, and via retro-activation of caspase-3, resulting in the amplification of its own proteolytic activation. PMID:12238950

  7. Pediatric Arm Reconstruction after Shot-gun Injury Using Peroneal Free-flap and Pedicled Latissimus Dorsi Muscle Flap: Late Follow-up.

    PubMed

    Olvera-Caballero, Carlos; Ortiz-Dominguez, Abel

    2016-08-01

    A 15-year-old patient harmed himself upon firing a shotgun that he was carrying when he slipped and fell, causing a destructive wound in the right arm with a medial entry hole and a posterolateral exit hole. The biceps, coracobrachialis, triceps, deltoids, skin cover, and humerus were injured; however, the blood vessels and major nerves of the area were surprisingly not affected. The residual skin muscle defect after debridements was 16 × 5 cm medially and posteriorly, and the bone loss was 7 cm. The wound was reconstructed during a single surgery with a free fibula flap and a pedicled flap of latissimus dorsi. Ten years after surgery, the patient presents neither functional deficit of the injured limb (shoulder, arm, forearm, and hand) nor sequelae in the donor areas; he performs his daily activities without any limitations. This case confirms that the use of free bone flaps and pedicled muscle flaps in pediatric patients can provide excellent long-term results. PMID:27622112

  8. Pediatric Arm Reconstruction after Shot-gun Injury Using Peroneal Free-flap and Pedicled Latissimus Dorsi Muscle Flap: Late Follow-up

    PubMed Central

    Ortiz-Dominguez, Abel

    2016-01-01

    Summary: A 15-year-old patient harmed himself upon firing a shotgun that he was carrying when he slipped and fell, causing a destructive wound in the right arm with a medial entry hole and a posterolateral exit hole. The biceps, coracobrachialis, triceps, deltoids, skin cover, and humerus were injured; however, the blood vessels and major nerves of the area were surprisingly not affected. The residual skin muscle defect after debridements was 16 × 5 cm medially and posteriorly, and the bone loss was 7 cm. The wound was reconstructed during a single surgery with a free fibula flap and a pedicled flap of latissimus dorsi. Ten years after surgery, the patient presents neither functional deficit of the injured limb (shoulder, arm, forearm, and hand) nor sequelae in the donor areas; he performs his daily activities without any limitations. This case confirms that the use of free bone flaps and pedicled muscle flaps in pediatric patients can provide excellent long-term results.

  9. Pediatric Arm Reconstruction after Shot-gun Injury Using Peroneal Free-flap and Pedicled Latissimus Dorsi Muscle Flap: Late Follow-up

    PubMed Central

    Ortiz-Dominguez, Abel

    2016-01-01

    Summary: A 15-year-old patient harmed himself upon firing a shotgun that he was carrying when he slipped and fell, causing a destructive wound in the right arm with a medial entry hole and a posterolateral exit hole. The biceps, coracobrachialis, triceps, deltoids, skin cover, and humerus were injured; however, the blood vessels and major nerves of the area were surprisingly not affected. The residual skin muscle defect after debridements was 16 × 5 cm medially and posteriorly, and the bone loss was 7 cm. The wound was reconstructed during a single surgery with a free fibula flap and a pedicled flap of latissimus dorsi. Ten years after surgery, the patient presents neither functional deficit of the injured limb (shoulder, arm, forearm, and hand) nor sequelae in the donor areas; he performs his daily activities without any limitations. This case confirms that the use of free bone flaps and pedicled muscle flaps in pediatric patients can provide excellent long-term results. PMID:27622112

  10. Circulating FGF21 proteolytic processing mediated by fibroblast activation protein

    PubMed Central

    Zhen, Eugene Y.; Jin, Zhaoyan; Ackermann, Bradley L.; Thomas, Melissa K.; Gutierrez, Jesus A.

    2015-01-01

    Fibroblast growth factor 21 (FGF21), a hormone implicated in the regulation of glucose homoeostasis, insulin sensitivity, lipid metabolism and body weight, is considered to be a promising therapeutic target for the treatment of metabolic disorders. Despite observations that FGF21 is rapidly proteolysed in circulation rending it potentially inactive, little is known regarding mechanisms by which FGF21 protein levels are regulated. We systematically investigated human FGF21 protein processing using mass spectrometry. In agreement with previous reports, circulating human FGF21 was found to be cleaved primarily after three proline residues at positions 2, 4 and 171. The extent of FGF21 processing was quantified in a small cohort of healthy human volunteers. Relative abundance of FGF21 proteins cleaved after Pro-2, Pro-4 and Pro-171 ranged from 16 to 30%, 10 to 25% and 10 to 34%, respectively. Dipeptidyl peptidase IV (DPP-IV) was found to be the primary protease responsible for N-terminal cleavages after residues Pro-2 and Pro-4. Importantly, fibroblast activation protein (FAP) was implicated as the protease responsible for C-terminal cleavage after Pro-171, rendering the protein inactive. The requirement of FAP for FGF21 proteolysis at the C-terminus was independently demonstrated by in vitro digestion, immunodepletion of FAP in human plasma, administration of an FAP-specific inhibitor and by human FGF21 protein processing patterns in FAP knockout mouse plasma. The discovery that FAP is responsible for FGF21 inactivation extends the FGF21 signalling pathway and may enable novel approaches to augment FGF21 actions for therapeutic applications. PMID:26635356

  11. Activation of G protein by opioid receptors: role of receptor number and G-protein concentration.

    PubMed

    Remmers, A E; Clark, M J; Alt, A; Medzihradsky, F; Woods, J H; Traynor, J R

    2000-05-19

    The collision-coupling model for receptor-G-protein interaction predicts that the rate of G-protein activation is dependent on receptor density, but not G-protein levels. C6 cells expressing mu- or delta-opioid receptors, or SH-SY5Y cells, were treated with beta-funaltrexamine (mu) or naltrindole-5'-isothiocyanate (delta) to decrease receptor number. The time course of full or partial agonist-stimulated ¿35SGTPgammaS binding did not vary in C6 cell membranes containing <1-25 pmol/mg mu-opioid receptor, or 1. 4-4.3 pmol/mg delta-opioid receptor, or in SHSY5Y cells containing 0. 16-0.39 pmol/mg receptor. The association of ¿35SGTPgammaS binding was faster in membranes from C6mu cells than from C6delta cells. A 10-fold reduction in functional G-protein, following pertussis toxin treatment, lowered the maximal level of ¿35SGTPgammaS binding but not the association rate. These data indicate a compartmentalization of opioid receptors and G protein at the cell membrane. PMID:10822058

  12. Switch Flap for Upper Eyelid Reconstruction—How Soon Should the Flap Be Divided?

    PubMed Central

    Yanai, Tetsu; Yasuta, Masato; Kawano, Hiroshige; Ishihara, Yasuhiro; Kikuchi, Mamoru

    2016-01-01

    Background: The results of a cohort of patients treated at one institution for upper eyelid reconstruction with the switch flap method after a defect due to excision of malignant tumor were reviewed. Methods: A retrospective data file review of all patients who had undergone total upper eyelid reconstruction with the switch flap method was conducted at the Saga University Hospital between April 2000 and October 2014. The follow-up lasted for varying periods during which the preoperative and postoperative photographs were compared as well. Results: A total of 10 patients with upper eyelid tumors, that is, 7 sebaceous carcinoma, 2 squamous cell carcinoma, and 1 basal cell carcinoma, underwent reconstructive surgery. With the switch flap technique, the defects resulting from tumor excision were completely covered in all cases. The mean of defect widths after tumor excision (A) was 18.8 mm (range, 15–25 mm), the mean of widths of switch flaps (B) was 13.3 mm (range, 8–22 mm), and the mean of B/A ratios was 0.69 (range, 0.5–0.88). When the switch flap was divided at 7 to 14 days, there was no flap loss, trichiasis, or corneal ulcer. Conclusion: Our protocol managed to make flaps with a B/A ratio of 0.5–0.7, and the flaps were divided at 7 to 14 days after surgery, the timing of which was much earlier than in the conventional method, lessening the possibility of complications. PMID:27200257

  13. Tunnelled tensor fascia lata flap for complex abdominal wall reconstruction

    PubMed Central

    Wang, Frederick; Buonocore, Samuel; Narayan, Deepak

    2011-01-01

    This study describes the treatment of two patients with recurrent, infected abdominal wall defects using bilateral delayed and tunnelled pedicled tensor fascia lata (TFL) myofascial flaps. TFL flaps were elevated and delayed for 4 weeks in both cases. In the second case, Parietex Composite mesh was positioned underneath the TFL flap and allowed to incorporate. After a delay of 4 weeks, the flaps were harvested and tunnelled subcutaneously to repair the abdominal wall defect. Both patients have stable repairs but had donor site seromas requiring drainage. Cadaver dissection was also performed to identify structures related to TFL flap harvest. We identified a variant of lateral femoral cutaneous nerve that traversed the TFL flap, necessitating meticulous dissection during surgery. In summary, we describe a new technique of incorporating mesh into the TFL prior to flap harvest for reconstruction of complex abdominal wall. PMID:22707661

  14. Protein kinase C directly phosphorylates the insulin receptor in vitro and reduces its protein-tyrosine kinase activity.

    PubMed Central

    Bollag, G E; Roth, R A; Beaudoin, J; Mochly-Rosen, D; Koshland, D E

    1986-01-01

    The beta subunit of purified insulin receptor is phosphorylated on a serine residue by purified preparations of protein kinase C (ATP: protein phosphotransferase, EC 2.7.1.37). This phosphorylation is inhibited by antibodies to protein kinase C and stimulated by phospholipids, diacylglycerol, and Ca2+. The phosphorylation of the receptor by protein kinase C does not affect its insulin-binding activity but does inhibit by 65% the receptor's intrinsic tyrosine-specific protein kinase activity (ATP: protein-tyrosine O-phosphotransferase, EC 2.7.1.112). These results indicate that activators of protein kinase C, such as phorbol esters, desensitize cells to insulin by direct protein kinase C action on the insulin receptor. Images PMID:3526339

  15. Activated G Protein Gαs Samples Multiple Endomembrane Compartments.

    PubMed

    Martin, Brent R; Lambert, Nevin A

    2016-09-23

    Heterotrimeric G proteins are localized to the plasma membrane where they transduce extracellular signals to intracellular effectors. G proteins also act at intracellular locations, and can translocate between cellular compartments. For example, Gαs can leave the plasma membrane and move to the cell interior after activation. However, the mechanism of Gαs translocation and its intracellular destination are not known. Here we use bioluminescence resonance energy transfer (BRET) to show that after activation, Gαs rapidly associates with the endoplasmic reticulum, mitochondria, and endosomes, consistent with indiscriminate sampling of intracellular membranes from the cytosol rather than transport via a specific vesicular pathway. The primary source of Gαs for endosomal compartments is constitutive endocytosis rather than activity-dependent internalization. Recycling of Gαs to the plasma membrane is complete 25 min after stimulation is discontinued. We also show that an acylation-deacylation cycle is important for the steady-state localization of Gαs at the plasma membrane, but our results do not support a role for deacylation in activity-dependent Gαs internalization. PMID:27528603

  16. Activity of lactoperoxidase when adsorbed on protein layers.

    PubMed

    Haberska, Karolina; Svensson, Olof; Shleev, Sergey; Lindh, Liselott; Arnebrant, Thomas; Ruzgas, Tautgirdas

    2008-09-15

    Lactoperoxidase (LPO) is an enzyme, which is used as an antimicrobial agent in a number of applications, e.g., food technology. In the majority of applications LPO is added to a homogeneous product phase or immobilised on product surface. In the latter case, however, the measurements of LPO activity are seldom reported. In this paper we have assessed LPO enzymatic activity on bare and protein modified gold surfaces by means of electrochemistry. It was found that LPO rapidly adsorbs to bare gold surfaces resulting in an amount of LPO adsorbed of 2.9mg/m(2). A lower amount of adsorbed LPO is obtained if the gold surface is exposed to bovine serum albumin, bovine or human mucin prior to LPO adsorption. The enzymatic activity of the adsorbed enzyme is in general preserved at the experimental conditions and varies only moderately when comparing bare gold and gold surface pretreated with the selected proteins. The measurement of LPO specific activity, however, indicate that it is about 1.5 times higher if LPO is adsorbed on gold surfaces containing a small amount of preadsorbed mucin in comparison to the LPO directly adsorbed on bare gold.

  17. Superoxide dismutase activity of Cu-bound prion protein

    NASA Astrophysics Data System (ADS)

    Hodak, Miroslav; Lu, Wenchang; Bernholc, Jerry

    2009-03-01

    Misfolding of the prion protein, PrP, has been linked to a group of neurodegenerative diseases, including the mad cow disease in cattle and the Creutzfeldt-Jakob disease in humans. The normal function of PrP is still unknown, but it was found that the PrP can efficiently bind Cu(II) ions. Early experiments suggested that Cu-PrP complex possesses significant superoxide dismutase (SOD) activity, but later experiments failed to confirm it and at present this issue remains unresolved. Using a recently developed hybrid DFT/DFT method, which combines Kohn-Sham DFT for the solute and its first solvation shells with orbital-free DFT for the remainder of the solvent, we have investigated SOD activity of PrP. The PrP is capable of incorporating Cu(II) ions in several binding modes and our calculations find that each mode has a different SOD activity. The highest activity found is comparable to those of well-known SOD proteins, suggesting that the conflicting experimental results may be due to different bindings of Cu(II) in those experiments.

  18. [National evaluation of the diagnosis of activated protein C resistance].

    PubMed

    Montiel-Manzano, Guadalupe; de la Peña-Díaz, Aurora; Majluf-Cruz, Abraham; Cesarman-Maus, Gabriela; Corona-de la Peña, Norma; Cruz-Cruz, Donají; Gaminio, Elizabeth; Martínez-Murillo, Carlos; Mayagoitia, Teresa; Miranda-Peralta, Enrique; Poblete, Teresita; Quintana-Martínez, Sandra; Ramírez, Raúl; Razo, Daniel; Ruiz de Chávez-Ochoa, Adriana; Reyes-Núñez, Virginia Adriana; Salazar, Rosario; Vicencio-Santiago, Guadalupe Virginia; Villa, Rosario; Reyes-Núñez, Aurelia Virginia

    2003-01-01

    Thrombophilia or prothrombotic state appears when activation of blood hemostatic mechanisms overcomes the physiological anticoagulant capacity allowing a thrombotic event. Thrombosis is the leading worldwide mortality cause and due to its high associated morbidity and mortality, it should be insisted in the opportune identification of a thrombophilic state. The study of thrombophilia identifies individuals at high risk for thrombosis. This meeting was conceived first to analyze the current status of the diagnosis of thrombophilia in Mexico and second to create the base for a national consensus for thrombophilia screening and for the establishment of a national center for laboratory reference and quality control for thrombophilia. Since searching of activated protein C resistance (APCR) and FV Leiden seem to have priority either in the clinical setting and in public health services, it was decided to start with these two abnormalities as a model to analyze the current status of thrombophilia diagnosis in the clinical laboratory. At this time, several thrombophilic abnormalities have been described however, APCR remains the most important cause of thrombophilia, accounting for as much as 20% to 60% of all venous thrombosis. APCR is a consequence of the resistance of activated FV to be inactivated by activated protein C. Procoagulant activity of activated FV increases the risk of thrombosis. Hereditary APCR is almost always due to a point mutation at the nucleotide 1691 of the FV gen inducing an Arg506Glu substitution in FV molecule. This mutation is better known as FV Leiden. Heterocygous carriers of FV Leiden have a thrombotic risk 5 to 10 times higher than general population while the risk for the homocygote state is increased 50 to 100-fold. When activated PC is added to plasma from patients with FV Leiden, this last resists the anticoagulant effect of activated PC. Therefore, thrombin production is not inhibited. This phenomenon is called APCR. The functional

  19. Schistosoma mansoni secretes a chemokine binding protein with antiinflammatory activity.

    PubMed

    Smith, Philip; Fallon, Rosie E; Mangan, Niamh E; Walsh, Caitriona M; Saraiva, Margarida; Sayers, Jon R; McKenzie, Andrew N J; Alcami, Antonio; Fallon, Padraic G

    2005-11-21

    The coevolution of humans and infectious agents has exerted selective pressure on the immune system to control potentially lethal infections. Correspondingly, pathogens have evolved with various strategies to modulate and circumvent the host's innate and adaptive immune response. Schistosoma species are helminth parasites with genes that have been selected to modulate the host to tolerate chronic worm infections, often for decades, without overt morbidity. The modulation of immunity by schistosomes has been shown to prevent a range of immune-mediated diseases, including allergies and autoimmunity. Individual immune-modulating schistosome molecules have, therefore, therapeutic potential as selective manipulators of the immune system to prevent unrelated diseases. Here we show that S. mansoni eggs secrete a protein into host tissues that binds certain chemokines and inhibits their interaction with host chemokine receptors and their biological activity. The purified recombinant S. mansoni chemokine binding protein (smCKBP) suppressed inflammation in several disease models. smCKBP is unrelated to host proteins and is the first described chemokine binding protein encoded by a pathogenic human parasite and may have potential as an antiinflammatory agent.

  20. BATMAV: a 2-DOF bio-inspired flapping flight platform

    NASA Astrophysics Data System (ADS)

    Bunget, Gheorghe; Seelecke, Stefan

    2010-04-01

    Due to the availability of small sensors, Micro-Aerial Vehicles (MAVs) can be used for detection missions of biological, chemical and nuclear agents. Traditionally these devices used fixed or rotary wings, actuated with electric DC motortransmission, a system which brings the disadvantage of a heavier platform. The overall objective of the BATMAV project is to develop a biologically inspired bat-like MAV with flexible and foldable wings for flapping flight. This paper presents a flight platform that features bat-inspired wings which are able to actively fold their elbow joints. A previous analysis of the flight physics for small birds, bats and large insects, revealed that the mammalian flight anatomy represents a suitable flight platform that can be actuated efficiently using Shape Memory Alloy (SMA) artificial-muscles. A previous study of the flight styles in bats based on the data collected by Norberg [1] helped to identify the required joint angles as relevant degrees of freedom for wing actuation. Using the engineering theory of robotic manipulators, engineering kinematic models of wings with 2 and 3-DOFs were designed to mimic the wing trajectories of the natural flier Plecotus auritus. Solid models of the bat-like skeleton were designed based on the linear and angular dimensions resulted from the kinematic models. This structure of the flight platform was fabricated using rapid prototyping technologies and assembled to form a desktop prototype with 2-DOFs wings. Preliminary flapping test showed suitable trajectories for wrist and wingtip that mimic the flapping cycle of the natural flyer.

  1. Results of instrumental and operative implementation of T-Flap

    NASA Astrophysics Data System (ADS)

    Marcelli, M.; Madonia, A.; Piermattei, V.; Mainardi, U.

    2012-04-01

    The development of new technologies for the study of the ocean processes is one of the most innovative aspects of oceanographic research that increasingly requires a large amount of data for an integrated approach to in situ observations, forecasting models and remotely sensed data. At now the marine measurement technologies are too expensive for an extensive utilization. The T-Flap technology (Temperature-Fluorescence LAunchable Probe) meets these needs by providing low cost and user-friendly in situ measuring of physical and bio-optical variables of water bodies, as opposed to traditional methods. Many experimental laboratory tests were carried out in order to increase the instrumental sensitivity of the fluorimetric sensor for the detection of chlorophyll a concentration. To test the selected components (LEDs, diodes, filters) and their new configurations, an optical circuit has been realized consisting on three prototypes of the measuring cell. Fluorescence excitation and detection efficiency of the new electro-optical assemblages were also tested by the comparison between the voltage output signals and spectroscopic in vivo fluorescence measures both of natural seawater samples and marine cultures at different concentrations. The upgraded sensors were tested during different oceanographic surveys performed in the last years both in the Tyrrhenian Sea and Adriatic Sea. The technological achievements were applied in different operative conditions: profiler along the water column (not expendable) and stand alone (in continuous surface acquisitions along tracks, in continuous acquisition on a buoy). In this work we present the results of the latest research activities on T-Flap evolution. In addition preliminary results about new sensors currently under development are shown, such as CDOM (Cromophoric Dissolved Organic Matter) fluorimetric sensor, based on T-Flap fluorescence technology, and conductivity sensor.

  2. Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis

    PubMed Central

    Kohansal-Nodehi, Mahdokht; Chua, John JE; Urlaub, Henning; Jahn, Reinhard; Czernik, Dominika

    2016-01-01

    Neurotransmitter release is mediated by the fast, calcium-triggered fusion of synaptic vesicles with the presynaptic plasma membrane, followed by endocytosis and recycling of the membrane of synaptic vesicles. While many of the proteins governing these processes are known, their regulation is only beginning to be understood. Here we have applied quantitative phosphoproteomics to identify changes in phosphorylation status of presynaptic proteins in resting and stimulated nerve terminals isolated from the brains of Wistar rats. Using rigorous quantification, we identified 252 phosphosites that are either up- or downregulated upon triggering calcium-dependent exocytosis. Particularly pronounced were regulated changes of phosphosites within protein constituents of the presynaptic active zone, including bassoon, piccolo, and RIM1. Additionally, we have mapped kinases and phosphatases that are activated upon stimulation. Overall, our study provides a snapshot of phosphorylation changes associated with presynaptic activity and provides a foundation for further functional analysis of key phosphosites involved in presynaptic plasticity. DOI: http://dx.doi.org/10.7554/eLife.14530.001 PMID:27115346

  3. Hepatitis B virus x protein induces autophagy via activating death-associated protein kinase.

    PubMed

    Zhang, H-T; Chen, G G; Hu, B-G; Zhang, Z-Y; Yun, J-P; He, M-L; Lai, P B S

    2014-01-01

    Hepatitis B virus x protein (HBX), a product of hepatitis B virus (HBV), is a multifunctional protein that regulates viral replication and various cellular functions. Recently, HBX has been shown to induce autophagy; however, the responsible mechanism is not fully known. In this study, we established stable HBX-expressing epithelial Chang cells as the platform to study how HBX induced autophagy. The results showed that the overexpression of HBX resulted in starvation-induced autophagy. HBX-induced autophagy was related to its ability to dephosphorylate/activate death-associated protein kinase (DAPK). The block of DAPK by its siRNA significantly counteracted HBX-mediated autophagy, confirming the positive role of DAPK in this process. HBX also induced Beclin 1, which functions at the downstream of the DAPK-mediated autophagy pathway. Although HBX could activate JNK, a kinase known to participate in autophagy in certain conditions, the change in JNK failed to influence HBX-induced autophagy. In conclusion, HBX induces autophagy via activating DAPK in a pathway related to Beclin 1, but not JNK. This new finding should help us to understand the role of autophagy in HBX-mediated pathogenesis and thus may provide targets for intervening HBX-related disorders.

  4. TALE factors poise promoters for activation by Hox proteins.

    PubMed

    Choe, Seong-Kyu; Ladam, Franck; Sagerström, Charles G

    2014-01-27

    Hox proteins form complexes with TALE cofactors from the Pbx and Prep/Meis families to control transcription, but it remains unclear how Hox:TALE complexes function. Examining a Hoxb1b:TALE complex that regulates zebrafish hoxb1a transcription, we find maternally deposited TALE proteins at the hoxb1a promoter already during blastula stages. These TALE factors recruit histone-modifying enzymes to promote an active chromatin profile at the hoxb1a promoter and also recruit RNA polymerase II (RNAPII) and P-TEFb. However, in the presence of TALE factors, RNAPII remains phosphorylated on serine 5 and hoxb1a transcription is inefficient. By gastrula stages, Hoxb1b binds together with TALE factors to the hoxb1a promoter. This triggers P-TEFb-mediated transitioning of RNAPII to the serine 2-phosphorylated form and efficient hoxb1a transcription. We conclude that TALE factors access promoters during early embryogenesis to poise them for activation but that Hox proteins are required to trigger efficient transcription.

  5. Review of Skull Base Reconstruction Using Locoregional Flaps and Free Flaps in Children and Adolescents

    PubMed Central

    Yano, Tomoyuki; Tanaka, Kentarou; Kishimoto, Seiji; Iida, Hideo; Okazaki, Mutsumi

    2011-01-01

    Tumors of the skull base are rare in children, and reconstruction in such patients has rarely been reported. We reviewed 16 cases of skull base reconstruction in patients under 18 years. The study group consisted of 10 boys and 6 girls, whose ages ranged from 2 to 17 years. Of the 16 cases, eight tumors were benign and eight were malignant. Defects were anterior in six cases, lateral in eight cases, and anterolateral in two cases. Reconstruction was performed with locoregional flaps in 11 cases and with free flaps in 5 cases. No significant difference was found between locoregional flaps and free flaps in total operative time, intraoperative blood loss, or postoperative hospital stay. However, in some cases, total operative time, reconstruction time, and blood loss increased to a degree unacceptable for pediatrics. Minor complications occurred in three patients and a major complication occurred in one case. Of four patients, three patients with postoperative complications had undergone chemoradiotherapy. Because of the physical weakness of pediatric patients, complicated reconstructive procedure should be avoided. We believe locoregional flaps will become the first choice for reconstruction. However, if patients have large, complex defects and have received radiotherapy, appropriate free flaps should be used to avoid postoperative complications. PMID:22547961

  6. Abscisic Acid Induces Mitogen-Activated Protein Kinase Activation in Barley Aleurone Protoplasts.

    PubMed

    Knetsch, MLW.; Wang, M.; Snaar-Jagalska, B. E.; Heimovaara-Dijkstra, S.

    1996-06-01

    Abscisic acid (ABA) induces a rapid and transient mitogen-activated protein (MAP) kinase activation in barley aleurone protoplasts. MAP kinase activity, measured as myelin basic protein phosphorylation by MAP kinase immunoprecipitates, increased after 1 min, peaked after 3 min, and decreased to basal levels after ~5 min of ABA treatment in vivo. Antibodies recognizing phosphorylated tyrosine residues precipitate with myelin basic protein kinase activity that has identical ABA activation characteristics and demonstrate that tyrosine phosphorylation of MAP kinase occurs during activation. The half-maximal concentration of ABA required for MAP kinase activation, 3 x 10-7 M, is very similar to that required for ABA-induced rab16 gene expression. The tyrosine phosphatase inhibitor phenylarsine oxide can completely block ABA-induced MAP kinase activation and rab16 gene expression. These results lead us to conclude that ABA activates MAP kinase via a tyrosine phosphatase and that these steps are a prerequisite for ABA induction of rab16 gene expression.

  7. Abscisic Acid Induces Mitogen-Activated Protein Kinase Activation in Barley Aleurone Protoplasts.

    PubMed Central

    Knetsch, MLW.; Wang, M.; Snaar-Jagalska, B. E.; Heimovaara-Dijkstra, S.

    1996-01-01

    Abscisic acid (ABA) induces a rapid and transient mitogen-activated protein (MAP) kinase activation in barley aleurone protoplasts. MAP kinase activity, measured as myelin basic protein phosphorylation by MAP kinase immunoprecipitates, increased after 1 min, peaked after 3 min, and decreased to basal levels after ~5 min of ABA treatment in vivo. Antibodies recognizing phosphorylated tyrosine residues precipitate with myelin basic protein kinase activity that has identical ABA activation characteristics and demonstrate that tyrosine phosphorylation of MAP kinase occurs during activation. The half-maximal concentration of ABA required for MAP kinase activation, 3 x 10-7 M, is very similar to that required for ABA-induced rab16 gene expression. The tyrosine phosphatase inhibitor phenylarsine oxide can completely block ABA-induced MAP kinase activation and rab16 gene expression. These results lead us to conclude that ABA activates MAP kinase via a tyrosine phosphatase and that these steps are a prerequisite for ABA induction of rab16 gene expression. PMID:12239411

  8. Repair of Fingertip Defect Using an Anterograde Pedicle Flap Based on the Dorsal Perforator

    PubMed Central

    Wei, Peng; Chen, Weiwei; Mei, Jin; Ding, Maochao; Yu, Yaling; Xi, Shanshan; Zhou, Renpeng

    2016-01-01

    Background: The purposes of this article are to introduce and assess the results of a long-term follow-up of using anterograde pedicle flap based on the dorsal branches of proper digital neurovascular bundles from the dorsum of the middle phalanx for the fingertip defect. Methods: Between February 2011 and December 2012, 31 patients underwent reconstruction of fingertip defects using a homodigital flap based on the dorsal perforator in the middle phalanx. The defect size ranged from 1.3 cm × 1.5 cm to 2.4 cm × 3.0 cm. During surgery, the flap was designed on the dorsal middle phalangeal region. The pedicle was a neurovascular bundle consisting of an artery, vein, and sensory nerve; the rotation of pedicle was <90 degrees. Results: The clinical results were satisfactory after 3 to 9 months of follow-up. The flaps were considered cosmetically acceptable by both patients and doctors. The sensory recovery was excellent, 2-point discrimination was 4.96 ± 1.47 mm, and the recovery of range of motion of the interphalangeal joints was very good. Conclusions: The anterograde island flap based on the dorsal branches of proper digital neurovascular bundles is an ideal aesthetic reconstruction method for fingertip defect. A 90-degree rotated island pedicle flap was very versatile, easy to design, and had good survival. This technique is simple with less damage to the donor site, without sacrificing the branch of the digital artery and nerve. The reliable source of blood supply and satisfactory recovery of sensation can be achieved without affecting the interphalangeal joint activity. PMID:27482478

  9. Fluctuation driven active molecular transport in passive channel proteins

    NASA Astrophysics Data System (ADS)

    Kosztin, Ioan

    2006-03-01

    Living cells interact with their extracellular environment through the cell membrane, which acts as a protective permeability barrier for preserving the internal integrity of the cell. However, cell metabolism requires controlled molecular transport across the cell membrane, a function that is fulfilled by a wide variety of transmembrane proteins, acting as either passive or active transporters. In this talk it is argued that, contrary to the general belief, in active cell membranes passive and spatially asymmetric channel proteins can act as active transporters by consuming energy from nonequilibrium fluctuations fueled by cell metabolism. This assertion is demonstrated in the case of the E. coli aquaglyceroporin GlpF channel protein, whose high resolution crystal structure is manifestly asymmetric. By calculating the glycerol flux through GlpF within the framework of a stochastic model, it is found that, as a result of channel asymmetry, glycerol uptake driven by a concentration gradient is enhanced significantly in the presence of non-equilibrium fluctuations. Furthermore, the enhancement caused by a ratchet-like mechanism is larger for the outward, i.e., from the cytoplasm to the periplasm, flux than for the inward one, suggesting that the same non-equilibrium fluctuations also play an important role in protecting the interior of the cell against poisoning by excess uptake of glycerol. Preliminary data on water and sugar transport through aquaporin and maltoporin channels, respectively, are indicative of the universality of the proposed nonequilibrium-fluctuation-driven active transport mechanism. This work was supported by grants from the Univ. of Missouri Research Board, the Institute for Theoretical Sciences and the Department of Energy (DOE Contract W-7405-ENG-36), and the National Science Foundation (FIBR-0526854).

  10. Activated protein C mediates a healing phenotype in cultured tenocytes.

    PubMed

    Xue, Meilang; Smith, Margaret M; Little, Christopher B; Sambrook, Philip; March, Lyn; Jackson, Christopher J

    2009-04-01

    Tendon injuries cause considerable morbidity in the general adult population. The tenocytes within the tendon have the full capacity to heal the tendon intrinsically. Activated protein C (APC) plays an important role in coagulation and inflammation and more recently has been shown to promote cutaneous wound healing. In this study we examined whether APC can induce a wound healing phenotype in tenocytes. Sheep tenocytes were treated with APC, endothelial protein C receptor (EPCR) blocking antibody (RCR252) and/or EPCR small interfering (si)RNA. Cell proliferation and migration were measured by crystal violet assay and a scratch wounding assay, respectively. The expression of EPCR, matrix metalloproteinase (MMP)-2, type I collagen and MAP kinase activity were detected by real time PCR, zymography, immunofluorescence, immunohistochemistry and Western blotting. APC stimulated proliferation, MMP-2 activity and type I collagen deposition in a dose-dependent manner and promoted migration of cultured tenocytes. APC dose-dependently stimulated phosphorylated (P)-ERK2 and inhibited P-p38. Interestingly, tenocytes expressed EPCR protein, which was up-regulated by APC. When tenocytes were pre-treated with RCR252 or EPCR siRNA the effect of APC on proliferation, MMP-2 and type 1 collagen synthesis and MAP kinases was blocked. APC promotes the growth, MMP-2 activity, type I collagen deposition and migration of tenocytes. Furthermore, EPCR is expressed by tenocytes and mediates the actions of APC, at least partly by signalling through selective MAP kinases. These data implicate APC as a potential healing agent for injured tendons.

  11. Flapping wing aerodynamics: from insects to vertebrates.

    PubMed

    Chin, Diana D; Lentink, David

    2016-04-01

    More than a million insects and approximately 11,000 vertebrates utilize flapping wings to fly. However, flapping flight has only been studied in a few of these species, so many challenges remain in understanding this form of locomotion. Five key aerodynamic mechanisms have been identified for insect flight. Among these is the leading edge vortex, which is a convergent solution to avoid stall for insects, bats and birds. The roles of the other mechanisms - added mass, clap and fling, rotational circulation and wing-wake interactions - have not yet been thoroughly studied in the context of vertebrate flight. Further challenges to understanding bat and bird flight are posed by the complex, dynamic wing morphologies of these species and the more turbulent airflow generated by their wings compared with that observed during insect flight. Nevertheless, three dimensionless numbers that combine key flow, morphological and kinematic parameters - the Reynolds number, Rossby number and advance ratio - govern flapping wing aerodynamics for both insects and vertebrates. These numbers can thus be used to organize an integrative framework for studying and comparing animal flapping flight. Here, we provide a roadmap for developing such a framework, highlighting the aerodynamic mechanisms that remain to be quantified and compared across species. Ultimately, incorporating complex flight maneuvers, environmental effects and developmental stages into this framework will also be essential to advancing our understanding of the biomechanics, movement ecology and evolution of animal flight. PMID:27030773

  12. Flapping wing aerodynamics: from insects to vertebrates.

    PubMed

    Chin, Diana D; Lentink, David

    2016-04-01

    More than a million insects and approximately 11,000 vertebrates utilize flapping wings to fly. However, flapping flight has only been studied in a few of these species, so many challenges remain in understanding this form of locomotion. Five key aerodynamic mechanisms have been identified for insect flight. Among these is the leading edge vortex, which is a convergent solution to avoid stall for insects, bats and birds. The roles of the other mechanisms - added mass, clap and fling, rotational circulation and wing-wake interactions - have not yet been thoroughly studied in the context of vertebrate flight. Further challenges to understanding bat and bird flight are posed by the complex, dynamic wing morphologies of these species and the more turbulent airflow generated by their wings compared with that observed during insect flight. Nevertheless, three dimensionless numbers that combine key flow, morphological and kinematic parameters - the Reynolds number, Rossby number and advance ratio - govern flapping wing aerodynamics for both insects and vertebrates. These numbers can thus be used to organize an integrative framework for studying and comparing animal flapping flight. Here, we provide a roadmap for developing such a framework, highlighting the aerodynamic mechanisms that remain to be quantified and compared across species. Ultimately, incorporating complex flight maneuvers, environmental effects and developmental stages into this framework will also be essential to advancing our understanding of the biomechanics, movement ecology and evolution of animal flight.

  13. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control... airplanes, it must be designed to account for the unsummetrical loads resulting from flight with the engines... independent of the flap drive system; or by an approved equivalent means; or (2) Be designed so that...

  14. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control... airplanes, it must be designed to account for the unsummetrical loads resulting from flight with the engines... independent of the flap drive system; or by an approved equivalent means; or (2) Be designed so that...

  15. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control... airplanes, it must be designed to account for the unsummetrical loads resulting from flight with the engines... independent of the flap drive system; or by an approved equivalent means; or (2) Be designed so that...

  16. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control... airplanes, it must be designed to account for the unsummetrical loads resulting from flight with the engines... independent of the flap drive system; or by an approved equivalent means; or (2) Be designed so that...

  17. Role of Deleted in Breast Cancer 1 (DBC1) Protein in SIRT1 Deacetylase Activation Induced by Protein Kinase A and AMP-activated Protein Kinase*

    PubMed Central

    Nin, Veronica; Escande, Carlos; Chini, Claudia C.; Giri, Shailendra; Camacho-Pereira, Juliana; Matalonga, Jonathan; Lou, Zhenkun; Chini, Eduardo N.

    2012-01-01

    The NAD+-dependent deacetylase SIRT1 is a key regulator of several aspects of metabolism and aging. SIRT1 activation is beneficial for several human diseases, including metabolic syndrome, diabetes, obesity, liver steatosis, and Alzheimer disease. We have recently shown that the protein deleted in breast cancer 1 (DBC1) is a key regulator of SIRT1 activity in vivo. Furthermore, SIRT1 and DBC1 form a dynamic complex that is regulated by the energetic state of the organism. Understanding how the interaction between SIRT1 and DBC1 is regulated is therefore essential to design strategies aimed to activate SIRT1. Here, we investigated which pathways can lead to the dissociation of SIRT1 and DBC1 and consequently to SIRT1 activation. We observed that PKA activation leads to a fast and transient activation of SIRT1 that is DBC1-dependent. In fact, an increase in cAMP/PKA activity resulted in the dissociation of SIRT1 and DBC1 in an AMP-activated protein kinase (AMPK)-dependent manner. Pharmacological AMPK activation led to SIRT1 activation by a DBC1-dependent mechanism. Indeed, we found that AMPK activators promote SIRT1-DBC1 dissociation in cells, resulting in an increase in SIRT1 activity. In addition, we observed that the SIRT1 activation promoted by PKA and AMPK occurs without changes in the intracellular levels of NAD+. We propose that PKA and AMPK can acutely activate SIRT1 by inducing dissociation of SIRT1 from its endogenous inhibitor DBC1. Our experiments provide new insight on the in vivo mechanism of SIRT1 regulation and a new avenue for the development of pharmacological SIRT1 activators targeted at the dissociation of the SIRT1-DBC1 complex. PMID:22553202

  18. Refolding techniques for recovering biologically active recombinant proteins from inclusion bodies.

    PubMed

    Yamaguchi, Hiroshi; Miyazaki, Masaya

    2014-02-20

    Biologically active proteins are useful for studying the biological functions of genes and for the development of therapeutic drugs and biomaterials in a biotechnology industry. Overexpression of recombinant proteins in bacteria, such as Escherichia coli, often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. As inclusion bodies contain relatively pure and intact proteins, protein refolding is an important process to obtain active recombinant proteins from inclusion bodies. However, conventional refolding methods, such as dialysis and dilution, are time consuming and, often, recovered yields of active proteins are low, and a trial-and-error process is required to achieve success. Recently, several approaches have been reported to refold these aggregated proteins into an active form. The strategies largely aim at reducing protein aggregation during the refolding procedure. This review focuses on protein refolding techniques using chemical additives and laminar flow in microfluidic chips for the efficient recovery of active proteins from inclusion bodies.

  19. Regulation of mitogen-activated protein kinases by a calcium/calmodulin-dependent protein kinase cascade.

    PubMed Central

    Enslen, H; Tokumitsu, H; Stork, P J; Davis, R J; Soderling, T R

    1996-01-01

    Membrane depolarization of NG108 cells gives rapid (< 5 min) activation of Ca2+/calmodulin-dependent protein kinase IV (CaM-KIV), as well as activation of c-Jun N-terminal kinase (JNK). To investigate whether the Ca2+-dependent activation of mitogen-activated protein kinases (ERK, JNK, and p38) might be mediated by the CaM kinase cascade, we have transfected PC12 cells, which lack CaM-KIV, with constitutively active mutants of CaM kinase kinase and/or CaM-KIV (CaM-KKc and CaM-KIVc, respectively). In the absence of depolarization, CaM-KKc transfection had no effect on Elk-dependent transcription of a luciferase reporter gene, whereas CaM-KIVc alone or in combination with CaM-KKc gave 7- to 10-fold and 60- to 80-fold stimulations, respectively, which were blocked by mitogen-activated protein (MAP) kinase phosphatase cotransfection. When epitope-tagged constructs of MAP kinases were co-transfected with CaM-KKc plus CaM-KIVc, the immunoprecipitated MAP kinases were activated 2-fold (ERK-2) and 7- to 10-fold (JNK-1 and p38). The JNK and p38 pathways were further investigated using specific c-Jun or ATF2-dependent transcriptional assays. We found that c-Jun/ATF2-dependent transcriptions were enhanced 7- to 10-fold by CaM-KIVc and 20- to 30-fold by CaM-KKc plus CaM-KIVc. In the case of the Jun-dependent transcription, this effect was not due to direct phosphorylation of c-Jun by activated CaM-KIV, since transcription was blocked by a dominant-negative JNK and by two MAP kinase phosphatases. Mutation of the phosphorylation site (Thr196) in CaM-KIV, which mediates its activation by CaM-KIV kinase, prevented activation of Elk-1, c-Jun, and ATF2 by the CaM kinase cascade. These results establish a new Ca2+-dependent mechanism for regulating MAP kinase pathways and resultant transcription. Images Fig. 1 Fig. 3 Fig. 4 PMID:8855261

  20. Mycobacteriophage putative GTPase-activating protein can potentiate antibiotics.

    PubMed

    Yan, Shuangquan; Xu, Mengmeng; Duan, Xiangke; Yu, Zhaoxiao; Li, Qiming; Xie, Longxiang; Fan, Xiangyu; Xie, Jianping

    2016-09-01

    The soaring incidences of infection by antimicrobial resistant (AR) pathogens and shortage of effective antibiotics with new mechanisms of action have renewed interest in phage therapy. This scenario is exemplified by resistant tuberculosis (TB), caused by resistant Mycobacterium tuberculosis. Mycobacteriophage SWU1 A321_gp67 encodes a putative GTPase-activating protein. Mycobacterium smegmatis with gp67 overexpression showed changed colony formation and biofilm morphology and supports the efficacy of streptomycin and capreomycin against Mycobacterium. gp67 down-regulated the transcription of genes involved in cell wall and biofilm development. To our knowledge, this is the first report to show that phage protein in addition to lysin or recombination components can synergize with existing antibiotics. Phage components might represent a promising new clue for better antibiotic potentiators. PMID:27345061

  1. Metals in the active site of native protein phosphatase-1.

    PubMed

    Heroes, Ewald; Rip, Jens; Beullens, Monique; Van Meervelt, Luc; De Gendt, Stefan; Bollen, Mathieu

    2015-08-01

    Protein phosphatase-1 (PP1) is a major protein Ser/Thr phosphatase in eukaryotic cells. Its activity depends on two metal ions in the catalytic site, which were identified as manganese in the bacterially expressed phosphatase. However, the identity of the metal ions in native PP1 is unknown. In this study, total reflection X-ray fluorescence (TXRF) was used to detect iron and zinc in PP1 that was purified from rabbit skeletal muscle. Metal exchange experiments confirmed that the distinct substrate specificity of recombinant and native PP1 is determined by the nature of their associated metals. We also found that the iron level associated with native PP1 is decreased by incubation with inhibitor-2, consistent with a function of inhibitor-2 as a PP1 chaperone. PMID:25890482

  2. Influence of inflow angle on flexible flap aerodynamic performance

    NASA Astrophysics Data System (ADS)

    Y Zhao, H.; Ye, Z.; Li, Z. M.; Li, C.

    2013-12-01

    Large scale wind turbines have larger blade lengths and weights, which creates new challenges for blade design. This paper selects NREL S809 airfoil, and uses the parameterized technology to realize the flexible trailing edge deformation, researches the dynamic aerodynamic characteristics in the process of continuous flexible deformation, analyses the influence of inflow angle on flexible flap aerodynamic performance, in order to further realize the flexible wind turbine blade design and provides some references for the active control scheme. The results show that compared with the original airfoil, proper trailing edge deformation can improve the lift coefficient, reduce the drag coefficient, and thereby more efficiently realize flow field active control. With inflow angle increases, dynamic lift-drag coefficient hysteresis loop shape deviation occurs, even turns into different shapes. Appropriate swing angle can improve the flap lift coefficient, but may cause early separation of flow. To improve the overall performance of wind turbine blades, different angular control should be used at different cross sections, in order to achieve the best performance.

  3. Cytoprotective-selective activated protein C therapy for ischemic stroke

    PubMed Central

    Mosnier, Laurent O.; Zlokovic, Berislav V.; Griffin, John H.

    2014-01-01

    Summary Despite years of research and efforts to translate stroke research to clinical therapy, ischemic stroke remains a major cause of death, disability, and diminished quality of life. Primary and secondary preventive measures combined with improved quality of care have made significant progress. However, no novel drug for ischemic stroke therapy has been approved in the past decade. Numerous studies have shown beneficial effects of activated protein C (APC) in rodent stroke models. In addition to its natural anticoagulant functions, APC conveys multiple direct cytoprotective effects on many different cell types that involve multiple receptors including protease activated receptor (PAR) 1, PAR3, and the endothelial protein C receptor (EPCR). Application of molecular engineered APC variants with altered selectivity profiles to rodent stroke models demonstrated that the beneficial effects of APC primarily require its cytoprotective activities but not its anticoagulant activities. Extensive basic, preclinical, and clinical research provided a compelling rationale based on strong evidence for translation of APC therapy that has led to the clinical development of the cytoprotective-selective APC variant, 3K3A-APC, for ischemic stroke. Recent identification of non-canonical PAR1 and PAR3 activation by APC that give rise to novel tethered-ligands capable of inducing biased cytoprotective signaling as opposed to the canonical signaling provides a mechanistic explanation for how APC-mediated PAR activation can selectively induce cytoprotective signaling pathways. Collectively, these paradigm-shifting discoveries provide detailed insights into the receptor targets and the molecular mechanisms for neuroprotection by cytoprotective-selective 3K3A-APC, which is currently a biologic drug in clinical trials for ischemic stroke. PMID:25230930

  4. Evidence for protein kinase C-dependent and -independent activation of mitogen-activated protein kinase in T cells: potential role of additional diacylglycerol binding proteins.

    PubMed

    Puente, L G; Stone, J C; Ostergaard, H L

    2000-12-15

    Activation of mitogen-activated protein kinases (MAPK) is a critical signal transduction event for CTL activation, but the signaling mechanisms responsible are not fully characterized. Protein kinase C (PKC) is thought to contribute to MAPK activation following TCR stimulation. We have found that dependence on PKC varies with the method used to stimulate the T cells. Extracellular signal-regulated kinase (ERK) activation in CTL stimulated with soluble cross-linked anti-CD3 is completely inhibited by the PKC inhibitor bisindolylmaleimide (BIM). In contrast, only the later time points in the course of ERK activation are sensitive to BIM when CTL are stimulated with immobilized anti-CD3, a condition that stimulates CTL degranulation. Surprisingly, MAPK activation in response to immobilized anti-CD3 is strongly inhibited at all time points by the diacylglycerol (DAG)-binding domain inhibitor calphostin C implicating the contribution of a DAG-dependent but PKC-independent pathway in the activation of ERK in CTL clones. Chronic exposure to phorbol ester down-regulates the expression of DAG-responsive PKC isoforms; however, this treatment of CTL clones does not inhibit anti-CD3-induced activation of MAPK. Phorbol ester-treated cells have reduced expression of several isoforms of PKC but still express the recently described DAG-binding Ras guanylnucleotide-releasing protein. These results indicate that the late phase of MAPK activation in CTL clones in response to immobilized anti-CD3 stimulation requires PKC while the early phase requires a DAG-dependent, BIM-resistant component.

  5. Protein kinase D activity controls endothelial nitric oxide synthesis.

    PubMed

    Aicart-Ramos, Clara; Sánchez-Ruiloba, Lucía; Gómez-Parrizas, Mónica; Zaragoza, Carlos; Iglesias, Teresa; Rodríguez-Crespo, Ignacio

    2014-08-01

    Vascular endothelial growth factor (VEGF) regulates key functions of the endothelium, such as angiogenesis or vessel repair in processes involving endothelial nitric oxide synthase (eNOS) activation. One of the effector kinases that become activated in endothelial cells upon VEGF treatment is protein kinase D (PKD). Here, we show that PKD phosphorylates eNOS, leading to its activation and a concomitant increase in NO synthesis. Using mass spectrometry, we show that the purified active kinase specifically phosphorylates recombinant eNOS on Ser1179. Treatment of endothelial cells with VEGF or phorbol 12,13-dibutyrate (PDBu) activates PKD and increases eNOS Ser1179 phosphorylation. In addition, pharmacological inhibition of PKD and gene silencing of both PKD1 and PKD2 abrogate VEGF signaling, resulting in a clear diminished migration of endothelial cells in a wound healing assay. Finally, inhibition of PKD in mice results in an almost complete disappearance of the VEGF-induced vasodilatation, as monitored through determination of the diameter of the carotid artery. Hence, our data indicate that PKD is a new regulatory kinase of eNOS in endothelial cells whose activity orchestrates mammalian vascular tone. PMID:24928905

  6. Amygdala kindling alters protein kinase C activity in dentate gyrus.

    PubMed

    Chen, S J; Desai, M A; Klann, E; Winder, D G; Sweatt, J D; Conn, P J

    1992-11-01

    Kindling is a use-dependent form of synaptic plasticity and a widely used model of epilepsy. Although kindling has been widely studied, the molecular mechanisms underlying induction of this phenomenon are not well understood. We determined the effect of amygdala kindling on protein kinase C (PKC) activity in various regions of rat brain. Kindling stimulation markedly elevated basal (Ca(2+)-independent) and Ca(2+)-stimulated phosphorylation of an endogenous PKC substrate (which we have termed P17) in homogenates of dentate gyrus, assayed 2 h after kindling stimulation. The increase in P17 phosphorylation appeared to be due at least in part to persistent PKC activation, as basal PKC activity assayed in vitro using an exogenous peptide substrate was increased in kindled dentate gyrus 2 h after the last kindling stimulation. A similar increase in basal PKC activity was observed in dentate gyrus 2 h after the first kindling stimulation. These results document a kindling-associated persistent PKC activation and suggest that the increased activity of PKC could play a role in the induction of the kindling effect.

  7. Protein glycation inhibitory activity and antioxidant capacity of clove extract.

    PubMed

    Suantawee, Tanyawan; Wesarachanon, Krittaporn; Anantsuphasak, Kanokphat; Daenphetploy, Tanuch; Thien-Ngern, Sroshin; Thilavech, Thavaree; Pasukamonset, Porntip; Ngamukote, Sathaporn; Adisakwattana, Sirichai

    2015-06-01

    Syzygium aromaticum (L.) (clove) is one of the most widely cultivated spices in many tropical countries. The aim of this study was to determine the phytochemical content, the antioxidant properties and the antiglycation properties of aqueous extract of clove against fructose-mediated protein glycation and oxidation. The result showed that the content of total phenolics and flavonoids in clove extract was 239.58 ± 0.70 mg gallic acid equivalents/g dried extract and 65.67 ± 0.01 mg catechin equivalents/g dried extract, respectively. In addition, clove exhibited antioxidant properties including DPPH radical scavenging activity (IC50 = 0.29 ± 0.01 mg/ml), Trolox equivalent antioxidant capacity (4.69 ± 0.03 μmol Trolox equivalents/mg dried extract), ferric reducing antioxidant power (20.55 ± 0.11 μmol ascorbic acid equivalents/mg dried extract), Oxygen radical absorbance capacity (31.12 ± 0.21 μmol Trolox equivalents/mg dried extract), hydroxyl radical scavenging activity (0.15 ± 0.04 mg Trolox equivalents/mg dried extract), and superoxide radical scavenging activity (18.82 ± 0.50 mg Trolox equivalents/mg dried extract). The aqueous extract of clove (0.25-1.00 mg/ml) significantly inhibited the formation of fluorescent advanced glycation end products (AGEs) and non-fluorescent AGEs (N(ɛ)-(carboxymethyl) lysine (CML)) in glycated BSA during 4 weeks of incubation. The extract also markedly prevented oxidation-induced protein damage by decreasing protein carbonyl formation and protecting against the loss of protein thiol group. These results clearly demonstrated that a polyphenol enriched clove extract, owing to its antioxidant, was capable to inhibit the formation of AGEs and protein glycation. The findings might lead to the possibility of using the clove extract for targeting diabetic complications.

  8. Egg Activation at Fertilization by a Soluble Sperm Protein.

    PubMed

    Swann, Karl; Lai, F Anthony

    2016-01-01

    The most fundamental unresolved issue of fertilization is to define how the sperm activates the egg to begin embryo development. Egg activation at fertilization in all species thus far examined is caused by some form of transient increase in the cytoplasmic free Ca(2+) concentration. What has not been clear, however, is precisely how the sperm triggers the large changes in Ca(2+) observed within the egg cytoplasm. Here, we review the studies indicating that the fertilizing sperm stimulates a cytosolic Ca(2+) increase in the egg specifically by delivering a soluble factor that diffuses into the cytosolic space of the egg upon gamete membrane fusion. Evidence is primarily considered in species of eggs where the sperm has been shown to elicit a cytosolic Ca(2+) increase by initiating Ca(2+) release from intracellular Ca(2+) stores. We suggest that our best understanding of these signaling events is in mammals, where the sperm triggers a prolonged series of intracellular Ca(2+) oscillations. The strongest empirical studies to date suggest that mammalian sperm-triggered Ca(2+) oscillations are caused by the introduction of a sperm-specific protein, called phospholipase C-zeta (PLCζ) that generates inositol trisphosphate within the egg. We will discuss the role and mechanism of action of PLCζ in detail at a molecular and cellular level. We will also consider some of the evidence that a soluble sperm protein might be involved in egg activation in nonmammalian species.

  9. A protein tyrosine phosphatase-like protein from baculovirus has RNA 5'-triphosphatase and diphosphatase activities.

    PubMed

    Takagi, T; Taylor, G S; Kusakabe, T; Charbonneau, H; Buratowski, S

    1998-08-18

    The superfamily of protein tyrosine phosphatases (PTPs) includes at least one enzyme with an RNA substrate. We recently showed that the RNA triphosphatase domain of the Caenorhabditis elegans mRNA capping enzyme is related to the PTP enzyme family by sequence similarity and mechanism. The PTP most similar in sequence to the capping enzyme triphosphatase is BVP, a dual-specificity PTP encoded by the Autographa californica nuclear polyhedrosis virus. Although BVP previously has been shown to have modest tyrosine and serine/threonine phosphatase activity, we find that it is much more potent as an RNA 5'-phosphatase. BVP sequentially removes gamma and beta phosphates from the 5' end of triphosphate-terminated RNA, leaving a 5'-monophosphate end. The activity was specific for polynucleotides; nucleotide triphosphates were not hydrolyzed. A mutant protein in which the active site cysteine was replaced with serine was inactive. Three other dual-specificity PTPs (VH1, VHR, and Cdc14) did not exhibit detectable RNA phosphatase activity. Therefore, capping enzyme and BVP are members of a distinct PTP-like subfamily that can remove phosphates from RNA. PMID:9707557

  10. Budding Yeast Silencing Complexes and Regulation of Sir2 Activity by Protein-Protein Interactions

    PubMed Central

    Tanny, Jason C.; Kirkpatrick, Donald S.; Gerber, Scott A.; Gygi, Steven P.; Moazed, Danesh

    2004-01-01

    Gene silencing in the budding yeast Saccharomyces cerevisiae requires the enzymatic activity of the Sir2 protein, a highly conserved NAD-dependent deacetylase. In order to study the activity of native Sir2, we purified and characterized two budding yeast Sir2 complexes: the Sir2/Sir4 complex, which mediates silencing at mating-type loci and at telomeres, and the RENT complex, which mediates silencing at the ribosomal DNA repeats. Analyses of the protein compositions of these complexes confirmed previously described interactions. We show that the assembly of Sir2 into native silencing complexes does not alter its selectivity for acetylated substrates, nor does it allow the deacetylation of nucleosomal histones. The inability of Sir2 complexes to deacetylate nucleosomes suggests that additional factors influence Sir2 activity in vivo. In contrast, Sir2 complexes show significant enhancement in their affinities for acetylated substrates and their sensitivities to the physiological inhibitor nicotinamide relative to recombinant Sir2. Reconstitution experiments showed that, for the Sir2/Sir4 complex, these differences stem from the physical interaction of Sir2 with Sir4. Finally, we provide evidence that the different nicotinamide sensitivities of Sir2/Sir4 and RENT in vitro could contribute to locus-specific differences in how Sir2 activity is regulated in vivo. PMID:15282295

  11. Immersion freezing of ice nucleation active protein complexes

    NASA Astrophysics Data System (ADS)

    Hartmann, S.; Augustin, S.; Clauss, T.; Wex, H.; Šantl-Temkiv, T.; Voigtländer, J.; Niedermeier, D.; Stratmann, F.

    2013-06-01

    Utilising the Leipzig Aerosol Cloud Interaction Simulator (LACIS), the immersion freezing behaviour of droplet ensembles containing monodisperse particles, generated from a Snomax™ solution/suspension, was investigated. Thereto ice fractions were measured in the temperature range between -5 °C to -38 °C. Snomax™ is an industrial product applied for artificial snow production and contains Pseudomonas syringae} bacteria which have long been used as model organism for atmospheric relevant ice nucleation active (INA) bacteria. The ice nucleation activity of such bacteria is controlled by INA protein complexes in their outer membrane. In our experiments, ice fractions increased steeply in the temperature range from about -6 °C to about -10 °C and then levelled off at ice fractions smaller than one. The plateau implies that not all examined droplets contained an INA protein complex. Assuming the INA protein complexes to be Poisson distributed over the investigated droplet populations, we developed the CHESS model (stoCHastic modEl of similar and poiSSon distributed ice nuclei) which allows for the calculation of ice fractions as function of temperature and time for a given nucleation rate. Matching calculated and measured ice fractions, we determined and parameterised the nucleation rate of INA protein complexes exhibiting class III ice nucleation behaviour. Utilising the CHESS model, together with the determined nucleation rate, we compared predictions from the model to experimental data from the literature and found good agreement. We found that (a) the heterogeneous ice nucleation rate expression quantifying the ice nucleation behaviour of the INA protein complex is capable of describing the ice nucleation behaviour observed in various experiments for both, Snomax™ and P. syringae bacteria, (b) the ice nucleation rate, and its temperature dependence, seem to be very similar regardless of whether the INA protein complexes inducing ice nucleation are attached

  12. Keap1-Independent Regulation of Nrf2 Activity by Protein Acetylation and a BET Bromodomain Protein

    PubMed Central

    Chatterjee, Nirmalya; Tian, Min; Spirohn, Kerstin; Boutros, Michael; Bohmann, Dirk

    2016-01-01

    Mammalian BET proteins comprise a family of bromodomain-containing epigenetic regulators with complex functions in chromatin organization and gene regulation. We identified the sole member of the BET protein family in Drosophila, Fs(1)h, as an inhibitor of the stress responsive transcription factor CncC, the fly ortholog of Nrf2. Fs(1)h physically interacts with CncC in a manner that requires the function of its bromodomains and the acetylation of CncC. Treatment of cultured Drosophila cells or adult flies with fs(1)h RNAi or with the BET protein inhibitor JQ1 de-represses CncC transcriptional activity and engages protective gene expression programs. The mechanism by which Fs(1)h inhibits CncC function is distinct from the canonical mechanism that stimulates Nrf2 function by abrogating Keap1-dependent proteasomal degradation. Consistent with the independent modes of CncC regulation by Keap1 and Fs(1)h, combinations of drugs that can specifically target these pathways cause a strong synergistic and specific activation of protective CncC- dependent gene expression and boosts oxidative stress resistance. This synergism might be exploitable for the design of combinatorial therapies to target diseases associated with oxidative stress or inflammation. PMID:27233051

  13. Keap1-Independent Regulation of Nrf2 Activity by Protein Acetylation and a BET Bromodomain Protein.

    PubMed

    Chatterjee, Nirmalya; Tian, Min; Spirohn, Kerstin; Boutros, Michael; Bohmann, Dirk

    2016-05-01

    Mammalian BET proteins comprise a family of bromodomain-containing epigenetic regulators with complex functions in chromatin organization and gene regulation. We identified the sole member of the BET protein family in Drosophila, Fs(1)h, as an inhibitor of the stress responsive transcription factor CncC, the fly ortholog of Nrf2. Fs(1)h physically interacts with CncC in a manner that requires the function of its bromodomains and the acetylation of CncC. Treatment of cultured Drosophila cells or adult flies with fs(1)h RNAi or with the BET protein inhibitor JQ1 de-represses CncC transcriptional activity and engages protective gene expression programs. The mechanism by which Fs(1)h inhibits CncC function is distinct from the canonical mechanism that stimulates Nrf2 function by abrogating Keap1-dependent proteasomal degradation. Consistent with the independent modes of CncC regulation by Keap1 and Fs(1)h, combinations of drugs that can specifically target these pathways cause a strong synergistic and specific activation of protective CncC- dependent gene expression and boosts oxidative stress resistance. This synergism might be exploitable for the design of combinatorial therapies to target diseases associated with oxidative stress or inflammation. PMID:27233051

  14. Interaction of the protein C activation peptide with platelets.

    PubMed

    Martínez-Cruz, Ruth; Canseco, María Del S Pina; Lopez-Martínez, Jael; Cruz, Pedro A Hernández; Pérez-Campos, Eduardo; Cruz, Margarito Martínez; Alva, Félix Córdoba; Majluf-Cruz, Abraham; Zenteno, Edgar; Ruiz-Argüelles, Alejandro

    2007-01-01

    The peptide NH(2)-DTEDQEDQVDPR-COOH is released during activation of protein C zymogen. We measured the effect of a synthetic peptide with an amino acid sequence similar to that of the natural peptide on platelets from healthy individuals using platelet aggregometry. We found that this synthetic peptide inhibits platelet aggregation induced by thrombin; furthermore, it diminishes mobilization of intraplatelet calcium. Molecular docking showed weak interaction between the synthetic peptide and thrombin. Our findings suggest that this synthetic peptide may interact with a receptor located on the platelet cell membrane.

  15. Activity-related redistribution of presynaptic proteins at the active zone.

    PubMed

    Tao-Cheng, J-H

    2006-09-01

    Immunogold labeling distributions of seven presynaptic proteins were quantitatively analyzed under control conditions and after high K+ depolarization in excitatory synapses from dissociated rat hippocampal cultures. Three parallel zones in presynaptic terminals were sampled: zones I and II, each about one synaptic vesicle wide extending from the active zone; and zone III, containing a distal pool of vesicles up to 200 nm from the presynaptic membrane. The distributions of SV2 and synaptophysin, two synaptic vesicle integral membrane proteins, generally followed the distribution of synaptic vesicles, which were typically evenly distributed under control conditions and had a notable depletion in zone III after stimulation. Labels of synapsin I and synuclein, two synaptic vesicle-associated proteins, were similar to each other; both were particularly sparse in zone I under control conditions but showed a prominent enrichment toward the active zone, after stimulation. Labels of Bassoon, Piccolo and RIM 1, three active zone proteins, had very different distribution profiles from one another under control conditions. Bassoon was enriched in zone II, Piccolo and RIM 1 in zone I. After stimulation, Bassoon and Piccolo remained relatively unchanged, but RIM 1 redistributed with a significant decrease in zone I, and increases in zones II and III. These results demonstrate that Bassoon and Piccolo are stable components of the active zone while RIM 1, synapsin I and synuclein undergo dynamic redistribution with synaptic activity.

  16. The innervated anterolateral thigh flap: anatomical study and clinical implications.

    PubMed

    Ribuffo, Diego; Cigna, Emanuele; Gargano, Francesco; Spalvieri, Cristina; Scuderi, Nicolò

    2005-02-01

    During the past 20 years, the neural anatomy of many flaps has been investigated, although no extensive studies have been reported yet on the anterolateral thigh flap. The goal of this study was to describe the sensory territories of the nerves supplying the anterolateral thigh flap with dissections on fresh cadavers and with local anesthetic injections in living subjects. The sensate anterolateral thigh flap is typically described as innervated by the lateral cutaneous femoral nerve. Two other well-known nerves, the superior perforator nerve and the median perforator nerve, which enter the flap at its medial border, might have a role in anterolateral thigh flap innervation. Twenty-nine anterolateral thigh flaps were elevated in 15 cadavers, and the lateral cutaneous femoral nerve, the superior perforator nerve, and median perforator nerve were dissected. In the injection study, the lateral cutaneous femoral nerve, superior perforator nerve, and median perforator nerve in 16 thighs of eight subjects were sequentially blocked. The resulting sensory deficit from each injection was mapped on the skin and superimposed on the marked anterolateral thigh flap territory. The study shows that the sensate anterolateral thigh flap is basically innervated by all three nerves. The lateral cutaneous femoral nerve was present in 29 of 29 thighs, whereas the superior perforator nerve was present in 25 of 29 and the median perforator nerve in 24 of 29 thighs. Furthermore, in the proximal half of the flap, the lateral cutaneous femoral nerve lies deep, whereas the superior perforator nerve and median perforator nerve lie more superficially. Whereas the lateral cutaneous femoral nerve innervates the entire flap, the superior perforator nerve innervates 25 percent of the flap and the median perforator nerve innervates 60 percent of the flap. Clinically, a small anterolateral thigh flap (7 x 5 cm) can be raised sparing the lateral cutaneous femoral nerve and using only the selective

  17. Interaction of Ca(2+)-dependent activator protein for secretion 1 (CAPS1) with septin family proteins in mouse brain.

    PubMed

    Hosono, Mayu; Shinoda, Yo; Hirano, Touko; Ishizaki, Yasuki; Furuichi, Teiichi; Sadakata, Tetsushi

    2016-03-23

    The Ca(2+)-dependent activator protein for secretion 1 (CAPS1) protein plays a regulatory role in the dense-core vesicle exocytosis pathway. To clarify the functions of this protein in the brain, we searched for novel interaction partners of CAPS1 by mass spectrometry. We identified a specific interaction of CAPS1 with septin family proteins. We also demonstrated that the C-terminal region of the CAPS1 protein binds to part of the deduced GTP-binding domain of septin proteins. It is possible that a tertiary complex of septin, CAPS, and syntaxin contributes to dense-core vesicle trafficking and exocytosis in neurons.

  18. SIRT1 Suppresses Activator Protein-1 Transcriptional Activity and Cyclooxygenase-2 Expression in Macrophages*

    PubMed Central

    Zhang, Ran; Chen, Hou-Zao; Liu, Jin-Jing; Jia, Yu-Yan; Zhang, Zhu-Qin; Yang, Rui-Feng; Zhang, Yuan; Xu, Jing; Wei, Yu-Sheng; Liu, De-Pei; Liang, Chih-Chuan

    2010-01-01

    SIRT1 (Sirtuin type 1), a mammalian orthologue of yeast SIR2 (silent information regulator 2), has been shown to mediate a variety of calorie restriction (CR)-induced physiological events, such as cell fate regulation via deacetylation of the substrate proteins. However, whether SIRT1 deacetylates activator protein-1 (AP-1) to influence its transcriptional activity and target gene expression is still unknown. Here we demonstrate that SIRT1 directly interacts with the basic leucine zipper domains of c-Fos and c-Jun, the major components of AP-1, by which SIRT1 suppressed the transcriptional activity of AP-1. This process requires the deacetylase activity of SIRT1. Notably, SIRT1 reduced the expression of COX-2, a typical AP-1 target gene, and decreased prostaglandin E2 (PGE2) production of peritoneal macrophages (pMΦs). pMΦs with SIRT1 overexpression displayed improved phagocytosis and tumoricidal functions, which are associated with depressed PGE2. Furthermore, SIRT1 protein level was up-regulated in CR mouse pMΦs, whereas elevated SIRT1 decreased COX-2 expression and improved PGE2-related macrophage functions that were reversed following inhibition of SIRT1 deacetylase activity. Thus, our results indicate that SIRT1 may be a mediator of CR-induced macrophage regulation, and its deacetylase activity contributes to the inhibition of AP-1 transcriptional activity and COX-2 expression leading to amelioration of macrophage function. PMID:20042607

  19. Early events of human T lymphocyte activation are associated with type I protein kinase A activity.

    PubMed Central

    Laxminarayana, D; Berrada, A; Kammer, G M

    1993-01-01

    Human T lymphocytes possess both the type I and II isozymes of protein kinase A (PKA). The type I (PKA-I) isozyme is predominantly associated with the plasma membrane, whereas the type II (PKA-II) isozyme is primarily localized to the cytosol. Because the functions of both PKA-I and PKA-II isozymes in the biochemical events of T lymphocyte activation have not been clearly elucidated, we tested the hypothesis that very early events of normal human T lymphocyte activation are mediated by the PKA-I and/or PKA-II isozyme(s). Fresh normal human T cells and a normal human CD4+ T cell line (GK606) activated with anti-CD3-epsilon and recombinant interleukin 1 alpha (rIL-1 alpha) exhibited a peak six- to sevenfold increase of PKA phosphotransferase activity at 5 min that returned to baseline by 60 min. Similarly, both fresh T cells and the T cell line activated by phorbol myristate acetate and ionomycin demonstrated a peak eightfold increase of PKA activity by 15 min that returned toward baseline by 60 min. Chromatographic separation of the PKA isozymes and quantification of phosphotransferase activities after T cell activation by either agonist pair showed preferential activation of the PKA-I isozyme, resulting in a significant reduction in the ratio of PKA-I to PKA-II isozyme activity from 3.1:1-6.2:1 to 1.1:1-3.2:1. PKA-I isozyme activation resulted in the release of free catalytic (C) subunit, an increase in C subunit phosphotransferase activity, and the phosphorylation of T cell plasma membrane-associated proteins, p14, p17, p20, p21, p38, and p48. However, activation of the PKA-I isozyme did not appear to be required for the transcription of IL-2 mRNA, an event necessary for mitosis. These data indicate that ligand-induced T cell activation is associated with rapid activation of the PKA-I, but not PKA-II, isozyme that results in phosphorylation of plasma membrane-associated proteins. The involvement of the PKA-I isozyme during the very early events of T cell

  20. Reconstruction of pressure sores with perforator-based propeller flaps.

    PubMed

    Jakubietz, Rafael G; Jakubietz, Danni F; Zahn, Robert; Schmidt, Karsten; Meffert, Rainer H; Jakubietz, Michael G

    2011-03-01

    Perforator flaps have been successfully used for reconstruction of pressure sores. Although V-Y advancement flaps approximate debrided wound edges, perforator-based propeller flaps allow rotation of healthy tissue into the defect. Perforator-based propeller flaps were planned in 13 patients. Seven pressure sores were over the sacrum, five over the ischial tuberosity, and one on the tip of the scapula. Three patients were paraplegic, six were bedridden, and five were ambulatory. In three patients, no perforators were found. In 10 patients, propeller flaps were transferred. In two patients, total flap necrosis occurred, which was reconstructed with local advancement flaps. In two cases, a wound dehiscence occurred and had to be revised. One hematoma required evacuation. No further complications were noted. No recurrence at the flap site occurred. Local perforator flaps allow closure of pressure sores without harvesting muscle. The propeller version has the added benefit of transferring tissue from a distant site, avoiding reapproximation of original wound edges. Twisting of the pedicle may cause torsion and venous obstruction. This can be avoided by dissecting a pedicle of at least 3 cm. Propeller flaps are a safe option for soft tissue reconstruction of pressure sores.

  1. Thermally activated charge transport in microbial protein nanowires

    PubMed Central

    Lampa-Pastirk, Sanela; Veazey, Joshua P.; Walsh, Kathleen A.; Feliciano, Gustavo T.; Steidl, Rebecca J.; Tessmer, Stuart H.; Reguera, Gemma

    2016-01-01

    The bacterium Geobacter sulfurreducens requires the expression of conductive protein filaments or pili to respire extracellular electron acceptors such as iron oxides and uranium and to wire electroactive biofilms, but the contribution of the protein fiber to charge transport has remained elusive. Here we demonstrate efficient long-range charge transport along individual pili purified free of metal and redox organic cofactors at rates high enough to satisfy the respiratory rates of the cell. Carrier characteristics were within the orders reported for organic semiconductors (mobility) and inorganic nanowires (concentration), and resistivity was within the lower ranges reported for moderately doped silicon nanowires. However, the pilus conductance and the carrier mobility decreased when one of the tyrosines of the predicted axial multistep hopping path was replaced with an alanine. Furthermore, low temperature scanning tunneling microscopy demonstrated the thermal dependence of the differential conductance at the low voltages that operate in biological systems. The results thus provide evidence for thermally activated multistep hopping as the mechanism that allows Geobacter pili to function as protein nanowires between the cell and extracellular electron acceptors. PMID:27009596

  2. Thermally activated charge transport in microbial protein nanowires.

    PubMed

    Lampa-Pastirk, Sanela; Veazey, Joshua P; Walsh, Kathleen A; Feliciano, Gustavo T; Steidl, Rebecca J; Tessmer, Stuart H; Reguera, Gemma

    2016-01-01

    The bacterium Geobacter sulfurreducens requires the expression of conductive protein filaments or pili to respire extracellular electron acceptors such as iron oxides and uranium and to wire electroactive biofilms, but the contribution of the protein fiber to charge transport has remained elusive. Here we demonstrate efficient long-range charge transport along individual pili purified free of metal and redox organic cofactors at rates high enough to satisfy the respiratory rates of the cell. Carrier characteristics were within the orders reported for organic semiconductors (mobility) and inorganic nanowires (concentration), and resistivity was within the lower ranges reported for moderately doped silicon nanowires. However, the pilus conductance and the carrier mobility decreased when one of the tyrosines of the predicted axial multistep hopping path was replaced with an alanine. Furthermore, low temperature scanning tunneling microscopy demonstrated the thermal dependence of the differential conductance at the low voltages that operate in biological systems. The results thus provide evidence for thermally activated multistep hopping as the mechanism that allows Geobacter pili to function as protein nanowires between the cell and extracellular electron acceptors. PMID:27009596

  3. Perivascular fat, AMP-activated protein kinase and vascular diseases

    PubMed Central

    Almabrouk, T A M; Ewart, M A; Salt, I P; Kennedy, S

    2014-01-01

    Perivascular adipose tissue (PVAT) is an active endocrine and paracrine organ that modulates vascular function, with implications for the pathophysiology of cardiovascular disease (CVD). Adipocytes and stromal cells contained within PVAT produce mediators (adipokines, cytokines, reactive oxygen species and gaseous compounds) with a range of paracrine effects modulating vascular smooth muscle cell contraction, proliferation and migration. However, the modulatory effect of PVAT on the vascular system in diseases, such as obesity, hypertension and atherosclerosis, remains poorly characterized. AMP-activated protein kinase (AMPK) regulates adipocyte metabolism, adipose biology and vascular function, and hence may be a potential therapeutic target for metabolic disorders such as type 2 diabetes mellitus (T2DM) and the vascular complications associated with obesity and T2DM. The role of AMPK in PVAT or the actions of PVAT have yet to be established, however. Activation of AMPK by pharmacological agents, such as metformin and thiazolidinediones, may modulate the activity of PVAT surrounding blood vessels and thereby contribute to their beneficial effect in cardiometabolic diseases. This review will provide a current perspective on how PVAT may influence vascular function via AMPK. We will also attempt to demonstrate how modulating AMPK activity using pharmacological agents could be exploited therapeutically to treat cardiometabolic diseases. PMID:24490856

  4. Comparative Analysis of Protein Tyrosine Phosphatases Regulating Microglial Activation

    PubMed Central

    Song, Gyun Jee; Kim, Jaehong; Kim, Jong-Heon; Song, Seungeun; Park, Hana; Zhang, Zhong-Yin

    2016-01-01

    Protein tyrosine phosphatases (PTPs) are key regulatory factors in inflammatory signaling pathways. Although PTPs have been extensively studied, little is known about their role in neuroinflammation. In the present study, we examined the expression of 6 different PTPs (PTP1B, TC-PTP, SHP2, MEG2, LYP, and RPTPβ) and their role in glial activation and neuroinflammation. All PTPs were expressed in brain and glia. The expression of PTP1B, SHP2, and LYP was enhanced in the inflamed brain. The expression of PTP1B, TC-PTP, and LYP was increased after treating microglia cells with lipopolysaccharide (LPS). To examine the role of PTPs in microglial activation and neuroinflammation, we used specific pharmacological inhibitors of PTPs. Inhibition of PTP1B, TC-PTP, SHP2, LYP, and RPTPβ suppressed nitric oxide production in LPS-treated microglial cells in a dose-dependent manner. Furthermore, intracerebroventricular injection of PTP1B, TC-PTP, SHP2, and RPTPβ inhibitors downregulated microglial activation in an LPS-induced neuroinflammation model. Our results indicate that multiple PTPs are involved in regulating microglial activation and neuroinflammation, with different expression patterns and specific functions. Thus, PTP inhibitors can be exploited for therapeutic modulation of microglial activation in neuroinflammatory diseases. PMID:27790059

  5. Physical activity and high-sensitivity C-reactive protein.

    PubMed

    Plaisance, Eric P; Grandjean, Peter W

    2006-01-01

    Cardiovascular disease (CVD) remains one of the leading causes of death and disability in developed countries around the world despite the documented success of lifestyle and pharmacological interventions. This illustrates the multifactorial nature of atherosclerosis and the use of novel inflammatory markers as an adjunct to risk factor reduction strategies. As evidence continues to accumulate that inflammation is involved in all stages of the development and progression of atherosclerosis, markers of inflammation such as high-sensitivity C-reactive protein (CRP) may provide additional information regarding the biological status of the atherosclerotic lesion. Recent investigations suggest that physical activity reduces CRP levels. Higher levels of physical activity and cardiorespiratory fitness are consistently associated with 6-35% lower CRP levels. Longitudinal training studies that have demonstrated reductions in CRP concentrations range from 16% to 41%, an effect that may be independent of baseline levels of CRP, body composition or weight loss. The average change in CRP associated with physical activity appears to be at least as good, if not better, than currently prescribed pharmacological interventions in similar populations. The primary purpose of this review will be to present evidence from both cross-sectional and longitudinal investigations that physical activity lowers CRP levels in a dose-response manner. Finally, this review will examine factors such as body composition, sex, blood sample timing, diet and smoking, which may influence the CRP response to physical activity. PMID:16646631

  6. Effects of Bezafibrate on the Survival of Random Skin Flaps in Rats.

    PubMed

    Lin, Bin; Lin, Yuting; Lin, Dingsheng; Cao, Bin

    2016-06-01

    Background Bezafibrate is widely used in clinics for its comparable angiogenic effect. Our research is to investigate the effect of bezafibrate on random skin flap survival. Materials and Methods The "McFarlane flap" rat models were established in 30 male Sprague-Dawley rats which were divided into two groups. The treatment group was given bezafibrate (400 mg/kg/day; gavage administration), and the control group received the vehicle. The flap surviving area was measured after 7 days, and the tissue samples were taken for histological analysis and edema measurement. Vascular endothelial growth factor (VEGF) was determined using immunohistochemical methods. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were examined with kits. Results Seven days after the operation, the surviving area in the treatment group was larger than in the control group (p < 0.01). The expression of VEGF was increased in the treatment group compared with that in the control group. And the activity of SOD was lower in the treatment group compared with those in the control group (p < 0.01). However, tissue edema, inflammatory cell infiltration, and MDA level were markedly lower in the treatment group than those in the control group (p < 0.01). Conclusion Bezafibrate improves the survival of random skin flaps effectively. PMID:26872027

  7. Structure and Activity of Tryptophan-rich TSPO Translocator Proteins

    PubMed Central

    Guo, Youzhong; Kalathur, Ravi C.; Liu, Qun; Kloss, Brian; Bruni, Renato; Ginter, Christopher; Kloppmann, Edda; Rost, Burkhard; Hendrickson, Wayne A.

    2015-01-01

    TSPO translocator proteins bind steroids and porphyrins, and they are implicated in many human diseases, for which they serve as biomarkers and therapeutic targets. TSPOs have tryptophan-rich sequences that are fhighly conserved from bacteria to mammals. We report crystal structures for Bacillus cereus TSPO (BcTSPO) down to 1.7Å resolution, including a complex with the benzodiazepine-like inhibitor PK11195. We also describe BcTSPO-mediated protoporphyrin IX (PpIX) reactions, including catalytic degradation to a previously undescribed heme derivative. We used structure-inspired mutations to investigate reaction mechanisms, and we showed that TSPOs from Xenopus and man have similar PpIX-directed activities. Although TSPOs have been regarded as transporters, the catalytic activity in PpIX degradation suggests physiological importance for TSPOs in protection against oxidative stress. PMID:25635100

  8. TALE proteins bind to both active and inactive chromatin.

    PubMed

    Scott, James N F; Kupinski, Adam P; Kirkham, Christopher M; Tuma, Roman; Boyes, Joan

    2014-02-15

    TALE (transcription activator-like effector) proteins can be tailored to bind to any DNA sequence of choice and thus are of immense utility for genome editing and the specific delivery of transcription activators. However, to perform these functions, they need to occupy their sites in chromatin. In the present study, we have systematically assessed TALE binding to chromatin substrates and find that in vitro TALEs bind to their target site on nucleosomes at the more accessible entry/exit sites, but not at the nucleosome dyad. We show further that in vivo TALEs bind to transcriptionally repressed chromatin and that transcription increases binding by only 2-fold. These data therefore imply that TALEs are likely to bind to their target in vivo even at inactive loci.

  9. Small Molecule Inhibitors Targeting Activator Protein 1 (AP-1)

    PubMed Central

    2015-01-01

    Activator protein 1 (AP-1) is a pivotal transcription factor that regulates a wide range of cellular processes including proliferation, apoptosis, differentiation, survival, cell migration, and transformation. Accumulating evidence supports that AP-1 plays an important role in several severe disorders including cancer, fibrosis, and organ injury, as well as inflammatory disorders such as asthma, psoriasis, and rheumatoid arthritis. AP-1 has emerged as an actively pursued drug discovery target over the past decade. Excitingly, a selective AP-1 inhibitor T-5224 (51) has been investigated in phase II human clinical trials. Nevertheless, no effective AP-1 inhibitors have yet been approved for clinical use. Despite significant advances achieved in understanding AP-1 biology and function, as well as the identification of small molecules modulating AP-1 associated signaling pathways, medicinal chemistry efforts remain an urgent need to yield selective and efficacious AP-1 inhibitors as a viable therapeutic strategy for human diseases. PMID:24831826

  10. Enhancing the Activity of a Protein by Stereospecific Unfolding

    PubMed Central

    Hua, Qing-xin; Xu, Bin; Huang, Kun; Hu, Shi-Quan; Nakagawa, Satoe; Jia, Wenhua; Wang, Shuhua; Whittaker, Jonathan; Katsoyannis, Panayotis G.; Weiss, Michael A.

    2009-01-01

    A central tenet of molecular biology holds that the function of a protein is mediated by its structure. An inactive ground-state conformation may nonetheless be enjoined by the interplay of competing biological constraints. A model is provided by insulin, well characterized at atomic resolution by x-ray crystallography. Here, we demonstrate that the activity of the hormone is enhanced by stereospecific unfolding of a conserved structural element. A bifunctional β-strand mediates both self-assembly (within β-cell storage vesicles) and receptor binding (in the bloodstream). This strand is anchored by an invariant side chain (PheB24); its substitution by Ala leads to an unstable but native-like analog of low activity. Substitution by d-Ala is equally destabilizing, and yet the protein diastereomer exhibits enhanced activity with segmental unfolding of the β-strand. Corresponding photoactivable derivatives (containing l- or d-para-azido-Phe) cross-link to the insulin receptor with higher d-specific efficiency. Aberrant exposure of hydrophobic surfaces in the analogs is associated with accelerated fibrillation, a form of aggregation-coupled misfolding associated with cellular toxicity. Conservation of PheB24, enforced by its dual role in native self-assembly and induced fit, thus highlights the implicit role of misfolding as an evolutionary constraint. Whereas classical crystal structures of insulin depict its storage form, signaling requires engagement of a detachable arm at an extended receptor interface. Because this active conformation resembles an amyloidogenic intermediate, we envisage that induced fit and self-assembly represent complementary molecular adaptations to potential proteotoxicity. The cryptic threat of misfolding poses a universal constraint in the evolution of polypeptide sequences. PMID:19321436

  11. Wake Vortex Alleviation Using Rapidly Actuated Segmented Gurney Flaps

    NASA Astrophysics Data System (ADS)

    Matalanis, Claude; Eaton, John

    2006-11-01

    A study to assess the potential for using rapidly actuated segmented Gurney flaps, also known as Miniature Trailing Edge Effectors (MiTEs), for active wake vortex alleviation is conducted using a half-span model wing with NACA 0012 shape and an aspect ratio of 4.1. All tests are performed with the wing at an 8.9 degree angle of attack and chord based Reynolds number around 350,000. The wing is equipped with an array of 13 MiTE pairs. Each MiTE has a flap that in the neutral position rests behind the blunt trailing edge of the wing, and in the down position extends 0.015 chord lengths perpendicular to the freestream on the pressure side of the wing. Dynamic PIV is used to measure the time dependent response of the vortex in the intermediate wake to various MiTE actuation schemes that deflect the vortex in both the spanwise and liftwise directions. A maximum spanwise deflection of 0.041 chord lengths is possible while nearly conserving lift. These intermediate wake results as well as pressure profile, five-hole probe, and static PIV measurements are used to form complete, experimentally-based initial conditions for vortex filament computations that are used to compute the far wake evolution. Results from these computations show that the perturbations created by MiTEs can be used to excite vortex instability.

  12. Inhibition of Type III Interferon Activity by Orthopoxvirus Immunomodulatory Proteins

    PubMed Central

    2010-01-01

    The type III interferon (IFN) family elicits an antiviral response that is nearly identical to that evoked by IFN-α/β. However, these cytokines (known as IFN-λ1, 2, and 3) signal through a distinct receptor, and thus may be resistant to the evasion strategies used by some viruses to avoid the IFN-α/β response. Orthopoxviruses are highly resistant to IFN-α/β because they encode well-characterized immunomodulatory proteins that inhibit IFN activity. These include a secreted receptor (B18R) that neutralizes IFN-α/β, and a cytoplasmic protein (E3L) that blocks IFN-α/β effector functions in infected cells. We therefore determined the ability of these immunomodulators to abrogate the IFN-λ–induced antiviral response. We found that (i) vaccinia virus (VACV) replication is resistant to IFN-λ antiviral activity; (ii) neither VACV B18R nor the variola virus homolog B20R neutralizes IFN-λ; (iii) VACV E3L inhibits the IFN-λ–mediated antiviral response through a PKR-dependent pathway; (iv) VACV infection inhibits IFN-λR–mediated signal transduction and gene expression. These results demonstrate differential sensitivity of IFN-λ to multiple distinct evasion mechanisms employed by a single virus. PMID:20038204

  13. Proteins with RNA Chaperone Activity: A World of Diverse Proteins with a Common Task—Impediment of RNA Misfolding

    PubMed Central

    Semrad, Katharina

    2011-01-01

    Proteins with RNA chaperone activity are ubiquitous proteins that play important roles in cellular mechanisms. They prevent RNA from misfolding by loosening misfolded structures without ATP consumption. RNA chaperone activity is studied in vitro and in vivo using oligonucleotide- or ribozyme-based assays. Due to their functional as well as structural diversity, a common chaperoning mechanism or universal motif has not yet been identified. A growing database of proteins with RNA chaperone activity has been established based on evaluation of chaperone activity via the described assays. Although the exact mechanism is not yet understood, it is more and more believed that disordered regions within proteins play an important role. This possible mechanism and which proteins were found to possess RNA chaperone activity are discussed here. PMID:21234377

  14. Biodegradable nanoparticles for protein delivery: analysis of preparation conditions on particle morphology and protein loading, activity and sustained release properties.

    PubMed

    Coleman, Jason; Lowman, Anthony

    2012-01-01

    PLGA particles have been extensively used as a sustained drug-delivery system, but there are multiple drawbacks when delivering proteins. The focus of this work is to address the most significant disadvantages to the W/O/W double emulsion procedure and demonstrate that simple changes to this procedure can have significant changes to particle size and dispersity and considerable improvements to protein loading, activity and sustained active protein release. A systematic approach was taken to analyze the effects of the following variables: solvent miscibility (dichloromethane (DCM), ethyl acetate, acetone), homogenization speed (10 000-25 000 rpm), PLGA concentration (10-30 mg/ml) and additives in both the organic (sucrose acetate isobutyrate (SAIB)) and aqueous (bovine serum albumin (BSA)) phases. Increasing solvent miscibility decreased particle size, dispersity and protein denaturation, while maintaining adequate protein loading. Increasing solvent miscibility also lowered the impact of homogenization on particle size and dispersity and protein activity. Changes to PLGA concentration demonstrated a minimum impact on particle size and dispersity, but showed an inverse relationship between protein encapsulation efficiency and particle protein weight percent. Most particles tested provided sustained release of active protein over 60 days. Increasing solvent miscibility resulted in increases in the percent of active protein released. When subjected to synthesis conditions with DCM as the solvent, BSA as a stabilizer resulted in the maximum stabilization of protein at a concentration of 100 mg/ml. At this concentration, BSA allowed for increases in the total amount of active protein delivered for all three solvents. The benefit of SAIB was primarily increased protein loading.

  15. Aerodynamic flight performance in flap-gliding birds and bats.

    PubMed

    Muijres, Florian T; Henningsson, Per; Stuiver, Melanie; Hedenström, Anders

    2012-08-01

    Many birds use a flight mode called undulating or flap-gliding flight, where they alternate between flapping and gliding phases, while only a few bats make use of such a flight mode. Among birds, flap-gliding is commonly used by medium to large species, where it is regarded to have a lower energetic cost than continuously flapping flight. Here, we introduce a novel model for estimating the energetic flight economy of flap-gliding animals, by determining the lift-to-drag ratio for flap-gliding based on empirical lift-to-drag ratio estimates for continuous flapping flight and for continuous gliding flight, respectively. We apply the model to flight performance data of the common swift (Apus apus) and of the lesser long-nosed bat (Leptonycteris yerbabuenae). The common swift is a typical flap-glider while-to the best of our knowledge-the lesser long-nosed bat does not use flap-gliding. The results show that, according to the model, the flap-gliding common swift saves up to 15% energy compared to a continuous flapping swift, and that this is primarily due to the exceptionally high lift-to-drag ratio in gliding flight relative to that in flapping flight for common swifts. The lesser long-nosed bat, on the other hand, seems not to be able to reduce energetic costs by flap-gliding. The difference in relative costs of flap-gliding flight between the common swift and the lesser long-nosed bat can be explained by differences in morphology, flight style and wake dynamics. The model presented here proves to be a valuable tool for estimating energetic flight economy in flap-gliding animals. The results show that flap-gliding flight that is naturally used by common swifts is indeed the most economic one of the two flight modes, while this is not the case for the non-flap-gliding lesser long-nosed bat. PMID:22726811

  16. Protein tyrosine kinase and mitogen-activated protein kinase signalling pathways contribute to differences in heterophil-mediated innate immune responsiveness between two lines of broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein tyrosine phosphorylation mediates signal transduction of cellular processes, with protein tyrosine kinases (PTKs) regulating virtually all signaling events. The mitogen-activated protein kinase (MAPK) super-family consists of three conserved pathways that convert receptor activation into ce...

  17. PPAR-β/δ activation promotes phospholipid transfer protein expression.

    PubMed

    Chehaibi, Khouloud; Cedó, Lídia; Metso, Jari; Palomer, Xavier; Santos, David; Quesada, Helena; Naceur Slimane, Mohamed; Wahli, Walter; Julve, Josep; Vázquez-Carrera, Manuel; Jauhiainen, Matti; Blanco-Vaca, Francisco; Escolà-Gil, Joan Carles

    2015-03-15

    The peroxisome proliferator-activated receptor (PPAR)-β/δ has emerged as a promising therapeutic target for treating dyslipidemia, including beneficial effects on HDL cholesterol (HDL-C). In the current study, we determined the effects of the PPAR-β/δ agonist GW0742 on HDL composition and the expression of liver HDL-related genes in mice and cultured human cells. The experiments were carried out in C57BL/6 wild-type, LDL receptor (LDLR)-deficient mice and PPAR-β/δ-deficient mice treated with GW0742 (10mg/kg/day) or a vehicle solution for 14 days. GW0742 upregulated liver phospholipid transfer protein (Pltp) gene expression and increased serum PLTP activity in mice. When given to wild-type mice, GW0742 significantly increased serum HDL-C and HDL phospholipids; GW0742 also raised serum potential to generate preβ-HDL formation. The GW0742-mediated effects on liver Pltp expression and serum enzyme activity were completely abolished in PPAR-β/δ-deficient mice. GW0742 also stimulated PLTP mRNA expression in mouse J774 macrophages, differentiated human THP-1 macrophages and human hepatoma Huh7. Collectively, our findings demonstrate a common transcriptional upregulation by GW0742-activated PPAR-β/δ of Pltp expression in cultured cells and in mouse liver resulting in enhanced serum PLTP activity. Our results also indicate that PPAR-β/δ activation may modulate PLTP-mediated preβ-HDL formation and macrophage cholesterol efflux.

  18. First web-space reconstruction by the anterolateral thigh flap.

    PubMed

    Adani, Roberto; Tarallo, Luigi; Marcoccio, Ignazio; Fregni, Umberto

    2006-04-01

    Four patients with severe contracture of the first web space were treated with an anterolateral thigh perforator flap. The flap size ranged from 10 to 13 cm in length and from 7 to 8 cm in width. The donor site was closed directly and thinning of the flap was performed in all cases. All flaps survived and there were no re-explorations. Web space opening was maintained over the follow-up period. There was an average postoperative increase of the angle of the first web space of 61 degrees. The thinned anterolateral thigh flap provides a pliable vascularized tissue for resurfacing the skin after release of severe contracture of the first web space and represents a reliable alternative to other flaps.

  19. Incidence of flap procedures in the management of burn patients.

    PubMed

    Lineaweaver, William C; Craft-Coffman, Beretta; Oswald, Tanya M

    2015-03-01

    Increased survival of burn patients presents opportunities for reconstructive strategies to improve outcomes in management of acute and secondary burn injuries. To assess one such strategy, namely flap reconstruction, we reviewed cases performed during the first 4.5 years of the JMS Burn and Reconstruction Center. We found that flap procedures accounted for 0.8% of acute cases (23 of 2723 procedures) and 33% of secondary cases (260 of 790 procedures). This initial finding shows that in this practice flap procedures are applied to a small number of acute problems while flap procedures comprise 33% of secondary procedures. Reconstructive flap surgery plays a measurable role in burn treatment at this center. Further study of outcomes and timing could lead to better understanding of optimal strategies for flap reconstruction in burns.

  20. Changes in quantitative norepinephrine levels in delayed pig flank flaps.

    PubMed

    Cutting, C; Bumsted, R; Bardach, J; Mooney, M; Johnson, S

    1982-04-01

    A quantitative norepinephrine assay was used to follow tissue norepinephrine concentrations serially in bipedicle delayed pig flank flaps. In contrast to a previous study by Palmer, norepinephrine concentration decreased significantly after 24 hr, but then gradually returned to normal at 10 days. This suggests that the pig flank flap maintains much of its adrenergic innervation following bipedicle delay. It appears unlikely that adrenergic denervation explains the delay phenomenon in this model. The combined results of this study and others suggest that the degree of adrenergic denervation of a flap is determined by the anatomic layout of the flap with respect to the underlying cutaneous adrenergic neural anatomy. This experiment suggests that the effectiveness of adrenergic blocking agents in improving flap survival is dependent on the degree of adrenergic denervation of the flap.

  1. Versatility of V-Y Flap in Gluteal Area

    PubMed Central

    El-Sabbagh, Ahmed

    2016-01-01

    BACKGROUND Lesions in perineal area are common. The usual treatment is coverage by skin graft and flaps which may be local, pedicled or free types. In this paper V-Y flaps were used to cover the defects. METHODS V-Y flaps were used in the gluteal area in 15 patients. Lesions were due to different causes. RESULTS Of 15 patients, 11 were males and 4 were females. Their ages ranged from 3 weeks to 52 years old. Two cases were due to myelomeningocele, necrotizing fasciitis (2 cases) and the rest were due to bed sores (11 cases). The flaps were located over the trochanter (3 cases), ischial (6 cases) and sacral (6 cases). Good healing and durable coverage were obtained in all cases except one case. CONCLUSION V-Y flap as a working horse flap is recommended in the gluteal area. PMID:27579271

  2. A Variable Light Domain Fluorogen Activating Protein Homodimerizes To Activate Dimethylindole Red

    SciTech Connect

    Senutovitch, Nina; Stanfield, Robyn L.; Bhattacharyya, Shantanu; Rule, Gordon S.; Wilson, Ian A.; Armitage, Bruce A.; Waggoner, Alan S.; Berget, Peter B.

    2012-07-11

    Novel fluorescent tools such as green fluorescent protein analogues and fluorogen activating proteins (FAPs) are useful in biological imaging for tracking protein dynamics in real time with a low fluorescence background. FAPs are single-chain variable fragments (scFvs) selected from a yeast surface display library that produce fluorescence upon binding a specific dye or fluorogen that is normally not fluorescent when present in solution. FAPs generally consist of human immunoglobulin variable heavy (V{sub H}) and variable light (V{sub L}) domains covalently attached via a glycine- and serine-rich linker. Previously, we determined that the yeast surface clone, V{sub H}-V{sub L} M8, could bind and activate the fluorogen dimethylindole red (DIR) but that the fluorogen activation properties were localized to the M8V{sub L} domain. We report here that both nuclear magnetic resonance and X-ray diffraction methods indicate the M8V{sub L} forms noncovalent, antiparallel homodimers that are the fluorogen activating species. The M8V{sub L} homodimers activate DIR by restriction of internal rotation of the bound dye. These structural results, together with directed evolution experiments with both V{sub H}-V{sub L} M8 and M8V{sub L}, led us to rationally design tandem, covalent homodimers of M8V{sub L} domains joined by a flexible linker that have a high affinity for DIR and good quantum yields.

  3. Sustained mitogen-activated protein kinase activation reprograms defense metabolism and phosphoprotein profile in Arabidopsis thaliana

    PubMed Central

    Lassowskat, Ines; Böttcher, Christoph; Eschen-Lippold, Lennart; Scheel, Dierk; Lee, Justin

    2014-01-01

    Mitogen-activated protein kinases (MAPKs) target a variety of protein substrates to regulate cellular signaling processes in eukaryotes. In plants, the number of identified MAPK substrates that control plant defense responses is still limited. Here, we generated transgenic Arabidopsis thaliana plants with an inducible system to simulate in vivo activation of two stress-activated MAPKs, MPK3, and MPK6. Metabolome analysis revealed that this artificial MPK3/6 activation (without any exposure to pathogens or other stresses) is sufficient to drive the production of major defense-related metabolites, including various camalexin, indole glucosinolate and agmatine derivatives. An accompanying (phospho)proteome analysis led to detection of hundreds of potential phosphoproteins downstream of MPK3/6 activation. Besides known MAPK substrates, many candidates on this list possess typical MAPK-targeted phosphosites and in many cases, the corresponding phosphopeptides were detected by mass spectrometry. Notably, several of these putative phosphoproteins have been reported to be associated with the biosynthesis of antimicrobial defense substances (e.g., WRKY transcription factors and proteins encoded by the genes from the “PEN” pathway required for penetration resistance to filamentous pathogens). Thus, this work provides an inventory of candidate phosphoproteins, including putative direct MAPK substrates, for future analysis of MAPK-mediated defense control. (Proteomics data are available with the identifier PXD001252 via ProteomeXchange, http://proteomecentral.proteomexchange.org). PMID:25368622

  4. Targeted Mutagenesis and Combinatorial Library Screening Enables Control of Protein Orientation on Surfaces and Increased Activity of Adsorbed Proteins.

    PubMed

    Cruz-Teran, Carlos A; Carlin, Kevin B; Efimenko, Kirill; Genzer, Jan; Rao, Balaji M

    2016-08-30

    While nonspecific adsorption is widely used for immobilizing proteins on solid surfaces, the random nature of protein adsorption may reduce the activity of immobilized proteins due to occlusion of the active site. We hypothesized that the orientation a protein assumes on a given surface can be controlled by systematically introducing mutations into a region distant from its active site, thereby retaining activity of the immobilized protein. To test this hypothesis, we generated a combinatorial protein library by randomizing six targeted residues in a binding protein derived from highly stable, nonimmunoglobulin Sso7d scaffold; mutations were targeted in a region that is distant from the binding site. This library was screened to isolate binders that retain binding to its cognate target (chicken immunoglobulin Y, cIgY) as well as exhibit adsorption on unmodified silica at pH 7.4 and high ionic strength conditions. A single mutant, Sso7d-2B5, was selected for further characterization. Sso7d-2B5 retained binding to cIgY with an apparent dissociation constant similar to that of the parent protein; both mutant and parent proteins saturated the surface of silica with similar densities. Strikingly, however, silica beads coated with Sso7d-2B5 could achieve up to 7-fold higher capture of cIgY than beads coated with the parent protein. These results strongly suggest that mutations introduced in Sso7d-2B5 alter its orientation relative to the parent protein, when adsorbed on silica surfaces. Our approach also provides a generalizable strategy for introducing mutations in proteins so as to improve their activity upon immobilization, and has direct relevance to development of protein-based biosensors and biocatalysts. PMID:27490089

  5. Stimulation of Periodontal Ligament Stem Cells by Dentin Matrix Protein 1 Activates Mitogen-Activated Protein Kinase and Osteoblast Differentiation

    PubMed Central

    Chandrasekaran, Sangeetha; Ramachandran, Amsaveni; Eapen, Asha; George, Anne

    2013-01-01

    Background Periodontitis can ultimately result in tooth loss. Many natural and synthetic materials have been tried to achieve periodontal regeneration, but the results remain variable and unpredictable. We hypothesized that exogenous treatment with dentin matrix protein 1 (DMP1) activates specific genes and results in phenotypic and functional changes in human periodontal ligament stem cells (hPDLSCs). Methods hPDLSCs were isolated from extracted teeth and cultured in the presence or absence of DMP1. Quantitative polymerase chain reactions were performed to analyze the expression of several genes involved in periodontal regeneration. hPDLSCs were also processed for immunocytochemical and Western blot analysis using phosphorylated extracellular signal-regulated kinase (pERK) and ERK antibodies. Alkaline phosphatase and von Kossa staining were performed to characterize the differentiation of hPDLSCs into osteoblasts. Field emission scanning electron microscopic analysis of the treated and control cell cultures were also performed. Results Treatment with DMP1 resulted in the upregulation of genes, such as matrix metalloproteinase-2, alkaline phosphatase, and transforming growth factor β1. Activation of ERK mitogen-activated protein kinase signaling pathway and translocation of pERK from the cytoplasm to the nucleus was observed. Overall, DMP1-treated cells showed increased expression of alkaline phosphatase, increased matrix, and mineralized nodule formation when compared with untreated controls. Conclusion DMP1 can orchestrate a coordinated expression of genes and phenotypic changes in hPDLSCs by activation of the ERK signaling pathway, which may provide a valuable strategy for tissue engineering approaches in periodontal regeneration. PMID:22612367

  6. Leishmania amazonensis: PKC-like protein kinase modulates the (Na++K+)ATPase activity.

    PubMed

    Almeida-Amaral, Elmo Eduardo de; Caruso-Neves, Celso; Lara, Lucienne Silva; Pinheiro, Carla Mônica; Meyer-Fernandes, José Roberto

    2007-08-01

    The present study aimed to identify the presence of protein kinase C-like (PKC-like) in Leishmania amazonensis and to elucidate its possible role in the modulation of the (Na(+)+K(+))ATPase activity. Immunoblotting experiments using antibody against a consensus sequence (Ac 543-549) of rabbit protein kinase C (PKC) revealed the presence of a protein kinase of 80 kDa in L. amazonensis. Measurements of protein kinase activity showed the presence of both (Ca(2+)-dependent) and (Ca(2+)-independent) protein kinase activity in plasma membrane and cytosol. Phorbol ester (PMA) activation of the Ca(2+)-dependent protein kinase stimulated the (Na(+)+K(+))ATPase activity, while activation of the Ca(2+)-independent protein kinase was inhibitory. Both effects of protein kinase on the (Na(+)+K(+))ATPase of the plasma membrane were lower than that observed in intact cells. PMA induced the translocation of protein kinase from cytosol to plasma membrane, indicating that the maximal effect of protein kinase on the (Na(+)+K(+))ATPase activity depends on the synergistic action of protein kinases from both plasma membrane and cytosol. This is the first demonstration of a protein kinase activated by PMA in L. amazonensis and the first evidence for a possible role in the regulation of the (Na(+)+K(+))ATPase activity in this trypanosomatid. Modulation of the (Na(+)+K(+))ATPase by protein kinase in a trypanosomatid opens up new possibilities to understand the regulation of ion homeostasis in this parasite. PMID:17475255

  7. [Fundamentals and principles of grafts and flaps].

    PubMed

    Cruz-Navarro, Natalio; León-Dueñas, Eduardo

    2014-01-01

    Reconstructive surgery of large urethral stenosis and the management of congenital anomalies such as hypospadias and epispadias require covering large cutaneous and mucosal defects with different techniques. The objective of this work is to define the main differences between tissues to be transferred and to study the principles that must govern the management of the various flaps and grafts used for these techniques. We analyze the anatomical and physiological features that may be key to understand the success and possible failures of these procedures, and we review technical details that must accompany in every case, not only during the operation, but also during the preoperative and postoperative period. We conclude stating that grafts (mainly oral and preputial mucosa) and flaps are increasingly used for the repair of urethral stenosis. Grafts must be prepared adequately in the back table and thinned to the maximum, and also be fixed properly, to guarantee their immobility until neovascularization is assured.

  8. Forward speed effects on blown flap noise

    NASA Technical Reports Server (NTRS)

    Pennock, A. P.

    1977-01-01

    The effects of forward speed on the noise of under-the-wing (externally blown flaps, EBF) and over-the-wing (upper surface blown, USB) blown flap configurations were measured in wind tunnel model tests with cold jets. The results are presented without correction for the effects (e.g., signal convection, shear layer refraction) associated with flight simulation in a wind tunnel or free jet facility. Noise decreases were generally observed at microphones forward of the wing. The reductions were larger at the low frequencies (below peak SPL) than at the high (above peak SPL). Noise increases of 10 dB or more were observed at the aft microphones, especially in the high frequency range.

  9. Aerosound from corner flow and flap flow

    NASA Technical Reports Server (NTRS)

    Meecham, W. C.

    1982-01-01

    Noise generation at the edge of a wing flap is analyzed. The phenomenon as a single vortex moving around a corner in an incompressible, potential flow is modelled. Vortex image retarding effects are proposed as an explanation for small Strouhal numbers. The model surface pressures, sound pressures (using Curle's theory), and Mach number dependencies agree with wind tunnel experiments. A double pressure peak is found in the model (credited to image action) which is qualitatively similar to measured sound correlations. Incompressible flow aerosound calculations are discussed. The effects of a series of vortices moving in the same idealized potential flow are also studied. The vortices are assumed to be statistically independent so their intensities can be added. The frequency of appearance of the vortices are determined from measurements. Diffraction effects caused by the presence of the wing near the dipole sound radiators on the flap surfaces are included.

  10. Combination of MS protein identification and bioassay of chromatographic fractions to identify biologically active substances from complex protein sources.

    PubMed

    Kuromitsu, Sadao; Yokota, Hiroyuki; Hiramoto, Masashi; Yuri, Masatoshi; Naitou, Masanori; Nakamura, Naoto; Kawabata, Shigeki; Kobori, Masato; Katoh, Masao; Furuchi, Kiyoshi; Mita, Haruhisa; Yamada, Tetsuo

    2009-06-01

    Purification of biologically active proteins from complex biological sources is a difficult task, usually requiring large amounts of sample and many separation steps. We found an active substance in a serum response element-dependent luciferase reporter gene bioassay in interstitial cystitis urine that we attempted to purify with column chromatography and the bioassay. With anion-exchange Mono Q and C4 reversed-phase columns, apparently sharp active peaks were obtained. However, more than 20 kinds of proteins were identified from the active fractions with MS, indicating that the purification was not complete. As further purification was difficult, we chose a candidate molecule by means of studying the correlation between MS protein identification scores and bioassay responses of chromatographic fractions near the active peaks. As a result, epidermal growth factor (EGF) was nominated as a candidate molecule among the identified proteins because the elution profile of EGF was consistent with that of the bioassay, and the correlation coefficient of EGF between MS protein identification scores and bioassay responses was the highest among all the identified proteins. With recombinant EGF and anti-EGF and anti-EGF receptor antibodies, EGF was confirmed to be the desired substance in interstitial cystitis urine. This approach required only 20 ml of urine sample and two column chromatographic steps. The combination of MS protein identification and bioassay of chromatographic fractions may be useful for identifying biologically active substances from complex protein sources.

  11. Extracellular protein analysis of activated sludge and their functions in wastewater treatment plant by shotgun proteomics

    PubMed Central

    Zhang, Peng; Shen, Yu; Guo, Jin-Song; Li, Chun; Wang, Han; Chen, You-Peng; Yan, Peng; Yang, Ji-Xiang; Fang, Fang

    2015-01-01

    In this work, proteins in extracellular polymeric substances extracted from anaerobic, anoxic and aerobic sludges of wastewater treatment plant (WWTP) were analyzed to probe their origins and functions. Extracellular proteins in WWTP sludges were identified using shotgun proteomics, and 130, 108 and 114 proteins in anaerobic, anoxic and aerobic samples were classified, respectively. Most proteins originated from cell and cell part, and their most major molecular functions were catalytic activity and binding activity. The results exhibited that the main roles of extracellular proteins in activated sludges were multivalence cations and organic molecules binding, as well as in catalysis and degradation. The catalytic activity proteins were more widespread in anaerobic sludge compared with those in anoxic and aerobic sludges. The structure difference between anaerobic and aerobic sludges could be associated with their catalytic activities proteins. The results also put forward a relation between the macro characteristics of activated sludges and micro functions of extracellular proteins in biological wastewater treatment process. PMID:26160685

  12. [Pedicled versus free TRAM flap for breast reconstruction].

    PubMed

    Galla, T J; Lukas, B; Feller, A M

    1999-03-01

    In breast reconstruction, the free TRAM-flap offers many advantages over the pedicled TRAM-flap. Due to its superior perfusion, the free flap rarely develops necrosis. Shaping of the flap is easier due to the lack of the thick muscle pedicle. Because the rectus muscle is spared, there is minimal donor site morbidity. However, the necessary microvascular anastomoses reduced the acceptance of the free TRAM-flap. During a 13-months period, 51 breast reconstructions were performed in 41 patients, 31 unilateral and ten bilateral. 45 flaps served for delayed reconstruction and six flaps for immediate reconstruction. The operations were performed by two teams working simultaneously. The average operating time was 3.9 hours for unilateral and 6.9 hours for bilateral delayed reconstruction. For immediate reconstruction, 6.2 and 6.3 hours were required for uni- and bilateral procedures, respectively. In 38 flaps, the thoracodorsal vessels served as recipient vessels; 13 flaps were anastomosed to the internal mammary artery and vein. Postoperative complications were observed in 13 patients. Three vessel anastomoses had to be revised. In one flap, a partial necrosis occurred; in two flaps hematoma evacuation was necessary. Two patients suffered from fat necroses at the abdomen and one umbilicus was lost. Skin irritations and seromas at the abdomen occurred in five patients. Pulmonary embolism was diagnosed in one patient three weeks postoperatively. Abdominal hernias or bulging in the epigastric area were not observed up to 15 months after reconstruction. These results reveal a low complication rate for breast reconstruction with the free TRAM-flap. The advantages of this technique as compared to the pedicled technique are discussed.

  13. USB noise reduction by nozzle and flap modifications

    NASA Technical Reports Server (NTRS)

    Hayden, R. E.

    1976-01-01

    The development of concepts for reducing upper surface blown flap noise at the source through flap modifications and special nozzles is reviewed. In particular, recent results obtained on the aerodynamic and acoustic performance of flaps with porous surfaces near the trailing edge and multi-slotted nozzles are reviewed. Considerable reduction (6-10 db) of the characteristic low frequency peak is shown. The aerodynamic performance is compared with conventional systems, and prospects for future improvements are discussed.

  14. Flight test pilot evaluation of a delayed flap approach procedure

    NASA Technical Reports Server (NTRS)

    Bull, J. S.; Edwards, F. G.; Foster, J. D.; Hegarty, D. M.; Drinkwater, F. J., III

    1977-01-01

    Using NASA's CV-990 aircraft, a delayed flap approach procedure was demonstrated to nine guest pilots from the air transport industry. Four demonstration flights and 37 approaches were conducted under VFR weather conditions. A limited pilot evaluation of the delayed flap procedure was obtained from pilot comments and from questionaires they completed. Pilot acceptability, pilot workload, and ATC compatibility were quantitatively rated. The delayed flap procedure was shown to be feasible, and suggestions for further development work were obtained.

  15. Integrins activate trimeric G proteins via the nonreceptor protein GIV/Girdin

    PubMed Central

    Leyme, Anthony; Marivin, Arthur; Perez-Gutierrez, Lorena; Nguyen, Lien T.

    2015-01-01

    Signal transduction via integrins and G protein–coupled receptors is critical to control cell behavior. These two receptor classes have been traditionally believed to trigger distinct and independent signaling cascades in response to extracellular cues. Here, we report a novel mechanism of integrin signaling that requires activation of the trimeric G protein Gαi by the nonreceptor guanine nucleotide exchange factor (GEF) GIV (also known as Girdin), a metastasis-associated protein. We demonstrate that GIV enhances integrin-dependent cell responses upon extracellular matrix stimulation and makes tumor cells more invasive. These responses include remodeling of the actin cytoskeleton and PI3K-dependent signaling, resulting in enhanced haptotaxis and invasion. We show that both GIV and its substrate Gαi3 are recruited to active integrin complexes and that tumor cells engineered to express GEF-deficient GIV fail to transduce integrin signals into proinvasive responses via a Gβγ-PI3K axis. Our discoveries delineate a novel mechanism by which integrin signaling is rewired during metastasis to result in increased tumor invasiveness. PMID:26391662

  16. The total protein content, protein fractions and proteases activities of drone prepupae of Apis mellifera due to varrosis.

    PubMed

    Zółtowska, Krystyna; Lipiński, Zbigniew; Dmitryjuk, Małgorzata

    2005-01-01

    The proteins level and activities of acid and alkaline proteases in whole body extracts of drone prepupae of Apis mellifera naturally infested with Varroa destructor were studied. The infested and a non-infested group did not differ significantly in their total protein content. However, some differences in protein profiles were found. A lack of three protein fractions of moderate and lower molecular weight in infested prepupae was noted. Moreover, some differences in the quantity of protein in most of the fractions were observed. The activity of acid proteases from infested prepupae was lower (p < 0.05) compared with the activity of these proteases from the non-infested one group. The infested drone had higher activity of alkaline proteases than non-infested but this difference was not statisticaly significant.

  17. The total protein content, protein fractions and proteases activities of drone prepupae of Apis mellifera due to varrosis.

    PubMed

    Zółtowska, Krystyna; Lipiński, Zbigniew; Dmitryjuk, Małgorzata

    2005-01-01

    The proteins level and activities of acid and alkaline proteases in whole body extracts of drone prepupae of Apis mellifera naturally infested with Varroa destructor were studied. The infested and a non-infested group did not differ significantly in their total protein content. However, some differences in protein profiles were found. A lack of three protein fractions of moderate and lower molecular weight in infested prepupae was noted. Moreover, some differences in the quantity of protein in most of the fractions were observed. The activity of acid proteases from infested prepupae was lower (p < 0.05) compared with the activity of these proteases from the non-infested one group. The infested drone had higher activity of alkaline proteases than non-infested but this difference was not statisticaly significant. PMID:16841690

  18. The Influence of Wing Loading on Turbofan Powered STOL Transports with and Without Externally Blown Flaps

    NASA Technical Reports Server (NTRS)

    Morris, R. L.; Hanke, C. R.; Pasley, L. H.; Rohling, W. J.

    1973-01-01

    The effects of wing loading on the design of short takeoff and landing (STOL) transports using (1) mechanical flap systems, and (2) externally blown flap systems are determined. Aircraft incorporating each high-lift method are sized for field lengths of 2,000 feet, 2,500 feet, and 3,500 feet, and for payloads of 40, 150, and 300 passengers, for a total of 18 point-design aircraft. An assumed 1975 level of technology is applied to both concepts in terms of propulsion, weights, active controls, supercritical wing methodology, and acoustics. Low-wing-loading STOL configurations with mechanical flaps are found to be competitive with externally blown flap STOL configurations over wide ranges of payload and field length for the airworthiness rules and technology improvements assumed. Because the results of design studies like this one are sensitive to the ground rules assumed, careful attention is paid to describing the assumptions. These assumptions must be understood before the results are compared with other STOL airplane studies.

  19. Annexin proteins PP4 and PP4-X. Comparative characterization of biological activities of placental and recombinant proteins.

    PubMed Central

    Römisch, J; Grote, M; Weithmann, K U; Heimburger, N; Amann, E

    1990-01-01

    The human placental proteins PP4 and PP4-X, belonging to the annexin protein family, were expressed in Escherichia coli at high yield. The proteins were purified to homogeneity. The physicochemical parameters of the recombinant proteins were determined and compared with those of their natural placental counterparts. Except for a minor change in the pI, the proteins appeared to be indistinguishable by several criteria. Both recombinant PP4 and recombinant PP4-X were biologically active in a thromboplastin inhibition test and in a phospholipase A2 inhibition test. Images Fig. 2. Fig. 3. Fig. 4. PMID:2148260

  20. Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants

    PubMed Central

    Kovtun, Yelena; Chiu, Wan-Ling; Tena, Guillaume; Sheen, Jen

    2000-01-01

    Despite the recognition of H2O2 as a central signaling molecule in stress and wounding responses, pathogen defense, and regulation of cell cycle and cell death, little is known about how the H2O2 signal is perceived and transduced in plant cells. We report here that H2O2 is a potent activator of mitogen-activated protein kinases (MAPKs) in Arabidopsis leaf cells. Using epitope tagging and a protoplast transient expression assay, we show that H2O2 can activate a specific Arabidopsis mitogen-activated protein kinase kinase kinase, ANP1, which initiates a phosphorylation cascade involving two stress MAPKs, AtMPK3 and AtMPK6. Constitutively active ANP1 mimics the H2O2 effect and initiates the MAPK cascade that induces specific stress-responsive genes, but it blocks the action of auxin, a plant mitogen and growth hormone. The latter observation provides a molecular link between oxidative stress and auxin signal transduction. Finally, we show that transgenic tobacco plants that express a constitutively active tobacco ANP1 orthologue, NPK1, display enhanced tolerance to multiple environmental stress conditions without activating previously described drought, cold, and abscisic acid signaling pathways. Thus, manipulation of key regulators of an oxidative stress signaling pathway, such as ANP1/NPK1, provides a strategy for engineering multiple stress tolerance that may greatly benefit agriculture. PMID:10717008

  1. Activation of AMP-activated protein kinase by tributyltin induces neuronal cell death

    SciTech Connect

    Nakatsu, Yusuke; Kotake, Yaichiro Hino, Atsuko; Ohta, Shigeru

    2008-08-01

    AMP-activated protein kinase (AMPK), a member of the metabolite-sensing protein kinase family, is activated by energy deficiency and is abundantly expressed in neurons. The environmental pollutant, tributyltin chloride (TBT), is a neurotoxin, and has been reported to decrease cellular ATP in some types of cells. Therefore, we investigated whether TBT activates AMPK, and whether its activation contributes to neuronal cell death, using primary cultures of cortical neurons. Cellular ATP levels were decreased 0.5 h after exposure to 500 nM TBT, and the reduction was time-dependent. It was confirmed that most neurons in our culture system express AMPK, and that TBT induced phosphorylation of AMPK. Compound C, an AMPK inhibitor, reduced the neurotoxicity of TBT, suggesting that AMPK is involved in TBT-induced cell death. Next, the downstream target of AMPK activation was investigated. Nitric oxide synthase, p38 phosphorylation and Akt dephosphorylation were not downstream of TBT-induced AMPK activation because these factors were not affected by compound C, but glutamate release was suggested to be controlled by AMPK. Our results suggest that activation of AMPK by TBT causes neuronal death through mediating glutamate release.

  2. Prion Protein Promotes Kidney Iron Uptake via Its Ferrireductase Activity*

    PubMed Central

    Haldar, Swati; Tripathi, Ajai; Qian, Juan; Beserra, Amber; Suda, Srinivas; McElwee, Matthew; Turner, Jerrold; Hopfer, Ulrich; Singh, Neena

    2015-01-01

    Brain iron-dyshomeostasis is an important cause of neurotoxicity in prion disorders, a group of neurodegenerative conditions associated with the conversion of prion protein (PrPC) from its normal conformation to an aggregated, PrP-scrapie (PrPSc) isoform. Alteration of iron homeostasis is believed to result from impaired function of PrPC in neuronal iron uptake via its ferrireductase activity. However, unequivocal evidence supporting the ferrireductase activity of PrPC is lacking. Kidney provides a relevant model for this evaluation because PrPC is expressed in the kidney, and ∼370 μg of iron are reabsorbed daily from the glomerular filtrate by kidney proximal tubule cells (PT), requiring ferrireductase activity. Here, we report that PrPC promotes the uptake of transferrin (Tf) and non-Tf-bound iron (NTBI) by the kidney in vivo and mainly NTBI by PT cells in vitro. Thus, uptake of 59Fe administered by gastric gavage, intravenously, or intraperitoneally was significantly lower in PrP-knock-out (PrP−/−) mouse kidney relative to PrP+/+ controls. Selective in vivo radiolabeling of plasma NTBI with 59Fe revealed similar results. Expression of exogenous PrPC in immortalized PT cells showed localization on the plasma membrane and intracellular vesicles and increased transepithelial transport of 59Fe-NTBI and to a smaller extent 59Fe-Tf from the apical to the basolateral domain. Notably, the ferrireductase-deficient mutant of PrP (PrPΔ51–89) lacked this activity. Furthermore, excess NTBI and hemin caused aggregation of PrPC to a detergent-insoluble form, limiting iron uptake. Together, these observations suggest that PrPC promotes retrieval of iron from the glomerular filtrate via its ferrireductase activity and modulates kidney iron metabolism. PMID:25572394

  3. Multifunctional antimicrobial proteins and peptides: natural activators of immune systems.

    PubMed

    Niyonsaba, François; Nagaoka, Isao; Ogawa, Hideoki; Okumura, Ko

    2009-01-01

    In addition to the physical barrier of the stratum corneum, cutaneous innate immunity also includes the release of various humoral mediators, such as cytokines and chemokines, recruitment and activation of phagocytes, and the production of antimicrobial proteins/peptides (AMPs). AMPs form an innate epithelial chemical shield, which provides a front-line component in innate immunity to inhibit microbial invasion; however, this might be an oversimplification of the diverse functions of these molecules. In fact, apart from exhibiting a broad spectrum of microbicidal properties, it is increasingly evident that AMPs display additional activities that are related to the stimulation and modulation of the cutaneous immune system. These diverse functions include chemoattraction and activation of immune and/or inflammatory cells, the production and release of cytokines and chemokines, acceleration of angiogenesis, promotion of wound healing, neutralization of harmful microbial products, and bridging of both innate and adaptive immunity. Thus, better understanding of the functions of AMPs in skin and identification of their signaling mechanisms may offer new strategies for the development of potential therapeutics for the treatment of infection- and/or inflammation-related skin diseases. Here, we briefly outline the structure, regulation of expression, and multifunctional roles of principal skin-derived AMPs.

  4. Allosteric activation of apicomplexan calcium-dependent protein kinases.

    PubMed

    Ingram, Jessica R; Knockenhauer, Kevin E; Markus, Benedikt M; Mandelbaum, Joseph; Ramek, Alexander; Shan, Yibing; Shaw, David E; Schwartz, Thomas U; Ploegh, Hidde L; Lourido, Sebastian

    2015-09-01

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7-kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes.

  5. MDMX exerts its oncogenic activity via suppression of retinoblastoma protein.

    PubMed

    Zhang, H; Hu, L; Qiu, W; Deng, T; Zhang, Y; Bergholz, J; Xiao, Z-X

    2015-10-29

    Inactivation of the retinoblastoma protein (RB) has a major role in the development of human malignancies. We have previously shown that MDM2, an ubiquitin E3 ligase and major negative regulator of p53, binds to and promotes proteasome-mediated degradation of RB. MDMX, a homolog of MDM2, also binds to and inhibits p53 transactivation activity, yet it does not possess intrinsic ubiquitin ligase activity. Here, we show that MDMX binds to and promotes RB degradation in an MDM2-dependent manner. Specifically, the MDMX C-terminal ring domain binds to the RB C-pocket and enhances MDM2-RB interaction. Silencing MDMX induces RB accumulation, cell cycle arrest and senescence-like phenotypes, which are reverted by simultaneous RB knockdown. Furthermore, MDMX ablation leads to significant retardation of xenograft tumor growth, concomitant with RB accumulation. These results demonstrate that MDMX exerts oncogenic activity via suppression of RB, and suggest that both MDM2 and MDMX could be chemotherapeutic targets. PMID:25703327

  6. Flapping instability of a liquid jet

    NASA Astrophysics Data System (ADS)

    Matas, Jean-Philippe; Cartellier, Alain

    2013-01-01

    We study the flapping instability observed when a liquid jet is incompletely atomized by a fast parallel gas stream: the remaining liquid jet is destabilized over a scale large compared with its radius, and breaks into liquid fragments. We characterize the symmetry of this instability and its frequency. The intact liquid length is measured as a function of gas and liquid velocity, and turns out to be longer than the one predicted by Raynal (1997) for a planar mixing layer. The frequency of the instability is measured with a spectral method, and is in agreement with the frequency observed for the planar shear instability, though slightly smaller. The planar, and not helical, symmetry of the instability makes it akin to a flapping instability, observed when a planar liquid sheet is atomized by two planar gas streams. We next measure drop sizes when the flapping instability is present, with a method based on image processing. Measured size distributions are in agreement with distributions observed in a mixing layer geometry for low gas velocities (long tail distribution). The mean drop diameter depends weakly on liquid velocity, and decreases as d10˜Ug0.9. On the contrary, Sauter diameter depends strongly on liquid velocity.

  7. Evaluation of flexible flapping wing concept

    NASA Astrophysics Data System (ADS)

    Rakotomamonjy, Thomas; Le Moing, Thierry; Danet, Brieuc; Gadoullet, Xavier; Osmont, Daniel; Dupont, Marc

    2009-03-01

    ONERA - The French Aerospace Lab - has launched an internal program on biologically-inspired Micro Air Vehicles (MAVs), covering many research topics such as unsteady aerodynamics, actuation, structural dynamics or control. The aim is to better understand the flapping flight performed in nature by insects, and to control state of the art technologies and applications in this field. For that purpose, a flight-dynamics oriented simulation model of a flapping-wing concept has been developed. This model, called OSCAB, features a body and two wings along which the aerodynamics efforts are integrated, so as to determine the global motion of the MAV. The model has been improved by taking into account the flexibility of the wings (flexion of the leading edge and passive torsion of the wings, induced by the flapping motion itself under wing inertia). Thus, it becomes possible to estimate the coupling between flexibility and the aerodynamic forces. Furthermore, the model shows that using elastic properties of the wings allows a diminution of the mechanical energy needed for wings motion, and a reduction of the number of actuators to be implanted into the MAV.

  8. Flapping inertia for selected rotor blades

    NASA Technical Reports Server (NTRS)

    Berry, John D.; May, Matthew J.

    1991-01-01

    Aerodynamics of helicopter rotor systems cannot be investigated without consideration for the dynamics of the rotor. One of the principal properties of the rotor which affects the rotor dynamics is the inertia of the rotor blade about its root attachment. Previous aerodynamic investigation have been performed on rotor blades with a variety of planforms to determine the performance differences due to blade planform. The blades tested for this investigation have been tested on the U.S. Army 2 meter rotor test system (2MRTS) in the NASA Langley 14 by 22 foot subsonic tunnel for hover performance. This investigation was intended to provide fundamental information on the flapping inertia of five rotor blades with differing planforms. The inertia of the bare cuff and the cuff with a blade extension were also measured for comparison with the inertia of the blades. Inertia was determined using a swing testing technique, using the period of oscillation to determine the effective flapping inertia. The effect of damping in the swing test was measured and described. A comparison of the flapping inertials for rectangular and tapered planform blades of approximately the same mass showed the tapered blades to have a lower inertia, as expected.

  9. Aerodynamics of high frequency flapping wings

    NASA Astrophysics Data System (ADS)

    Hu, Zheng; Roll, Jesse; Cheng, Bo; Deng, Xinyan

    2010-11-01

    We investigated the aerodynamic performance of high frequency flapping wings using a 2.5 gram robotic insect mechanism developed in our lab. The mechanism flaps up to 65Hz with a pair of man-made wing mounted with 10cm wingtip-to-wingtip span. The mean aerodynamic lift force was measured by a lever platform, and the flow velocity and vorticity were measured using a stereo DPIV system in the frontal, parasagittal, and horizontal planes. Both near field (leading edge vortex) and far field flow (induced flow) were measured with instantaneous and phase-averaged results. Systematic experiments were performed on the man-made wings, cicada and hawk moth wings due to their similar size, frequency and Reynolds number. For insect wings, we used both dry and freshly-cut wings. The aerodynamic force increase with flapping frequency and the man-made wing generates more than 4 grams of lift at 35Hz with 3 volt input. Here we present the experimental results and the major differences in their aerodynamic performances.

  10. Mys protein regulates protein kinase A activity by interacting with regulatory type Ialpha subunit during vertebrate development.

    PubMed

    Kotani, Tomoya; Iemura, Shun-ichiro; Natsume, Tohru; Kawakami, Koichi; Yamashita, Masakane

    2010-02-12

    During embryonic development, protein kinase A (PKA) plays a key role in cell fate specification by antagonizing the Hedgehog (Hh) signaling pathway. However, the mechanism by which PKA activity is regulated remains unknown. Here we show that the Misty somites (Mys) protein regulates the level of PKA activity during embryonic development in zebrafish. We isolate PKA regulatory type Ialpha subunit (Prkar1a) as a protein interacting with Mys by pulldown assay in HEK293 cells followed by mass spectrometry analysis. We show an interaction between endogenous Mys and Prkar1a in the zebrafish embryo. Mys binds to Prkar1a in its C terminus region, termed PRB domain, and activates PKA in vitro. Conversely, knockdown of Mys in zebrafish embryos results in reduction in PKA activity. We also show that knockdown of Mys induces ectopic activation of Hh target genes in the eyes, neural tube, and somites downstream of Smoothened, a protein essential for transduction of Hh signaling activity. The altered patterning of gene expression is rescued by activation of PKA. Together, our results reveal a molecular mechanism of regulation of PKA activity that is dependent on a protein-protein interaction and demonstrate that PKA activity regulated by Mys is indispensable for negative regulation of the Hh signaling pathway in Hh-responsive cells. PMID:20018846

  11. Vortex leading edge flap assembly for supersonic airplanes

    NASA Technical Reports Server (NTRS)

    Rudolph, Peter K. C. (Inventor)

    1997-01-01

    A leading edge flap (16) for supersonic transport airplanes is disclosed. In its stowed position, the leading edge flap forms the lower surface of the wing leading edge up to the horizontal center of the leading edge radius. For low speed operation, the vortex leading edge flap moves forward and rotates down. The upward curve of the flap leading edge triggers flow separation on the flap and rotational flow on the upper surface of the flap (vortex). The rounded shape of the upper fixed leading edge provides the conditions for a controlled reattachment of the flow on the upper wing surface and therefore a stable vortex. The vortex generates lift and a nose-up pitching moment. This improves maximum lift at low speed, reduces attitude for a given lift coefficient and improves lift to drag ratio. The mechanism (27) to move the vortex flap consists of two spanwise supports (24) with two diverging straight tracks (64 and 68) each and a screw drive mechanism (62) in the center of the flap panel (29). The flap motion is essentially normal to the airloads and therefore requires only low actuation forces.

  12. Rotational flaps in oncologic breast surgery. Anatomical and technical considerations.

    PubMed

    Acea Nebril, Benigno; Builes Ramírez, Sergio; García Novoa, Alejandra; Varela Lamas, Cristina

    2016-01-01

    Local flaps are a group of surgical procedures that can solve the thoracic closure of large defects after breast cancer surgery with low morbidity. Its use in skin necrosis complications after conservative surgery or skin sparing mastectomies facilitates the initiation of adjuvant treatments and reduces delays in this patient group. This article describes the anatomical basis for the planning of thoracic and abdominal local flaps. Also, the application of these local flaps for closing large defects in the chest and selective flaps for skin coverage by necrosis in breast conserving surgery.

  13. Rotational flaps in oncologic breast surgery. Anatomical and technical considerations.

    PubMed

    Acea Nebril, Benigno; Builes Ramírez, Sergio; García Novoa, Alejandra; Varela Lamas, Cristina

    2016-01-01

    Local flaps are a group of surgical procedures that can solve the thoracic closure of large defects after breast cancer surgery with low morbidity. Its use in skin necrosis complications after conservative surgery or skin sparing mastectomies facilitates the initiation of adjuvant treatments and reduces delays in this patient group. This article describes the anatomical basis for the planning of thoracic and abdominal local flaps. Also, the application of these local flaps for closing large defects in the chest and selective flaps for skin coverage by necrosis in breast conserving surgery. PMID:27140865

  14. Monocortical Deep Circumflex Iliac Artery Flap in Jaw Reconstruction.

    PubMed

    Moon, Seong-Yong

    2015-06-01

    Conventionally deep circumflex iliac artery (DCIA) flap had been harvested as bicortical form. However, several complications and adverse effects occurred such as abnormal hip contour, hernia, severe bleeding tendency, gait disturbance, and hypoesthesia. All the 9 patients required reconstruction of the jaw with microvascular free flaps after radical resection. Monocortical bone segment was harvested from the anterior iliac crest, and the amount of bone harvested was from 47 to 90 mm (mean, 63 ± 14.6). Monocortical deep circumflex iliac artery flap has sufficient advantages in donor-site morbidity, which is one of the factors to choose flap.

  15. A novel animal model for skin flap prelamination with biomaterials

    PubMed Central

    Zhou, Xianyu; Luo, Xusong; Liu, Fei; Gu, Chuan; Wang, Xi; Yang, Qun; Qian, Yunliang; Yang, Jun

    2016-01-01

    Several animal models of skin flap construction were reported using biomaterials in a way similar to prefabrication. However, there are few animal model using biomaterials similar to prelamination, another main way of clinical skin flap construction that has been proved to be reliable. Can biomaterials be added in skin flap prelamination to reduce the use of autogenous tissues? Beside individual clinical attempts, animal model is needed for randomized controlled trial to objectively evaluate the feasibility and further investigation. Combining human Acellular Dermal Matrix (hADM) and autologous skin graft, we prelaminated flaps based on inguinal fascia. One, two, three and four weeks later, hADM exhibited a sound revascularization and host cell infiltration. Prelaminated skin flaps were then raised and microsurgically transplanted back to groin region. Except for flaps after one week of prelamination, flaps from other subgroups successfully reconstructed defects. After six to sixteen weeks of transplantation, hADM was proved to being able to maintain its original structure, having a wealth of host tissue cells and achieving full revascularization.To our knowledge, this is the first animal model of prelaminating skin flap with biomaterials. Success of this animal model indicates that novel flap prelamination with biomaterials is feasible. PMID:27659066

  16. Numerical analysis of the V-Y shaped advancement flap.

    PubMed

    Remache, D; Chambert, J; Pauchot, J; Jacquet, E

    2015-10-01

    The V-Y advancement flap is a usual technique for the closure of skin defects. A triangular flap is incised adjacent to a skin defect of rectangular shape. As the flap is advanced to close the initial defect, two smaller defects in the shape of a parallelogram are formed with respect to a reflection symmetry. The height of the defects depends on the apex angle of the flap and the closure efforts are related to the defects height. Andrades et al. 2005 have performed a geometrical analysis of the V-Y flap technique in order to reach a compromise between the flap size and the defects width. However, the geometrical approach does not consider the mechanical properties of the skin. The present analysis based on the finite element method is proposed as a complement to the geometrical one. This analysis aims to highlight the major role of the skin elasticity for a full analysis of the V-Y advancement flap. Furthermore, the study of this technique shows that closing at the flap apex seems mechanically the most interesting step. Thus different strategies of defect closure at the flap apex stemming from surgeon's know-how have been tested by numerical simulations. PMID:26342442

  17. Lower extremity reconstruction with the anterolateral thigh flap.

    PubMed

    Nosrati, Naveed; Chao, Albert H; Chang, David W; Yu, Peirong

    2012-05-01

    The versatility and location of the anterolateral thigh (ALT) flap make it well suited for lower extremity reconstruction. The purpose of this study was to evaluate surgical and functional outcomes by specific anatomic regions in the lower extremity to better define the role of the ALT flap in lower extremity reconstruction. A retrospective review of patients undergoing lower extremity reconstruction with an ALT flap between July 2002 and December 2010 was performed. Total 46 patients underwent lower extremity reconstruction with an ALT flap, of whom 29 (63%) had a pedicled flap and 17 (37%) a microvascular free flap. Defects were located in the hip/buttocks (n = 8), groin (n = 13), thigh (n = 8), knee (n = 5), leg (n = 6), and foot/ankle (n = 6). The mean postoperative follow-up was 4 months. Total flap loss occurred in two patients (4%). There were 11 recipient site complications (24%). The most common complication was recipient site seroma, which occurred in five patients (11%), all of whom had hip/buttock or groin defects. Overall, 38 patients (83%) returned to their preoperative functional status. The ALT flap is an effective method of lower extremity reconstruction. It can be performed as a pedicled or free flap, with good surgical and functional outcomes. PMID:22399252

  18. Clinical Experiences with the Scapular Fascial Free Flap

    PubMed Central

    Park, Il Ho; Chang, Yong Joon; Kim, Jae Hyun

    2016-01-01

    Background The goal of reconstruction is to provide coverage of exposed vital structures with well-vascularized tissue for optimal restoration of form and function. Here, we present our clinical experience with the use of the scapular fascial free flap to correct facial asymmetry and to reconstruct soft tissue defects of the extremities. Methods We used a scapular fascial free flap in 12 cases for soft tissue coverage of the extremities or facial soft tissue augmentation. Results The flaps ranged in size from 3×12 to 13×23 cm. No cases of total loss of the flap occurred. Partial loss of the flap occurred in 1 patient, who was treated with a turnover flap using the adjacent scapular fascial flap and a skin graft. Partial loss of the skin graft occurred in 4 patients due to infection or hematoma beneath the graft, and these patients underwent another skin graft. Four cases of seroma at the donor site occurred, and these cases were treated with conservative management or capsulectomy and quilting sutures. Conclusions The scapular fascial free flap has many advantages, including a durable surface for restoration of form and contours, a large size with a constant pedicle, adequate surface for tendon gliding, and minimal donor-site scarring. We conclude that despite the occurrence of a small number of complications, the scapular fascial free flap should be considered to be a viable option for soft tissue coverage of the extremities and facial soft tissue augmentation. PMID:27689051

  19. Clinical Experiences with the Scapular Fascial Free Flap

    PubMed Central

    Park, Il Ho; Chang, Yong Joon; Kim, Jae Hyun

    2016-01-01

    Background The goal of reconstruction is to provide coverage of exposed vital structures with well-vascularized tissue for optimal restoration of form and function. Here, we present our clinical experience with the use of the scapular fascial free flap to correct facial asymmetry and to reconstruct soft tissue defects of the extremities. Methods We used a scapular fascial free flap in 12 cases for soft tissue coverage of the extremities or facial soft tissue augmentation. Results The flaps ranged in size from 3×12 to 13×23 cm. No cases of total loss of the flap occurred. Partial loss of the flap occurred in 1 patient, who was treated with a turnover flap using the adjacent scapular fascial flap and a skin graft. Partial loss of the skin graft occurred in 4 patients due to infection or hematoma beneath the graft, and these patients underwent another skin graft. Four cases of seroma at the donor site occurred, and these cases were treated with conservative management or capsulectomy and quilting sutures. Conclusions The scapular fascial free flap has many advantages, including a durable surface for restoration of form and contours, a large size with a constant pedicle, adequate surface for tendon gliding, and minimal donor-site scarring. We conclude that despite the occurrence of a small number of complications, the scapular fascial free flap should be considered to be a viable option for soft tissue coverage of the extremities and facial soft tissue augmentation.

  20. Antitussive and immunomodulating activities of instant coffee arabinogalactan-protein.

    PubMed

    Nosáľová, G; Prisenžňáková, L; Paulovičová, E; Capek, P; Matulová, M; Navarini, L; Liverani, F Suggi

    2011-11-01

    A low molecular mass arabinogalactan-protein (AGP) composed of galactose and arabinose with a low protein content, isolated from the instant coffee powder of Coffea arabica beans, has been tested on antitussive (in vivo) and immunomodulating (ex vivo) activities. The results of antitussive tests revealed a significant dose dependant cough-suppressive effect of coffee AGP. It was observed 30 or 60 min after AGP administration and its efficacy lasted during the entire experiment course. Immunological tests showed that AGP affected some mediators of immunocompetent cells of immune system as TNF-α, IFN-γ and IL-2 cytokines. It seems that coffee AGP is a good inductor of both pro-inflammatory cytokines TNF-α and IFN-γ, however, less potent in TNF-α induction in comparison with that of β-D-glucan. Evident induction of TNF-α, IL-2 and IFN-γ cytokines, pro-TH1 polarization supports our conclusion about bio-immunological efficacy of AGP with an emphasis on the cellular immunity.