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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. The novel benzimidazole derivative BRP-7 inhibits leukotriene biosynthesis in vitro and in vivo by targeting 5-lipoxygenase-activating protein (FLAP)

    PubMed Central

    Pergola, C; Gerstmeier, J; Mönch, B; Çalışkan, B; Luderer, S; Weinigel, C; Barz, D; Maczewsky, J; Pace, S; Rossi, A; Sautebin, L; Banoglu, E; Werz, O

    2014-01-01

    BACKGROUND AND PURPOSE Leukotrienes (LTs) are inflammatory mediators produced via the 5-lipoxygenase (5-LOX) pathway and are linked to diverse disorders, including asthma, allergic rhinitis and cardiovascular diseases. We recently identified the benzimidazole derivative BRP-7 as chemotype for anti-LT agents by virtual screening targeting 5-LOX-activating protein (FLAP). Here, we aimed to reveal the in vitro and in vivo pharmacology of BRP-7 as an inhibitor of LT biosynthesis. EXPERIMENTAL APPROACH We analysed LT formation and performed mechanistic studies in human neutrophils and monocytes, in human whole blood (HWB) and in cell-free assays. The effectiveness of BRP-7 in vivo was evaluated in rat carrageenan-induced pleurisy and mouse zymosan-induced peritonitis. KEY RESULTS BRP-7 potently suppressed LT formation in neutrophils and monocytes and this was accompanied by impaired 5-LOX co-localization with FLAP. Neither the cellular viability nor the activity of 5-LOX in cell-free assays was affected by BRP-7, indicating that a functional FLAP is needed for BRP-7 to inhibit LTs, and FLAP bound to BRP-7 linked to a solid matrix. Compared with the FLAP inhibitor MK-886, BRP-7 did not significantly inhibit COX-1 or microsomal prostaglandin E2 synthase-1, implying the selectivity of BRP-7 for FLAP. Finally, BRP-7 was effective in HWB and impaired inflammation in vivo, in rat pleurisy and mouse peritonitis, along with reducing LT levels. CONCLUSIONS AND IMPLICATIONS BRP-7 potently suppresses LT biosynthesis by interacting with FLAP and exhibits anti-inflammatory effectiveness in vivo, with promising potential for further development. PMID:24641614

  3. 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

  4. Correlation of Smart Active Flap Rotor Loads

    NASA Technical Reports Server (NTRS)

    Kottapalli, Sesi; Straub, Friedrich

    2009-01-01

    The ability to predict SMART active trailing edge flap rotor loads is explored in this study. Full-scale wind tunnel data recently acquired in the NASA Ames 40- by 80- Foot Wind Tunnel are compared with analytical results from CAMRAD II. For the 5-bladed rotor, two high-speed forward flight cases are considered, namely, a 0 deg flap deflection case and a 5P, 2 deg flap deflection case. Overall, the correlation is reasonable, with the following exceptions: the torsion moment frequency and the chordwise bending moment are under predicted. In general, the effect of the 5P, 2 deg flap motion is captured by the analysis, though there is over prediction in the neighborhood of the 105 deg and 120 deg azimuthal locations. Changes to the flexbeam torsion stiffness are also briefly considered in this study, as this stiffness will be updated in the future. Finally, the indication is that compressibility effects are important, and this suggests that computational fluid dynamics might improve the current correlation.

  5. Global optimization of actively morphing flapping wings

    NASA Astrophysics Data System (ADS)

    Ghommem, Mehdi; Hajj, Muhammad R.; Mook, Dean T.; Stanford, Bret K.; Beran, Philip S.; Snyder, Richard D.; Watson, Layne T.

    2012-08-01

    We consider active shape morphing to optimize the flight performance of flapping wings. To this end, we combine a three-dimensional version of the unsteady vortex lattice method (UVLM) with a deterministic global optimization algorithm to identify the optimal kinematics that maximize the propulsive efficiency under lift and thrust constraints. The UVLM applies only to incompressible, inviscid flows where the separation lines are known a priori. Two types of morphing parameterization are investigated here—trigonometric and spline-based. The results show that the spline-based morphing, which requires specification of more design variables, yields a significant improvement in terms of propulsive efficiency. Furthermore, we remark that the average value of the lift coefficient in the optimized kinematics remained equal to the value in the baseline case (without morphing). This indicates that morphing is most efficiently used to generate thrust and not to increase lift beyond the basic value obtained by flapping only. Besides, our study gives comparable optimal efficiencies to those obtained from previous studies based on gradient-based optimization, but completely different design points (especially for the spline-based morphing), which would indicate that the design space associated with the flapping kinematics is very complex.

  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. Active Flap Control of the SMART Rotor for Vibration Reduction

    NASA Technical Reports Server (NTRS)

    Hall, Steven R.; Anand, R. Vaidyanathan; Straub, Friedrich K.; Lau, Benton H.

    2009-01-01

    Active control methodologies were applied to a full-scale active flap rotor obtained during a joint Boeing/ DARPA/NASA/Army test in the Air Force National Full-Scale Aerodynamic Complex 40- by 80-foot anechoic wind tunnel. The active flap rotor is a full-scale MD 900 helicopter main rotor with each of its five blades modified to include an on-blade piezoelectric actuator-driven flap with a span of 18% of radius, 25% of chord, and located at 83% radius. Vibration control demonstrated the potential of active flaps for effective control of vibratory loads, especially normal force loads. Active control of normal force vibratory loads using active flaps and a continuous-time higher harmonic control algorithm was very effective, reducing harmonic (1-5P) normal force vibratory loads by 95% in both cruise and approach conditions. Control of vibratory roll and pitch moments was also demonstrated, although moment control was less effective than normal force control. Finally, active control was used to precisely control blade flap position for correlation with pretest predictions of rotor aeroacoustics. Flap displacements were commanded to follow specific harmonic profiles of 2 deg or more in amplitude, and the flap deflection errors obtained were less than 0.2 deg r.m.s.

  8. Rotor Flapping Response to Active Control

    NASA Technical Reports Server (NTRS)

    Nguyen, Khanh; Johnson, Wayne

    2004-01-01

    Rotor active control using higher harmonic blade pitch has been proposed as a means to reduce both rotor radiated noise and airframe vibration and to enhance rotor performance. The higher harmonic input, however, can affect rotor thrust and cyclic flapping - the basic trim characteristics of the rotor. Some of the trim changes can negate the active control benefits. For example, wind tunnel test results of a full scale BO-105 rotor with individual-blade control indicate some rotor performance improvements, accompanied with changes in rotor trim, using two-per-rev blade pitch input. The observed performance benefits could therefore be a simple manifestation of the trim change rather than an efficient redistribution of the rotor airloads. More recently, the flight test of the BO-105 helicopter equip,ped with individual-blade-control actuators also reported trim changes whenever the two-per-rev blade pitch for noise reduction was activated. The pilot had to adjust the trim control to maintain the aircraft under a constant flight path. These two cases highlight the, importance of trim considerations in the application of active control to rotorcraft.

  9. Wind Tunnel Test of the SMART Active Flap Rotor

    NASA Technical Reports Server (NTRS)

    Straub, Friedrich K.; Anand, Vaidyanthan R.; Birchette, Terrence S.; Lau, Benton H.

    2009-01-01

    Boeing and a team from Air Force, NASA, Army, DARPA, MIT, UCLA, and U. of Maryland have successfully completed a wind-tunnel test of the smart material actuated rotor technology (SMART) rotor in the 40- by 80-foot wind-tunnel of the National Full-Scale Aerodynamic Complex at NASA Ames Research Center. The Boeing SMART rotor is a full-scale, five-bladed bearingless MD 900 helicopter rotor modified with a piezoelectric-actuated trailing edge flap on each blade. The eleven-week test program evaluated the forward flight characteristics of the active-flap rotor at speeds up to 155 knots, gathered data to validate state-of-the-art codes for rotor aero-acoustic analysis, and quantified the effects of open and closed loop active flap control on rotor loads, noise, and performance. The test demonstrated on-blade smart material control of flaps on a full-scale rotor for the first time in a wind tunnel. The effectiveness of the active flap control on noise and vibration was conclusively demonstrated. Results showed significant reductions up to 6dB in blade-vortex-interaction and in-plane noise, as well as reductions in vibratory hub loads up to 80%. Trailing-edge flap deflections were controlled within 0.1 degrees of the commanded value. The impact of the active flap on control power, rotor smoothing, and performance was also demonstrated. Finally, the reliability of the flap actuation system was successfully proven in more than 60 hours of wind-tunnel testing.

  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. 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

  12. Effects of MK-886, a 5-lipoxygenase activating protein (FLAP) inhibitor, and 5-lipoxygenase deficiency on the forced swimming behavior of mice.

    PubMed

    Uz, Tolga; Dimitrijevic, Nikola; Imbesi, Marta; Manev, Hari; Manev, Radmila

    2008-05-01

    A common biological pathway may contribute to the comorbidity of atherosclerosis and depression. Increased activity of the enzymatic 5-lipoxygenase (5-LOX, 5LO) pathway is a contributing factor in atherosclerosis and a 5-LOX inhibitor, MK-886, is beneficial in animal models of atherosclerosis. In the brain, MK-886 increases phosphorylation of the glutamate receptor subunit GluR1, and the increased phosphorylation of this receptor has been associated with antidepressant treatment. In this work, we evaluated the behavioral effects of MK-886 in an automated assay of mouse forced swimming, which identifies antidepressant activity as increased climbing behavior and/or decreased rest time. Whereas a single injection of MK-886 (3 and 10 mg/kg) did not affect forced swimming behaviors assayed 30 min later, six daily injections of 3 mg/kg MK-886 slightly increased climbing and significantly reduced rest time in wild-type mice but not in 5-LOX-deficient mice. A diet delivery of MK-886, 4 micro/(100 mg(body-weight)day), required 3 weeks to affect forced swimming; it increased climbing behavior. Climbing behavior was also increased in naive 5-LOX-deficient mice compared to naive wild-type controls. These results suggest that 5-LOX inhibition and deficiency may be associated with antidepressant activity. Increased climbing in a forced swimming assay is a typical outcome of antidepressants that increase noradrenergic and dopaminergic activity. Interestingly, 5-LOX deficiency and MK-886 treatment have been shown to be capable of increasing the behavioral effects of a noradrenaline/dopamine-potentiating drug, cocaine. Future research is needed to evaluate the clinical relevance of our findings. PMID:18403121

  13. Metabolic adaptations in delayed skin flaps. Glucose utilization and hexokinase activity.

    PubMed

    Im, M J; Su, C T; Hoopes, J E

    1979-08-01

    The distribution of glucose and hexokinase activity was determined in the epithelial tissue of delayed bipedicled skin flaps in guinea pigs. The periods of "delay" were 1, 3, 7, 14, or 21 days. The flap survival was maximal (100% of the flap) when the flap elevation was performed either 7 or 14 days following the "delay" procedure. When the flap elevation was performed 1, 3, or 21 days following the "delay" procedure, the result was partial necrosis. A differential distribution of epithelial glucose was found within the bipedicled flaps. The lowest glucose level (30% of normal) was at a distance of 2 to 3.5 cm from the end of the caudal pedicle during the first day after the "delay" procedure. This decreased glucose content recovered toward normal levels during the later part of the "delay" period. The bipedicled flaps exhibited increased hexokinase activity during the 3-week period of the "delay," and the responses of hexokinase activity and tissue glucose levels to the "delay" procedure were reciprocal in the caudal half of the flaps. PMID:377341

  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. The Effect of the Active Ingredient Thymoquinone on Flap Viability in Random Pattern Flaps in Rats.

    PubMed

    Kocak, Omer Faruk; Bozan, Nazim; Oksuz, Mustafa; Yuce, Serdar; Demir, Canser Yılmaz; Bulut, Gulay; Ragbetli, Murat Cetin

    2016-08-01

    Thymoquinone (TQ) is a plant extract that has been shown to have antioxidant, anti-inflammatory, angiogenic, antimicrobial, and anticarcinogenic effects. The aim of this study is to research how the use of TQ affects flap viability. 42 rats were placed into 6 groups, with 7 rats in each. A 3 × 10 cm McFarlane flap model was used on the test animals. The sham group had used neither surgical nor TQ treatment. The control group had surgery but no treatment afterwards. The preoperative TQ group was given oral doses of 2 mg/kg. TQ for 10 days preoperatively with no treatment after the surgical procedure. The postoperative TQ group received oral doses of 2 mg/kg TQ for 10 days after the surgical process. The preoperative + postoperative (pre + postoperative) TQ group was given oral doses of 2 mg/kg TQ for 10 days both preoperatively and postoperatively. Finally, the dimethylsulfoxide group received 10 mg/kg dimethylsulfoxide (DMSO) for 10 days both preoperatively and postoperatively. Ten days after surgery the findings were evaluated. The average rates of necrosis were found to be 29.7 % in the control group, 19.18 % in the preoperative TQ group, 13.05 % in the postoperative TQ group, 8.42 % in the pre + postoperative TQ group, and 29.03 % in the DMSO group. The experimental groups had better area measurement, histopathological, and electron microscopic results than the control group (All; p < 0.05). We believe that, because of its antioxidant, anti-inflammatory, and angiogenic properties, thymoquinone is an agent that can prevent ischemia-reperfusion damage and, therefore, prevent necrosis. PMID:27072137

  17. 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

  18. 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

  19. Development and whirl tower test of the SMART active flap rotor

    NASA Astrophysics Data System (ADS)

    Straub, Friedrich K.; Kennedy, Dennis K.; Stemple, Alan D.; Anand, V. R.; Birchette, Terry S.

    2004-07-01

    A full scale Smart Material Actuated Rotor Technology (SMART) system with piezoelectric actuated blade flaps was developed and whirl tower tested. The development effort included design, fabrication, and component testing of rotor blades, trailing edge flaps, piezoelectric actuators, switching power amplifiers, and the data/power system. Simulations and model scale wind tunnel tests have shown that this system can provide 80% vibration reduction, 10dB noise reduction for a helicopter passing overhead, and substantial aerodynamic performance gains. Whirl tower testing of the 34-foot diameter rotor demonstrated the functionality, robustness, and required authority of the active flap system. The program involved extensive development work and risk reduction tests which resulted in a robust, high performance actuator and a tightly integrated actuator, flap, and blade system. The actuator demonstrated excellent performance during bench testing and has accumulated over 60 million cycles under a spectrum of loading conditions. The flight worthy active flap rotor blades were based on a modified design of the FAA certified MD900 Explorer production rotor blade. Whirl tower testing was conducted with full rotor instrumentation and a 5-component balance. The rotor was tested for 13 hours under a range of conditions, including 7 hours of flap operation. Flap inputs included open loop static and dynamic commands. The flaps showed excellent authority with oscillatory thrust greater than 10% of the steady baseline thrust. Various flap actuation frequency sweeps were run to investigate the dynamics of the rotor and the flap system. Limited closed loop tests used hub accelerations and hub loads for feedback. Proving the integration, robust operation, and authority of the flap system were the key objectives met by the whirl tower test. This success depended on tailoring the piezoelectric materials and actuator to the application and meeting actuator/blade integration requirements

  20. Design and development of an active Gurney flap for rotorcraft

    NASA Astrophysics Data System (ADS)

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

    2013-03-01

    The EU's Green Rotorcraft programme will develop an Active Gurney Flap (AGF) for a full-scale helicopter main rotor blade as part of its `smart adaptive rotor blade' technology demonstrators. AGFs can be utilized to provide a localized and variable lift enhancement on the rotor, enabling a redistribution of loading on the rotor blade around the rotor azimuth. Further advantages include the possibility of using AGFs to allow a rotor speed reduction, which subsequently provides acoustic benefits. Designed to be integrable into a commercial helicopter blade, and thereby capable of withstanding real in-flight centrifugal loading, blade vibrations and aerodynamic loads, the demonstrator is expected to achieve a high technology readiness level (TRL). The AGF will be validated initially by a constant blade section 2D wind tunnel test and latterly by full blade 3D whirl tower testing. This paper presents the methodology adopted for the AGF concept topology selection, based on a series of both qualitative and quantitative performance criteria. Two different AGF candidate mechanisms are compared, both powered by a small commercial electromagnetic actuator. In both topologies, the link between the actuator and the control surface consists of two rotating torque bars, pivoting on flexure bearings. This provides the required reliability and precision, while making the design virtually frictionless. The engineering analysis presented suggests that both candidates would perform satisfactorily in a 2D wind tunnel test, but that equally, both have design constraints which limit their potential to be further taken into a whirl tower test under full scale centrifugal and inertial loads.

  1. Myeloid Cell 5-Lipoxygenase Activating Protein Modulates the Response to Vascular Injury

    PubMed Central

    Yu, Zhou; Ricciotti, Emanuela; Miwa, Takashi; Liu, Shulin; Ihida-Stansbury, Kaori; Landersberg, Gavin; Jones, Peter L.; Scalia, Rosario; Song, Wenchao; Assoian, Richard K.; FitzGerald, Garret A.

    2013-01-01

    Rationale Human genetics have implicated the 5- lipoxygenase (5-LO) enzyme in the pathogenesis of cardiovascular disease and an inhibitor of the 5-LO activating protein (FLAP) is in clinical development for asthma. Objective Here we determined whether FLAP deletion modifies the response to vascular injury. Methods and Results Vascular remodeling was characterized 4 weeks after femoral arterial injury in FLAP knockout (FLAP KO) mice and wild type (WT) controls. Both neointimal hyperplasia and the intima/media ratio of the injured artery were significantly reduced in the FLAP KOs while endothelial integrity was preserved. Lesional myeloid cells were depleted and vascular smooth muscle cell (VSMC) proliferation, as reflected by bromodeoxyuridine (BrdU) incorporation, was markedly attenuated by FLAP deletion. Inflammatory cytokine release from FLAP KO macrophages was depressed and their restricted ability to induce VSMC migration ex vivo was rescued with leukotriene B4 (LTB4). FLAP deletion restrained injury and attenuated upregulation of the extracellular matrix protein, tenascin C (TNC), which affords a scaffold for VSMC migration. Correspondingly, the phenotypic modulation of VSMC to a more synthetic phenotype, reflected by morphological change, loss of α-smooth muscle cell actin and upregulation of vascular cell adhesion molecule (VCAM) -1 was also suppressed in FLAP KO mice. Transplantation of FLAP replete myeloid cells rescued the proliferative response to vascular injury. Conclusion Expression of lesional FLAP in myeloid cells promotes LTB4 dependent VSMC phenotypic modulation, intimal migration and proliferation. PMID:23250985

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. Tanshinone IIA Pretreatment Renders Free Flaps against Hypoxic Injury through Activating Wnt Signaling and Upregulating Stem Cell-Related Biomarkers

    PubMed Central

    Xu, Zihan; Zhang, Zhenxin; Wu, Lijun; Sun, Yaowen; Guo, Yadong; Qin, Gaoping; Mu, Shengzhi; Fan, Ronghui; Wang, Benfeng; Gao, Wenjie

    2014-01-01

    Partial or total flap necrosis after flap transplantation is sometimes clinically encountered in reconstructive surgery, often as a result of a period of hypoxia that exceeds the tolerance of the flap tissue. In this study, we determine whether tanshinone IIA (TSA) pretreatment can protect flap tissue against hypoxic injury and improve its viability. Primary epithelial cells isolated from the dorsal skin of mice were pretreated with TSA for two weeks. Cell counting kit-8 and Trypan Blue assays were carried out to examine the proliferation of TSA-pretreated cells after exposure to cobalt chloride. Then, Polymerase chain reaction and Western blot analysis were used to determine the expression of β-catenin, GSK-3β, SOX2, and OCT4 in TSA-treated cells. In vivo, after mice were pretreated with TSA for two weeks, a reproducible ischemic flap model was implemented, and the area of surviving tissue in the transplanted flaps was measured. Immunohistochemistry was also conducted to examine the related biomarkers mentioned above. Results show that epidermal cells, pretreated with TSA, showed enhanced resistance to hypoxia. Activation of the Wnt signaling pathway in TSA-pretreated cells was characterized by the upregulation of β-catenin and the downregulation of GSK-3β. The expression of SOX2 and OCT4 controlled by Wnt signaling were also found higher in TSA pretreated epithelial cells. In the reproducible ischaemic flap model, pretreatment with TSA enhanced resistance to hypoxia and increased the area of surviving tissue in transplanted flaps. The expression of Wnt signaling pathway components, stem-cell related biomarkers, and CD34, which are involved in the regeneration of blood vessels, was also upregulated in TSA-pretreated flap tissue. The results show that TSA pretreatment protects free flaps against hypoxic injury and increases the area of surviving tissue by activating Wnt signaling and upregulating stem cell-related biomarkers. PMID:25302618

  7. 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

  8. 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.

  9. Pitching-motion-activated flapping foil near solid walls for power extraction: A numerical investigation

    NASA Astrophysics Data System (ADS)

    Wu, J.; Qiu, Y. L.; Shu, C.; Zhao, N.

    2014-08-01

    A numerical investigation on the power extraction of a pitching-motion-activated flapping foil near solid walls is performed by using an immersed boundary-lattice Boltzmann method in this study. The flapping motions of the foil include a forced pitching component and an induced plunging component. The foil is placed either near a solid wall or between two parallel plane walls. Compared to previous work on the flapping foil for power extraction, the effect of the walls is first considered in this work. At a Reynolds number of 1100 and with the position of the foil pitching axis at third chord, the influences of the mechanical parameters (such as damping coefficient and spring constant) of the foil, the amplitude and frequency of the pitching motion and the clearance between the foil pitching axis and the wall on the power extraction performance of the flapping foil are systematically evaluated. Compared to the situation of free stream, the power extraction performance of the foil near the wall is improved. For given amplitude and frequency, as the clearance decreases the net power extraction efficiency improves. Moreover, as the foil is placed near one wall, there is a transverse shift to the plunging motion that consequently weakens the improvement of net power extraction efficiency. In contrast, the shift can be significantly eliminated as the foil is placed between two walls, which can further improve the net power extraction efficiency. In addition, it is found that the efficiency improvement is essentially from the increased power extraction, which is due to the generation of high lift force.

  10. 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

  11. Use of active control systems to improve bending and rotor flapping response of a tilt rotor VTOL airplane

    NASA Technical Reports Server (NTRS)

    Whitaker, H. P.; Cheng, Y.

    1975-01-01

    The results are summarized of an analytical study of the use of active control systems for the purpose of reducing the root mean square response of wing vertical bending and rotor flapping to atmospheric turbulence for a tilt-rotor VTOL airplane. Only the wing/rotor assembly was considered so that results of a wind tunnel test program would be applicable in a subsequent phase of the research. The capabilities and limitations of simple single feedback configurations were identified, and the most promising multiloop feedback configurations were then investigated. Design parameters were selected so as to minimize either wing bending or rotor flapping response. Within the constraints imposed by practical levels of feedback gains and complexity and by considerations of safety, reduction in response due to turbulence of the order of 30 to 50 percent is predicted using the rotor longitudinal cyclic and a trailing edge wing flap as control effectors.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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

  18. Reduction of urease activity by interaction with the flap covering the active site.

    PubMed

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

    2015-02-23

    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

  19. 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

  20. 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

  1. 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.

  2. Axial flow effects on robustness of vortical structures about actively deflected wings in flapping flight

    NASA Astrophysics Data System (ADS)

    Medina, Albert; Kweon, Jihoon; Choi, Haecheon; Eldredge, Jeff D.

    2012-11-01

    Flapping wing flight has garnered much attention in the past decade driven by our desire to understand capabilities observed in nature and to develop agile small-scale aerial vehicles. Nature has demonstrated the breadth of maneuverability achievable by flapping wing flight. However, despite recent advances the role of wing flexibility remains poorly understood. In an effort to develop a deeper understanding of wing deflection effects and to explore novel approaches to increasing leading-edge vortex robustness, this three-dimensional computational study explores the aerodynamics of low aspect ratio plates, in hovering kinematics, with isolated flexion lines undergoing prescribed deflection. Major flexion lines, recognized as the primary avenue for deflection in biological fliers, are isolated here in two distinct configurations, resulting in deflection about the wing root and the wing tip, respectively. Of interest is the interaction between axial flow along the span and the vortical structures about the wing. It is proposed that the modes of deflection explored may provide a means of axial flow control for favorably promoting LEV robustness over a broad range of flapping conditions, and provide insight into the nature of flexibility in flapping wing flight. National Science Foundation, National Research Foundation of Korea.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. Active control system for a rotor blade trailing-edge flap

    NASA Astrophysics Data System (ADS)

    Duvernier, Marc; Reithler, Livier; Guerrero, Jean Y.; Rossi, Rinaldo A.

    2000-06-01

    Reducing the external noise is becoming a major issue for helicopter manufacturers. The idea beyond this goal is to reduce or even avoid the blade vortex interaction (BVI), especially during descent and flights over inhabited areas. This can be achieved by changing locally the lift of the blade. Several strategies to reach this goal are under investigation at EUROCOPTER such as the control of the local incidence of the blade by a direct lift flap. AEROSPATIALE MATRA Corporate Research Centre and AEROSPATIALE MATRA MISSILES proposed an actuator system able to answer EUROCOPTER's needs for moving a direct lift flap. The present paper describes the definition, manufacturing and testing of this new actuator system. This actuator is based on an electromagnetic patented actuation system developed by AEROSPATIALE MATRA MISSILES for missile and aeronautic applications. The particularity of this actuator is its ability to produce the desired force on its whole range of stroke. The flap is designed to be fitted on a DAUPHIN type blade produced by EUROCOPTER and the actuator system was designed to fit the room available within the blade and to produce the right amount of stroke and force within the required frequency range. Other constraints such as centrifugal loading were also taken into account. This paper describes briefly the specifications and the major characteristics of the actuating system and presents some results of its behavior on a representative composite test-bed manufactured by EUROCOPTER when subjected to realistic mechanical loads.

  8. 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

  9. 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

  10. 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...

  11. Use of microbial transglutaminase and non-meat proteins to improve 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 a value-added low sodium chloride (NaCl), phosphate-free patty using minced channel catfish (Ictalurus punctatus) belly flap meat. The effect of microbial transglutaminase (MTGase) and non-meat proteins (isolated soy protein, ISP and whey protein concentrate, WPC) ...

  12. 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.

  13. 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 focused on 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 prote...

  14. 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

  15. Protein extraction from activated sludge.

    PubMed

    Denecke, M

    2006-01-01

    Two methods for the separation of protein originating from activated sludge were compared. In one method, the total protein was isolated out of the activated sludge (crude extract). These samples included all dissolved proteins originating from the bacterial cells and biofilm made up of extracellular polymeric substances (EPS). Every time polyacrylamide gel electrophoresis (PAGE) was done, the protein bands from samples of crude extract were covered by polymeric substances including carbohydrates, uronic acids or humic compounds. Using the immunoblot technique it was possible to demonstrate the presence of the heat shock protein HSP70 in crude extracts of activated sludge. The comparison of protein fingerprints required that clear and distinct bands appear on the PAGE analysis. To this end, a procedure to separates bacterial cells from the EPS was developed. Bacterial cells were separated by incubation with EDTA and subsequent filtration. The isolated cells were directly incubated in a sample buffer. PMID:16898150

  16. Segmented vortex flaps

    NASA Technical Reports Server (NTRS)

    Rao, D. M.

    1983-01-01

    Segmented vortex flaps were suggested as a means of delaying the vortex spill-over causing thrust loss over the outboard region of single-panel flaps. Also proposed was hinge-line setback for exploiting leading-edge suction in conjunction with vortex flaps to improve the overall thrust per unit flap area. These two concepts in combination were tested on a 60-deg cropped delta wing model. Significant improvement in flap efficiency was indicated by a reduction of the flap/wing area from 11.4% of single-panel flap to 6.3% of a two segment delta flap design, with no lift/drag penalty at lift coefficients between 0.5 and 0.7. The more efficient vortex flap arrangement of this study should benefit the performance attainable with flaps of given area on wings of moderate leading-edge sweep.

  17. Activation of an Alternative, Rec12 (Spo11)-Independent Pathway of Fission Yeast Meiotic Recombination in the Absence of a DNA Flap Endonuclease

    PubMed Central

    Farah, Joseph A.; Cromie, Gareth; Davis, Luther; Steiner, Walter W.; Smith, Gerald R.

    2005-01-01

    Spo11 or a homologous protein appears to be essential for meiotic DNA double-strand break (DSB) formation and recombination in all organisms tested. We report here the first example of an alternative, mutationally activated pathway for meiotic recombination in the absence of Rec12, the Spo11 homolog of Schizosaccharomyces pombe. Rad2, a FEN-1 flap endonuclease homolog, is involved in processing Okazaki fragments. In its absence, meiotic recombination and proper segregation of chromosomes were restored in rec12Δ mutants to nearly wild-type levels. Although readily detectable in wild-type strains, meiosis-specific DSBs were undetectable in recombination-proficient rad2Δ rec12Δ strains. On the basis of the biochemical properties of Rad2, we propose that meiotic recombination by this alternative (Rec*) pathway can be initiated by non-DSB lesions, such as nicks and gaps, which accumulate during premeiotic DNA replication in the absence of Okazaki fragment processing. We compare the Rec* pathway to alternative pathways of homologous recombination in other organisms. PMID:16118186

  18. Slotted variable camber flap

    NASA Technical Reports Server (NTRS)

    Andrews, D. G. (Inventor)

    1984-01-01

    Variable camber actuator assemblies broaden the range of speeds at which lift to drag performance is maximized for slotted flap wings. Lift is improved over a broader range of cruising speeds by varying wing camber with rotational flap movements that do not introduce wing slots and induced drag. Forward flaps are secured to forward flange links which extended from, and are a part of forward flap linkage assemblies. The forward flaps rotate about flap pivots with their rotational displacement controlled by variable camber actuator assemblies located between the forward flaps and the forward flange links. Rear flaps are held relative to the forward flaps by rear flap linkage assemblies which may act independently from the forward flap linkage assemblies and the variable camber actuator assemblies. Wing camber is varied by rotating the flaps with the variable camber actuator assemblies while the flaps are in a deployed or tucked position. Rotating the flaps in a tucked position does not introduce significant wing surface discontinuities, and reduces aircraft fuel consumption on most flight profiles.

  19. 5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation.

    PubMed

    Gerstmeier, Jana; Newcomer, Marcia E; Dennhardt, Sophie; Romp, Erik; Fischer, Jana; Werz, Oliver; Garscha, Ulrike

    2016-05-01

    Leukotrienes (LTs) are proinflammatory lipid mediators formed from arachidonic acid in a 2-step reaction catalyzed by 5-lipoxygenase (5-LOX) requiring the formation of 5-HPETE [5(S)-hydroperoxy-6-trans-8,11,14-cis-eicosatetraenoic acid] and its subsequent transformation to LTA4 5-LOX is thought to receive arachidonic acid from the nuclear membrane-embedded 5-LOX-activating protein (FLAP). The crystal structure of 5-LOX revealed an active site concealed by F177 and Y181 (FY cork). We examined the influence of the FY cork on 5-LOX activity and membrane binding in HEK293 cells in the absence and presence of FLAP. Uncapping the 5-LOX active site by mutation of F177 and/or Y181 to alanine (5-LOX-F177A, 5-LOX-Y181A, 5-LOX-F177/Y181A) resulted in delayed and diminished 5-LOX membrane association in A23187-stimulated cells. For 5-LOX-F177A and 5-LOX-F177/Y181A, formation of 5-LOX products was dramatically reduced relative to 5-LOX-wild type (wt). Strikingly, coexpression of FLAP in A23187-activated HEK293 cells effectively restored formation of 5-H(p)ETE (5-hydroxy- and 5-peroxy-6-trans-8,11,14-cis-eicosatetraenoic acid) by these same 5-LOX mutants (≈60-70% 5-LOX-wt levels) but not of LTA4 hydrolysis products. Yet 5-LOX-Y181A generated 5-H(p)ETE at levels comparable to 5-LOX-wt but reduced LTA4 hydrolysis products. Coexpression of FLAP partially restored LTA4 hydrolysis product formation by 5-LOX-Y181A. Together, the data suggest that the concealed FY cork impacts membrane association and that FLAP may help shield an uncapped active site.-Gerstmeier, J., Newcomer, M. E., Dennhardt, S., Romp, E., Fischer, J., Werz, O., Garscha, U. 5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation. PMID:26842853

  20. A comparative molecular dynamics study on BACE1 and BACE2 flap flexibility.

    PubMed

    Kumalo, H M; Soliman, Mahmoud E

    2016-10-01

    Beta-amyloid precursor protein cleavage enzyme1 (BACE1) and beta-amyloid precursor protein cleavage enzyme2 (BACE2), members of aspartyl protease family, are close homologs and have high similarity in their protein crystal structures. However, their enzymatic properties are different, which leads to different clinical outcomes. In this study, we performed sequence analysis and all-atom molecular dynamic (MD) simulations for both enzymes in their ligand-free states in order to compare their dynamical flap behaviors. This is to enhance our understanding of the relationship between sequence, structure and the dynamics of this protein family. Sequence analysis shows that in BACE1 and BACE2, most of the ligand-binding sites are conserved, indicative of their enzymatic property as aspartyl protease members. The other conserved residues are more or less unsystematically localized throughout the structure. Herein, we proposed and applied different combined parameters to define the asymmetric flap motion; the distance, d1, between the flap tip and the flexible region; the dihedral angle, φ, to account for the twisting motion and the TriCα angle, θ2 and θ1. All four combined parameters were found to appropriately define the observed "twisting" motion during the flaps different conformational states. Additional analysis of the parameters indicated that the flaps can exist in an ensemble of conformations, i.e. closed, semi-open and open conformations for both systems. However, the behavior of the flap tips during simulations is different between BACE1 and BACE2. The BACE1 active site cavity is more spacious as compared to that of BACE2. The analysis of 10S loop and 113S loop showed a similar trend to that of flaps, with the BACE1 loops being more flexible and less stable than those of BACE2. We believe that the results, methods and perspectives highlighted in this report would assist researchers in the discovery of BACE inhibitors as potential Alzheimer's disease therapies

  1. Bacterial Heat Shock Protein Activity

    PubMed Central

    Maleki, Farajollah; Khosravi, Afra; Nasser, Ahmad; Taghinejad, Hamid

    2016-01-01

    Bacteria are exposed to different types of stress in their growth conditions. They have developed appropriate responses, modulated by the re-modeling of protein complexes and by phosphorylation dependent signal transduction systems, to adapt and to survive in a variety range of nature. Proteins are essential components for biologic activity in the eukaryotic and prokaryotic cell. Heat Shock Proteins (HSP) have been identified from various organisms and have critical role in cell hemostasis. Chaperone can sense environment and have different potential role in the organism evolution. PMID:27134861

  2. Antiviral activities of whey proteins.

    PubMed

    Ng, Tzi Bun; Cheung, Randy Chi Fai; Wong, Jack Ho; Wang, Yan; Ip, Denis Tsz Ming; Wan, David Chi Cheong; Xia, Jiang

    2015-09-01

    Milk contains an array of proteins with useful bioactivities. Many milk proteins encompassing native or chemically modified casein, lactoferrin, alpha-lactalbumin, and beta-lactoglobulin demonstrated antiviral activities. Casein and alpha-lactalbumin gained anti-HIV activity after modification with 3-hydroxyphthalic anhydride. Many milk proteins inhibited HIV reverse transcriptase. Bovine glycolactin, angiogenin-1, lactogenin, casein, alpha-lactalbumin, beta-lactoglobulin, bovine lactoferrampin, and human lactoferrampin inhibited HIV-1 protease and integrase. Several mammalian lactoferrins prevented hepatitis C infection. Lactoferrin, methylated alpha-lactalbumin and methylated beta-lactoglobulin inhibited human cytomegalovirus. Chemically modified alpha-lactalbumin, beta-lactoglobulin and lysozyme, lactoferrin and lactoferricin, methylated alpha-lactalbumin, methylated and ethylated beta-lactoglobulins inhibited HSV. Chemically modified bovine beta-lactoglobulin had antihuman papillomavirus activity. Beta-lactoglobulin, lactoferrin, esterified beta-lactoglobulin, and esterified lactoferrindisplayed anti-avian influenza A (H5N1) activity. Lactoferrin inhibited respiratory syncytial virus, hepatitis B virus, adenovirus, poliovirus, hantavirus, sindbis virus, semliki forest virus, echovirus, and enterovirus. Milk mucin, apolactoferrin, Fe(3+)-lactoferrin, beta-lactoglobulin, human lactadherin, bovine IgG, and bovine kappa-casein demonstrated antihuman rotavirus activity. PMID:26198883

  3. Abrogation of fibroblast activation protein enzymatic activity attenuates tumor growth.

    PubMed

    Cheng, Jonathan D; Valianou, Matthildi; Canutescu, Adrian A; Jaffe, Eileen K; Lee, Hyung-Ok; Wang, Hao; Lai, Jack H; Bachovchin, William W; Weiner, Louis M

    2005-03-01

    Tumor-associated fibroblasts are functionally and phenotypically distinct from normal fibroblasts that are not in the tumor microenvironment. Fibroblast activation protein is a 95 kDa cell surface glycoprotein expressed by tumor stromal fibroblasts, and has been shown to have dipeptidyl peptidase and collagenase activity. Site-directed mutagenesis at the catalytic site of fibroblast activation protein, Ser624 --> Ala624, resulted in an approximately 100,000-fold loss of fibroblast activation protein dipeptidyl peptidase (DPP) activity. HEK293 cells transfected with wild-type fibroblast activation protein, enzymatic mutant (S624A) fibroblast activation protein, or vector alone, were inoculated subcutaneously into immunodeficient mouse to assess the contribution of fibroblast activation protein enzymatic activity to tumor growth. Overexpression of wild-type fibroblast activation protein showed growth potentiation and enhanced tumorigenicity compared with both fibroblast activation protein S624A and vector-transfected HEK293 xenografts. HEK293 cells transfected with fibroblast activation protein S624A showed tumor growth rates and tumorigenicity potential similar only to vector-transfected HEK293. In vivo assessment of fibroblast activation protein DPP activity of these tumors showed enhanced enzymatic activity of wild-type fibroblast activation protein, with only baseline levels of fibroblast activation protein DPP activity in either fibroblast activation protein S624A or vector-only xenografts. These results indicate that the enzymatic activity of fibroblast activation protein is necessary for fibroblast activation protein-driven tumor growth in the HEK293 xenograft model system. This establishes the proof-of-principle that the enzymatic activity of fibroblast activation protein plays an important role in the promotion of tumor growth, and provides an attractive target for therapeutics designed to alter fibroblast activation protein-induced tumor growth by targeting

  4. The platysma myocutaneous flap.

    PubMed

    Baur, Dale A; Williams, Jonathan; Alakaily, Xena

    2014-08-01

    Reconstructing defects of the oral mucosa or skin of the lower one-third of the face can be accomplished by a variety of techniques. This article presents two versions of the platysma myocutaneous flap, which is a reliable, axial pattern, pedicled flap capable of providing excellent one-stage reconstruction of such defects. As discussed herein, the superiorly based and posteriorly based versions of the flap have wide application in the oral and facial region. Also provided is a review of other uses of this flap in head and neck surgery. PMID:24958382

  5. Keyhole Flap Nipple Reconstruction

    PubMed Central

    Cash, Camille G.; Iman, Al-Haj; Spiegel, Aldona J.; Cronin, Ernest D.

    2016-01-01

    Summary: 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.

  6. 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

  7. Protein Needs of Physically Active Children.

    PubMed

    Volterman, Kimberly A; Atkinson, Stephanie A

    2016-05-01

    Current Dietary Reference Intakes (DRI) for protein for children and youth require revision as they were derived primarily on nitrogen balance data in young children or extrapolated from adult values; did not account for the possible influence of above average physical activity; and did not set an upper tolerable level of intake. Revision of the protein DRIs requires new research that investigates: 1) long-term dose-response to identify protein and essential amino acid requirements of both sexes at various pubertal stages and under differing conditions of physical activity; 2) the acute protein needs (quantity and timing) following a single bout of exercise; 3) the potential adverse effects of chronic high intakes of protein; and 4) new measurement techniques (i.e., IAAO or stable isotope methodologies) to improve accuracy of protein needs. While active individuals may require protein in excess of current DRIs, most active Canadian children and youth have habitual protein intakes that exceed current recommendations. PMID:27137165

  8. 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.

  9. Effects of a synovial flap and gelatin/β-tricalcium phosphate sponges loaded with mesenchymal stem cells, bone morphogenetic protein-2, and platelet rich plasma on equine osteochondral defects.

    PubMed

    Seo, Jong-Pil; Kambayashi, Yoshinori; Itho, Megumi; Haneda, Shingo; Yamada, Kazutaka; Furuoka, Hidefumi; Tabata, Yasuhiko; Sasaki, Naoki

    2015-08-01

    This study aimed to evaluate the efficacy of a synovial flap and gelatin/β-tricalcium phosphate (GT) sponge loaded with mesenchymal stem cells (MSCs), bone morphogenetic protein-2 (BMP-2), and platelet rich plasma (PRP) for repairing of osteochondral defects in horses. Osteochondral defects were created on the medial condyle of both femurs (n=5). In the test group, a GT sponge loaded with MSCs, BMP-2, and PRP (GT/MSCs/BMP-2/PRP) was inserted into the defect and then covered with a synovial flap. In the control group, the defect was treated only with the GT/MSCs/BMP-2/PRP. The test group showed significantly higher macroscopic scores than the control group. In addition, hyaline cartilaginous tissue was detected in the test group in areas larger than those in the control group. This study demonstrated that the combination of a synovial flap and GT sponge loaded with MSCs, BMP-2, and PRP promoted osteochondral regeneration in an equine model. PMID:26267104

  10. 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.

  11. Oral reconstruction with submental flap

    PubMed Central

    Rahpeyma, Amin; Khajehahmadi, Saeedeh

    2013-01-01

    Background: Submental flap is a useful technique for reconstruction of medium to large oral cavity defects. Hair bearing nature of this flap in men makes it less appropriate. Therefore, deepithelialized variant is introduced to overcome the problem of hair with this flap. Recently, application of this flap has been introduced in maxillofacial trauma patients. Materials and Methods: Deepithelialized orthograde submental flap is used for the reconstruction of oral cavity mucosal defects. Results: Four cases including two trauma patients and two squamous cell carcinomas (SCCs) of oral cavity were treated using deepithelialized orthograde submental flap. There were no complications in all four patients and secondary epithelialization occurred in raw surface of the flap which was exposed to oral cavity. Conclusion: Deepithelialized orthograde submental flap is very effective in reconstruction of oral cavity in men. The problem of hair is readily solved using this technique without jeopardizing flap blood supply. PMID:24205473

  12. Long Wavelength Monitoring of Protein Kinase Activity

    PubMed Central

    Oien, Nathan P.; Nguyen, Luong T.; Jernigan, Finith E.; Priestman, Melanie A.

    2014-01-01

    A family of long wavelength protein kinase fluorescent reporters is described in which the probing wavelength is pre-programmed using readily available fluorophores. These agents can assess protein kinase activity within the optical window of tissue, as exemplified by monitoring endogenous cAMP-dependent protein kinase activity (1) in erythrocyte lysates and (2) in intact erythrocytes using a light-activatable reporter. PMID:24604833

  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. 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.

  15. Free craniotomy versus osteoplastic craniotomy, assessment of flap viability using 99mTC MDP SPECT.

    PubMed

    Shelef, Ilan; Golan, Haim; Merkin, Vladimir; Melamed, Israel; Benifla, Mony

    2016-09-01

    There are currently two accepted neurosurgical methods to perform a bony flap. In an osteoplastic flap, the flap is attached to surrounding muscle. In a free flap, the flap is not attached to adjacent tissues. The former is less common due to its complexity and the extensive time required for the surgery; yet the rate of infection is significantly lower, a clear explanation for which is unknown. The objective of this study was to test the hypothesis that the osteoplastic flap acts as a live implant that resumes its blood flow and metabolic activity; contrasting with the free flap, which does not have sufficient blood flow, and therefore acts as a foreign body. Seven patients who underwent craniotomy with osteoplastic flaps and five with free flaps had planar bone and single photon emission computed tomography (SPECT) scans of the skull at 3-7days postoperative, after injection of the radioisotope, 99m-technetium-methylene diphosphonate (99m-Tc-MDP). We compared radioactive uptake as a measure of metabolic activity between osteoplastic and free flaps. Mean normalized radioactive uptakes in the centers of the flaps, calculated as the ratios of uptakes in the flap centers to uptakes in normal contralateral bone, were [mean: 1.7 (SD: 0.8)] and [0.6 (0.1)] for the osteoplastic and free flap groups respectively and were [2.4 (0.8)] and [1.3 (0.4)] in the borders of the flaps. Our analyses suggest that in craniotomy, the use of an osteoplastic flap, in contrast to free flap, retains bone viability. PMID:27068014

  16. Externally blown flap impingement noise

    NASA Technical Reports Server (NTRS)

    Lasagna, P. L.; Putnam, T. W.

    1972-01-01

    Tests of the noise produced by the impingement of the jet exhaust on the wing and flap for an externally blown flap system were conducted with a CF700 turbofan engine and an F-111B wing panel. The noise produced with a daisy nozzle installed on the engine was greater than that produced by a conical nozzle at the same thrust. The presence of the wing next to the test nozzles increased the noise, as did increasing the flap deflection angle. Compared with the conical nozzle, the daisy nozzle produced slightly less noise at a flap deflection of 60 deg but produced more noise at the lower flap deflections tested. Tests showed that the single-slotted flap deflected 60 deg, produced less noise than the double-slotted flaps. Also, maintaining the maximum distance between the exit nozzle and flap system resulted in a minor reduction in noise.

  17. Dietary protein considerations to support active aging.

    PubMed

    Wall, Benjamin T; Cermak, Naomi M; van Loon, Luc J C

    2014-11-01

    Given our rapidly aging world-wide population, the loss of skeletal muscle mass with healthy aging (sarcopenia) represents an important societal and public health concern. Maintaining or adopting an active lifestyle alleviates age-related muscle loss to a certain extent. Over time, even small losses of muscle tissue can hinder the ability to maintain an active lifestyle and, as such, contribute to the development of frailty and metabolic disease. Considerable research focus has addressed the application of dietary protein supplementation to support exercise-induced gains in muscle mass in younger individuals. In contrast, the role of dietary protein in supporting the maintenance (or gain) of skeletal muscle mass in active older persons has received less attention. Older individuals display a blunted muscle protein synthetic response to dietary protein ingestion. However, this reduced anabolic response can largely be overcome when physical activity is performed in close temporal proximity to protein consumption. Moreover, recent evidence has helped elucidate the optimal type and amount of dietary protein that should be ingested by the older adult throughout the day in order to maximize the skeletal muscle adaptive response to physical activity. Evidence demonstrates that when these principles are adhered to, muscle maintenance or hypertrophy over prolonged periods can be further augmented in active older persons. The present review outlines the current understanding of the role that dietary protein occupies in the lifestyle of active older adults as a means to increase skeletal muscle mass, strength and function, and thus support healthier aging. PMID:25355192

  18. Recent advances for FLAP inhibitors.

    PubMed

    Pettersen, Daniel; Davidsson, Öjvind; Whatling, Carl

    2015-07-01

    A number of FLAP inhibitors have been progressed to clinical trials for respiratory and other inflammatory indications but so far no drug has reached the market. With this Digest we assess the opportunity to develop FLAP inhibitors for indications beyond respiratory disease, and in particular for atherosclerotic cardiovascular disease. We also show how recently disclosed FLAP inhibitors have structurally evolved from the first generation FLAP inhibitors paving the way for new compound classes. PMID:26004579

  19. 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.

  20. 7-flap perineal urethrostomy

    PubMed Central

    Parker, Daniel C.; Morey, Allen F.

    2015-01-01

    Perineal urethrostomy (PU) has been performed with success for the treatment of refractory and advanced urethral stricture disease for at least the past six decades. Here, we review the indications and outcomes of PU for indications such as complex hypospadias repair and urethral stricture disease resulting from trauma, infection, and failure of prior urethroplasty. We also describe the role of 7-flap PU, a novel alternative to the conventional approach that offers the surgeon added flexibility in tailoring urethrostomy creation based on intraoperative findings. The authors’ updated experience with 7-flap PU demonstrates a comparable 95% success rate in patients with a wide variety of stricture etiology. PU through either a conventional approach or a 7-flap technique is a valuable option for improving the quality of life in patients with debilitating urethral stricture disease. PMID:26816809

  1. A novel five-lipoxygenase activity protein inhibitor labeled with carbon-14 and deuterium.

    PubMed

    Latli, Bachir; Hrapchak, Matt; Gao, Joe J; Busacca, Carl A; Senanayake, Chris H

    2015-07-01

    2-[4-(3-{(1R)-1-[4-(2-Aminopyrimidin-5-yl)phenyl]-1-cyclopropylethyl}-1,2,4-oxadiazol-5-yl)-1H-pyrazol-1-yl]-N,N-dimethylacetamide (1), is a novel and selective five-lipoxygenase activity protein (FLAP) inhibitor with excellent pharmacokinetics properties. The availability of a key chiral intermediate allowed the synthesis of [(14) C]-(1) in six radiochemical steps and in 47% overall radiochemical yield with a specific activity of 51 mCi/mmol using carbon-14 zinc cyanide. 2-Chloro-N,N-dimethyl-(2)H6-acetamide was prepared and condensed with a penultimate intermediate to give [(2)H6]-(1) in very high yield and in more than 99% isotopic enrichment. PMID:26190342

  2. Externally blown flap impingement noise.

    NASA Technical Reports Server (NTRS)

    Putnam, T. W.; Lasagna, P. L.

    1972-01-01

    An investigation of externally blown flap impingement noise was conducted using a full-scale turbofan engine and aircraft wing. The noise produced with a daisy nozzle installed on the engine exhaust system was greater than that produced by a conical nozzle at the same thrust. The daisy nozzle caused the jet velocity to decay about 35 percent at the flap. The presence of the wing next to the conical nozzle increased the noise, as did increasing the flap deflection. Compared with the conical nozzle, the daisy nozzle produced slightly less noise at a flap deflection of 60 deg but produced more noise at the lower flap deflections tested.

  3. 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.

  4. 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

  5. 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

  6. DNA-based control of protein activity.

    PubMed

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

    2016-03-01

    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

  7. Uncommon Flaps for Chest Wall Reconstruction

    PubMed Central

    Matros, Evan; Disa, Joseph J.

    2011-01-01

    The omentum, external oblique musculocutaneous, and thoracoepigastric flaps are uncommonly used for chest wall reconstruction. Nevertheless, awareness and knowledge of these flaps is essential for reconstructive surgeons because they fill specific niche indications or serve as lifeboats when workhorse flaps are unavailable. The current report describes the anatomic basis, technical aspects of flap elevation, and indications for these unusual flaps. PMID:22294943

  8. 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

  9. Heart rate and estimated energy expenditure of flapping and gliding in black-browed albatrosses.

    PubMed

    Sakamoto, Kentaro Q; Takahashi, Akinori; Iwata, Takashi; Yamamoto, Takashi; Yamamoto, Maki; Trathan, Philip N

    2013-08-15

    Albatrosses are known to expend only a small amount of energy during flight. The low energy cost of albatross flight has been attributed to energy-efficient gliding (soaring) with sporadic flapping, although little is known about how much time and energy albatrosses expend in flapping versus gliding during cruising flight. Here, we examined the heart rates (used as an instantaneous index of energy expenditure) and flapping activities of free-ranging black-browed albatrosses (Thalassarche melanophrys) to estimate the energy cost of flapping as well as time spent in flapping activities. The heart rate of albatrosses during flight (144 beats min(-1)) was similar to that while sitting on the water (150 beats min(-1)). In contrast, heart rate was much higher during takeoff and landing (ca. 200 beats min(-1)). Heart rate during cruising flight was linearly correlated with the number of wing flaps per minute, suggesting an extra energy burden of flapping. Albatrosses spend only 4.6±1.4% of their time flapping during cruising flight, which was significantly lower than during and shortly after takeoff (9.8±3.5%). Flapping activity, which amounted to just 4.6% of the time in flight, accounted for 13.3% of the total energy expenditure during cruising flight. These results support the idea that albatrosses achieve energy-efficient flight by reducing the time spent in flapping activity, which is associated with high energy expenditure. PMID:23661772

  10. Towards an advanced vortex flap system: The cavity flap

    NASA Technical Reports Server (NTRS)

    Rao, D. M.

    1986-01-01

    An extension of the vortex flap concept was explored with the aim of providing high-alpha flight control capability coupled with maneuver drag reduction for highly swept wing configurations. A retractable lower surface flap mounted on a translating hinge is proposed, allowing chordwise extension as well as deflection, the two movements being independently controlled. The frontal cavity formed by the partially extended and deflected flap captures a vortex above a certain angle of attack. The cavity vortex downwash alleviates the effective incidence of the wing leading edge, thus modulating vortex lift; at the same time, the induced suction in the cavity generates thrust. These postulated aerodynamic features of the cavity flap were validated through low speed tunnel pressure and visualization tests on a 65 deg swept oblique wing model, which also provided initial trends of the leading edge vortex alleviation and cavity suction with respect to flap extension, deflection and angle of attack. Force tests on a 60 deg delta model further showed the cavity flap L/D performance to compare favorably with the conventional vortex flap. A two segment flap arrangement with independently control led segments was envisaged for exploiting the vortex modulation capability of the cavity flap for pitch, roll and yaw control, in addition to drag reduction at high angles of attack.

  11. 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

  12. 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

  13. The shunt from the cyclooxygenase to lipoxygenase pathway in human osteoarthritic subchondral osteoblasts is linked with a variable expression of the 5-lipoxygenase-activating protein.

    PubMed

    Maxis, Kelitha; Delalandre, Aline; Martel-Pelletier, Johanne; Pelletier, Jean-Pierre; Duval, Nicolas; Lajeunesse, Daniel

    2006-01-01

    Osteoarthritis (OA) is characterized by articular cartilage degradation and hypertrophic bone changes with osteophyte formation and abnormal bone remodeling. Two groups of OA patients were identified via the production of variable and opposite levels of prostaglandin E2 (PGE2) or leukotriene B4 (LTB4) by subchondral osteoblasts, PGE2 levels discriminating between low and high subgroups. We studied whether the expression of 5-lipoxygenase (5-LO) or 5-LO-activating protein (FLAP) is responsible for the shunt from prostaglandins to leukotrienes. FLAP mRNA levels varied in low and high OA groups compared with normal, whereas mRNA levels of 5-LO were similar in all osteoblasts. Selective inhibition of cyclooxygenase-2 (COX-2) with NS-398-stimulated FLAP expression in the high OA osteoblasts subgroup, whereas it was without effect in the low OA osteoblasts subgroup. The addition of PGE2 to the low OA osteoblasts subgroup decreased FLAP expression but failed to affect it in the high OA osteoblasts subgroup. LTB4 levels in OA osteoblasts were stimulated about twofold by 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) plus transforming growth factor-beta (TGF-beta), a situation corresponding to their effect on FLAP mRNA levels. Treatments with 1,25(OH)2D3 and TGF-beta also modulated PGE2 production. TGF-beta stimulated PGE2 production in both OA osteoblast groups, whereas 1,25(OH)2D3 alone had a limited effect but decreased the effect of TGF-beta in the low OA osteoblasts subgroup. This modulation of PGE2 production was mirrored by the synthesis of COX-2. IL-18 levels were only slightly increased in a subgroup of OA osteoblasts compared with normal; however, no relationship was observed overall between IL-18 and PGE2 levels in normal and OA osteoblasts. These results suggest that the shunt from the production of PGE2 to LTB4 is through regulation of the expression of FLAP, not 5-LO, in OA osteoblasts. The expression of FLAP in OA osteoblasts is also modulated differently by 1,25(OH

  14. Superficial ulnar artery perforator flap.

    PubMed

    Schonauer, Fabrizio; Marlino, Sergio; Turrà, Francesco; Graziano, Pasquale; Dell'Aversana Orabona, Giovanni

    2014-09-01

    Superficial ulnar artery is a rare finding but shows significant surgical implications. Its thinness and pliability make this flap an excellent solution for soft tissue reconstruction, especially in the head and neck region. We hereby report a successful free superficial ulnar artery perforator forearm flap transfer for tongue reconstruction. A 64-year-old man presenting with a squamous cell carcinoma of the left tongue underwent a wide resection of the tumor, left radical neck dissection, and reconstruction of the tongue and the left tonsillar pillar with the mentioned flap. No complications were observed postoperatively. The flap survived completely; no recurrence at 6 months of follow-up was detected. Superficial ulnar artery perforator flap has shown to be a safe alternative to other free tissue flaps in specific forearm anatomic conditions. PMID:25102397

  15. Skin flaps and grafts - self-care

    MedlinePlus

    ... Regional flaps - self-care; Distant flaps - self-care; Free flap - self-care; Skin autografting - self-care ... the dressing and area around it clean and free from dirt or sweat. Don't let the ...

  16. Externally blown flap noise research

    NASA Technical Reports Server (NTRS)

    Dorsch, R. G.

    1974-01-01

    The Lewis Research Center cold-flow model externally blown flap (EBF) noise research test program is summarized. Both engine under-the-wing and over-the-wing EBF wing section configurations were studied. Ten large scale and nineteen small scale EBF models were tested. A limited number of forward airspeed effect and flap noise suppression tests were also run. The key results and conclusions drawn from the flap noise tests are summarized and discussed.

  17. The vortex flap

    NASA Astrophysics Data System (ADS)

    Buerge, Brandon T.

    The Vortex Flap is a new type of mechanically driven high-lift device consisting of a rotating cylinder placed underneath and near the trailing edge of an airfoil. Wind tunnel tests were designed and conducted in the Washington University Low-Speed Wind Tunnel. Wind tunnel tests indicate that the Vortex Flap produces notable lift coefficient increments and increases maximum lift coefficients, particularly for the low Reynolds number range tested. The best configurations of the configurations investigated (not necessarily optimal) produce lift increments of 300-900% at low-to-moderate angles of attack, and increase the maximum lift coefficient on the order of 200%. The large lift increments found, particularly at low angles of attack, underscore the ability to drive the airfoil to high lift coefficients even at low angles of attack, a potentially useful characteristic for certain flight maneuvers. Regions of fairly high L/D (on the order of 10) as well as low L/D performance were identified. The nondimensional cylinder rotation speed was found to be the most important experimental parameter. Methods for correcting wind tunnel data were developed and outlined, and a Response Surface Method was applied to the corrected data for ease of interpretation. Performance comparisons between the Vortex Flap and other trailing-edge high-lift devices are included. To demonstrate the potential of the device, a Navy mission specification for a VTOL ship-borne UAV, currently filled by a rotary-wing aircraft, is analyzed using a hypothetical fixed wing aircraft and the Vortex Flap. It is demonstrated that, under certain reasonable wind-over-deck conditions, such an aircraft could hypothetically fill a VTOL mission.

  18. 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.

  19. Petunia nectar proteins have ribonuclease activity.

    PubMed

    Hillwig, Melissa S; Liu, Xiaoteng; Liu, Guangyu; Thornburg, Robert W; Macintosh, Gustavo C

    2010-06-01

    Plants requiring an insect pollinator often produce nectar as a reward for the pollinator's visitations. This rich secretion needs mechanisms to inhibit microbial growth. In Nicotiana spp. nectar, anti-microbial activity is due to the production of hydrogen peroxide. In a close relative, Petunia hybrida, limited production of hydrogen peroxide was found; yet petunia nectar still has anti-bacterial properties, suggesting that a different mechanism may exist for this inhibition. The nectar proteins of petunia plants were compared with those of ornamental tobacco and significant differences were found in protein profiles and function between these two closely related species. Among those proteins, RNase activities unique to petunia nectar were identified. The genes corresponding to four RNase T2 proteins from Petunia hybrida that show unique expression patterns in different plant tissues were cloned. Two of these enzymes, RNase Phy3 and RNase Phy4 are unique among the T2 family and contain characteristics similar to both S- and S-like RNases. Analysis of amino acid patterns suggest that these proteins are an intermediate between S- and S-like RNases, and support the hypothesis that S-RNases evolved from defence RNases expressed in floral parts. This is the first report of RNase activities in nectar. PMID:20460362

  20. Petunia nectar proteins have ribonuclease activity

    PubMed Central

    Hillwig, Melissa S.; Liu, Xiaoteng; Liu, Guangyu; Thornburg, Robert W.; MacIntosh, Gustavo C.

    2010-01-01

    Plants requiring an insect pollinator often produce nectar as a reward for the pollinator's visitations. This rich secretion needs mechanisms to inhibit microbial growth. In Nicotiana spp. nectar, anti-microbial activity is due to the production of hydrogen peroxide. In a close relative, Petunia hybrida, limited production of hydrogen peroxide was found; yet petunia nectar still has anti-bacterial properties, suggesting that a different mechanism may exist for this inhibition. The nectar proteins of petunia plants were compared with those of ornamental tobacco and significant differences were found in protein profiles and function between these two closely related species. Among those proteins, RNase activities unique to petunia nectar were identified. The genes corresponding to four RNase T2 proteins from Petunia hybrida that show unique expression patterns in different plant tissues were cloned. Two of these enzymes, RNase Phy3 and RNase Phy4 are unique among the T2 family and contain characteristics similar to both S- and S-like RNases. Analysis of amino acid patterns suggest that these proteins are an intermediate between S- and S-like RNases, and support the hypothesis that S-RNases evolved from defence RNases expressed in floral parts. This is the first report of RNase activities in nectar. PMID:20460362

  1. 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

  2. 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.

  3. Ex Vivo Prefabricated Rat Skin Flap Using Cell Sheets and an Arteriovenous Vascular Bundle

    PubMed Central

    Fujisawa, Daisuke; Sekine, Hidekazu; Okano, Teruo; Sakurai, Hiroyuki

    2015-01-01

    Background: Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades. However, a regenerative skin flap has not been developed that could provide immediate blood flow after transplantation. Here, we make a regenerative skin flap ex vivo that is potentially suitable for microsurgical transplantation in future clinical applications. Methods: In rats, for preparing a stable vascular carrier, a femoral vascular pedicle was sandwiched between collagen sponges and inserted into a porous chamber in the abdomen. The vascular bed was harvested 3 weeks later, and extracorporeal perfusion was performed. A green fluorescent protein positive epidermal cell sheet was placed on the vascular bed. After perfusion culture, the whole construct was harvested and fixed for morphological analyses. Results: After approximately 10 days perfusion, the epidermal cell sheet cornified sufficiently. The desquamated corneum was positive for filaggrin. The basement membrane protein laminin 332 and type 4 collagen were deposited on the interface area between the vascular bed and the epidermal cell sheet. Moreover, an electron microscopic image showed anchoring junctions and keratohyalin granules. These results show that we were able to produce native-like skin. Conclusions: We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future. PMID:26180725

  4. 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

  5. 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.

  6. 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

  7. Deep circumflex iliac perforator flap.

    PubMed

    Kimata, Yoshihiro

    2003-07-01

    The increased freedom of the DCIP flap from the harvested iliac crest facilitates correct positioning. To ensure that the DCIP flap can be safely elevated, however, the presence of perforators (approximately 1 cm in diameter) must be confirmed preoperatively and intraoperatively. PMID:12916597

  8. 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

  9. 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.

  10. 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

  11. [Mitogen-activated protein kinases in atherosclerosis].

    PubMed

    Bryk, Dorota; Olejarz, Wioletta; Zapolska-Downar, Danuta

    2014-01-01

    Intracellular signalling cascades, in which MAPK (mitogen-activated protein kinases) intermediate, are responsible for a biological response of a cell to an external stimulus. MAP kinases, which include ERK1/2 (extracellular signalling-regulated kinase), JNK (c-Jun N-terminal kinase) and p 38 MAPK, regulate the activity of many proteins, enzymes and transcription factors and thus have a wide spectrum of biological effects. Many basic scientific studies have defined numerous details of their pathway organization and activation. There are also more and more studies suggesting that individual MAP kinases probably play an important role in the pathogenesis of atherosclerosis. They may mediate inflammatory processes, endothelial cell activation, monocyte/macrophage recruitment and activation, smooth muscle cell proliferation and T-lymphocyte differentiation, all of which represent crucial mechanisms involved in pathogenesis of atherosclerosis. The specific inhibition of an activity of the respective MAP kinases may prove a new therapeutic approach to attenuate atherosclerotic plaque formation in the future. In this paper, we review the current state of knowledge concerning MAP kinase-dependent cellular and molecular mechanisms underlying atherosclerosis. PMID:24491891

  12. 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

  13. Theory of flapping flight

    NASA Technical Reports Server (NTRS)

    Lippisch, Alexander

    1925-01-01

    Before attempting to construct a human-powered aircraft, the aviator will first try to post himself theoretically on the possible method of operating the flapping wings. This report will present a graphic and mathematical method, which renders it possible to determine the power required, so far as it can be done on the basis of the wing dimensions. We will first consider the form of the flight path through the air. The simplest form is probably the curve of ordinary wave motion. After finding the flight curve, we must next determine the change in the angle of attack while passing through the different phases of the wave.

  14. 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

  15. Detection of protein C activation in humans.

    PubMed Central

    Bauer, K A; Kass, B L; Beeler, D L; Rosenberg, R D

    1984-01-01

    We have developed a radioimmunoassay (RIA) for the dodecapeptide that is liberated from protein C when this zymogen is activated by thrombin bound to thrombomodulin present on the vascular endothelium. The protein C activation peptide (PCP) was synthesized using the solid-phase method of Merrifield. Antisera were raised in rabbits to the synthetic analogue coupled to bovine serum albumin with glutaraldehyde. The antibody population obtained was used together with a 125I-labeled tyrosinated ligand and various concentrations of unlabeled PCP to construct a double antibody RIA capable of measuring as little as 10 pM of this component. We have established that the synthetic dodecapeptide has the same immunoreactivity as the native peptide and that the reactivity of protein C is less than 1/2,000 that of PCP on a molar basis. The extremely low levels of peptide in normal individuals as well as the nonspecific contributions of plasma constituents to the immunoreactive signal, necessitated the development of a procedure by which the PCP could be reproducibly extracted from plasma and concentrated approximately 20-fold. This methodology permitted us to demonstrate that the plasma PCP levels in 17 normal donors averaged 6.47 pM, and that elevations up to 180 pM were observed in individuals with evidence of disseminated intravascular coagulation. The validity of these measurements of protein C activation is supported by the fact that, in both of these situations, the RIA signal migrates on reverse-phase high pressure liquid chromatography in a manner identical to that of the native dodecapeptide. We have also noted that the mean PCP concentration in seven patients fully anticoagulated with warfarin averaged 2.61 pM. Our studies also show that PCP is cleared from the plasma of primates with a t1/2 of approximately 5 min. Given that the t1/2 of activated protein C is estimated to be 10-15 min, the latter enzyme appears to exert its effects on the activated cofactors of the

  16. Edmund Prince Fowler Award Thesis. Evaluation of random skin flap survival in a porcine model.

    PubMed

    Pratt, M F

    1996-06-01

    The pathophysiology of random skin flap necrosis in the pig model was studied the effects of several drugs on skin flap survival were examined. The investigated drugs included acetylsalicylic acid (ASA), pentoxifylline (PTX), prostaglandin E2 (PGE2), and an experimental 21-aminosteroid, U-74389G. Each drug altered different parameters known to be associated with tissue necrosis. Demonstrated mechanisms of skin flap failure included the alteration of erythrocyte flexibility and platelet function and the activation of neutrophils with resultant accumulation of damaging oxygen-free radicals. Random skin flap survival did not improve with ASA but did improve significantly with PTX, PGE2, and U-74389G. The results of this study underscore the importance of neutrophil-mediated necrosis in the pathophysiology of skin flap failure. The data further demonstrate the need to develop drugs aimed at reversing or preventing the tissue damage from oxygen-free radicals in order to enhance the survival of random skin flaps. PMID:8656954

  17. 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.

  18. Flap Dynamics in Aspartic Proteases: A Computational Perspective.

    PubMed

    Mahanti, Mukul; Bhakat, Soumendranath; Nilsson, Ulf J; Söderhjelm, Pär

    2016-08-01

    Recent advances in biochemistry and drug design have placed proteases as one of the critical target groups for developing novel small-molecule inhibitors. Among all proteases, aspartic proteases have gained significant attention due to their role in HIV/AIDS, malaria, Alzheimer's disease, etc. The binding cleft is covered by one or two β-hairpins (flaps) which need to be opened before a ligand can bind. After binding, the flaps close to retain the ligand in the active site. Development of computational tools has improved our understanding of flap dynamics and its role in ligand recognition. In the past decade, several computational approaches, for example molecular dynamics (MD) simulations, coarse-grained simulations, replica-exchange molecular dynamics (REMD) and metadynamics, have been used to understand flap dynamics and conformational motions associated with flap movements. This review is intended to summarize the computational progress towards understanding the flap dynamics of proteases and to be a reference for future studies in this field. PMID:26872937

  19. 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

  20. Traumatic flap dislocation 10 years after LASIK. Case report and literature review.

    PubMed

    Khoueir, Z; Haddad, N M; Saad, A; Chelala, E; Warrak, E

    2013-01-01

    We report a case of traumatic partial flap dislocation 10 years after uneventful laser in situ keratomileusis (LASIK). The patient was treated bilaterally for hyperopia and astigmatism with LASIK. A superior-hinged corneal flap was created using the Moria M2 microkeratome (Moria SA, Antony, France) and the surgery was uneventful. Ten years later, partial flap dislocation was diagnosed after mild trauma. This case suggests that flap dislocations can occur during recreational activities up to 10 years after surgery. Full visual recovery is achievable if the case is managed promptly. Further studies should evaluate the potential protective role of an inferior hinge during LASIK. PMID:23219507

  1. 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.

  2. Heat dissipation guides activation in signaling proteins.

    PubMed

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

    2015-08-18

    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

  3. 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.

  4. Dancing girl flap: a new flap suitable for web release.

    PubMed

    Shinya, K

    1999-12-01

    To create a deep web, a flap must be designed to have a high elongation effect in one direction along the mid-lateral line of the finger and also to have a shortening effect in the other direction, crossing at a right angle to the mid-lateral line. The dancing girl flap is a modification of a four-flap Z-plasty with two additional Z-plasties. It has a high elongation effect in one direction (>550%) and a shortening effect in the other direction at a right angle (<33%), creating a deep, U-shaped surface. This new flap can be used to release severe scar contracture with a web, and is most suitable for incomplete syndactyly with webs as high as the proximal interphalangeal joint. PMID:10597822

  5. Activation and activities of the p53 tumour suppressor protein

    PubMed Central

    Bálint, É; Vousden, K H

    2001-01-01

    The p53 tumour suppressor protein inhibits malignant progression by mediating cell cycle arrest, apoptosis or repair following cellular stress. One of the major regulators of p53 function is the MDM2 protein, and multiple forms of cellular stress activate p53 by inhibiting the MDM2-mediated degradation of p53. Mutations in p53, or disruption of the pathways that allow activation of p53, seem to be a general feature of all cancers. Here we review recent advances in our understanding of the pathways that regulate p53 and the pathways that are induced by p53, as well as their implications for cancer therapy. © 2001 Cancer Research Campaign http://www.bjcancer.com PMID:11747320

  6. Keystone Flap: Versatile Flap for Reconstruction of Limb Defects

    PubMed Central

    Janna, Rakesh K.

    2015-01-01

    Background: There is always a constant search for a new solution to tackle defects in the limbs. The technique has to be simple, easily reproducible and performed within a short duration. The answer is keystone island flap keystone flap is a simple, less time consuming, durable and easily reproducible option to reconstruct most of the limb defects. Aim: The aim of this article is to study the usefulness of keystone flap in reconstruction of various upper and lower limb defects. Materials and Methods: This retrospective review involves study of 20 patients undergoing keystone flap reconstruction for various defects from 2012 to 2014. Patient demographic data, medical histories, comorbidities, surgical indications, defect characteristics and locations, hospitalization, complications and follow-up were evaluated and are presented as uncontrolled case series. Results: Ages of the patients were ranging from 18 to 65 y with an average of 38.75y. Among the defects, 10 were following trauma (50%), 5 were due to tumour resection (25%), 3 followed debridement of abscess (15%) and another 2 defects were due to surgical wound dehiscence (10%). The largest defect covered by this flap in our study measured 45 x 18 cm and the smallest defect was 8 x4 cm. The average intra-operative time was 45.5 min (range 20-90 min). Fourteen flaps were done to cover lower limb defects (70%), 4 for upper limb defects and 2 were for defects in the axilla. Partial flap necrosis was observed in one case. The average duration of hospital stay of patients was 3.45 d. All patients were followed until they achieved stable, healed wound.The overall success rate was 95%. Conclusion: Keystone flap can be safely used to cover various limb defects with minimal pain, a sensate cover and excellent cosmetic outcome, minimizing the need for microsurgical techniques or prolonged operative time. PMID:25954659

  7. The Role of Mitogen-Activated Protein Kinase-Activated Protein Kinases (MAPKAPKs) in Inflammation

    PubMed Central

    Moens, Ugo; Kostenko, Sergiy; Sveinbjørnsson, Baldur

    2013-01-01

    Mitogen-activated protein kinase (MAPK) pathways are implicated in several cellular processes including proliferation, differentiation, apoptosis, cell survival, cell motility, metabolism, stress response and inflammation. MAPK pathways transmit and convert a plethora of extracellular signals by three consecutive phosphorylation events involving a MAPK kinase kinase, a MAPK kinase, and a MAPK. In turn MAPKs phosphorylate substrates, including other protein kinases referred to as MAPK-activated protein kinases (MAPKAPKs). Eleven mammalian MAPKAPKs have been identified: ribosomal-S6-kinases (RSK1-4), mitogen- and stress-activated kinases (MSK1-2), MAPK-interacting kinases (MNK1-2), MAPKAPK-2 (MK2), MAPKAPK-3 (MK3), and MAPKAPK-5 (MK5). The role of these MAPKAPKs in inflammation will be reviewed. PMID:24705157

  8. 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.

  9. 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.

  10. 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

  11. Monooxygenase activity of type 3 copper proteins.

    PubMed

    Itoh, Shinobu; Fukuzumi, Shunichi

    2007-07-01

    The molecular mechanism of the monooxygenase (phenolase) activity of type 3 copper proteins has been examined in detail both in the model systems and in the enzymatic systems. The reaction of a side-on peroxo dicopper(II) model compound ( A) and neutral phenols proceeds via a proton-coupled electron-transfer (PCET) mechanism to generate phenoxyl radical species, which collapse each other to give the corresponding C-C coupling dimer products. In this reaction, a bis(mu-oxo)dicopper(III) complex ( B) generated by O-O bond homolysis of A is suggested to be a real active species. On the other hand, the reaction of lithium phenolates (deprotonated form of phenols) with the same side-on peroxo dicopper(II) complex proceeds via an electrophilic aromatic substitution mechanism to give the oxygenated products (catechols). The mechanistic difference between these two systems has been discussed on the basis of the Marcus theory of electron transfer and Hammett analysis. Mechanistic details of the monooxygenase activity of tyrosinase have also been examined using a simplified enzymatic reaction system to demonstrate that the enzymatic reaction mechanism is virtually the same as that of the model reaction, that is, an electrophilic aromatic substitution mechanism. In addition, the monooxygenase activity of the oxygen carrier protein hemocyanin has been explored for the first time by employing urea as an additive in the reaction system. In this case as well, the ortho-hydroxylation of phenols to catechols has been demonstrated to involve the same ionic mechanism. PMID:17461541

  12. 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

  13. 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.

  14. Perforator Flaps in Head and Neck Reconstruction

    PubMed Central

    Chana, Jagdeep S.; Odili, Joy

    2010-01-01

    Free tissue transfer has revolutionized the management of complex head and neck defects. Perforator flaps represent the most recent advance in the development of free flap surgery. These flaps are based on perforating vessels and can be harvested without significant damage to associated muscles, thereby reducing the postoperative morbidity associated with muscle-based flaps. Elevation of perforator flaps requires meticulous technique and can be more challenging than raising muscle-based flaps. Use of a Doppler device enables reliable identification of the perforating vessels and aids in the design of free-style free flaps, where the flaps are designed purely according to the perforator located. The major advantage of free-style free flaps is that an unlimited number of flaps can potentially be designed on much shorter pedicles. The anterolateral thigh flap is the most commonly used perforator flap in head and neck reconstruction. Its use is described in detail, as is use of other less common perforator flaps. This article also describes head and neck reconstruction in a region-specific manner and gives a short-list of suitable flaps based on the location of the defect. PMID:22550446

  15. Modified osteomyocutaneous iliac crest flaps transplantation.

    PubMed

    Liu, Jun; Song, Dajiang; Li, Jinsong; Xu, Jian; Lv, Hongbin

    2015-04-01

    The paper aims to improve the operative technique of osteomyocutaneous iliac crest flap harvesting, further minimise morbidity of donor site, and improve the effect of recipient site reconstruction. From March 2005 to March 2011, 55 cases of osteomyocutaneous iliac crest flap harvested by different methods were performed to reconstruct the defects of the extremities. Twenty-nine cases were reconstructed with a traditional deep circumflex iliac artery osteomusculocutaneous flap. Twenty-six cases were repaired with modified osteomyocutaneous iliac crest flaps. In 29 cases with a traditional DCIA osteomusculocutaneous flap, two cases showed the injured lateral femoral cutaneous nerve. Flapnecrosis was significant in two cases. Arterial compromise occurred in one case 5 days after operation completion and led to flap failure. Three flaps developed postoperative venous congestion, but only one flap received re-exploration. In the other two cases, some stitches were removed for decompression. All three flaps survived. In two cases, marginal flap necrosis occurred, but no secondary skin grafting was required. In 26 cases with modified flap transplantation, one case showed the injured lateral femoral cutaneous nerve. All flaps survived totally. Osseous integration was achieved in all 55 cases in 3 ∼ 9 months after operation. The modified osteomyocutaneous iliac crest flap technique enhances flap safety, provides the additional advantages of reducing donor-site morbidity, and improves the recipient-site contour. PMID:25001367

  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. Protein-protein interactions in plant mitogen-activated protein kinase cascades.

    PubMed

    Zhang, Tong; Chen, Sixue; Harmon, Alice C

    2016-02-01

    Mitogen-activated protein kinases (MAPKs) form tightly controlled signaling cascades that play essential roles in plant growth, development, and defense. However, the molecular mechanisms underlying MAPK cascades are still elusive, due largely to our poor understanding of how they relay the signals. Extensive effort has been devoted to characterization of MAPK-substrate interactions to illustrate phosphorylation-based signaling. The diverse MAPK substrates identified also shed light on how spatiotemporal-specific protein-protein interactions function in distinct MAPK cascade-mediated biological processes. This review surveys various technologies used for characterizing MAPK-substrate interactions and presents case studies of MPK4 and MPK6, highlighting the multiple functions of MAPKs. Mass spectrometry-based approaches in identifying MAPK-interacting proteins are emphasized due to their increasing utility and effectiveness. The potential for using MAPKs and their substrates in enhancing plant stress tolerance is also discussed. PMID:26646897

  18. 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

  19. Experience in Reconstruction for Small Digital Defects With Free Flaps.

    PubMed

    Hung, Min-Hsiang; Huang, Kuo-Feng; Chiu, Haw-Yen; Chao, Wai-Nang

    2016-03-01

    Traumatic injuries to the digits resulting in soft tissue or bone loss require reconstruction. Traditionally, local flaps, such as homodigital flaps, heterodigital flaps, pedicled flaps, or distant flaps, are used for digital resurfacing. However, free tissue transfers can be used in selected patients. In this study, we present the use of different free flaps including groin skin flaps, groin osteocutaneous flaps, groin chimeric flaps, second dorsal metacarpal artery flaps, and partial toe flaps for digital reconstruction. A total of 19 digits were treated with 16 free flaps in our hospital. Of the flaps used, 5 were free groin skin flaps, 4 were free partial toe flaps, 3 were free groin chimeric flaps, 2 were free groin osteocutaneous flaps, and 2 were free second dorsal metacarpal artery flaps. The average flap size was 4.7 × 2.0 cm (range, 1.5 × 1 to 5 × 4 cm), and the average operative time was 6.0 hours (range, 4-9 hours). All flaps survived without partial or total necrosis. In conclusion, the free flap is a reliable and safe alternative for digital reconstruction. Moreover, the free groin flap provides not only a chimeric pattern for multiple fingers coverage but also an osteocutaneous pattern for thumb lengthening. The free second dorsal metacarpal artery flap provides a tenocutaneous pattern for tendon reconstruction and soft tissue coverage simultaneously, and the free partial toe flap is an excellent alternative for pulp reconstruction in terms of aesthetic appearance and functional outcome. PMID:26808771

  20. Treatment of degloving injury involving multiple fingers with combined abdominal superficial fascial flap, dorsalis pedis flap, dorsal toe flap, and toe-web flap

    PubMed Central

    Han, Fengshan; Wang, Guangnan; Li, Gaoshan; Ping, Juan; Mao, Zhi

    2015-01-01

    Background Our aim was to summarize the treatment of degloving injury involving multiple fingers using combined abdominal superficial fascial flap, dorsalis pedis flap, dorsal toe flap, and toe-web flap. Patients and methods Each degloved finger was debrided under microscopic guidance and embedded in the superficial layer of the abdominal fascia. The abdominal skin was sutured to the skin on the back and side of the hand to promote circumferential healing. After removal, the only remaining injured region was on the flexor surface, and this was repaired by multiple dorsal toe flaps, toe-web flaps, and dorsalis pedis flaps to provide blood vessels and sensory nerves. All fingers had proper flap thickness 3–6 months after surgery, and required only lateral Z-plasty modification with web deepening and widening to narrow the fingers and extend their relative length. Results We completed flap-graft and finger narrowing for 25 fingers in eight patients. Abdominal skin flaps and dorsal toe flaps were grafted, and resulted in both firmness and softness, providing finger flexibility. The dorsal toe flap provided good blood circulation and sensory nerves, and was used to cover the finger-flexor surface to regain sensation and stability when holding objects. During the 1–8 years of follow-up, sensation on the finger-flexor side recovered to the S3–4 level, and patient satisfaction based on the Michigan Hand Outcomes Questionnaire was 4–5. Flap ulcers or bone/tendon necrosis were not observed. Conclusion Treatment of degloving injury involving multiple fingers with combined abdominal superficial fascial flap, dorsalis pedis flap, dorsal toe flap, and toe-web flap was effective and reliable. PMID:26229479

  1. 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.

  2. 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.

  3. Chlorogenic Acid Enhances Abdominal Skin Flap Survival Based on Epigastric Artery in Nondiabetic and Diabetic Rats.

    PubMed

    Bagdas, Deniz; Etoz, Betul Cam; Gul, Zulfiye; Ozyigit, Musa Ozgur; Cinkilic, Nilufer; Inan, Sevda; Buyukcoskun, Naciye Isbil; Ozluk, Kasim; Gurun, Mine Sibel

    2016-08-01

    Previous studies showed that chlorogenic acid (CGA) accelerates wound healing via its antioxidant activity. We aimed to investigate the effect of CGA in an experimental epigastric abdominal skin flap model in nondiabetic and diabetic rats. Rats were firstly divided into 2 groups: nondiabetic and diabetic. Diabetes was induced by streptozotocin. Then, 4 subgroups were created for each group: vehicle as well as 0.2 mg/0.5 mL, 1 mg/0.5 mL, and 5 mg/0.5 mL CGA treatments. Right epigastric artery-based abdominal skin flaps were elevated and sutured back into their original position. Chlorogenic acid or vehicle was injected once into the femoral arteries by leaving the epigastric artery as the single artery feeding the flaps during the injection. On postoperative day 7, flap survivals were evaluated, and the rats were killed. Distal flap tissues were collected for histopathological and biochemical assays. Chlorogenic acid showed greater flap survival in both nondiabetic and diabetic rats. Capillary density was increased, and necrosis was reduced in the CGA-treated rats. Chlorogenic acid decreased malondialdehyde levels as well as increased reduced glutathione and superoxide dismutase levels in the flap tissues. This study showed that CGA significantly improved flap survival by its antioxidant activities with intra-arterial local injections. PMID:25356637

  4. Free thin paraumbilical perforator-based flaps.

    PubMed

    Koshima, I; Moriguchi, T; Soeda, S; Tanaka, H; Umeda, N

    1992-07-01

    A free paraumbilical perforator-based flap fed by a muscle perforator from the inferior deep epigastric artery and with no muscle was used in 13 patients. Among them, a free thin paraumbilical perforator-based flap with a thin layer of fat, to protect the subdermal plexus of the vessels, was used in seven patients. The dominant pedicle perforator of this thin flap is usually located around the umbilicus and a large flap can be obtained. Its critical length-to-breath ratio is considered to be 4:3. The advantages of this flap are a long and large vascular pedicle, rare postoperative abdominal herniation, little bulkiness of the flap, and a relatively large skin territory. The disadvantages are technical difficulties in dissection of the perforator and anatomical variation in the location of the perforator. We believe this flap largely overcomes the problems of the conventional rectus abdominis musculocutaneous flap. PMID:1386718

  5. Nucleolar protein B23 has molecular chaperone activities.

    PubMed Central

    Szebeni, A.; Olson, M. O.

    1999-01-01

    Protein B23 is an abundant, multifunctional nucleolar phosphoprotein whose activities are proposed to play a role in ribosome assembly. Szebeni et al. (1997) showed stimulation of nuclear import in vitro by protein B23 and suggested that this effect was due to a molecular chaperone-like activity. Protein B23 was tested for chaperone activities using several protein substrates. The temperature-dependent and -independent aggregation of the HIV-1 Rev protein was measured using a zero angle light scattering (turbidity) assay. Protein B23 inhibited the aggregation of the Rev protein, with the amount of inhibition proportional to the concentration of B23 added. This activity was saturable with nearly complete inhibition when the molar ratio of B23:Rev was slightly above one. Protein B23 also protected liver alcohol dehydrogenase (LADH), carboxypeptidase A, citrate synthase, and rhodanese from aggregation during thermal denaturation and preserved the enzyme activity of LADH under these conditions. In addition, protein B23 was able to promote the restoration of activity of LADH previously denatured with guanidine-HCl. Protein B23 preferentially bound denatured substrates and exposed hydrophobic regions when complexed with denatured proteins. Thus, by several criteria, protein B23 behaves like a molecular chaperone; these activities may be related to its role in ribosome biogenesis. PMID:10211837

  6. Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities

    PubMed Central

    Shen, Hsin-Hui; Lithgow, Trevor; Martin, Lisandra L.

    2013-01-01

    The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity. We also discuss examples where the same membrane proteins have been successfully reconstituted into two or more model membrane systems with comparison of the observed activity in each system. Understanding of the behavioral changes for proteins in model membrane systems after membrane reconstitution is often a prerequisite to protein research. It is essential to find better solutions for retaining membrane protein activities for measurement and characterization in vitro. PMID:23344058

  7. 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

  8. 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. PMID:25841702

  9. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Flap interconnection. 23.701 Section 23.701... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control Systems § 23.701 Flap interconnection. (a) The main wing flaps and related movable surfaces as a...

  10. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Flap interconnection. 23.701 Section 23.701... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control Systems § 23.701 Flap interconnection. (a) The main wing flaps and related movable surfaces as a...

  11. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Flap interconnection. 23.701 Section 23.701... STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control Systems § 23.701 Flap interconnection. (a) The main wing flaps and related movable surfaces as a...

  12. The bilobed flap for popliteal defect reconstruction

    PubMed Central

    Kelahmetoglu, Osman; Yagmur, Caglayan; Aslan, Ozan; Firinciogullari, Remzi

    2014-01-01

    Abstract Bilobed flaps were first introduced to close small nasal defects. We reconstructed a defect of the popliteal fossa using a random-pattern bilobed flap. We recommend the use of random-pattern bilobed flaps as a reliable technique for covering defects of the popliteal fossa.

  13. Designing Mimics of Membrane Active Proteins

    PubMed Central

    Sgolastra, Federica; deRonde, Brittany M.; Sarapas, Joel M.; Som, Abhigyan; Tew, Gregory N.

    2014-01-01

    CONSPECTUS As a semi-permeable barrier that controls the flux of biomolecules in and out the cell, the plasma membrane is critical in cell function and survival. Many proteins interact with the plasma membrane and modulate its physiology. Within this large landscape of membrane-active molecules, researchers have focused significant attention on two specific classes of peptides, antimicrobial peptides (AMPs) and cell penetrating peptides (CPPs) because of their unique properties. In this account, we describe our efforts over the last decade to build and understand synthetic mimics of antimicrobial peptides (SMAMPs). These endeavors represent one specific example of a much larger effort to understand how synthetic molecules interact with and manipulate the plasma membrane. Using both defined molecular weight oligomers and easier to produce, but heterogeneous, polymers, it has been possible to generate scaffolds with biological potency superior to the natural analogs. In one case, a compound has progressed through a phase II clinical trial for pan)staph infections. Modern biophysical assays highlighted the interplay between the synthetic scaffold and lipid composition leading to negative Gaussian curvature, a requirement for both pore formation and endosomal escape. The complexity of this interplay between lipids, bilayer components, and the scaffolds remains to be better resolved, but significant new insight has been provided. It is worthwhile to consider the various aspects of permeation and how these are related to ‘pore formation.’ More recently, our efforts have expanded toward protein transduction domains, or cell penetrating peptide, mimics. The combination of unique molecular scaffolds and guanidinium) rich side chains has produced an array of polymers with robust transduction (and delivery) activity. Being a new area, the fundamental interactions between these new scaffolds and the plasma membrane are just beginning to be understood. Negative Gaussian

  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. 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

  16. Endoplasmic reticulum stress activates transglutaminase 2 leading to protein aggregation

    PubMed Central

    LEE, JIN-HAENG; JEONG, JAEHO; JEONG, EUI MAN; CHO, SUNG-YUP; KANG, JEONG WOOK; LIM, JISUN; HEO, JINBEOM; KANG, HYUNSOOK; KIM, IN-GYU; SHIN, DONG-MYUNG

    2014-01-01

    Aberrant activation of transglutaminase 2 (TGase2) contributes to a variety of protein conformational disorders such as neurodegenerative diseases and age-related cataracts. The accumulation of improperly folded proteins in the endoplasmic reticulum (ER) triggers the unfolded protein response (UPR), which promotes either repair or degradation of the damaged proteins. Inadequate UPR results in protein aggregation that may contribute to the development of age-related degenerative diseases. TGase2 is a calcium-dependent enzyme that irreversibly modifies proteins by forming cross-linked protein aggregates. Intracellular TGase2 is activated by oxidative stress which generates large quantities of unfolded proteins. However, the relationship between TGase2 activity and UPR has not yet been established. In the present study, we demonstrated that ER stress activated TGase2 in various cell types. TGase2 activation was dependent on the ER stress-induced increase in the intracellular calcium ion concentration but not on the TGase2 protein expression level. Enzyme substrate analysis revealed that TGase2-mediated protein modification promoted protein aggregation concurrently with decreasing water solubility. Moreover, treatment with KCC009, a TGase2 inhibitor, abrogated ER stress-induced TGase2 activation and subsequent protein aggregation. However, TGase2 activation had no effect on ER stress-induced cell death. These results demonstrate that the accumulation of misfolded proteins activates TGase2, which further accelerates the formation of protein aggregates. Therefore, we suggest that inhibition of TGase2 may be a novel strategy by which to prevent the protein aggregation in age-related degenerative diseases. PMID:24481335

  17. 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.

  18. The effect of platelet rich plasma on angiogenesis in ischemic flaps in VEGFR2-luc mice.

    PubMed

    Sönmez, Tolga Taha; Vinogradov, Alexandra; Zor, Fatih; Kweider, Nisreen; Lippross, Sebastian; Liehn, Elisa Anamaria; Naziroglu, Mustafa; Hölzle, Frank; Wruck, Christoph; Pufe, Thomas; Tohidnezhad, Mersedeh

    2013-04-01

    To improve skin flap healing, one promising strategy in reconstructive surgery might be to optimize platelet rich plasma (PRP) bioactivity and the ischemia-altered expression of genes. We studied both the effect of PRP on ischemic flaps, and whether in vivo bioluminescence imaging (BLI) is a suitable method for the longitudinal monitoring of angiogenesis in surgical wounds. Axial murine skin flaps were created in four experimental groups. In vivo measurements of VEGFR2 expression levels were made every other day until the 14th day. The local VEGF level and microvessel density were quantified on the 14th day via ELISA and immunohistochemistry, and flap survival rates were measured. We demonstrated that PRP and induced ischemia have a beneficial influence on angiogenesis and flap healing. Combining the two resulted in a significantly robust increase in angiogenesis and flap survival rate that was corroborated by bioluminescence imaging of VEGFR2 activity. This study shows that angiogenic effects of PRP may be potentialized by the stimulus of induced ischemia during free flap harvesting, and thus the two procedures appear to have a synergistic effect on flap healing. This study further demonstrates that BLI of modulated genes in reconstructive surgery is a valuable model for longitudinal in vivo evaluation of angiogenesis. PMID:23352038

  19. 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

  20. BCL-2 and Bax Expression in Skin Flaps Treated with Finasteride or Azelaic Acid

    PubMed Central

    Ayatollahi, Seyyed Abdulmajid; Ajami, Marjan; Reyhanfard, Hamed; Asadi, Yasin; Nassiri-Kashani, Mansour; Rashighi Firoozabadi, Mehdi; Davoodi, Sayed Hossein; Habibi, Esmaeil; Pazoki-Toroudi, Hamidreza

    2012-01-01

    Despite all modern surgical techniques, skin flap that is considered as the main method in most reconstructive surgeries puts the skin tissue at danger of necrosis and apoptosis derived from ischemia. Therefore, finding a treatment for decreasing the apoptosis derived from flap ischemia will be useful in clinic. In present study, we evaluated the effect of azelaic acid 20% and finasteride on expression of BCL-2 and bax proteins after the skin flap surgery. For this purpose, 21 rats were entered in three groups including control, azelaic acid 20% and finasteride, all experienced skin flap surgery and then flap tissue was assessed for determining the expression of proteins in 5 slices prepared from each rat that were graded between – to +++ scales. Both azelaic acid and finasteride increased the expression of BCL-2 protein (p < 0.05) and decrease the expression of bax protein (p < 0.05). These results suggested an antiapoptotic role for finasteride and azelaic acid in preserving the flap after the ischemia reperfusion insult. PMID:24250563

  1. Four-flap Breast Reconstruction: Bilateral Stacked DIEP and PAP Flaps

    PubMed Central

    Mayo, James L.; Allen, Robert J.

    2015-01-01

    Background: In cases of bilateral breast reconstruction when the deep inferior epigastric perforator (DIEP) free flap alone does not provide sufficient volume for body-specific reconstruction, stacking each DIEP flap with a second free flap will deliver added volume and maintain a purely autologous reconstruction. Stacking the profunda artery perforator (PAP) flap with the DIEP flap offers favorable aesthetics and ideal operative efficiency. We present the indications, technique, and outcomes of our experience with 4-flap breast reconstruction using stacked DIEP/PAP flaps. Methods: The authors performed 4-flap DIEP/PAP breast reconstruction in 20 patients who required bilateral reconstruction without adequate single donor flap volume. The timing of reconstruction, average mastectomy/flap weights, and operative time are reported. Complications reviewed include fat necrosis, dehiscence, hematoma, seroma, mastectomy flap necrosis, and flap loss. Results: Twenty patients underwent 4-flap DIEP/PAP breast reconstruction. Surgical time averaged 7 hours and 20 minutes. The primary recipient vessels were the antegrade and retrograde internal mammary vessels. No flap losses occurred. Complications included 1 hematoma, 1 incidence of arterial and venous thrombosis successfully treated with anastomotic revision, 1 incidence of thigh donor site dehiscence, and 3 episodes of minor mastectomy skin flap necrosis. Conclusions: Four-flap breast reconstruction is a favorable autologous reconstructive option for patients requiring bilateral reconstruction without adequate single donor flap volume. Stacking DIEP/PAP flaps as described is both safe and efficient. Furthermore, this combination provides superior aesthetics mirroring the natural geometry of the breast. Bilateral stacked DIEP/PAP flaps represent our first choice for breast reconstruction in this patient population. PMID:26090273

  2. Induced-shear piezoelectric actuators for rotor blade trailing edge flaps

    NASA Astrophysics Data System (ADS)

    Centolanza, Louis R.; Smith, Edward C.; Munsky, Brian

    2002-02-01

    Much of the current rotorcraft research is focused on improving performance by reducing unwanted helicopter noise and vibration. One of the most promising active rotorcraft vibration control systems is an active trailing edge flap. In this paper, an induced-shear piezoelectric tube actuator is used in conjunction with a simple lever-cusp hinge amplification device to generate a useful combination of trailing edge flap deflections and hinge moments. A finite-element model of the actuator tube and trailing edge flap (including aerodynamic and inertial loading) was used to guide the design of the actuator-flap system. A full-scale induced shear tube actuator flap system was fabricated and bench top testing was conducted to validate the analysis. Hinge moments corresponding to various rotor speeds were applied to the actuator using mechanical springs. The testing demonstrated that for an applied electric field of 3 kV cm-1, the tube actuator deflected a representative full-scale 12 inch flap +/-2.8° at 0 rpm and +/-1.4° for a hinge moment simulating a 400 rpm condition. The per cent error between the predicted and experimental full-scale flap deflections ranged from 4% (low rpm) to 12.5% (large rpm). Increasing the electric field to 4 kV cm-1 results in +/-2.5° flap deflection at a rotation speed of 400 rpm, according to the design analysis. A trade study was conducted to compare the performance of the piezoelectric tube actuator to the state of the art in trailing edge flap actuators and indicated that the induced-shear tube actuator shows promise as a trailing edge flap actuator.

  3. 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

  4. 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.

  5. Cost-effectiveness of monitoring free flaps.

    PubMed

    Subramaniam, Shiva; Sharp, David; Jardim, Christopher; Batstone, Martin D

    2016-06-01

    Methods of free flap monitoring have become more sophisticated and expensive. This study aims to determine the cost of free flap monitoring and examine its cost effectiveness. We examined a group of patients who had had free flaps to the head and neck over a two-year period, and combined these results with costs obtained from business managers and staff. There were 132 free flaps with a success rate of 99%. The cost of monitoring was Aus $193/flap. Clinical monitoring during this time period cost Aus$25 476 and did not lead to the salvage of any free flaps. Cost equivalence is reached between monitoring and not monitoring only at a failure rate of 15.8%. This is to our knowledge the first study to calculate the cost of clinical monitoring of free flaps, and to examine its cost-effectiveness. PMID:27015730

  6. Scrotal reconstruction with modified pudendal thigh flaps.

    PubMed

    Mopuri, Nabil; O'Connor, Edmund Fitzgerald; Iwuagwu, Fortune C

    2016-02-01

    Scrotal skin loss can occur following trauma, Fournier's gangrene, post tumour excision, burns, etc. There are many techniques described in the literature including residual scrotal skin mobilization, skin grafts, pedicled and free flaps. The management is complex and challenging shown by the multiplicity of flaps and techniques described in the literature. We used a modified pudendal thigh flap to reconstruct scrotal defects in five patients. This study describes the vascularity of the flap, technique of elevation and the inset of the flap. The elevation and particularly the insetting make it different from other flaps raised on this vascular network for scrotal reconstruction. This pedicled flap is robust, reliable, resilient and produces a neo-scrotum that looks natural in appearance, offers good-quality skin cover and cushion to the testes as well as protective sensation. PMID:26774357

  7. 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?

  8. Flap--edge flowfield measurements

    NASA Astrophysics Data System (ADS)

    Pye, John D.; Cantwell, Brian J.

    1997-11-01

    Recent studies of airframe noise suggest that the wing and flap trailing--edges as well as the flap side--edge are areas of significant noise generation. To identify the fluid dynamic processes associated with these noise sources, we are examining the flow--field around a NACA 63--215 Mod B main element airfoil configured with a half--span Fowler flap. The tests are performed in a low--speed wind tunnel at a Reynolds number of ~ 6.0×10^5. A hot wire traverse system is used to map the mean velocities and turbulence intensities in the near wake region of the flow. Measurements of the pressure fluctuations along the flap side--edge and in the cove of the airfoil configuration are made with pressure transducers mounted inside the airfoil. The experimental data are in good qualitative agreement with the numerical simulation of a slightly higher Reynolds number flow ( ~ 1.5×10^6) around a geometrically similar airfoil configuration.

  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. Force Generation by Flapping Foils

    NASA Astrophysics Data System (ADS)

    Bandyopadhyay, P. R.; Donnelly, M.

    1996-11-01

    Aquatic animals like fish use flapping caudal fins to produce axial and cross-stream forces. During WW2, German scientists had built and tested an underwater vehicle powered by similar flapping foils. We have examined the forces produced by a pair of flapping foils. We have examined the forced produced by a pair of flapping foils attached to the tail end of a small axisymmetric cylinder. The foils operate in-phase (called waving), or in anti-phase (called clapping). In a low-speed water tunnel, we have undertaken time-dependent measurements of axial and cross-stream forces and moments that are exerted by the vortex shedding process over the entire body. Phase-matched LDV measurements of vorticity-velocity vectors, as well as limited flow visualization of the periodic vortex shedding process have also been carried out. The direction of the induced velocity within a pair of shed vortices determines the nature of the forces produced, viz., thrust or drag or cross-stream forces. The clapping mode produces a widely dispersed symmetric array of vortices which results in axial forces only (thrust and rag). On the other hand, the vortex array is staggered in the waving mode and cross-stream (maneuvering) forces are then generated.

  11. 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.

  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. DIETARY PROTEIN AND LACTOSE INCREASE TRANSLATION INITIATION FACTOR ACTIVATION AND TISSUE PROTEIN SYNTHESIS IN NEONATAL PIGS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein synthesis and eukaryotic initiation factor (eIF) activation are increased in muscle and liver of pigs parenterally infused with amino acids and insulin. To examine the effects of enteral protein and carbohydrate on protein synthesis, pigs (n = 42, 1.7 kg body wt) were fed isocaloric milk die...

  15. 14 CFR 23.697 - Wing flap controls.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Wing flap controls. 23.697 Section 23.697... Systems § 23.697 Wing flap controls. (a) Each wing flap control must be designed so that, when the flap... with § 23.145(b)(3) necessitates wing flap retraction to positions that are not fully retracted,...

  16. 14 CFR 23.697 - Wing flap controls.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Wing flap controls. 23.697 Section 23.697... Systems § 23.697 Wing flap controls. (a) Each wing flap control must be designed so that, when the flap... with § 23.145(b)(3) necessitates wing flap retraction to positions that are not fully retracted,...

  17. 14 CFR 23.697 - Wing flap controls.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Wing flap controls. 23.697 Section 23.697... Systems § 23.697 Wing flap controls. (a) Each wing flap control must be designed so that, when the flap... with § 23.145(b)(3) necessitates wing flap retraction to positions that are not fully retracted,...

  18. 14 CFR 23.697 - Wing flap controls.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Wing flap controls. 23.697 Section 23.697... Systems § 23.697 Wing flap controls. (a) Each wing flap control must be designed so that, when the flap... with § 23.145(b)(3) necessitates wing flap retraction to positions that are not fully retracted,...

  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. Flapping propulsion with tip pitch control

    NASA Astrophysics Data System (ADS)

    Huera-Huarte, Francisco; Gharib, Morteza

    2014-11-01

    The effect of flexibility in the propulsion performance and efficiency of oscillating pitching foils has received a large amount of attention in the past years. Scientists have used simplified robotic models that mimic the kinematics of flying and swimming animals, in order to get inspiration to build more efficient engineering systems. Compliance is one of the aspects that has received more attention, as it seems to be a common feature in nature's flyers and swimmers. Active or passive control elements are also common in nature. We will show how thrust generation in a pitching fin, can be greatly affected by controlling the tip pitch motion dynamically and independently of the fin itself. This is in fact a controlled local change of curvature of the end of the fin. A robotic system has been designed in a way that not only flapping amplitudes and frequencies can be controlled, but also the amplitudes and frequencies of the tip and the phase difference between the tip and the fin. We measured thrust forces and the vortex dynamics in the near wake of the system, by using planar DPIV (Digital Particle Image Velocimetry) in a wide variety of flapping situations with tip control. Funding from Spanish Ministry of Science through Grant DPI2012-37904 is gratefully acknowledged.

  1. Poliovirus protein 2C has ATPase and GTPase activities.

    PubMed

    Rodríguez, P L; Carrasco, L

    1993-04-15

    Poliovirus protein 2C belongs to an expanding group of proteins containing a nucleotide binding motif in their sequence. We present evidence that poliovirus 2C has nucleoside triphosphatase (NTPase) activity and binds to RNA. Poliovirus 2C was expressed in Escherichia coli cells as a fusion protein with the maltose binding protein (MBP). The fusion protein MBP-2C is efficiently cut by protease Xa within the 2C region. Thus, the fusion protein as such was used to assay for the putative activities of poliovirus 2C. Deletion mutants were constructed which lacked different portions of the 2C carboxyl terminus: mutant 2C delta 1 lacked the last 169 amino acids, whereas mutant 2C delta 2 had the last 74 amino acids deleted. The fusion proteins MBP-2C, MBP-2BC, and the mutant MBP-2C delta 2 that contained the first 255 amino acids of 2C had NTPase activity. Both ATPase and GTPase activities are inhibited by antibodies directed against the MBP-2C protein. Analysis of the ability of the different proteins to bind to labeled RNA indicates that MBP-2C and MBP-2BC form a complex, whereas none of the mutants interacted with RNA, indicating that the RNA binding domain lies beyond amino acid 255. None of the fusion proteins had detectable helicase activity. We suggest that poliovirus protein 2C shows similarities to the GTPases group involved in vesicular traffic and transports the viral RNA replication complexes. These results provide the first experimental evidence that poliovirus protein 2C is an NTPase and that this protein has affinity for nucleic acids. PMID:8385138

  2. Protein determination in seeds by proton activation

    NASA Astrophysics Data System (ADS)

    Morales, J. R.; Dinator, M. I.; Cerda, P.

    1989-04-01

    A proton beam of 6.6 MeV has been used to produce 11C and 13N in Araucaria Araucana seeds. Their positron decay allows determination of the N/C ratio. In seeds the nitrogen content is associated to proteins while carbon is spread in the organic material. Samples were irradiated for about 10 min with a beam intensity of 5 nA on areas of 1 mm 2. Slices of the seed were radially explored, showing a larger concentration of protein in the center.

  3. Monitoring G protein activation in cells with BRET

    PubMed Central

    Masuho, Ikuo; Martemyanov, Kirill A.; Lambert, Nevin A.

    2016-01-01

    Summary Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using standard laboratory equipment. PMID:26260597

  4. 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

  5. Pressure Distribution Over Airfoils with Fowler Flaps

    NASA Technical Reports Server (NTRS)

    Wenzinger, Carl J; Anderson, Walter B

    1938-01-01

    Report presents the results of tests made of a Clark y airfoil with a Clark y Fowler flap and of an NACA 23012 airfoil with NACA Fowler flaps. Some of the tests were made in the 7 by 10-foot wind tunnel and others in the 5-foot vertical wind tunnel. The pressures were measured on the upper and lower surfaces at one chord section both on the main airfoils and on the flaps for several angles of attack with the flaps located at the maximum-lift settings. A test installation was used in which the model was mounted in the wind tunnel between large end planes so that two-dimensional flow was approximated. The data are given in the form of pressure-distribution diagrams and as plots of calculated coefficients for the airfoil-and-flap combinations and for the flaps alone.

  6. New constitutive latex osmotin-like proteins lacking antifungal activity.

    PubMed

    Freitas, Cleverson D T; Silva, Maria Z R; Bruno-Moreno, Frederico; Monteiro-Moreira, Ana C O; Moreira, Renato A; Ramos, Márcio V

    2015-11-01

    Proteins that share similar primary sequences to the protein originally described in salt-stressed tobacco cells have been named osmotins. So far, only two osmotin-like proteins were purified and characterized of latex fluids. Osmotin from Carica papaya latex is an inducible protein lacking antifungal activity, whereas the Calotropis procera latex osmotin is a constitutive antifungal protein. To get additional insights into this subject, we investigated osmotins in latex fluids of five species. Two potential osmotin-like proteins in Cryptostegia grandiflora and Plumeria rubra latex were detected by immunological cross-reactivity with polyclonal antibodies produced against the C. procera latex osmotin (CpOsm) by ELISA, Dot Blot and Western Blot assays. Osmotin-like proteins were not detected in the latex of Thevetia peruviana, Himatanthus drasticus and healthy Carica papaya fruits. Later, the two new osmotin-like proteins were purified through immunoaffinity chromatography with anti-CpOsm immobilized antibodies. Worth noting the chromatographic efficiency allowed for the purification of the osmotin-like protein belonging to H. drasticus latex, which was not detectable by immunoassays. The identification of the purified proteins was confirmed after MS/MS analyses of their tryptic digests. It is concluded that the constitutive osmotin-like proteins reported here share structural similarities to CpOsm. However, unlike CpOsm, they did not exhibit antifungal activity against Fusarium solani and Colletotrichum gloeosporioides. These results suggest that osmotins of different latex sources may be involved in distinct physiological or defensive events. PMID:26231325

  7. 14 CFR 25.701 - Flap and slat interconnection.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... flaps or slats retracted on one side and extended on the other, the motion of flaps or slats on opposite...) For airplanes with flaps or slats that are not subjected to slipstream conditions, the structure...

  8. 14 CFR 25.701 - Flap and slat interconnection.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... flaps or slats retracted on one side and extended on the other, the motion of flaps or slats on opposite...) For airplanes with flaps or slats that are not subjected to slipstream conditions, the structure...

  9. 14 CFR 25.701 - Flap and slat interconnection.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... flaps or slats retracted on one side and extended on the other, the motion of flaps or slats on opposite...) For airplanes with flaps or slats that are not subjected to slipstream conditions, the structure...

  10. Four Flaps Technique for Neoumbilicoplasty

    PubMed Central

    Lee, Young Taek; Kwon, Chan; Rhee, Seung Chul; Cho, Sang Hun

    2015-01-01

    The absence or disfigurement of the umbilicus is both cosmetically and psychologically distressing to patients. The goal of aesthetically pleasing umbilical reconstruction is to create a neoumbilicus with sufficient depth and good morphology, with natural-looking superior hooding and minimal scarring. Although many reports have presented techniques for creating new and attractive umbilici, we developed a technique that we term the "four flaps technique" for creating a neoumbilicus in circumstances such as the congenital absence of the umbilicus or the lack of remaining umbilical tissue following the excision of a hypertrophic or scarred umbilicus. This method uses the neighboring tissue by simply elevating four flaps and can yield sufficient depth and an aesthetically pleasing shape with appropriate superior hooding. PMID:26015893

  11. Axial pattern skin flaps in cats.

    PubMed

    Remedios, A M; Bauer, M S; Bowen, C V; Fowler, J D

    1991-01-01

    The major direct cutaneous vessels identified in the cat include the omocervical, thoracodorsal, deep circumflex iliac, and caudal superficial epigastric arteries. Axial pattern skin flaps based on the thoracodorsal and caudal superficial epigastric arteries have been developed in cats. Rotation of these flaps as islands allows skin coverage to the carpus and metatarsus, respectively. The thoracodorsal and caudal superficial epigastric flaps provide a practical, one-step option in the reconstruction of large skin defects involving the distal extremities of cats. PMID:2011063

  12. Craniotomy flap osteomyelitis: a diagnostic approach

    SciTech Connect

    Blumenkopf, B.; Hartshorne, M.F.; Bauman, J.M.; Cawthon, M.A.; Patton, J.A.; Friedman, A.H.

    1987-01-01

    Nine cases of suspected craniotomy flap osteomyelitis evaluated by combined bone and gallium scanning are presented. In six cases, the clinical data were inconclusive and evaluation by radionuclide imaging provided an accurate negative diagnosis. The other three cases considered positive by this technique were proven infected at subsequent exploration and flap removal. The use of radionuclide bone and gallium imaging should be considered in cases of possible craniotomy flap osteomyelitis.

  13. Wing flapping with minimum energy

    NASA Technical Reports Server (NTRS)

    Jones, R. T.

    1980-01-01

    A technique employed by Prandtl and Munk is adapted for the case of a wing in flapping motion to determine its lift distribution. The problem may be reduced to one of minimizing induced drag for a specified and periodically varying bending moment at the wing root. It is concluded that two wings in close tandem arrangement, moving in opposite phase, would eliminate the induced aerodynamic losses calculated

  14. Protein kinase activity associated with pancreatic zymogen granules.

    PubMed

    Burnham, D B; Munowitz, P; Thorn, N; Williams, J A

    1985-05-01

    Purified zymogen granules were prepared from rat pancreas by using an iso-osmotic Percoll gradient. In the presence of [gamma-32P]ATP, phosphorylation of several granule proteins was induced by Ca2+, most notably a Mr-13 000 protein, whereas addition of cyclic AMP was without effect. When phosphatidylserine was also added, Ca2+ increased the phosphorylation of additional proteins, with the largest effect on a protein of Mr 62 000. Purified granules were also able to phosphorylate exogenous substrates. Ca2+-induced phosphorylation of lysine-rich histone was enhanced over 3-fold in the presence of phosphatidylserine, and cyclic AMP-activated protein kinase activity was revealed with mixed histone as substrate. The concentrations of free Ca2+ and cyclic AMP required for half-maximal phosphorylation of both endogenous and exogenous proteins were 1-3 microM and 57 nM respectively. Treatment of granules with 0.25 M-KCl resulted in the release of phosphatidylserine-dependent kinase activity into a high-speed granule supernatant. In contrast, granule-protein substrates of Ca2+-activated kinase activity were resistant to KCl extraction, and in fact were present in purified granule membranes. Kinase activity activated by cyclic AMP was not extracted by KCl treatment. It is concluded that phosphorylation of integral membrane proteins in the zymogen granule can be induced by one or more Ca2+-activated protein kinases. Such a reaction is a potential mechanism by which exocytosis may be regulated in the exocrine pancreas by Ca2+-mediated secretagogues. PMID:4004796

  15. Protein kinase activity associated with pancreatic zymogen granules.

    PubMed Central

    Burnham, D B; Munowitz, P; Thorn, N; Williams, J A

    1985-01-01

    Purified zymogen granules were prepared from rat pancreas by using an iso-osmotic Percoll gradient. In the presence of [gamma-32P]ATP, phosphorylation of several granule proteins was induced by Ca2+, most notably a Mr-13 000 protein, whereas addition of cyclic AMP was without effect. When phosphatidylserine was also added, Ca2+ increased the phosphorylation of additional proteins, with the largest effect on a protein of Mr 62 000. Purified granules were also able to phosphorylate exogenous substrates. Ca2+-induced phosphorylation of lysine-rich histone was enhanced over 3-fold in the presence of phosphatidylserine, and cyclic AMP-activated protein kinase activity was revealed with mixed histone as substrate. The concentrations of free Ca2+ and cyclic AMP required for half-maximal phosphorylation of both endogenous and exogenous proteins were 1-3 microM and 57 nM respectively. Treatment of granules with 0.25 M-KCl resulted in the release of phosphatidylserine-dependent kinase activity into a high-speed granule supernatant. In contrast, granule-protein substrates of Ca2+-activated kinase activity were resistant to KCl extraction, and in fact were present in purified granule membranes. Kinase activity activated by cyclic AMP was not extracted by KCl treatment. It is concluded that phosphorylation of integral membrane proteins in the zymogen granule can be induced by one or more Ca2+-activated protein kinases. Such a reaction is a potential mechanism by which exocytosis may be regulated in the exocrine pancreas by Ca2+-mediated secretagogues. Images Fig. 1. Fig. 2. Fig. 7. Fig. 8. PMID:4004796

  16. The kinetics of ER fusion protein activation in vivo

    PubMed Central

    Wilson, Catherine H.; Gamper, Ivonne; Perfetto, Alessandra; Auw, Jeremy; Littlewood, Trevor D.; Evan, Gerard I.

    2014-01-01

    Reversibly switchable proteins are powerful tools with which to explore protein function in vitro and in vivo. For example, the activity of many proteins fused to the hormone-binding domain of the modified estrogen receptor (ERTAM) can be regulated by provision or removal of 4-hydroxytamoxifen (4-OHT). Despite the widespread use of ERTAM fusions in vivo, inadequate data are available as to the most efficacious routes for systemic tamoxifen delivery. In this study, we have used two well-characterised ERTAM fusion proteins, both reversibly activated by 4-OHT, to compare the effectiveness and kinetics of 4-OHT delivery in mice in vivo by either tamoxifen in food or by intraperitoneal injection. Our data indicate that dietary tamoxifen offers an effective, facile and ethically preferable means for long term activation of ERTAM fusion proteins in vivo. PMID:24662815

  17. A water tunnel study of Gurney flaps

    NASA Technical Reports Server (NTRS)

    Neuhart, Dan H.; Pendergraft, Odis C., Jr.

    1988-01-01

    Several Gurney flap configurations were tested in the NASA Langley 16 x 24 inch Water Tunnel. These devices provided an increased region of attached flow on a wing upper surface relative to the wing without the flaps. The recirculation region behind the flap was visualized and shown to be consistent with hypotheses stated in previous research. Although the test Reynolds number for this study was several orders of magnitude below those in previous investigations, the effect of the Gurney flaps is in qualitative agreement with them. This is as would be expected from first order effects for high lift devices.

  18. An investigation of the flap edge flowfield

    NASA Astrophysics Data System (ADS)

    Pye, John David

    To identify and understand the fluid dynamic processes associated with flow in the region of a flap side edge, a NACA 63-215 Mod B main element with a half-span Fowler flap was tested in the JIAA Low Speed Wind Tunnel at Stanford University. Measurements were made using a variety of techniques to capture the effects of the flap edge vortex. Pressure sensitive paint was applied to the upper surface of both the flap and main element, as well as to the flap side edge. Fast response pressure transducers were mounted interior to the model to measure surface pressure fluctuations on the flap side edge. Single component hotwire data was taken in the near wake region of the flap edge. In addition to the data experimentally obtained, a computational data set of a geometrically similar model at a flight Reynolds number was used for comparison. The data indicates the presence of a dual vortex structure along the flap side edge. This structure is seen to grow, merge, and ultimately become a single symmetric vortex as it progresses downstream. Surface pressure fluctuations on the side edge scale as three power laws with free stream velocity as different flow regions are encountered. By varying the model rigging, indications of a confined source region for the pressure fluctuations were observed. A spatial survey of the correlation between flap side edge surface pressure fluctuations and the near-wake fluctuating velocity field shows increased correlation coefficients for the region surrounding the vortex core.

  19. Microsurgical free flaps: Controversies in maxillofacial reconstruction

    PubMed Central

    George, Rinku K.; Krishnamurthy, Arvind

    2013-01-01

    Reconstructive microsurgery for oral and maxillofacial (OMF) defects is considered as a niche specialty and is performed regularly only in a handful of centers. Till recently the pectoralis major myocutaneous flap (PMMC) was considered to be the benchmark for OMF reconstruction. This philosophy is changing fast with rapid advancement in reconstructive microsurgery. Due to improvement in instrumentation and the development of finer techniques of flap harvesting we can positively state that microsurgery has come of age. Better techniques, microscopes and micro instruments enable us to do things previously unimaginable. Supramicrosurgery and ultrathin flaps are a testimony to this. Years of innovation in reconstructive microsurgery have given us a reasonably good number of very excellent flaps. Tremendous work has been put into producing some exceptionally brilliant research articles, sometimes contradicting each other. This has led to the need for clarity in some areas in this field. This article will review some controversies in reconstructive microsurgery and analyze some of the most common microvascular free flaps (MFF) used in OMF reconstruction. It aims to buttress the fact that three flaps-the radial forearm free flap (RFFF), anterolateral thigh flap (ALT) and fibula are the ones most expedient in the surgeon's arsenal, since they can cater to almost all sizeable defects we come across after ablative surgery in the OMF region. They can thus aptly be titled as the workhorses of OMF reconstruction with regard to free flaps. PMID:23662264

  20. 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

  1. 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.

  2. A new classification of spreader flap techniques.

    PubMed

    Wurm, Jochen; Kovacevic, Milos

    2013-12-01

    The objective of this study was to introduce various spreader flap technique modifications to adjust the width of the middle nasal vault in patients who underwent rhinoplasties with humpectomy. Decisive modifications of current spreader flap techniques were performed to allow a more natural restoration of the middle nasal vault and the internal nasal valve after humpectomy. Additional steps provide tools to adjust the width and shape of the middle nasal vault according to patients' requirements. The techniques were categorized into "basic spreader flaps," "flaring spreader flaps," "support spreader flaps," and "interrupted spreader flaps." The various spreader flap techniques were used during 576 primary septorhinoplasties in patients with hump noses, hump/crooked noses, or hump/tension noses. The average follow-up was 19 months. Patients who received basic spreader flaps or a flaring spreader flaps tended to show a slightly too wide middle nasal vault, revision surgery was necessary in four of these cases. All other patients showed an appropriate width in the middle nasal vault and an aesthetically pleasing course of the dorsal aesthetic lines. No signs of inverted v deformities or collapse of the internal nasal valve were observed in any of the patients. Patients who had reported impaired nasal breathing preoperatively described clearly improved subjective symptoms. The described techniques appear to be appropriate and highly promising as a supplement to existing procedures for reconstructing the middle nasal vault and internal nasal valves. No additional cartilage grafts are needed. PMID:24327250

  3. 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

  4. Arterialized Venous Bone Flaps: An Experimental Investigation.

    PubMed

    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

  5. 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.

  6. 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

  7. Evidence for a receptor protein of activated carcinogen

    PubMed Central

    Mainigi, Kumar D.; Sorof, Sam

    1977-01-01

    During carcinogenesis in rat liver by 3′-methyl-4-dimethylaminoazobenzene, the two moieties of the principal liver carcinogen-protein complex have considerably different turnover rates. With continued ingestion of the azocarcinogen by rats, the bound azo dye in the complex has a half-life of 2.5 ± 0.25 (SD) days, while the protein moiety has a half-life of 8.7 ± 1.6 days (probability of identity <0.001). In addition, the interaction of the azocarcinogen with the principal target protein in vivo appears to extend the half-life of the protein itself from 3.3 ± 0.2 days in normal liver to 8.7 ± 1.6 days (P < 0.001). The slowing of the turnover of the protein by the carcinogen appears to be readily reversible, since soon after the cessation of azocarcinogen feeding, the half-life of the protein returns to that of the target protein in normal liver. The considerable difference in turnover rates of the two moieties of the complex and the reversible effects of the carcinogen in slowing the turnover of the protein moiety suggest that the two moieties of the native azoprotein are noncovalently linked and that they have different biological activities. The native complex appears to contain azo dye in an activated state that is capable of yielding a reactive electrophile, because after protein denaturation the bound azo dye was previously found to have properties that are indicative of covalent linkage to the protein. The retardation in the biological turnover rate of the protein moiety, apparently resulting from interaction with azocarcinogen, is in agreement with the known ligand-induced stabilization in vitro and reduced rate of proteolytic degradation in vivo of other proteins that result from conformational change to a more compact configuration. Our evidence is consistent with the hypothesis that the principal liver carcinogen-protein complex contains hydrophobically bound activated azocarcinogen, whose specificity of reaction with critical macromolecule(s) in

  8. 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

  9. Pestivirus NS3 (p80) protein possesses RNA helicase activity.

    PubMed Central

    Warrener, P; Collett, M S

    1995-01-01

    The pestivirus bovine viral diarrhea virus (BVDV) p80 protein (referred to here as the NS3 protein) contains amino acid sequence motifs predictive of three enzymatic activities: serine proteinase, nucleoside triphosphatase, and RNA helicase. We have previously demonstrated that the former two enzymatic activities are associated with this protein. Here, we show that a purified recombinant BVDV NS3 protein derived from baculovirus-infected insect cells possesses RNA helicase activity. BVDV NS3 RNA helicase activity was specifically inhibited by monoclonal antibodies to the p80 protein. The activity was dependent on the presence of nucleoside triphosphate and divalent cation, with a preference for ATP and Mn2+. Hydrolysis of the nucleoside triphosphate was necessary for strand displacement. The helicase activity required substrates with an un-base-paired region on the template strand 3' of the duplex region. As few as three un-base-paired nucleotides were sufficient for efficient oligonucleotide displacement. However, the enzyme did not act on substrates having a single-stranded region only to the 5' end of the duplex or on substrates lacking single-stranded regions altogether (blunt-ended duplex substrates), suggesting that the directionality of the BVDV RNA helicase was 3' to 5' with respect to the template strand. The BVDV helicase activity was able to displace both RNA and DNA oligonucleotides from RNA template strands but was unable to release oligonucleotides from DNA templates. The possible role of this activity in pestivirus replication is discussed. PMID:7853509

  10. 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

  11. Iron-chelating activity of chickpea protein hydrolysate peptides.

    PubMed

    Torres-Fuentes, Cristina; Alaiz, Manuel; Vioque, Javier

    2012-10-01

    Chickpea-chelating peptides were purified and analysed for their iron-chelating activity. These peptides were purified after affinity and gel filtration chromatography from a chickpea protein hydrolysate produced with pepsin and pancreatin. Iron-chelating activity was higher in purified peptide fractions than in the original hydrolysate. Histidine contents were positively correlated with the iron-chelating activity. Hence fractions with histidine contents above 20% showed the highest chelating activity. These results show that iron-chelating peptides are generated after chickpea protein hydrolysis with pepsin plus pancreatin. These peptides, through metal chelation, may increase iron solubility and bioavailability and improve iron absorption. PMID:25005984

  12. Inclusion bodies and purification of proteins in biologically active forms.

    PubMed

    Mukhopadhyay, A

    1997-01-01

    Even though recombinant DNA technology has made possible the production of valuable therapeutic proteins, its accumulation in the host cell as inclusion body poses serious problems in the recovery of functionally active proteins. In the last twenty years, alternative techniques have been evolved to purify biologically active proteins from inclusion bodies. Most of these remain only as inventions and very few are commercially exploited. This review summarizes the developments in isolation, refolding and purification of proteins from inclusion bodies that could be used for vaccine and non-vaccine applications. The second section involves a discussion on inclusion bodies, how they are formed, and their physicochemical properties. In vivo protein folding in Escherichia coli and kinetics of in vitro protein folding are the subjects of the third and fourth sections respectively. The next section covers the recovery of bioactive protein from inclusion bodies: it includes isolation of inclusion body from host cell debris, purification in denatured state alternate refolding techniques, and final purification of active molecules. Since purity and safety are two important issues in therapeutic grade proteins, the following three sections are devoted to immunological and biological characterization of biomolecules, nature, and type of impurities normally encountered, and their detection. Lastly, two case studies are discussed to demonstrate the sequence of process steps involved. PMID:8939059

  13. Activators of G Protein Signaling in the Kidney

    PubMed Central

    2015-01-01

    Heterotrimeric G proteins play a crucial role in regulating signal processing to maintain normal cellular homeostasis, and subtle perturbations in its activity can potentially lead to the pathogenesis of renal disorders or diseases. Cell-surface receptors and accessory proteins, which normally modify and organize the coupling of individual G protein subunits, contribute to the regulation of heterotrimeric G protein activity and their convergence and/or divergence of downstream signaling initiated by effector systems. Activators of G protein signaling (AGS) are a family of accessory proteins that intervene at multiple distinct points during the activation–inactivation cycle of G proteins, even in the absence of receptor stimulation. Perturbations in the expression of individual AGS proteins have been reported to modulate signal transduction pathways in a wide array of diseases and disorders within the brain, heart, immune system, and more recently, the kidney. This review will provide an overview of the expression profile, localization, and putative biologic role of the AGS family in the context of normal and diseased states of the kidney. PMID:25628392

  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 PAGESBeta

    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. Design and analysis of rotor systems with multiple trailing edge flaps and resonant actuators

    NASA Astrophysics Data System (ADS)

    Kim, Jun-Sik

    The purpose of this thesis is to develop piezoelectric resonant actuation systems and new active control methods utilizing the multiple trailing-edge flaps' configuration for rotorcraft vibration suppression and blade loads control. An aeroelastic model is developed for a composite rotor blade with multiple trailing-edge flaps. The rotor blade airloads are calculated using quasi-steady blade element aerodynamics with a free wake model for rotor inflow. A compressible unsteady aerodynamics model is employed to accurately predict the incremental trailing edge flap airloads. Both the finite wing effect and actuator saturation for trailing-edge flaps are also included in an aeroelastic analysis. For a composite articulated rotor, a new active blade loads control method is developed and tested numerically. The concept involves straightening the blade by introducing dual trailing edge flaps. The objective function, which includes vibratory hub loads, bending moment harmonics and active flap control inputs, is minimized by an integrated optimal control/optimization process. A numerical simulation is performed for the steady-state forward flight of an advance ratio of 0.35. It is demonstrated that through straightening the rotor blade, which mimics the behavior of a rigid blade, both the bending moments and vibratory hub loads can be significantly reduced by 32% and 57%, respectively. An active vibration control method is developed and analyzed for a hingeless rotor. The concept involves deflecting each individual trailing-edge flap using a compact resonant actuation system. Each resonant actuation system could yield high authority, while operating at a single frequency. Parametric studies are conducted to explore the finite wing effect of trailing-edge flaps and actuator saturation. A numerical simulation has been performed for the steady-state forward flight (mu = 0.15 ˜ 0.35). It is demonstrated that multiple trailing-edge flap configuration with the resonant actuation

  17. Development of an improved aeroelastic model for the investigation of vibration reduction in helicopter rotors using trailing edge flaps

    NASA Astrophysics Data System (ADS)

    Myrtle, Timothy Fitzgerald

    1998-12-01

    This dissertation describes the development of an aeroelastic model of a helicopter rotor incorporating partial span trailing edge flaps on the blade and its application to the investigation of vibration reduction using active control. A new two-dimensional unsteady aerodynamic model for an airfoil/flap combination is described that includes compressibility and unsteady freestream effects. This new aerodynamic model is based on a rational function approximation (RFA) approach. In this approach, oscillatory response data obtained for a selected set of generalized airfoil and flap motions is used to generate an approximate aerodynamic transfer function which can be transformed to the time domain to form a state space aerodynamic model. In this dissertation, a method is described for adapting the conventional RFA approach to include unsteady freestream effects. Excellent agreement is demonstrated between the response of the new aerodynamic model and an exact incompressible solution to the unsteady freestream case. This model provides a complete description of the unsteady flap hinge moments due to airfoil and flap motion, allowing a complete and accurate characterization of control actuation requirements. The structural model utilizes an elastic blade model which includes fully coupled flap-lag-torsional dynamics and includes the effects of moderate deflections. The aeroelastic model is formulated in the time domain, with the coupled trim/response solution obtained using direct numerical integration in combination with autopilot type controller. A conventional higher harmonic control approach is used to investigate vibration reduction. Vibration control studies are performed which compare results using the new aerodynamic model and incompressible quasisteady Theodorsen aerodynamics. Significant differences were observed in the required deflections and control moments, indicating that compressibility and unsteady effects are necessary to properly characterize the

  18. 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

  19. 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

  20. [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

  1. Secondary onlay free flap reconstruction of glossectomy defects following initial successful flap restoration.

    PubMed

    Rihani, Jordan; Lee, Thomas; Ducic, Yadranko

    2013-08-01

    Patients who undergo tongue reconstruction over time may develop gradual worsening of dysarthria and dysphagia secondary to flap atrophy. At our institution, these patients undergo a secondary flap onlay procedure for augmentation of the neotongue. We review a total of 11 patients with total glossectomy defect who underwent secondary tongue augmentation with secondary onlay free flap consisting of radial forearm free flap (n = 6) and rectus free flap (n = 5). There was improvement in swallowing in 7 of 11 patients. Five (45.4%) patients achieved gastric tube independence. Seven (63.6%) patients achieved a varying degree of oral intake. All patients achieved tracheostomy independence. Dysarthria was improved in all patients. There were no flap failures. Therefore, a secondary onlay flap technique is feasible and may improve dysphagia and dysarthria to achieve gastric tube and tracheostomy independence in total glossectomy patients with delayed tongue atrophy. PMID:23625797

  2. Distally based sural artery flap without sural nerve.

    PubMed

    Motamed, Sadrollah; Yavari, Masood; Mofrad, Hamid Reza Hallaj; Rafiee, Reza; Shahraki, Feaz Niazi

    2010-01-01

    The distal third of the tibia, ankle and heel area is difficult to reconstruct. For small to medium size defects, local flaps are often an easier alternative than free flap. In lower limb surgery, the sural flap is based on this principle and this flap is becoming increasingly popular. The distally based superficial sural artery flap, first described as a distally based neuro skin flap by masquelet et al., is a skin island flap supplied by the vascular axis of the sural nerve. The main disadvantage of distally based sural artery flap is sacrifice of the sural nerve because it is described the concept of neurocutaneus island flap. We describe one case of reverse sural flap without sural nerve .The aim of this paper is to establish the reliability of this flap even without sural nerve. PMID:21133008

  3. 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

  4. 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.

  5. 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

  6. Activation of a human chromosomal replication origin by protein tethering

    PubMed Central

    Chen, Xiaomi; Liu, Guoqi; Leffak, Michael

    2013-01-01

    The specification of mammalian chromosomal replication origins is incompletely understood. To analyze the assembly and activation of prereplicative complexes (pre-RCs), we tested the effects of tethered binding of chromatin acetyltransferases and replication proteins on chromosomal c-myc origin deletion mutants containing a GAL4-binding cassette. GAL4DBD (DNA binding domain) fusions with Orc2, Cdt1, E2F1 or HBO1 coordinated the recruitment of the Mcm7 helicase subunit, the DNA unwinding element (DUE)-binding protein DUE-B and the minichromosome maintenance (MCM) helicase activator Cdc45 to the replicator, and restored origin activity. In contrast, replication protein binding and origin activity were not stimulated by fusion protein binding in the absence of flanking c-myc DNA. Substitution of the GAL4-binding site for the c-myc replicator DUE allowed Orc2 and Mcm7 binding, but eliminated origin activity, indicating that the DUE is essential for pre-RC activation. Additionally, tethering of DUE-B was not sufficient to recruit Cdc45 or activate pre-RCs formed in the absence of a DUE. These results show directly in a chromosomal background that chromatin acetylation, Orc2 or Cdt1 suffice to recruit all downstream replication initiation activities to a prospective origin, and that chromosomal origin activity requires singular DNA sequences. PMID:23658226

  7. 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

  8. Understanding the Role of Chord-wise Flexibility in Flapping Wing Flight

    NASA Astrophysics Data System (ADS)

    Gaston, Zachary; Dong, Haibo; Wan, Hui; Ol, Michael

    2011-11-01

    Aerodynamic performance of flapping hinged plates is numerically studied to explore the effects of chord-wise flexibility in flapping wing flight. The plate with chord-wise flexibility is modeled as a two-link mechanism with a torsional spring hinge in between. The upper-link of the plate is controlled by prescribed motion and the rest of body is subjected to passive deflection due to fluid-body interaction. The effect of forced to natural frequency ratio is studied first for a flapping hinged-plate, on which prescribed hovering motion is actively applied. The effects of torsional stiffness and chord-wise flexibility are further explored for pitching and plunging plates, observing the flow phenomena and lift production as a result of this change. Comparisons between rigid plates, free-to-pivot hinged plates, and the torsional spring hinged plates are made, identifying a more optimal model for promoting lift production in flapping plates.

  9. Free flap pulse oximetry utilizing reflectance photoplethysmography.

    PubMed

    Zaman, T; Kyriacou, P A; Pal, S K

    2013-01-01

    The successful salvage of a free flap is dependent on the continuous monitoring of perfusion. To date there is no widely accepted and readily available post-operative monitoring technique to reliably assess the viability of free flaps by continuously monitoring free flap blood oxygen saturation. In an attempt to overcome the limitations of the current techniques a reflectance photoplethysmographic (PPG) processing system has been developed with the capability of real-time estimation of arterial oxygen saturation (SpO2) levels implemented in LabVIEW. This system was evaluated in clinical measurements on five patients undergoing breast reconstruction using Deep Inferior Epigastric Perforator (DIEP) flap. Good quality PPG signals were obtained from the flaps and fingers simultaneously. The estimated free flap SpO2 values were in broad agreement with the oxygen saturation readings from the commercial pulse oximeter. The results suggest that reflectance free flap photoplethysmography can be used as a continuous monitoring technique to non-invasively monitor the perfusion of free flaps. PMID:24110620

  10. 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 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control Systems § 23.701 Flap...

  11. 14 CFR 23.701 - Flap interconnection.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Flap interconnection. 23.701 Section 23.701 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: NORMAL, UTILITY, ACROBATIC, AND COMMUTER CATEGORY AIRPLANES Design and Construction Control Systems § 23.701 Flap...

  12. Use of free flaps in burn trauma.

    PubMed

    Tvrdek, M; Nejedlý, A; Kletenský, J; Pros, Z; Broz, L

    1994-01-01

    The present currently used method of treating and reconstructing injuries caused by electric current and deep thermal injuries is compared with the method of early reconstruction using free flaps. Brief summary of present experience substantiating this method is presented. The three presented clinical cases show subacute reconstruction in this type of injury by free flaps transfer. PMID:7618395

  13. 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....

  14. 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....

  15. 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....

  16. 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....

  17. 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....

  18. The posterior calf fascial free flap.

    PubMed

    Walton, R L; Matory, W E; Petry, J J

    1985-12-01

    Six posterior calf fascial free flaps were employed to reconstruct defects of the upper and lower extremities. One flap failed due to a constricting dressing. Two flaps sustained partial loss secondary to bleeding and hematoma formation. One flap dehisced at the distal suture line due to mobility of an underlying fracture. All surviving flaps eventually healed and resulted in stable, thin coverage. Donor-site morbidity has been minimal. Shortcomings of this flap model have been defined in the peculiarities of its thinness, diffuse vascular oozing, the extent of the vascular territory, and in postoperative monitoring. These problems are analyzed and recommendations for their resolution are presented. Fascia represents a unique tissue which offers an exciting new dimension in the reconstruction of certain defects--particularly those in which thinness is a desirable option. In the posterior calf model, the inclusion of fat represents an alternative modification that allows the surgeon to tailor the design to a variety of problems where fascia alone is too thin and a cutaneous flap is too thick. This concept may find its greatest application in wounds involving the hand or foot. We believe that this and other fascial flap prototypes may offer an ideal solution for reconstruction of major wounds of the extremities. PMID:3906718

  19. Thumb Reconstruction Using Foucher’s Flap

    PubMed Central

    Kola, Nardi

    2016-01-01

    BACKGROUND: Extensive pulp defects of the thumb, with the exposure of tendon or bone, are challenging reconstructive problems. Surgical treatment includes the use of local, regional, and free flaps. AIM: This paper is focused in Foucher’s neuro vascular flap. First DMCA or Foucher’s pedicle flap is a successful thumb reconstruction method, especially in patients not disturbed by its cosmetic appearance. MATERIAL AND METHODS: The first dorsal metacarpal artery (FDMCA) arises from the radial artery in the first intermetacarpal space, just distal to the tendon of the extensor pollicis longus. Pulp area of the thumb is the area where Foucher’s flap is more utilizable. This technique has other applications such as first web reconstruction, thumb lengthening, and following resection of tumors on the dorsum of the hand. RESULTS: We have in study 7 cases with work related trauma in two years period of time, between 2012 and 2014. We had only one partial flap survival and all the other flaps survived entirely. We have also taken in consideration subjective satisfaction with a range score from 4 to 10, cold intolerance, flap area and donor site sensibility with a range score from low to medium to normal. CONCLUSION: Careful pedicle discovery, secured elevation, pedicle strangulation prevention are very important for flap survival.

  20. G Protein Activation without a GEF in the Plant Kingdom

    PubMed Central

    Wang, Hao; Matthews, Melissa; Bradford, William; Bennetzen, Jeffrey L.; Jones, Alan M.

    2012-01-01

    Animal heterotrimeric G proteins are activated by guanine nucleotide exchange factors (GEF), typically seven transmembrane receptors that trigger GDP release and subsequent GTP binding. In contrast, the Arabidopsis thaliana G protein (AtGPA1) rapidly activates itself without a GEF and is instead regulated by a seven transmembrane Regulator of G protein Signaling (7TM-RGS) protein that promotes GTP hydrolysis to reset the inactive (GDP-bound) state. It is not known if this unusual activation is a major and constraining part of the evolutionary history of G signaling in eukaryotes. In particular, it is not known if this is an ancestral form or if this mechanism is maintained, and therefore constrained, within the plant kingdom. To determine if this mode of signal regulation is conserved throughout the plant kingdom, we analyzed available plant genomes for G protein signaling components, and we purified individually the plant components encoded in an informative set of plant genomes in order to determine their activation properties in vitro. While the subunits of the heterotrimeric G protein complex are encoded in vascular plant genomes, the 7TM-RGS genes were lost in all investigated grasses. Despite the absence of a Gα-inactivating protein in grasses, all vascular plant Gα proteins examined rapidly released GDP without a receptor and slowly hydrolyzed GTP, indicating that these Gα are self-activating. We showed further that a single amino acid substitution found naturally in grass Gα proteins reduced the Gα-RGS interaction, and this amino acid substitution occurred before the loss of the RGS gene in the grass lineage. Like grasses, non-vascular plants also appear to lack RGS proteins. However, unlike grasses, one representative non-vascular plant Gα showed rapid GTP hydrolysis, likely compensating for the loss of the RGS gene. Our findings, the loss of a regulatory gene and the retention of the “self-activating” trait, indicate the existence of divergent

  1. Influence of hinge point on flexible flap aerodynamic performance

    NASA Astrophysics Data System (ADS)

    Y Zhao, H.; Ye, Z.; Wu, P.; Li, C.

    2013-12-01

    Large scale wind turbines lead to increasing blade lengths and weights, which presents new challenges for blade design. This paper selects NREL S809 airfoil, uses the parameterized technology to realize the flexible trailing edge deformation, researches the static aerodynamic characteristics of wind turbine blade airfoil with flexible deformation, and the dynamic aerodynamic characteristics in the process of continuous deformation, analyses the influence of hinge point position 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 hinge point moving forward, total aerodynamic performance of flexible flap improves. Positive swing angle can push the transition point backward, thus postpones the occurrence of the transition phenomenon.

  2. 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

  3. 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. PMID:27080133

  4. 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

  5. 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

  6. The clinical application of anterolateral thigh flap.

    PubMed

    Lee, Yao-Chou; Chiu, Haw-Yen; Shieh, Shyh-Jou

    2011-01-01

    The anterolateral thigh flap can provide a large skin paddle nourished by a long and large-caliber pedicle and can be harvested by two-team work. Most importantly, the donor-site morbidity is minimal. However, the anatomic variations decreased its popularity. By adapting free-style flap concepts, such as preoperative mapping of the perforators and being familiar with retrograde perforator dissection, this disadvantage had been overcome gradually. Furthermore, several modifications widen its clinical applications: the fascia lata can be included for sling or tendon reconstruction, the bulkiness could be created by including vastus lateralis muscle or deepithelization of skin flap, the pliability could be increased by suprafascial dissection or primary thinning, the pedicle length could be lengthening by proximally eccentric placement of the perforator, and so forth. Combined with these technical and conceptual advancements, the anterolateral thigh flap has become the workhorse flap for soft-tissue reconstructions from head to toe. PMID:22567234

  7. Fasciocutaneous flap for vaginal and perineal reconstruction

    SciTech Connect

    Wang, T.N.; Whetzel, T.; Mathes, S.J.; Vasconez, L.O.

    1987-07-01

    A skin and fascia flap from the medial thigh is proposed for vaginal and perineal reconstruction. Dissection, vascular injection, and radiographs of 20 fresh cadaver limbs uniformly demonstrated the presence of a communicating suprafascial vascular plexus in the medial thigh. Three to four nonaxial vessels were consistently found to enter the proximal plexus from within 5 cm of the perineum. Preservation of these vessels permitted reliable elevation of a 9 X 20 cm fasciocutaneous flap without using the gracilis muscle as a vascular carrier. Fifteen flaps in 13 patients were used for vaginal replacement and coverage of vulvectomy, groin, and ischial defects. Depending on the magnitude of the defect, simultaneous and independent elevation of the gracilis muscle provided additional vascularized coverage as needed. Our experience indicates that the medial thigh fasciocutaneous flap is a durable, less bulky, and potentially sensate alternative to the gracilis musculocutaneous flap for vaginal and perineal reconstruction.

  8. 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.

  9. 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.

  10. 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. PMID:24037925

  11. 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.

  12. Liposomal packaging generates Wnt protein with in vivo biological activity.

    PubMed

    Morrell, Nathan T; Leucht, Philipp; Zhao, Ludan; Kim, Jae-Beom; ten Berge, Derk; Ponnusamy, Karthik; Carre, A Lyonel; Dudek, Henryk; Zachlederova, Marie; McElhaney, Michael; Brunton, Shirley; Gunzner, Janet; Callow, Marinella; Polakis, Paul; Costa, Mike; Zhang, Xiaoyan M; Helms, Jill A; Nusse, Roel

    2008-01-01

    Wnt signals exercise strong cell-biological and regenerative effects of considerable therapeutic value. There are, however, no specific Wnt agonists and no method for in vivo delivery of purified Wnt proteins. Wnts contain lipid adducts that are required for activity and we exploited this lipophilicity by packaging purified Wnt3a protein into lipid vesicles. Rather than being encapsulated, Wnts are tethered to the liposomal surface, where they enhance and sustain Wnt signaling in vitro. Molecules that effectively antagonize soluble Wnt3a protein but are ineffective against the Wnt3a signal presented by a cell in a paracrine or autocrine manner are also unable to block liposomal Wnt3a activity, suggesting that liposomal packaging mimics the biological state of active Wnts. When delivered subcutaneously, Wnt3a liposomes induce hair follicle neogenesis, demonstrating their robust biological activity in a regenerative context. PMID:18698373

  13. Activation of protein kinase C induces mitogen-activated protein kinase dephosphorylation and pronucleus formation in rat oocytes.

    PubMed

    Lu, Qing; Smith, Gary D; Chen, Da-Yuan; Han, Zhi-Ming; Sun, Qing-Yuan

    2002-07-01

    Mammalian oocytes are arrested at metaphase of the second meiotic division (MII) before fertilization. When oocytes are stimulated by spermatozoa, they exit MII stage and complete meiosis. It has been suggested that an immediate increase in intracellular free calcium concentration and inactivation of maturation promoting factor (MPF) are required for oocyte activation. However, the underlying mechanism is still unclear. In the present study, we investigated the role of protein kinase C (PKC) and mitogen-activated protein (MAP) kinase, and their interplay in rat oocyte activation. We found that MAP kinase became dephosphorylated in correlation with pronucleus formation after fertilization. Protein kinase C activators, phorbol 12-myriatate 13-acetate (PMA) and 1,2-dioctanoyl-rac-glycerol (diC8), triggered dephosphorylation of MAP kinase and pronucleus formation in a dose-dependent and time-dependent manner. Dephosphorylation of MAP kinase was also correlated with pronucleus formation when oocytes were treated with PKC activators. Effects of PKC activators were abolished by the PKC inhibitors, calphostin C and staurosporine, as well as a protein phosphatase blocker, okadaic acid (OA). These results suggest that PKC activation may cause rat oocyte pronucleus formation via MAP kinase dephosphorylation, which is probably mediated by OA-sensitive protein phosphatases. We also provide evidence supporting the involvement of such a process in fertilization. PMID:12080000

  14. 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.

  15. 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

  16. 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

  17. Activities of the Sex-lethal protein in RNA binding and protein:protein interactions.

    PubMed Central

    Samuels, M; Deshpande, G; Schedl, P

    1998-01-01

    The Drosophila sex determination gene Sex-lethal (Sxl) controls its own expression, and the expression of downstream target genes such as transformer , by regulating pre-mRNA splicing and mRNA translation. Sxl codes an RNA-binding protein that consists of an N-terminus of approximately 100 amino acids, two 90 amino acid RRM domains, R1 and R2, and an 80 amino acid C-terminus. In the studies reported here we have examined the functional properties of the different Sxl protein domains in RNA binding and in protein:protein interactions. The two RRM domains are responsible for RNA binding. Specificity in the recognition of target RNAs requires both RRM domains, and proteins which consist of the single domains or duplicated domains have anomalous RNA recognition properties. Moreover, the length of the linker between domains can affect RNA recognition properties. Our results indicate that the two RRM domains mediate Sxl:Sxl protein interactions, and that these interactions probably occur both in cis and trans. We speculate that cis interactions between R1 and R2 play a role in RNA recognition by the Sxl protein, while trans interactions stabilize complex formation on target RNAs that contain two or more closely spaced binding sites. Finally, we show that the interaction of Sxl with the snRNP protein Snf is mediated by the R1 RRM domain. PMID:9592147

  18. 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.

  19. 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.

  20. Probing heterotrimeric G protein activation: applications to biased ligands

    PubMed Central

    Denis, Colette; Saulière, Aude; Galandrin, Ségolène; Sénard, Jean-Michel; Galés, Céline

    2012-01-01

    Cell surface G protein-coupled receptors (GPCRs) drive numerous signaling pathways involved in the regulation of a broad range of physiologic processes. Today, they represent the largest target for modern drugs development with potential application in all clinical fields. Recently, the concept of “ligand-directed trafficking” has led to a conceptual revolution in pharmacological theory, thus opening new avenues for drug discovery. Accordingly, GPCRs do not function as simple on-off switch but rather as filters capable of selecting activation of specific signals and thus generating textured responses to ligands, a phenomenon often referred to as ligand-biased signaling. Also, one challenging task today remains optimization of pharmacological assays with increased sensitivity so to better appreciate the inherent texture of ligand responses. However, considering that a single receptor has pleiotropic signalling properties and that each signal can crosstalk at different levels, biased activity remains thus difficult to evaluate. One strategy to overcome these limitations would be examining the initial steps following receptor activation. Even if some G protein-independent functions have been recently described, heterotrimeric G protein activation remains a general hallmark for all GPCRs families and the first cellular event subsequent to agonist binding to the receptor. Herein, we review the different methodologies classically used or recently developed to monitor G protein activation and discuss them in the context of G protein biased -ligands. PMID:22229559

  1. Bryostatins activate protein kinase C in intact human platelets

    SciTech Connect

    Smith, J.B.; Tallant, E.A.; Pettit, G.R.; Wallace, R.W.

    1986-05-01

    Bryostatins, macrocyclic lactones isolated from a marine bryozoan, have antineoplastic activity in the P388 lymphocytic leukemia system. These compounds also stimulate growth in Swiss 3T3 cells, induce secretion in leukocytes, inhibit phorbol dibutyrate binding to a high affinity receptor, and activate the C-kinase in vitro. In human platelets, phorbol esters induce aggregation and activate protein kinase C, resulting in phosphorylation of a 47K protein and the 20K myosin light chain. The authors now show that bryostatin 7 (B-7) triggers platelet aggregation to the same rate and extent as phorbol 12-myristate 13-acetate (PMA). B-7 also causes the in vivo activation of the C-kinase, resulting in phosphorylation of both the 47K and the 20K proteins; the time courses and dose-responses of these B-7-induced phosphorylations were similar to those found with PMA. In addition, B-7 increases the level of /sup 32/P-incorporation into the platelet polyphosphoinositides, which also occurs in response to PMA. Bryostatin 3 (B-3), which has been shown to be much less potent than B-7 in mimicking other PMA effects, was much less effective than PMA or B-7 in inducing platelet aggregation and in stimulating /sup 32/P-incorporation into both proteins and the phosphoinositides. These results demonstrate that, intact human platelets, bryostatins mimic the phorbol esters tumor promoters and directly activate protein kinase C.

  2. Probing heterotrimeric G protein activation: applications to biased ligands.

    PubMed

    Denis, Colette; Saulière, Aude; Galandrin, Segolene; Sénard, Jean-Michel; Galés, Céline

    2012-01-01

    Cell surface G protein-coupled receptors (GPCRs) drive numerous signaling pathways involved in the regulation of a broad range of physiologic processes. Today, they represent the largest target for modern drugs development with potential application in all clinical fields. Recently, the concept of "ligand-directed trafficking" has led to a conceptual revolution in pharmacological theory, thus opening new avenues for drug discovery. Accordingly, GPCRs do not function as simple on-off switch but rather as filters capable of selecting the activation of specific signals and thus generating texture responses to ligands, a phenomenon often referred to as ligand-biased signaling. Also, one challenging task today remains optimization of pharmacological assays with increased sensitivity so to better appreciate the inherent texture of ligands. However, considering that a single receptor has pleiotropic signaling properties and that each signal can crosstalk at different levels, biased activity remains thus difficult to evaluate. One strategy to overcome these limitations would be examining the initial steps following receptor activation. Even, if some G protein independent functions have been recently described, heterotrimeric G protein activation remains a general hallmark for all GPCRs families and the first cellular event subsequent to agonist binding to the receptor. Herein, we review the different methodologies classically used or recently developed to monitor G protein activation and discussed them in the context of G protein biased-ligands. PMID:22229559

  3. NALP3 inflammasome activation in protein misfolding diseases.

    PubMed

    Shi, Fushan; Kouadir, Mohammed; Yang, Yang

    2015-08-15

    Protein-misfolding diseases, such as Alzheimer's disease, type 2 diabetes, Prion diseases, and Parkinson's disease, are characterized by inflammatory reactions. In all these diseases, IL-1β (Interlukine-1β) has been shown to be an important regulator, and the misfolded proteins are proved to be triggers of the release of IL-1β. Recently, several reports demonstrated that the inflammasome activation is involved in the progress of the misfolded protein diseases, and that the inflammasome can recognize pathogenic proteins leading to the release of IL-1β. In this review, we discuss the role of inflammasome in the pathogenesis of misfolded protein diseases and the potential of inflammasome-targeting therapeutic interventions in the management of these diseases. PMID:26037399

  4. Pudendal thigh flap for repair of rectovaginal fistula.

    PubMed

    Sathappan, S; Rica, M A I

    2006-08-01

    The pudendal thigh flap or the Singapore flap is a versatile flap that can be used in the repair of recto-vaginal fistulae. Apart from the potential problem of hair growth, this neurovascular flap proves to be surprisingly simple in technique, robust and has a high potential for normal or near-normal function. PMID:17240589

  5. Peptides and proteins with antimicrobial activity

    PubMed Central

    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. PMID:21264153

  6. Early wound healing of laser in situ keratomileusis–like flaps after treatment with human corneal stromal stem cells

    PubMed Central

    Morgan, Siân R.; Dooley, Erin P.; Kamma-Lorger, Christina; Funderburgh, James L.; Funderburgh, Martha L.; Meek, Keith M.

    2016-01-01

    Purpose To use a well-established organ culture model to investigate the effects of corneal stromal stem cells on the optical and biomechanical properties of corneal wounds after laser in situ keratomileusis (LASIK)–like flap creation. Setting School of Optometry and Vision Sciences, Cardiff University, Cardiff, Wales, United Kingdom. Design Experimental study. Methods The LASIK-like flaps were produced in sheep corneas. The flap beds were treated with corneal stromal stem cells and were then replaced and allowed to heal for different periods of up to 3 weeks in organ culture. The optical transmission of the cornea, the force required to detach the flap, and the presence of myofibroblasts near the flap bed were measured. Results Corneal stromal stem cell–treated flap beds were statistically significantly more transparent after 3 weeks in culture than the untreated controls. At 3 weeks, the mean force necessary to detach the flap was more than twice the force required for the respective control samples. Concurrently, there were 44% activated cells immediately below the flap margin of the controls compared with 29% in the same region of the corneal stromal stem cell–treated flaps. Conclusions In this system, the presence of corneal stromal stem cells at the wound margin significantly increased the adherence of LASIK-like flaps while maintaining corneal transparency. It is postulated that this is achieved by the deposition of extracellular connective tissue similar to that found in the normal cornea and by the paucity of activated keratocytes (myofibroblasts), which are known to scatter a significant amount of the incident light. Financial Disclosure No author has a financial or proprietary interest in any material or method mentioned. PMID:27026456

  7. Enhancer-specific modulation of E protein activity.

    PubMed

    Markus, Maurice; Du, Zhimei; Benezra, Robert

    2002-02-22

    Homodimeric complexes of members of the E protein family of basic helix-loop-helix (bHLH) transcription factors are important for tissue-specific activation of genes in B lymphocytes (Bain, G., Gruenwald, S., and Murre, C. (1993) Mol. Cell Biol. 13, 3522-3529; Shen, C. P., and Kadesch, T. (1995) Mol. Cell Biol. 15, 4518-4524; Jacobs, Y., et al. (1994) Mol. Cell Biol. 14, 4087-4096; Wilson, R. B., et al. (1991) Mol. Cell Biol. 11, 6185-6191). These homodimers, however, have little activity on myogenic enhancers (Weintraub, H., Genetta, T., and Kadesch, T. (1994) Genes Dev. 8, 2203-2211). We report here the identification of a novel cis-acting transcriptional repression domain in the E protein family of bHLH transcription factors. This domain, the Rep domain, is present in each of the known vertebrate E proteins. Extensive mapping analysis demonstrates that this domain is an acidic region of 30 amino acids with a predicted loop structure. Fusion studies indicate that the Rep domain can repress both of the E protein transactivation domains (AD1 and AD2). Physiologically, the Rep domain plays a key role in maintaining E protein homodimers in an inactive state on myogenic enhancers. In addition, we demonstrate that Rep domain mediated repression of AD1 is a necessary for the function of MyoD-E protein heterodimeric complexes. These studies demonstrate that the Rep domain is important for modulating the transcriptional activity of E proteins and provide key insights into both the selectivity and mechanism of action of E protein containing bHLH protein complexes. PMID:11724804

  8. A Novel Native Derived Coronary Artery Tissue-Flap Model

    PubMed Central

    Kranz, Alexander; Hülsmann, Jörn; Pinto, Antonio; Barth, Mareike; Fomin, Andrey; Lichtenberg, Artur; Akhyari, Payam

    2013-01-01

    Although tissue-engineering approaches have led to significant progress in the quest of finding a viable substitute for dysfunctional myocardium, the vascularization of such bioartificial constructs still remains a major challenge. Hence, there is a need for model systems that allow us to study and better understand cardiac and vascular biology to overcome current limitations. Therefore, in this study, in toto decellularized rat hearts with a patent vessel system were processed into standardized coronary artery tissue flaps adherent to the ascending aorta. Protein diffusivity analysis and blood perfusion of the coronary arteries showed proper sealing of the de-endothelialized vessels. Retrograde aortic perfusion allowed for selective seeding of the coronary artery system, while surface seeding of the tissue flaps allowed for additional controlled coculture with cardiac cells. The coronary artery tissue-flap model offers a patent and perfusable coronary vascular architecture with a preserved cardiac extracellular matrix, therefore mimicking nature's input to the highest possible degree. This offers the possibility to study re-endothelialization and endothelial function of different donor cell types and their interaction with cardiac cells in a standardized biologically derived cardiac in vitro model, while establishing a platform that could be used for in vitro drug testing and stem cell differentiation studies. PMID:23631507

  9. Chemical labelling of active serum thioester proteins for quantification.

    PubMed

    Holm, Lotta; Ackland, Gareth L; Edwards, Mark R; Breckenridge, Ross A; Sim, Robert B; Offer, John

    2012-02-01

    The complement serum proteins C3 and C4 and the protease inhibitor α-2 macroglobulin are all members of the C3/α-2M thioester protein family, an evolutionarily ancient and conserved family that contains an intrachain thioester bond. The chemistry of the thioester bond is a key to the function of the thioester proteins. All these proteins function by covalently linking to their target by acyl transfer of the protein via the thioester moiety. We show that the signature thioester bond can be targeted with nucleophiles linked to a bioreporter molecule, site-specifically modifying the whole, intact thioester protein. Conditions were optimised to label selectively and efficiently pull-down unprocessed thioester-containing proteins from serum. We demonstrated pull-down of full-length C3, α-2M and C4 from sera in high salt, using a biotinylated nucleophile and streptavidin-coated resin, confirmed by MALDI-TOF MS identification of the gel bands. The potential for the development of a quantitative method for measuring active C3 in serum was investigated in patient sera pre and post operation. Quantifying active C3 in clinical assays using current methods is difficult. Methods based on antibody detection (e.g. nephelometry) do not distinguish between active C3 and inactive breakdown products. C3-specific haemolytic assays can be used, but these require use of relatively unstable reagents. The current work represents a promising robust, enzyme- and antibody-free chemical method for detecting active thioester proteins in blood, plasma or serum. PMID:21852021

  10. 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

  11. Hydrodynamic schooling of flapping swimmers.

    PubMed

    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

  12. Hydrodynamic schooling of flapping swimmers

    DOE PAGESBeta

    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

  13. 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.

  14. 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

  15. 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.

  16. Apoptosis in extracorporeal preserved inguinal fat flaps of the rat.

    PubMed

    Herold, Christian; Rennekampff, Hans O; Ohm, Lea; Strauss, Sarah; Linkner, Jörn; Reimers, Kerstin; Allmeling, Christina; Vaske, Bernhard; Vogt, Peter M

    2012-04-01

    Fat cells are fragile cells with a short life span outside the body. Ways to reduce cell death in a biochemical way are almost unknown due to scarce information on the type of cellular death that is induced in fat tissue. This study was designed to investigate the apoptotic pathways of fat tissue in a permanent perfusion bioreactor system with the Hannover preservation solution and the Eurocollins solution in fat flaps of rats. In Lewis rats, the inguinal adipofascial flaps were elevated bilaterally and placed in a bioreactor at 37°C. To detect caspases 3, 8, 9 and 12, immunofluorescence stains of fat tissue specimen were analysed at several time points after preservation of flaps were placed in Hannover solution and Eurocollins solution for 10 days. An additional visual assessment of viability by a calcein based life/dead test was performed. It revealed a superior viability of the adipose tissue preserved in Hannover solution. Immunofluorescence staining demonstrated that apoptotic pathways via mitochondria, endoplasmatic reticulum and death receptors were activated, as Caspases 8, 9 and 12 were detected. Caspase 3 as an effector in the common apoptotic pathway was detected as well. Adipose tissue preserved at 37°C ex vivo in a bioreactor system undergoes apoptosis. Immunofluorescence examination of the fat tissue preserved ex vivo revealed that apoptotic pathways via mitochondria, endoplasmatic reticulum and death receptors are being activated. Significantly less activation of Caspase 3, 8, 9 and 12 in flaps preserved in Hannover solution in comparison to Eurocollins was found, supporting the anti apoptotic characteristics of Hannover solution. Based on these findings, further research to modify the apoptotic pathways to ameliorate viability of fat tissue can be performed. PMID:22223358

  17. 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

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

    PubMed Central

    Gaestel, Matthias

    2016-01-01

    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

  19. 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

  20. 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

  1. Proteins of the ETS family with transcriptional repressor activity.

    PubMed

    Mavrothalassitis, G; Ghysdael, J

    2000-12-18

    ETS proteins form one of the largest families of signal-dependent transcriptional regulators, mediating cellular proliferation, differentiation and tumorigenesis. Most of the known ETS proteins have been shown to activate transcription. However, four ETS proteins (YAN, ERF, NET and TEL) can act as transcriptional repressors. In three cases (ERF, NET and TEL) distinct repression domains have been identified and there are indications that NET and TEL may mediate transcription via Histone Deacetylase recruitment. All four proteins appear to be regulated by MAPKs, though for YAN and ERF this regulation seems to be restricted to ERKs. YAN, ERF and TEL have been implicated in cellular proliferation although there are indications suggesting a possible involvement of YAN and TEL in differentiation as well. Other ETS-domain proteins have been shown to repress transcription in a context specific manner, and there are suggestions that the ETS DNA-binding domain may act as a transcriptional repressor. Transcriptional repression by ETS domain proteins adds an other level in the orchestrated regulation by this diverse family of transcription factors that often recognize similar if not identical binding sites on DNA and are believed to regulate critical genes in a variety of biological processes. Definitive assessment of the importance of this novel regulatory level will require the identification of ETS proteins target genes and the further analysis of transcriptional control and biological function of these proteins in defined pathways. PMID:11175368

  2. Signal peptides are allosteric activators of the protein translocase.

    PubMed

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

    2009-11-19

    Extra-cytoplasmic polypeptides are usually synthesized as 'preproteins' carrying amino-terminal, cleavable signal peptides and secreted across membranes by translocases. The main bacterial translocase comprises the SecYEG protein-conducting channel and the peripheral ATPase motor SecA. Most proteins destined for the periplasm and beyond are exported post-translationally by SecA. Preprotein targeting to SecA is thought to involve signal peptides and chaperones like SecB. Here we show that signal peptides have a new role beyond targeting: they are essential allosteric activators of the translocase. On 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; second, 'trapping' that engages non-native preprotein mature domains docked with high affinity on the secretion apparatus; and third, 'secretion' during which trapped mature domains undergo several turnovers of translocation in segments. 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

  3. Numerical Analysis of the EXPERT Open Flap Assembly in Plasma Wind Tunnel Conditions

    NASA Astrophysics Data System (ADS)

    Di Clemente, M.; Marini, M.; Di Benedetto, S.

    2009-01-01

    In the frame of the EXPERT program a number of tests to be performed in the CIRA Plasma Wind Tunnel (PWT) "Scirocco" is scheduled, [1]. Among these, one is dedicated to the Open Flap Assembly (OFA) with the aim to address some technical issues related to scientific objectives of Payload 6 (under DLR responsibility, [2]), related with the analysis of flow reattachment on the flap due to the shock wave boundary layer interaction, and Payload 8 (under RUAG responsibility, [3]) whose main objective is the measurement of the heat flux in the rear surface of the flap through an infrared camera mounted in the cavity, and to consolidate the ground-to- flight extrapolation relations for the characterization of the convective heating on the flap and the reattachment and radiative heating in the cavity. In the present work, the numerical activities carried out before the test for the definition of the experimental conditions, starting from the requirements to be reproduced on the model, will be shown. Once the preliminary model configuration, its position inside the test chamber and the preliminary PWT operating conditions have been defined, a series of facility regulations (reservoir pressure P0 and reservoir enthalpy H0) has been simulated in terms of nozzle axi-symmetric flow, flow around the PWT hemi-spherical calibration probe and two-dimensional flow around the flap model. These simulations have allowed the assessment of the typical flow features and the analysis of some effects as those related to the flow angularity at nozzle exit, the wall temperature on the flap and the gap/step in front of the flap itself, as well as an estimation of radiative heating between the flap rear surface and the cavity.

  4. 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.

  5. G Protein Activation Stimulates Phospholipase D Signaling in Plants.

    PubMed Central

    Munnik, T.; Arisz, S. A.; De Vrije, T.; Musgrave, A.

    1995-01-01

    We provide direct evidence for phospholipase D (PLD) signaling in plants by showing that this enzyme is stimulated by the G protein activators mastoparan, ethanol, and cholera toxin. An in vivo assay for PLD activity in plant cells was developed based on the use of a "reporter alcohol" rather than water as a transphosphatidylation substrate. The product was a phosphatidyl alcohol, which, in contrast to the normal product phosphatidic acid, is a specific measure of PLD activity. When 32P-labeled cells were treated with 0.1% n-butanol, 32P-phosphatidyl butanol (32P-PtdBut) was formed in a time-dependent manner. In cells treated with any of the three G protein activators, the production of 32P-PtdBut was increased in a dose-dependent manner. The G protein involved was pertussis toxin insensitive. Ethanol could activate PLD but was itself consumed by PLD as transphosphatidylation substrate. In contrast, secondary alcohols (e.g., sec-butyl alcohol) activated PLD but did not function as substrate, whereas tertiary alcohols did neither. Although most of the experiments were performed with the green alga Chlamydomonas eugametos, the relevance for higher plants was demonstrated by showing that PLD in carnation petals could also be activated by mastoparan. The results indicate that PLD activation must be considered as a potential signal transduction mechanism in plants, just as in animals. PMID:12242371

  6. A Theileria parva type 1 protein phosphatase activity.

    PubMed

    Cayla, X; Garcia, A; Baumgartner, M; Ozon, R; Langsley, G

    2000-09-01

    The protozoan parasite Theileria (spp. parva and annulata) infects bovine leukocytes and provokes a leukaemia-like disease in vivo. In this study, we have detected a type 1 serine/threonine phosphatase activity with phosphorylase a as a substrate, in protein extracts of parasites purified from infected B lymphocytes. In contrast to this type 1 activity, dose response experiments with okadaic acid (OA), a well characterised inhibitor of type 1 and 2A protein phosphatases, indicated that type 2A is the predominant activity detected in host B cells. Furthermore, consistent with polycation-specific activation of the type 2A phosphatase, protamine failed to activate the parasite-associated phosphorylase a phosphatase activity. Moreover, inhibition of phosphorylase a dephosphorylation by phospho-DARPP-32, a specific type 1 inhibitor, clearly demonstrated that a type 1 phosphatase is specifically associated with the parasite, while the type 2A is predominantly expressed in the host lymphocyte. Since an antibody against bovine catalytic protein phosphatase 1 (PP1) subunit only recognised the PP1 in B cells, but not in parasite extracts, we conclude that in parasites the PP1 activity is of parasitic origin. Intriguingly, since type 1 OA-sensitive phosphatase activity has been recently described in Plasmodium falciparum, we can conclude that these medically important parasites produce their one PP1. PMID:10989153

  7. Epidural Blood Patch Using Manometry for Sinking Skin Flap Syndrome.

    PubMed

    Turner, James D; Farmer, Justin L; Dobson, Sean W

    2016-06-01

    We describe here a 55-year-old male patient with a medical history significant for chronic back pain and substance abuse with cocaine who sustained a traumatic subarachnoid hemorrhage after a fall from a roof while acutely intoxicated on cocaine requiring decompressive hemicraniectomy and cranioplasty that was complicated by an epidural abscess requiring a repeat craniectomy. He was diagnosed with sinking skin flap syndrome consistent with altered mental status and a sunken skin flap with increased midline shift. Despite treatment with Trendelenburg positioning and appropriate fluid management, the patient continued to decline, and an epidural blood patch was requested for treatment. After placement of the epidural blood patch using manometry in the epidural space, the patient's neurologic status improved allowing him to ultimately receive a cranioplasty. The patient is now able to perform several of his activities of daily living and communicate effectively. PMID:27075425

  8. 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

  9. 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.

  10. Contractions Activate Hormone-Sensitive Lipase in Rat Muscle by Protein Kinase C and Mitogen-Activated Protein Kinase

    PubMed Central

    Donsmark, Morten; Langfort, Jozef; Holm, Cecilia; Ploug, Thorkil; Galbo, Henrik

    2003-01-01

    Intramuscular triacylglycerol is an important energy store and is also related to insulin resistance. The mobilization of fatty acids from this pool is probably regulated by hormone-sensitive lipase (HSL), which has recently been shown to exist in muscle and to be activated by both adrenaline and contractions. Adrenaline acts via cAMP-dependent protein kinase (PKA). The signalling mediating the effect of contractions is unknown and was explored in this study. Incubated soleus muscles from 70 g male rats were electrically stimulated to perform repeated tetanic contractions for 5 min. The contraction-induced activation of HSL was abolished by the protein kinase C (PKC) inhibitors bisindolylmaleimide I and calphostin C and reduced 50 % by the mitogen-activated protein kinase kinase (MEK) inhibitor U0126, which also completely blocked extracellular signal-regulated kinase (ERK) 1 and 2 phosphorylation. None of the inhibitors reduced adrenaline-induced HSL activation in soleus muscle. Both phorbol-12-myristate-13-acetate (PMA), which activates PKC and, in turn, ERK, and caffeine, which increases intracellular Ca2+ without eliciting contraction, increased HSL activity. Activated ERK increased HSL activity in supernatant from basal but not from electrically stimulated muscle. In conclusion, in muscle, PKC can stimulate HSL through ERK. Contractions and adrenaline enhance muscle HSL activity by different signalling mechanisms. The effect of contractions is mediated by PKC, at least partly via the ERK pathway. PMID:12794177

  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. 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.

  13. Platysma flap using dual skin paddles.

    PubMed

    Mustafa, El Muiz A; Bhatia, Sunil K; Hammond, Doug E

    2016-07-01

    The platysma myocutaneous flap is rarely reported as an option for reconstruction in the head and neck. In our experience it is a reliable flap that is easy to raise, has a limited risk of morbidity at the donor site, and allows for early recommencement of oral function. We report a patient with an unusually aggressive orocutaneous fistula that complicated the simple extraction of teeth. We modified the platysma flap to provide two independent skin paddles so we could reconstruct both the intraoral and cutaneous defects simultaneously with a good clinical outcome. PMID:26947107

  14. New drag laws for flapping flight

    NASA Astrophysics Data System (ADS)

    Agre, Natalie; Zhang, Jun; Ristroph, Leif

    2014-11-01

    Classical aerodynamic theory predicts that a steadily-moving wing experiences fluid forces proportional to the square of its speed. For bird and insect flight, however, there is currently no model for how drag is affected by flapping motions of the wings. By considering simple wings driven to oscillate while progressing through the air, we discover that flapping significantly changes the magnitude of drag and fundamentally alters its scaling with speed. These measurements motivate a new aerodynamic force law that could help to understand the free-flight dynamics, control, and stability of insects and flapping-wing robots.

  15. Evaluation of antioxidant activities of zein protein fractions.

    PubMed

    Tang, Ning; Zhuang, Hong

    2014-11-01

    Zein protein was extracted from the by-product corn gluten meal. The obtained zein protein was 1st hydrolyzed by 4 different proteases. The antioxidant activities of the hydrolysates or peptides were evaluated by free radical scavenging activity, metal ion chelating activity, and lipid peroxidation inhibitory capacity. Among hydrolysates produced, alkaline protease hydrolysates exhibited the highest antioxidant activity. A regression model was established by uniform design to optimize the alkaline protease hydrolysis conditions. The hydrolysates with molecular weight < 3 kDa obtained from ultrafiltration showed the highest antioxidant activities in all relevant assays. The hydrolysates with molecular weight <3 kDa were subsequently purified by gel filtration chromatography, and fraction F3 exhibited the highest antioxidant activities. Two peptides were identified from fraction F3 using LC-ESI-Q-TOF MS/MS as Pro-Phe (263.13 Da) and Leu-Pro-Phe (375.46 Da). These peptides exhibited good free radical scavenging activity and lipid peroxidation inhibitory effect. The results clearly indicated that zein protein fractions are good sources for the development of natural antioxidants for the food industry. PMID:25350353

  16. Antioxidant activity of whey protein hydrolysates in milk beverage system.

    PubMed

    Mann, Bimlesh; Kumari, Anuradha; Kumar, Rajesh; Sharma, Rajan; Prajapati, Kishore; Mahboob, Shaik; Athira, S

    2015-06-01

    The aim of the present study was to evaluate the antioxidant activity of flavoured milk enriched with antioxidative whey protein hydrolysates (WPHs) by radical scavenging method. Whey protein concentrate (WPC) was hydrolyzed by using three commercial proteases; flavouzyme, alcalase and corolase PP and these WPHs were analyzed for degree of hydrolysis and antioxidant activity. The antioxidant activities of these WPHs were evaluated using ABTS method. Trolox equivalent antioxidant activity of all the hydrolysates i.e. flavourzyme (0.81 ± 0.04), alcalase (1.16 ± 0.05) and corolase (1.42 ± 0.12) was higher than the WPC (0.19 ± 0.01). Among these, whey protein hydrolysates prepared using corolase showed maximum antioxidant activity. Total 15 β-lactoglobulin, 1 α-lactoalbumin, and 6 β-casein derived peptide fragments were identified in the WPHs by LC-MS/MS. Due to their size and characteristic amino acid composition, all the identified peptides may contribute for the antioxidant activity. The strawberry and chocolate flavoured milk was supplemented with WPC and WPHs and 2 % addition has shown increase in antioxidant activity upto 42 %. The result suggests that WPH could be used as natural biofunctional ingredients in enhancing antioxidant properties of food products. PMID:26028704

  17. Oxidation of methionine residues in proteins of activated human neutrophils.

    PubMed Central

    Fliss, H; Weissbach, H; Brot, N

    1983-01-01

    A simple assay for the detection of 35S-labeled methionine sulfoxide residues in proteins is described. The assay, which is based on the ability of CNBr to react with methionine but not with methionine sulfoxide, requires the prelabeling of cellular proteins with [35S]methionine. The assay was used to study the extent of methionine oxidation in newly synthesized proteins of both activated and quiescent human neutrophils. In cells undergoing a phorbol 12-myristate 13-acetate-induced respiratory burst, about 66% of all methionine residues in newly synthesized proteins were oxidized to the sulfoxide derivative, as compared with 9% in cells not treated with the phorbol ester. In contrast, quantitation of methionine sulfoxide content in the total cellular protein by means of amino acid analysis showed that only 22% of all methionine residues were oxidized in activated cells as compared with 9% in quiescent cells. It is proposed that methionine residues in nascent polypeptide chains are more susceptible to oxidation than those in completed proteins. PMID:6580633

  18. Blended Cutout Flap for Reduction of Jet-Flap Interaction Noise

    NASA Technical Reports Server (NTRS)

    Czech, Michael J (Inventor); Thomas, Russell H. (Inventor)

    2014-01-01

    An aircraft system includes a wing and a trailing edge device coupled to the wing. The trailing edge device is movable relative to the wing, and includes a leading edge and a trailing edge having a center flap portion and a plurality of outer edge portions integrally combined with the center flap portion such that the center flap portion is shorter in width than that of outer edge portions.

  19. DNA binding specificity and sequence of Xanthomonas campestris catabolite gene activator protein-like protein.

    PubMed Central

    Dong, Q; Ebright, R H

    1992-01-01

    The Xanthomonas campestris catabolite gene activator protein-like protein (CLP) can substitute for the Escherichia coli catabolite gene activator protein (CAP) in transcription activation at the lac promoter (V. de Crecy-Lagard, P. Glaser, P. Lejeune, O. Sismeiro, C. Barber, M. Daniels, and A. Danchin, J. Bacteriol. 172:5877-5883, 1990). We show that CLP has the same DNA binding specificity as CAP at positions 5, 6, and 7 of the DNA half site. In addition, we show that the amino acids at positions 1 and 2 of the recognition helix of CLP are identical to the amino acids at positions 1 and 2 of the recognition helix of CAP:i.e., Arg at position 1 and Glu at position 2. PMID:1322886

  20. Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis.

    PubMed

    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. PMID:27115346

  1. 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.

  2. Ubiquitously expressed transcript is a novel interacting protein of protein inhibitor of activated signal transducer and activator of transcription 2

    PubMed Central

    KONG, XIANG; MA, SHIKUN; GUO, JIAQIAN; MA, YAN; HU, YANQIU; WANG, JIANJUN; ZHENG, YING

    2015-01-01

    Protein inhibitor of activated signal transducer and activator of transcription 2 (PIAS2) is a member of the PIAS protein family. This protein family modulates the activity of several transcription factors and acts as an E3 ubiquitin ligase in the sumoylation pathway. To improve understanding of the physiological roles of PIAS2, the current study used a yeast two-hybrid system to screen mouse stem cell cDNA libraries for proteins that interact with PIAS2. The screening identified an interaction between PIAS2 and ubiquitously expressed transcript (UXT). UXT, also termed androgen receptor trapped clone-27, is an α-class prefoldin-type chaperone that acts as a coregulator for various transcription factors, including nuclear factor-κB and androgen receptor (AR). A direct interaction between PIAS2 and UXT was confirmed by direct yeast two-hybrid analysis. In vitro evidence of the association of UXT with PIAS2 was obtained by co-immunoprecipitation. Colocalization between PIAS2 and UXT was identified in the nucleus and cytoplasm of HEK 293T and human cervical carcinoma HeLa cells. The results of the current study suggested that UXT is a binding protein of PIAS2, and interaction between PIAS2 and UXT may be important for the transcriptional activation of AR. PMID:25434787

  3. 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).

  4. Activities at the Universal Protein Resource (UniProt)

    PubMed Central

    2014-01-01

    The mission of the Universal Protein Resource (UniProt) (http://www.uniprot.org) is to provide the scientific community with a comprehensive, high-quality and freely accessible resource of protein sequences and functional annotation. It integrates, interprets and standardizes data from literature and numerous resources to achieve the most comprehensive catalog possible of protein information. The central activities are the biocuration of the UniProt Knowledgebase and the dissemination of these data through our Web site and web services. UniProt is produced by the UniProt Consortium, which consists of groups from the European Bioinformatics Institute (EBI), the SIB Swiss Institute of Bioinformatics (SIB) and the Protein Information Resource (PIR). UniProt is updated and distributed every 4 weeks and can be accessed online for searches or downloads. PMID:24253303

  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. 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. PMID:26405422

  7. Locoregional use of lateral thoracic artery perforator flap as a propeller flap.

    PubMed

    Baghaki, Semih; Cevirme, Mirza; Diyarbakirli, Murat; Tatar, Cihad; Aydin, Yagmur

    2015-05-01

    Although thoracodorsal system is a fundamental source of various flaps, lateral thoracic region has not been a popular flap donor area. There is limited data on the use of lateral intercostal artery perforator flap and lateral thoracic artery perforator flap. In this case series, lateral thoracic artery perforator flap has been used in locoregional (axilla, pectoral region, and arm) reconstruction as an island or propeller flap.Eighteen patients have been operated on between September 2010 and January 2013. The age of the patients ranged between 16 and 68 years with a median of 38 years. A thorough chart review has been performed with preoperative and postoperative photographs. Duration of hospitalization, complication rate and long term results have been documented.Nine patients had severe burn contracture of axilla, 7 patients had axillary hidradenitis suppurativa, 1 patient had giant neurofibroma of arm, and 1 patient had malignant peripheral nerve sheath tumor of pectoral area. Seventeen flaps survived totally, and in 1 flap, distal superficial slough of skin has been observed. No recurrence in hidradenitis or peripheral nerve sheath tumor has been observed. Donor site scar is well hidden in anatomical position. The range of motion of affected extremities returned to normal after reconstruction.Lateral thoracic area provides a reliable flap option with a wide arc of rotation when lateral thoracic artery perforators are used. PMID:25875722

  8. ["Separation delay" on random flap: an experimental and clinical practice on delayed flap].

    PubMed

    Zhao, S Q

    1990-08-01

    A new technique of delayed flap, that is called "Separation delay" by the authors, has succeeded in animal experiment and clinical practice. In the years of 1985-1989, 11 cases of random skin flaps on the patients had been performed with the new method. All the flaps look like table tennis rackets. It's pedicle is very narrow, simultaneously, the flap itself is very large. So it is quite suitable for repairing a neighbouring wound surface. It can be rotated 180 degrees. "The Separation delay" is a handy way without microsurgical technique. It is also an useful and reliable technique for resurfacing wound on plastic and reconstructive surgery. PMID:2086104

  9. 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

  10. 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

  11. Streptococcal Surface Proteins Activate the Contact System and Control Its Antibacterial Activity*

    PubMed Central

    Wollein Waldetoft, Kristofer; Svensson, Lisbeth; Mörgelin, Matthias; Olin, Anders I.; Nitsche-Schmitz, D. Patric; Björck, Lars; Frick, Inga-Maria

    2012-01-01

    Group G streptococci (GGS) are important bacterial pathogens in humans. Here, we investigated the interactions between GGS and the contact system, a procoagulant and proinflammatory proteolytic cascade that, upon activation, also generates antibacterial peptides. Two surface proteins of GGS, protein FOG and protein G (PG), were found to bind contact system proteins. Experiments utilizing contact protein-deficient human plasma and isogenic GGS mutant strains lacking FOG or PG showed that FOG and PG both activate the procoagulant branch of the contact system. In contrast, only FOG induced cleavage of high molecular weight kininogen, generating the proinflammatory bradykinin peptide and additional high molecular weight kininogen fragments containing the antimicrobial peptide NAT-26. On the other hand, PG protected the bacteria against the antibacterial effect of NAT-26. These findings underline the significance of the contact system in innate immunity and demonstrate that GGS have evolved surface proteins to exploit and modulate its effects. PMID:22648411

  12. Gi/o proteins: expression for direct activation enquiry.

    PubMed

    Di Cesare Mannelli, Lorenzo; Pacini, Alessandra; Toscano, Annarita; Fortini, Martina; Berti, Debora; Ghelardini, Carla; Galeotti, Nicoletta; Baglioni, Piero; Bartolini, Alessandro

    2006-05-01

    G protein-mediated pathways are fundamental mechanisms of cell signaling. In this paper, the expression and the characterization of the alphai1, alphai3, alphao1, beta1, and gamma2 subunits of the human G protein are described. This approach was developed to evaluate the G protein activation profile of new compounds. pCR-TOPO T7 vectors, engineered to contain the target sequences, were used to transform Escherichia coli competent cells. Subunits were over-expressed in a preparative scale as fusion proteins with a six-histidine tag, and subsequently purified by metal chelate chromatography. Afterward, the His-tag was removed by enterokinase digestion, and the secondary structures of the recombinant subunits were analyzed by circular dichroism. To assess the functionality of the subunits, the rate of GTP hydrolysis and GTPgammaS binding were evaluated both in the absence and in the presence of two modulators: the peptidic activator Mastoparan and the non-peptidic activator N-dodecyl-lysinamide (ML250). Tests were conducted on isolated alpha-subunit and on heterotrimeric alphabetagamma complex, alone or reconstituted in phospholipidic vesicles. Our results show that recombinant subunits are stable, properly folded and, fully active, which makes them suitable candidates for functional studies. PMID:16364655

  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. 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

  15. 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

  16. Estrogen receptor α inhibitor activates the unfolded protein response, blocks protein synthesis, and induces tumor regression.

    PubMed

    Andruska, Neal D; Zheng, Xiaobin; Yang, Xujuan; Mao, Chengjian; Cherian, Mathew M; Mahapatra, Lily; Helferich, William G; Shapiro, David J

    2015-04-14

    Recurrent estrogen receptor α (ERα)-positive breast and ovarian cancers are often therapy resistant. Using screening and functional validation, we identified BHPI, a potent noncompetitive small molecule ERα biomodulator that selectively blocks proliferation of drug-resistant ERα-positive breast and ovarian cancer cells. In a mouse xenograft model of breast cancer, BHPI induced rapid and substantial tumor regression. Whereas BHPI potently inhibits nuclear estrogen-ERα-regulated gene expression, BHPI is effective because it elicits sustained ERα-dependent activation of the endoplasmic reticulum (EnR) stress sensor, the unfolded protein response (UPR), and persistent inhibition of protein synthesis. BHPI distorts a newly described action of estrogen-ERα: mild and transient UPR activation. In contrast, BHPI elicits massive and sustained UPR activation, converting the UPR from protective to toxic. In ERα(+) cancer cells, BHPI rapidly hyperactivates plasma membrane PLCγ, generating inositol 1,4,5-triphosphate (IP3), which opens EnR IP3R calcium channels, rapidly depleting EnR Ca(2+) stores. This leads to activation of all three arms of the UPR. Activation of the PERK arm stimulates phosphorylation of eukaryotic initiation factor 2α (eIF2α), resulting in rapid inhibition of protein synthesis. The cell attempts to restore EnR Ca(2+) levels, but the open EnR IP3R calcium channel leads to an ATP-depleting futile cycle, resulting in activation of the energy sensor AMP-activated protein kinase and phosphorylation of eukaryotic elongation factor 2 (eEF2). eEF2 phosphorylation inhibits protein synthesis at a second site. BHPI's novel mode of action, high potency, and effectiveness in therapy-resistant tumor cells make it an exceptional candidate for further mechanistic and therapeutic exploration. PMID:25825714

  17. 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. PMID:2514798

  18. 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

  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. In vitro antithrombotic activities of peanut protein hydrolysates.

    PubMed

    Zhang, Shao Bing

    2016-07-01

    The antithrombotic activities of peanut protein hydrolysates were investigated using a microplates assay. When peanut proteins were hydrolyzed to a limited extent by various enzymes, their thrombin inhibitory abilities were significantly enhanced. However, the resultant hydrolysates showed significantly different activities even at the same degrees of hydrolysis. The hydrolysates generated by Alcalase 2.4L displayed the best antithrombotic activities and the hydrolysis process was further optimized by response surface methodology. The antithrombotic activities were increased to 86% based on a protein concentration of 50mg/ml under the optimal conditions: pH 8.5, enzyme concentration of 5000IU/g of peanut proteins, and 2h hydrolysis time at 50°C. The Alcalase 2.4L crude hydrolysates were then fractionated successively by preparative and semi-preparative reverse-phase high-performance liquid chromatography (RP-HPLC). The peptide fraction collected inhibited thrombin-catalyzed coagulation of fibrinogen completely at a concentration of 0.4mg/ml, with an antithrombotic activity close to that of heparin at quite a low concentration (0.2mg/ml). This peptide fraction was further analyzed by online reverse-phase ultra-performance liquid chromatography (RP-UPLC) coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and three new peptides were identified as Ser-Trp-Ala-Gln-Leu, Gly-Asn-His-Glu-Ala-Gly-Glu and Cys-Phe-Asn-Glu-Tyr-Glu, respectively. This research provided an effective way to produce antithrombotic peptides from peanut proteins, and also helped to elucidate the structure-function relationships of peanut peptides. PMID:26920259

  1. P-47 Thunderbolt with dive recovery flaps

    NASA Technical Reports Server (NTRS)

    1946-01-01

    Caption: 'The dive recovery flaps on this P-47 Thunderbolt are barely visible underneath the wings. Photograph and caption published in Winds of Change, 75th Anniversary NASA publication (pages 52-53 and 130), by James Schultz.

  2. Skin flaps and grafts - self-care

    MedlinePlus

    ... skin infection Surgery for skin cancer Venous ulcers , pressure ulcers , or diabetic ulcers that do not heal After mastectomy or amputation Donor sites for grafts and flaps are chosen ... surgery than the wound due to newly exposed nerve endings.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. Design of a Split Intein with Exceptional Protein Splicing Activity.

    PubMed

    Stevens, Adam J; Brown, Zachary Z; Shah, Neel H; Sekar, Giridhar; Cowburn, David; Muir, Tom W

    2016-02-24

    Protein trans-splicing (PTS) by split inteins has found widespread use in chemical biology and biotechnology. Herein, we describe the use of a consensus design approach to engineer a split intein with enhanced stability and activity that make it more robust than any known PTS system. Using batch mutagenesis, we first conduct a detailed analysis of the difference in splicing rates between the Npu (fast) and Ssp (slow) split inteins of the DnaE family and find that most impactful residues lie on the second shell of the protein, directly adjacent to the active site. These residues are then used to generate an alignment of 73 naturally occurring DnaE inteins that are predicted to be fast. The consensus sequence from this alignment (Cfa) demonstrates both rapid protein splicing and unprecedented thermal and chaotropic stability. Moreover, when fused to various proteins including antibody heavy chains, the N-terminal fragment of Cfa exhibits increased expression levels relative to other N-intein fusions. The durability and efficiency of Cfa should improve current intein based technologies and may provide a platform for the development of new protein chemistry techniques. PMID:26854538

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

    PubMed

    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

  9. 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

  10. Protein kinase domain of twitchin has protein kinase activity and an autoinhibitory region.

    PubMed

    Lei, J; Tang, X; Chambers, T C; Pohl, J; Benian, G M

    1994-08-19

    Twitchin is a 753-kDa polypeptide located in the muscle A-bands of the nematode, Caenorhabditis elegans. It consists of multiple copies of both fibronectin III and immunoglobulin C2 domains and, near the C terminus, a protein kinase domain with greatest homology to the catalytic domains of myosin light chain kinases. We have expressed and purified from Escherichia coli twitchin's protein kinase catalytic core and flanking sequences that do not include fibronectin III and immunoglobulin C2 domains. The protein was shown to phosphorylate a model substrate and to undergo autophosphorylation. The autophosphorylation occurs at a slow rate, attaining a maximum at 3 h with a stoichiometry of about 1.0 mol of phosphate/mol of protein, probably through an intramolecular mechanism. Sequence analysis of proteolytically derived phosphopeptides revealed that autophosphorylation occurred N-terminal to the catalytic core, predominantly at Thr-5910, with possible minor sites at Ser5912 and/or Ser-5913. This portion of twitchin (residues 5890-6268) was also phosphorylated in vitro by protein kinase C in the absence of calcium and phosphotidylserine, but not by cAMP-dependent protein kinase. By comparing the activities of three twitchin segments, the enzyme appears to be inhibited by the 60-amino acid residues lying just C-terminal to the kinase catalytic core. Thus, like a number of other protein kinases including myosin light chain kinases, the twitchin kinase appears to be autoregulated. PMID:8063727

  11. NRIP, a novel calmodulin binding protein, activates calcineurin to dephosphorylate human papillomavirus E2 protein.

    PubMed

    Chang, Szu-Wei; Tsao, Yeou-Ping; Lin, Chia-Yi; Chen, Show-Li

    2011-07-01

    Previously, we found a gene named nuclear receptor interaction protein (NRIP) (or DCAF6 or IQWD1). We demonstrate that NRIP is a novel binding protein for human papillomavirus 16 (HPV-16) E2 protein. HPV-16 E2 and NRIP can directly associate into a complex in vivo and in vitro, and the N-terminal domain of NRIP interacts with the transactivation domain of HPV-16 E2. Only full-length NRIP can stabilize E2 protein and induce HPV gene expression, and NRIP silenced by two designed small interfering RNAs (siRNAs) decreases E2 protein levels and E2-driven gene expression. We found that NRIP can directly bind with calmodulin in the presence of calcium through its IQ domain, resulting in decreased E2 ubiquitination and increased E2 protein stability. Complex formation between NRIP and calcium/calmodulin activates the phosphatase calcineurin to dephosphorylate E2 and increase E2 protein stability. We present evidences for E2 phosphorylation in vivo and show that NRIP acts as a scaffold to recruit E2 and calcium/calmodulin to prevent polyubiquitination and degradation of E2, enhancing E2 stability and E2-driven gene expression. PMID:21543494

  12. 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

  13. Bilateral pedicled gracilis flap for scrotal reconstruction.

    PubMed

    Daigeler, Adrien; Behr, Björn; Mikhail, Bassem Daniel; Lehnhardt, Marcus; Wallner, Christoph

    2016-09-01

    Trauma, infection and cancer can cause severe scrotal defects. The demand to a definitive reconstruction in terms of aesthetics and function is high. Primary closure, skin grafts and local fasciocutaneous flaps do not meet these high demands in most cases. The authors treated a series of patients requiring scrotal reconstruction with bilateral pedicled gracilis muscle flaps and split thickness skin grafts, resulting in satisfying aesthetic and functional outcomes. PMID:27318782

  14. Dynamic responses of a two-dimensional flapping foil motion

    NASA Astrophysics Data System (ADS)

    Lu, Xi-Yun; Liao, Qin

    2006-09-01

    The investigation of a flapping foil, which is used as a basic mode of the flapping-based locomotion in insects, birds, and fish, is performed by solving the Navier-Stokes equations numerically. In this Brief Communication we provide insight into the understanding of dynamics of a flapping foil. A critical flapping Reynolds number based on the flapping frequency and amplitude, above which a forward flapping movement occurs, is predicted. The dynamics of the flapping foil are analyzed in two dynamic responses, i.e., an oscillatory movement and a steady movement, which depend on the density ratio between the foil and the surrounded fluid. The steady movement response is related to the forward flapping motion. The Strouhal number that governs a vortex shedding for the forward flapping foil is calculated and lies in the range where flying and swimming animals will be likely to tune for high propulsive efficiency.

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

    PubMed

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

    2015-06-12

    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

  16. Mitogen Activated Protein kinase signal transduction pathways in the prostate

    PubMed Central

    Maroni, Paul D; Koul, Sweaty; Meacham, Randall B; Koul, Hari K

    2004-01-01

    The biochemistry of the mitogen activated protein kinases ERK, JNK, and p38 have been studied in prostate physiology in an attempt to elucidate novel mechanisms and pathways for the treatment of prostatic disease. We reviewed articles examining mitogen-activated protein kinases using prostate tissue or cell lines. As with other tissue types, these signaling modules are links/transmitters for important pathways in prostate cells that can result in cellular survival or apoptosis. While the activation of the ERK pathway appears to primarily result in survival, the roles of JNK and p38 are less clear. Manipulation of these pathways could have important implications for the treatment of prostate cancer and benign prostatic hypertrophy. PMID:15219238

  17. 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

  18. The Effect of Epigallocatechin Gallate on Flap Viability of Rat Perforator Abdominal Flaps.

    PubMed

    Aksakal, İbrahim Alper; Küçüker, İsmail; Önger, Mehmet Emin; Engin, Murat Sinan; Keleş, Musa Kemal; Demir, Ahmet

    2016-05-01

    Background Epigallocatechin gallate (EGCG) is a substance abundant in green tea. In this study, the effects of EGCG on perforator flap viability were investigated. Methods A total of 40 rats were assigned to four groups of 10 each. In each subject, a 4 × 6 cm abdominal skin flap was raised and adapted back onto its place. In the control group, no further procedures were taken. In the flap group, 40 mg/kg/d EGCG was injected into the flap. In the gavage group, 100 mg/kg/d EGCG was given through a feeding tube. In the intraperitoneal group, 50 mg/kg/d EGCG was injected intraperitoneally. On the 7th postoperative day, flaps were photographed and the viable areas were measured and compared via a one-way analysis of variance. Results The ratios of viable and contracted flap area were 9.15/12.01, 4.59/16.46, 11.56/11.20, and 11.65/10.77 cm(2) for the control, flap group, gavage group, and intraperitoneal group, respectively. While the flap group yielded the worst results in the sense of flap contraction and viability (p < 0.001), the gavage and intraperitoneal groups were significantly better than those of the control group (p = 0.03). Histologically, epidermal, papillary dermal, and capillary tissue volumes were evaluated. In comparison to the control group, the flap group yielded significantly increased epidermal and dermal volumes (p = 0.03), however, these values were significantly decreased (p = 0.04) in the gavage and intraperitoneal groups. Capillary volumes were significantly decreased in EGCG treatment groups (p < 0.01). Conclusion Our experiment has shown that oral and intraperitoneal administration of EGCG increases the perforator flap viability when compared with controls, while direct injection decreases the viability. PMID:26919381

  19. A GTPase-activating protein for the G protein Galphaz. Identification, purification, and mechanism of action.

    PubMed

    Wang, J; Tu, Y; Woodson, J; Song, X; Ross, E M

    1997-02-28

    A GTPase-activating protein (GAP) specific for Galphaz was identified in brain, spleen, retina, platelet, C6 glioma cells, and several other tissues and cells. Gz GAP from bovine brain is a membrane protein that is refractory to solubilization with most detergents but was solubilized with warm Triton X-100 and purified up to 50,000-fold. Activity is associated with at least two separate proteins of Mr approximately 22,000 and 28,000, both of which have similar specific activities. In an assay that measures the rate of hydrolysis of GTP pre-bound to detergent-soluble Galphaz, the GAP accelerates hydrolysis over 200-fold, from 0.014 to 3 min -1 at 15 degrees C, or to >/=20 min-1 at 30 degrees C. It does not alter rates of nucleotide association or dissociation. When co-reconstituted into phospholipid vesicles with trimeric Gz and m2 muscarinic receptor, Gz GAP accelerates agonist-stimulated steady-state GTP hydrolysis as predicted by its effect on the hydrolytic reaction. In the single turnover assay, the Km of the GAP for Galphaz-GTP is 2 nM. Its activity is inhibited by Galphaz-guanosine 5'-O-thiotriphosphate (Galphaz-GTPgammaS) or by Galphaz-GDP/AlF4 with Ki approximately 1.5 nM for both species; Galphaz-GDP does not inhibit. G protein betagamma subunits inhibit Gz GAP activity, apparently by forming a GTP-Galphazbetagamma complex that is a poor GAP substrate. Gz GAP displays little GAP activity toward Galphai1 or Galphao, but its activity with Galphaz is competitively inhibited by both Galphai1 and Galphao at nanomolar concentrations when they are bound to GTPgammaS but not to GDP. Neither phospholipase C-beta1 (a Gq GAP) nor several adenylyl cyclase isoforms display Gz GAP activity. PMID:9038185

  20. 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. PMID:25865799

  1. Preliminary Design and Evaluation of an Airfoil with Continuous Trailing-Edge Flap

    NASA Technical Reports Server (NTRS)

    Shen, Jinwei; Thornburgh, Robert P.; Kreshock, Andrew R.; Wilbur, Matthew L.; Liu, Yi

    2012-01-01

    This paper presents the preliminary design and evaluation of an airfoil with active continuous trailing-edge flap (CTEF) as a potential rotorcraft active control device. The development of structural cross-section models of a continuous trailing-edge flap airfoil is described. The CTEF deformations with MFC actuation are predicted by NASTRAN and UM/VABS analyses. Good agreement is shown between the predictions from the two analyses. Approximately two degrees of CTEF deflection, defined as the rotation angle of the trailing edge, is achieved with the baseline MFC-PZT bender. The 2D aerodynamic characteristics of the continuous trailing-edge flap are evaluated using a CFD analysis. The aerodynamic efficiency of a continuous trailing-edge flap is compared to that of a conventional discrete trailing-edge flap (DTEF). It is found that the aerodynamic characteristics of a CTEF are equivalent to those of a conventional DTEF with the same deflection angle but with a smaller flap chord. A fluid structure interaction procedure is implemented to predict the deflection of the continuous trailingedge flap under aerodynamic pressure. The reductions in CTEF deflection are overall small when aerodynamic pressure is applied: 2.7% reduction is shown with a CTEF deflection angle of two degrees and at angle of attack of six degrees. In addition, newly developed MFC-PMN actuator is found to be a good supplement to MFC-PZT when applied as the bender outside layers. A mixed MFC-PZT and MFC-PMN bender generates 3% more CTEF deformation than an MFC-PZT only bender and 5% more than an MFC-PMN only bender under aerodynamic loads.

  2. 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.

  3. 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.

  4. 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

  5. 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)

  6. Crystal structure of type I 3-dehydroquinate dehydratase of Aquifex aeolicus suggests closing of active site flap is not essential for enzyme action.

    PubMed

    Devi, Aribam Swarmistha; Ebihara, Akio; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Kumarevel, Thirumananseri; Ponnuraj, Karthe

    2013-03-01

    Structural analyses of enzymes involved in biosynthetic pathways that are present in micro-organisms, but absent from mammals (for example Shikimate pathway) are important in developing anti-microbial drugs. Crystal structure of the Shikimate pathway enzyme, type I 3-dehydroquinate dehydratase (3-DHQase) from the hyperthermophilic bacterium Aquifex aeolicus was solved both as an apo form and in complex with a ligand. The complex structure revealed an interesting structural difference when compared to other ligand-bound type I 3-DHQases suggesting that closure of the active site loop is not essential for catalysis. This provides new insights into the catalytic mechanism of type I 3-DHQases. PMID:23396056

  7. 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

  8. Circadian Regulation of Sucrose Phosphate Synthase Activity in Tomato by Protein Phosphatase Activity.

    PubMed Central

    Jones, T. L.; Ort, D. R.

    1997-01-01

    Sucrose phosphate synthase (SPS), a key enzyme in sucrose biosynthesis, is regulated by protein phosphorylation and shows a circadian pattern of activity in tomato. SPS is most active in its dephosphorylated state, which normally coincides with daytime. Applying okadaic acid, a potent protein phosphatase inhibitor, prevents SPS activation. More interesting is that a brief treatment with cycloheximide, a cytoplasmic translation inhibitor, also prevents the light activation of SPS without any effect on the amount of SPS protein. Cordycepin, an inhibitor of transcript synthesis and processing, has the same effect. Both of these inhibitors also prevent the activation phase of the circadian rhythm in SPS activity. Conversely, cycloheximide and cordycepin do not prevent the decline in circadian SPS activity that normally occurs at night. These observations indicate that SPS phosphatase activity but not SPS kinase activity is controlled, directly or indirectly, at the level of gene expression. Taken together, these data imply that there is a circadian rhythm controlling the transcription of a protein phosphatase that subsequently dictates the circadian rhythm in SPS activity via effects on this enzyme's phosphorylation state. PMID:12223667

  9. 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

  10. Site–Specific Monoubiquitination Activates Ras by Impeding GTPase Activating Protein Function

    PubMed Central

    Baker, Rachael; Lewis, Steven M.; Sasaki, Atsuo T.; Wilkerson, Emily M.; Locasale, Jason W.; Cantley, Lewis C.; Kuhlman, Brian; Dohlman, Henrik G.; Campbell, Sharon L.

    2012-01-01

    SUMMARY Cell growth and differentiation are controlled by growth factor receptors coupled to the GTPase Ras. Oncogenic mutations disrupt GTPase activity leading to persistent Ras signaling and cancer progression. Recent evidence indicates that monoubiquitination of Ras leads to Ras activation. Mutation of the primary site of monoubiquitination impairs the ability of activated K–Ras to promote tumor growth. To determine the mechanism of human Ras activation we chemically ubiquitinated the protein and analyzed its function by NMR, computational modeling, and biochemical activity measurements. We established that monoubiquitination has little effect on Ras GTP binding, GTP hydrolysis, or exchange factor activation, but severely abrogates the response to GTPase activating proteins in a site–specific manner. These findings reveal a new mechanism by which Ras can trigger persistent signaling in the absence of receptor activation or an oncogenic mutation. PMID:23178454

  11. 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

  12. 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.

  13. 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.

  14. Unwinding activity of cold shock proteins and RNA metabolism.

    PubMed

    Phadtare, Sangita

    2011-01-01

    Temperature downshift from 37 °C to 15 °C results in the exertion of cold shock response in Escherichia coli, which induces cold shock proteins, such as CsdA. Previously, we showed that the helicase activity of CsdA is critical for its function in the cold acclimation of cells and its primary role is mRNA degradation. Only RhlE (helicase), CspA (RNA chaperone) and RNase R (exoribonuclease) were found to complement the cold shock function of CsdA. RNase R has two independent activities, helicase and ribonuclease, only helicase being essential for the functional complementation of CsdA. Here, we discuss the significance of above findings as these emphasize the importance of the unwinding activity of cold-shock-inducible proteins in the RNA metabolism at low temperature, which may be different than that at 37 °C. It requires assistance of proteins to destabilize the secondary structures in mRNAs that are stabilized upon temperature downshift, hindering the activity of ribonucleases. PMID:21445001

  15. Protein kinase activity associated with simian virus 40 T antigen.

    PubMed Central

    Griffin, J D; Spangler, G; Livingston, D M

    1979-01-01

    Incubation of simian virus 40 (SV40) tumor (T) antigen-containing immunoprecipitates with [gamma-32P]ATP results in the incorporation of radioactive phosphate into large T antigen. Highly purified preparations of large T antigen from a SV40-transformed cell line, SV80, are able to catalyze the phosphorylation of a known phosphate acceptor, casein. The kinase activity migrates with large T antigen through multiple purification steps. Sedimentation analysis under non-T-antigen-aggregating conditions reveals that kinase activity and the immunoreactive protein comigrate as a 6S structure. The kinase activity of purified preparations of large T antigen can be specifically adsorbed to solid-phase anti-T IgG, and partially purified T antigen from a SV40 tsA transformation is thermolabile in its ability to phosphorylate casein when compared to comparably purified wild-type T antigen. These observations indicate that the SV40 large T antigen is closely associated with protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) activity. Images PMID:223152

  16. Maxillofacial reconstruction with nasolabial and facial artery musculomucosal flaps.

    PubMed

    Braasch, Daniel Cameron; Lam, Din; Oh, Esther S

    2014-08-01

    The nasolabial and facial artery musculomucosal (FAMM) flaps are predictable methods to reconstruct perioral and intraoral defects with vascularized tissue. The nasolabial flap can be harvested as an axial or random patterned flap, whereas the FAMM flap is truly an axial patterned flap, with either a superior or an inferior base. Both flaps have been widely used to provide predictable results, with low morbidity. Future studies are needed to further prove their use in compromised patients, including patients with a history of head and neck radiation and neck dissections. PMID:25086694

  17. Dynamical Theory of Activated Processes in Globular Proteins

    NASA Astrophysics Data System (ADS)

    Northrup, Scott H.; Pear, Michael R.; Lee, Chyuan-Yih; McCammon, J. Andrew; Karplus, Martin

    1982-07-01

    A methos is described for calculating the reaction rate in globular proteins of activated processes such as ligand binding or enzymatic catalysis. The method is based on the determination of the probability that the system is in the transition state and of the magnitude of the reactive flux for transition-state systems. An ``umbrella sampling'' simulation procedure is outlined for evaluating the transition-state probability. The reactive flux is obtained from an approach described previously for calculating the dynamics of transition-state trajectories. An application to the rotational isomerization of an aromatic ring in the bovine pancreatic trypsin inhibitor is presented. The results demonstrate the feasibility of calculating rate constants for reactions in proteins and point to the importance of solvent effects for reactions that occur near the protein surface.

  18. Receptor Activity-Modifying Proteins (RAMPs): New Insights and Roles.

    PubMed

    Hay, Debbie L; Pioszak, Augen A

    2016-01-01

    It is now recognized that G protein-coupled receptors (GPCRs), once considered largely independent functional units, have a far more diverse molecular architecture. Receptor activity-modifying proteins (RAMPs) provide an important example of proteins that interact with GPCRs to modify their function. RAMPs are able to act as pharmacological switches and chaperones, and they can regulate signaling and/or trafficking in a receptor-dependent manner. This review covers recent discoveries in the RAMP field and summarizes the known GPCR partners and functions of RAMPs. We also discuss the first peptide-bound structures of RAMP-GPCR complexes, which give insight into the molecular mechanisms that enable RAMPs to alter the pharmacology and signaling of GPCRs. PMID:26514202

  19. Dynamical theory of activated processes in globular proteins.

    PubMed Central

    Northrup, S H; Pear, M R; Lee, C Y; McCammon, J A; Karplus, M

    1982-01-01

    A method is described for calculating the reaction rate in globular proteins of activated processes such as ligand binding or enzymatic catalysis. The method is based on the determination of the probability that the system is in the transition state and of the magnitude of the reactive flux for transition-state systems. An "umbrella sampling" simulation procedure is outlined for evaluating the transition-state probability. The reactive flux is obtained from an approach described previously for calculating the dynamics of transition-state trajectories. An application to the rotational isomerization of an aromatic ring in the bovine pancreatic trypsin inhibitor is presented. The results demonstrate the feasibility of calculating rate constants for reactions in proteins and point to the importance of solvent effects for reactions that occur near the protein surface. PMID:6955788

  20. 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

  1. Molecular architecture of the active mitochondrial protein gate.

    PubMed

    Shiota, Takuya; Imai, Kenichiro; Qiu, Jian; Hewitt, Victoria L; Tan, Khershing; Shen, Hsin-Hui; Sakiyama, Noriyuki; Fukasawa, Yoshinori; Hayat, Sikander; Kamiya, Megumi; Elofsson, Arne; Tomii, Kentaro; Horton, Paul; Wiedemann, Nils; Pfanner, Nikolaus; Lithgow, Trevor; Endo, Toshiya

    2015-09-25

    Mitochondria fulfill central functions in cellular energetics, metabolism, and signaling. The outer membrane translocator complex (the TOM complex) imports most mitochondrial proteins, but its architecture is unknown. Using a cross-linking approach, we mapped the active translocator down to single amino acid residues, revealing different transport paths for preproteins through the Tom40 channel. An N-terminal segment of Tom40 passes from the cytosol through the channel to recruit chaperones from the intermembrane space that guide the transfer of hydrophobic preproteins. The translocator contains three Tom40 β-barrel channels sandwiched between a central α-helical Tom22 receptor cluster and external regulatory Tom proteins. The preprotein-translocating trimeric complex exchanges with a dimeric isoform to assemble new TOM complexes. Dynamic coupling of α-helical receptors, β-barrel channels, and chaperones generates a versatile machinery that transports about 1000 different proteins. PMID:26404837

  2. Combined posterior flap and anterior suspended flap dacryocystorhinostomy: A modification of external dacryocystorhinostomy

    PubMed Central

    Deka, Amarendra; Saikia, S. P.; Bhuyan, S. K.

    2010-01-01

    Background: External dacryocystorhinostomy (DCR) remains a reliable surgical technique for the treatment of obstruction of lacrimal drainage system beyond the common canalicular opening. Aim: To describe a simple modified double flap external DCR technique. Materials and Methods: Ninety six consecutive cases of chronic dacryocystitis with or without mucocele were selected irrespective of age and sex. In a modification to routine external DCR, a modified technique was followed, where both anterior and posterior flaps of lacrimal sac and nasal mucosa are created and sutured. Two double armed sutures were used to join the edges of anterior flaps, and elevate them anteriorly to avoid adhesion or apposition with underlying sutured posterior flaps, and to approximate the deep plane of the wound. Results: At the end of average follow-up period of 13 months, we observed 98.9% objective and 96.8% subjective success rates. The average operation time was 45 minutes. No significant intraoperative or postoperative complications were noticed. Conclusion: We believe that combined posterior flap and anterior suspended flap DCR technique is simple to perform and has the advantage of both double flap DCR and anterior suspension of anterior flaps. The results of the study showed the efficacy of this simple modification. PMID:20606867

  3. The evolving breast reconstruction: from latissimus dorsi musculocutaneous flap to a propeller thoracodorsal fasciocutaneous flap

    PubMed Central

    Gunnarsson, Gudjon Leifur

    2014-01-01

    The aim of this editorial is to give an update on the use of the propeller thoracodorsal artery perforator flap (TAP/TDAP-flap) within the field of breast reconstruction. The TAP-flap can be dissected by a combined use of a monopolar cautery and a scalpel. Microsurgical instruments are generally not needed. The propeller TAP-flap can be designed in different ways, three of these have been published: (I) an oblique upwards design; (II) a horizontal design; (III) an oblique downward design. The latissimus dorsi-flap is a good and reliable option for breast reconstruction, but has been criticized for morbidity and complications. The TAP-flap does not seem to impair the function of the shoulder or arm and the morbidity appears to be scarce. However, an implant is often needed in combination with the TAP-flap, which results in implant related morbidity over time. The TAP-flap seems to be a promising tool for oncoplastic and reconstructive breast surgery and will certainly become an invaluable addition to breast reconstructive methods. PMID:25207206

  4. Modulation of the protein kinase activity of mTOR.

    PubMed

    Lawrence, J C; Lin, T A; McMahon, L P; Choi, K M

    2004-01-01

    mTOR is a founding member of a family of protein kinases having catalytic domains homologous to those in phosphatidylinositol 3-OH kinase. mTOR participates in the control by insulin of the phosphorylation of lipin, which is required for adipocyte differentiation, and the two translational regulators, p70S6K and PHAS-I. The phosphorylation of mTOR, itself, is stimulated by insulin in Ser2448, a site that is also phosphorylated by protein kinase B (PKB) in vitro and in response to activation of PKB activity in vivo. Ser2448 is located in a short stretch of amino acids not found in the two TOR proteins in yeast. A mutant mTOR lacking this stretch exhibited increased activity, and binding of the antibody, mTAb-1, to this region markedly increased mTOR activity. In contrast, rapamycin-FKBP12 inhibited mTOR activity towards both PHAS-I and p70S6K, although this complex inhibited the phosphorylation of some sites more than that of others. Mutating Ser2035 to Ile in the FKBP12-rapamycin binding domain rendered mTOR resistant to inhibition by rapamycin. Unexpectedly, this mutation markedly decreased the ability of mTOR to phosphorylate certain sites in both PHAS-I and p70S6K. The results support the hypotheses that rapamycin disrupts substrate recognition instead of directly inhibiting phosphotransferase activity and that mTOR activity in cells is controlled by the phosphorylation of an inhibitory regulatory domain containing the mTAb-1 epitope. PMID:14560959

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

    PubMed Central

    Burnier, Laurent

    2013-01-01

    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

  6. 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

  7. Hepatitis B virus X protein activates transcription factor NF-kappa B without a requirement for protein kinase C.

    PubMed Central

    Lucito, R; Schneider, R J

    1992-01-01

    The hepatitis B virus X protein stimulates transcription from a variety of promoter elements, including those activated by transcription factor NF-kappa B. A diverse group of extra- and intracellular agents, including growth factors and the human immunodeficiency virus tat protein, have been shown to require a functional protein kinase C (PKC) system to achieve activation of NF-kappa B. In this study we have investigated the molecular mechanism by which X protein activates NF-kappa B. We demonstrate that in hepatocytes, X protein induces a maximal activation of NF-kappa B corresponding to the sequestered pool of factor, which is also activated by phorbol esters. To determine whether X protein requires activation of PKC to stimulate transcription by NF-kappa B, we attempted to prevent transactivation by X protein in the presence of the PKC inhibitors calphostin C and H7. We show that PKC inhibitors do not block X protein activation of NF-kappa B, whereas they largely impair activation by phorbol esters. In addition, activation of PKC is correlated with its translocation from the cytoplasm to the plasma membrane. The subcellular distribution of PKC was investigated by introducing X protein from a replication-defective adenovirus vector, followed by immunochemical detection of PKC in cell fractions. These data also indicate that X protein stimulates transcription by NF-kappa B without the activation and translocation of PKC. Images PMID:1309924

  8. 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

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

    PubMed Central

    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

  10. 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

  11. Design and experimental results for a turbine with jet flap stator and jet flap

    NASA Technical Reports Server (NTRS)

    Bettner, J. L.; Blessing, J. O.

    1973-01-01

    The overall performance and detailed stator performance of a negative hub reaction turbine design featuring a moderately low solidity jet flap stator and a jet flap rotor were determined. Testing was conducted over a range of turbine expansion ratios at design speed. At each expansion ratio, the stator jet flow and rotor jet flow ranged up to about 7 and 8 percent, respectively, of the turbine inlet flow. The performance of the jet flap stator/jet flap rotor turbine was compared with that of a turbine which used the same jet flap rotor and a conventional, high solidity plan stator. The effect on performance of increased axial spacing between the jet stator and rotor was also investigated.

  12. 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

  13. Complement activation and cytokine response by BioProtein, a bacterial single cell protein.

    PubMed

    Sikkeland, L I B; Thorgersen, E B; Haug, T; Mollnes, T E

    2007-04-01

    The bacterial single cell protein (BSCP), BioProtein, is dried bacterial mass derived from fermentation of the gram negative bacteria Methylococcus capsulatus, used for animal and fish feed. Workers in this industry suffer frequently from pulmonary and systemic symptoms which may be induced by an inflammatory reaction. The aim of the present study was to examine the effect of BSCP on inflammation in vitro as evaluated by complement activation and cytokine production. Human serum was incubated with BSCP and complement activation products specific for all pathways were detected by enzyme-linked immunosorbent assay (ELISA). Human whole blood anti-coagulated with lepirudin was incubated with BSCP and a panel of 27 biological mediators was measured using multiplex technology. BSCP induced a dose-dependent complement activation as revealed by a pronounced increase in alternative and terminal pathway activation (fivefold and 20-fold, respectively) at doses from 1 microg BSCP/ml serum and a similar, but less extensive (two- to fourfold) increase in activation of the lectin and classical pathways at doses from 100 and 1000 microg BSCP/ml serum, respectively. Similarly, BSCP induced a dose-dependent production of a number of cytokines, chemokines and growth factors in human whole blood. At doses as low as 0 x 05-0 x 5 microg BSCP/ml blood a substantial increase was seen for tumour necrosis factor (TNF)-alpha, interleukin (IL)-1-beta, IL-6, interferon (IFN)-gamma, IL-8, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, IL-4, IL-9, IL-17, IL-1Ra, granulocyte-colony-stimulating factor (G-CSF) and vascular endothelial growth factor (VEGF). Thus, BSCP induced a substantial activation of all three initial complement pathways as well as a pronounced cytokine response in vitro, indicating a potent inflammatory property of this agent. PMID:17302729

  14. 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

  15. Regulation of Orange Carotenoid Protein Activity in Cyanobacterial Photoprotection.

    PubMed

    Thurotte, Adrien; Lopez-Igual, Rocio; Wilson, Adjélé; Comolet, Léa; Bourcier de Carbon, Céline; Xiao, Fugui; Kirilovsky, Diana

    2015-09-01

    Plants, algae, and cyanobacteria have developed mechanisms to decrease the energy arriving at reaction centers to protect themselves from high irradiance. In cyanobacteria, the photoactive Orange Carotenoid Protein (OCP) and the Fluorescence Recovery Protein are essential elements in this mechanism. Absorption of strong blue-green light by the OCP induces carotenoid and protein conformational changes converting the orange (inactive) OCP into a red (active) OCP. Only the red orange carotenoid protein (OCP(r)) is able to bind to phycobilisomes, the cyanobacterial antenna, and to quench excess energy. In this work, we have constructed and characterized several OCP mutants and focused on the role of the OCP N-terminal arm in photoactivation and excitation energy dissipation. The N-terminal arm largely stabilizes the closed orange OCP structure by interacting with its C-terminal domain. This avoids photoactivation at low irradiance. In addition, it slows the OCP detachment from phycobilisomes by hindering fluorescence recovery protein interaction with bound OCP(r). This maintains thermal dissipation of excess energy for a longer time. Pro-22, at the beginning of the N-terminal arm, has a key role in the correct positioning of the arm in OCP(r), enabling strong OCP binding to phycobilisomes, but is not essential for photoactivation. Our results also show that the opening of the OCP during photoactivation is caused by the movement of the C-terminal domain with respect to the N-terminal domain and the N-terminal arm. PMID:26195570

  16. 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.

  17. Protein A-like activity and streptococcal cross-reactions.

    PubMed Central

    Kingston, D

    1981-01-01

    Recognition of the protein A-like activity of some strains of group A streptococci has thrown doubt on much previous work suggesting antigenic cross-reactions between these streptococci and mammalian tissues. The strains used in our previous studies have now been examined by the mixed reverse passive antiglobulin reaction (MRPAH) for the 'non-specific' absorption of purified Fc portion of human IgG. They were found to have only traces of activity. The strain of Staphylococcus aureus used to control 'non-specific' absorption by bacterial cell walls was strongly positive. Protein A-like material as detected in this way was not therefore responsible for our earlier results. PMID:7039880

  18. 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. PMID:23981650

  19. 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

  20. 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...

  1. Wounding systemically activates a mitogen-activated protein kinase in forage and turf grasses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Forage and turf grasses are continually cut and grazed by livestock, however very little is known concerning the perception or molecular responses to wounding. Mechanical wounding rapidly activated a 46 kDa and a 44 kDa mitogen-activated protein kinase (MAPK) in six different grass species. In the m...

  2. 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.

  3. 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

  4. 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.

  5. [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

  6. Proapoptotic Activities of Protein Disulfide Isomerase (PDI) and PDIA3 Protein, a Role of the Bcl-2 Protein Bak*

    PubMed Central

    Zhao, Guoping; Lu, Huayi; Li, Chi

    2015-01-01

    Protein disulfide isomerase (PDI) family proteins are classified as enzymatic chaperones for reconstructing misfolded proteins. Previous studies have shown that several PDI members possess potential proapoptotic functions. However, the detailed molecular mechanisms of PDI-mediated apoptosis are not completely known. In this study, we investigated how two members of PDI family, PDI and PDIA3, modulate apoptotic signaling. Inhibiting PDI and PDIA3 activities pharmacologically alleviates apoptosis induced by various apoptotic stimuli. Although a decrease of PDIA3 expression alleviates apoptotic responses, overexpression of PDIA3 exacerbates apoptotic signaling. Importantly, Bak, but not Bax, is essential for PDIA3-induced proapoptotic signaling. Furthermore, both purified PDI and PDIA3 proteins induce Bak-dependent, but not Bax-dependent, mitochondrial outer membrane permeabilization in vitro, probably through triggering Bak oligomerization on mitochondria. Our results suggest that both of PDI and PDIA3 possess Bak-dependent proapoptotic function through inducing mitochondrial outer membrane permeabilization, which provides a new mechanism linking ER chaperone proteins and apoptotic signaling. PMID:25697356

  7. 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

  8. 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

  9. 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

  10. Antioxidative activity of protein hydrolysates prepared from alkaline-aided channel catfish protein isolates.

    PubMed

    Theodore, Ann E; Raghavan, Sivakumar; Kristinsson, Hordur G

    2008-08-27

    Antioxidative activity of hydrolyzed protein prepared from alkali-solubilized catfish protein isolates was studied. The isolates were hydrolyzed to 5, 15, and 30% degree of hydrolysis using the protease enzyme, Protamex. Hydrolyzed protein was separated into hydrolysates and soluble supernatants, and both of these fractions were studied for their metal chelating ability, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and their ability to inhibit the formation of thiobarbituric acid reactive substances (TBARS) in washed tilapia muscle containing tilapia hemolysate. Both hydrolysates and supernatants were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results showed that DPPH radical scavenging ability and reducing power of catfish protein hydrolysates decreased, whereas the ORAC value, metal chelating ability, and ability to inhibit TBARS increased, with an increase in the degree of hydrolysis. Hydrolysate samples showed higher DPPH radical scavenging ability and Fe(3+) reducing ability, and supernatant samples had higher metal chelating ability. In general, low molecular weight (MW) peptides had high ORAC values and high metal chelating ability, and high MW peptides had a higher reducing power (FRAP) and were more effective in scavenging DPPH radicals. In a washed muscle model system, the ability of catfish protein hydrolysates and their corresponding supernatants to inhibit the formation of TBARS increased with an increase in the degree of hydrolysis. PMID:18662014

  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. 14 CFR 25.701 - Flap and slat interconnection.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... equivalent means. (b) If a wing flap or slat interconnection or equivalent means is used, it must be designed... be designed for the loads imposed when the wing flaps or slats on one side are carrying the...

  13. Reconstruction Using Locoregional Flaps for Large Skull Base Defects.

    PubMed

    Hatano, Takaharu; Motomura, Hisashi; Ayabe, Shinobu

    2015-06-01

    We present a modified locoregional flap for the reconstruction of large anterior skull base defects that should be reconstructed with a free flap according to Yano's algorithm. No classification of skull base defects had been proposed for a long time. Yano et al suggested a new classification in 2012. The lb defect of Yano's classification extends horizontally from the cribriform plate to the orbital roof. According to Yano's algorithm for subsequent skull base reconstructive procedures, a lb defect should be reconstructed with a free flap such as an anterolateral thigh free flap or rectus abdominis myocutaneous free flap. However, our modified locoregional flap has also enabled reconstruction of lb defects. In this case series, we used a locoregional flap for lb defects. No major postoperative complications occurred. We present our modified locoregional flap that enables reconstruction of lb defects. PMID:26225296

  14. Flapping wing PIV and force measurements

    NASA Astrophysics Data System (ADS)

    Cameron, Benjamin H.

    Flapping wing aerodynamics has been of interest to engineers recently due in part to the DARPA (Defense Advanced Research Projects Agency) MAV (Micro-Aerial Vehicle) initiative. MAVs are small unmanned aerial vehicles with length scales similar to birds and insects. Flapping wing MAVs would serve as mobile and stealthy sensing platforms capable of gathering intelligence in hazardous and physically inaccessible locations. Traditional means of lift and thrust generation become inefficient when scaled to these sizes, therefore a flapping wing propulsion system will be necessary. The design of a flapping wing MAV requires the ability to measure forces and velocities around the wing. Three components of velocity were measured in the wake of a two dimensional (2D) flapping airfoil model using a novel application of stereoscopic DPIV (Digital Particle Image Velocimetry). One component of force was measured using a newly proposed method outlined in the dissertation. The force measurement technique relies on a specific sequence of data acquisition, which has the benefit of reducing measurement uncertainty and noise. No experiments of this type have been conducted, and no direct aerodynamic force data exists for the low Reynolds numbers applicable to flapping wing MAVs. The well-established stereoscopic DPIV technique produces relatively low uncertainties while the new force measurement technique has not been previously tested. Theoretical analysis and experimental results show that aerodynamic forces are attainable for chord Reynolds numbers as low as 1,000, which is significantly lower than previous studies. PIV measurements reveal symmetric and asymmetric wake topologies for a NACA 0012 and flat plate airfoil. A sinusoidally heaving flat plate airfoil produces highly deflected wakes for a wider range of flapping conditions than a NACA 0012 airfoil. Deflected wakes are of potentially interest since both lift and thrust components of force are developed. The flat plate also

  15. 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

  16. 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

  17. Caspase processing activates atypical protein kinase C zeta by relieving autoinhibition and destabilizes the protein.

    PubMed Central

    Smith, Lucinda; Wang, Zhi; Smith, Jeffrey B

    2003-01-01

    Treatment of HeLa cells with tumour necrosis factor alpha (TNFalpha) induced caspase processing of ectopic PKC (protein kinase C) zeta, which converted most of the holoenzyme into the freed kinase domain and increased immune-complex kinase activity. The goal of the present study was to determine the basis for the increased kinase activity that is associated with caspase processing of PKC zeta. Atypical PKC iota is largely identical with PKC zeta, except for a 60-amino-acid segment that lacks the caspase-processing sites of the zeta isoform. Replacement of this segment of PKC zeta with the corresponding segment of PKC iota prevented caspase processing and activation of the kinase function. Processing of purified recombinant PKC zeta by caspase 3 in vitro markedly increased its kinase activity. Caspase processing activated PKC zeta in vitro or intracellularly without increasing the phosphorylation of Thr410 of PKC zeta, which is required for catalytic competency. The freed kinase domain of PKC zeta had a much shorter half-life than the holoenzyme in transfected HeLa cells and in non-transfected kidney epithelial cells. Treatment with TNF-alpha shortened the half-life of the kinase domain protein, and proteasome blockade stabilized the protein. Studies of kinase-domain mutants indicate that a lack of negative charge at Thr410 can shorten the half-life of the freed kinase domain. The present findings indicate that the freed kinase domain has substantially higher kinase activity and a much shorter half-life than the holoenzyme because of accelerated degradation by the ubiquitin-proteasome system. PMID:12887331

  18. Centromeric binding and activity of Protein Phosphatase 4

    PubMed Central

    Lipinszki, Zoltan; Lefevre, Stephane; Savoian, Matthew S.; Singleton, Martin R.; Glover, David M.; Przewloka, Marcin R.

    2015-01-01

    The cell division cycle requires tight coupling between protein phosphorylation and dephosphorylation. However, understanding the cell cycle roles of multimeric protein phosphatases has been limited by the lack of knowledge of how their diverse regulatory subunits target highly conserved catalytic subunits to their sites of action. Phosphoprotein phosphatase 4 (PP4) has been recently shown to participate in the regulation of cell cycle progression. We now find that the EVH1 domain of the regulatory subunit 3 of Drosophila PP4, Falafel (Flfl), directly interacts with the centromeric protein C (CENP-C). Unlike other EVH1 domains that interact with proline-rich ligands, the crystal structure of the Flfl amino-terminal EVH1 domain bound to a CENP-C peptide reveals a new target-recognition mode for the phosphatase subunit. We also show that binding of Flfl to CENP-C is required to bring PP4 activity to centromeres to maintain CENP-C and attached core kinetochore proteins at chromosomes during mitosis. PMID:25562660

  19. Covalent agonists for studying G protein-coupled receptor activation

    PubMed Central

    Weichert, Dietmar; Kruse, Andrew C.; Manglik, Aashish; Hiller, Christine; Zhang, Cheng; Hübner, Harald; Kobilka, Brian K.; Gmeiner, Peter

    2014-01-01

    Structural studies on G protein-coupled receptors (GPCRs) provide important insights into the architecture and function of these important drug targets. However, the crystallization of GPCRs in active states is particularly challenging, requiring the formation of stable and conformationally homogeneous ligand-receptor complexes. Native hormones, neurotransmitters, and synthetic agonists that bind with low affinity are ineffective at stabilizing an active state for crystallogenesis. To promote structural studies on the pharmacologically highly relevant class of aminergic GPCRs, we here present the development of covalently binding molecular tools activating Gs-, Gi-, and Gq-coupled receptors. The covalent agonists are derived from the monoamine neurotransmitters noradrenaline, dopamine, serotonin, and histamine, and they were accessed using a general and versatile synthetic strategy. We demonstrate that the tool compounds presented herein display an efficient covalent binding mode and that the respective covalent ligand-receptor complexes activate G proteins comparable to the natural neurotransmitters. A crystal structure of the β2-adrenoreceptor in complex with a covalent noradrenaline analog and a conformationally selective antibody (nanobody) verified that these agonists can be used to facilitate crystallogenesis. PMID:25006259

  20. 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.

  1. [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

  2. 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

  3. Protective effect of hydrogen-rich saline on ischemia/reperfusion injury in rat skin flap*

    PubMed Central

    Zhao, Ling; Wang, You-bin; Qin, Shi-rui; Ma, Xue-mei; Sun, Xue-jun; Wang, Ming-lian; Zhong, Ru-gang

    2013-01-01

    Objective: Skin damage induced by ischemia/reperfusion (I/R) is a multifactorial process that often occurs in plastic surgery. The mechanisms of I/R injury include hypoxia, inflammation, and oxidative damage. Hydrogen gas has been reported to alleviate cerebral I/R injury by acting as a free radical scavenger. Here, we assessed the protective effect of hydrogen-rich saline (HRS) on skin flap I/R injury. Methods: Abdominal skin flaps of rats were elevated and ischemia was induced for 3 h; subsequently, HRS or physiological saline was administered intraperitoneally 10 min before reperfusion. On postoperative Day 5, flap survival, blood perfusion, the accumulation of reactive oxygen species (ROS), and levels of cytokines were evaluated. Histological examinations were performed to assess inflammatory cell infiltration. Results: Skin flap survival and blood flow perfusion were improved by HRS relative to the controls. The production of malondialdehyde (MDA), an indicator of lipid peroxidation, was markedly reduced. A multiplex cytokine assay revealed that HRS reduced the elevation in the levels of inflammatory cytokines, chemokines and growth factors, with the exception of RANTES (regulated on activation, normal T-cell expressed and secreted) growth factor. HRS treatment also reduced inflammatory cell infiltration induced by I/R injury. Conclusions: Our findings suggest that HRS mitigates I/R injury by decreasing inflammation and, therefore, has the potential for application as a therapy for improving skin flap survival. PMID:23645175

  4. Design Of A Scientific Experiment On Expert Flap At CIRA SCIROCCO Plasma Wind Tunnel

    NASA Astrophysics Data System (ADS)

    Trifoni, E.; Del Vecchio, A.; Di Clemente, M.; De Simone, V.; Martucci, A.; Purpura, C.; Savino, L.; Cipullo, A.

    2011-05-01

    A scientific experiment at CIRA SCIROCCO Plasma Wind Tunnel was conceived to assess the design of ESA EXPERT Open Flap PL6/8 under flight representative heat flux and load conditions. A specific and dedicated model holder was designed to test the Qualification Model of the instrumented CMC Flap in real size and equipped with the same flight instrumentation, on the basis of the structural architecture (actively cooled Leading Edge, TPS and cold structure) developed and successfully tested in the frame of the EXPERT PL7 SWBLI qualification test in SCIROCCO PWT facility. The Test conditions were designed in order to get significant heat flux and temperature on the Flap symmetry plane. The Test duration has been established with the aim at realizing the radiative equilibrium condition on the Flap and with respect to the duration of the entire re-entry of the Expert capsule The maximum total heat load over the Flap experienced in flight conditions during the re-entry trajectory was also evaluated. In the present paper an overview of the test design parameters, expected outcomes and preliminary experimental results are reported.

  5. 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...

  6. 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...

  7. 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...

  8. Unexpected flap thickness in laser in situ keratomileusis.

    PubMed

    Giledi, Osama; Daya, Sheraz M

    2003-09-01

    We report a case of an unexpected thick flap during laser in situ keratomileusis (LASIK) that led to abandonment of surgery. This report illustrates the importance of stromal bed measurements after flap creation in LASIK. A thicker-than-expected flap can lead to a thinner-than-anticipated residual cornea and subsequent ectasia or even perforation during laser ablation. It is possible that reports of ectasia in normal thickness corneas reflect thicker-than-anticipated flaps. PMID:14522308

  9. 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...

  10. 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...

  11. 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

  12. 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.

  13. 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).

  14. 4-Phenylbutyrate protects rat skin flaps against ischemia-reperfusion injury and apoptosis by inhibiting endoplasmic reticulum stress

    PubMed Central

    YUE, ZHEN-SHUANG; ZENG, LIN-RU; QUAN, REN-FU; TANG, YANG-HUA; ZHENG, WEN-JIE; QU, GANG; XU, CAN-DA; ZHU, FANG-BING; HUANG, ZHONG-MING

    2016-01-01

    4-phenylbutyrate (4-PBA) is a low molecular weight fatty acid, which has been demonstrated to regulate endoplasmic reticulum (ER) stress. ER stress-induced cell apoptosis has an important role in skin flap ischemia; however, a pharmacological approach for treating ischemia-induced ER dysfunction has yet to be reported. In the present study, the effects of 4-PBA-induced ER stress inhibition on ischemia-reperfusion injury were investigated in the skin flap of rats, and transcriptional regulation was examined. 4-PBA attenuated ischemia-reperfusion injury and inhibited cell apoptosis in the skin flap. Furthermore, 4-PBA reversed the increased expression levels of two ER stress markers: CCAAT/enhancer-binding protein-homologous protein and glucose-regulated protein 78. These results suggested that 4-PBA was able to protect rat skin flaps against ischemia-reperfusion injury and apoptosis by inhibiting ER stress marker expression and ER stress-mediated apoptosis. The beneficial effects of 4-PBA may prove useful in the treatment of skin flap ischemia-reperfusion injury. PMID:26648447

  15. Mitogen-activated protein kinase (MAPK) in cardiac tissues.

    PubMed

    Page, C; Doubell, A F

    Mitogen-activated protein kinase (MAPK) has recently emerged as a prominent role player in intracellular signalling in the ventricular myocyte with attention being focussed on its possible role in the development of ventricular hypertrophy. It is becoming clear that MAPK is also active in other cells of cardiac origin such as cardiac fibroblasts and possible functions of this signalling pathway in the heart have yet to be explored. In this report the mammalian MAPK pathway is briefly outlined, before reviewing current knowledge of the MAPK pathway in cardiac tissue (ventricular myocytes, vascular smooth muscle cells and cardiac fibroblasts). New data is also presented on the presence and activity of MAPK in two additional cardiac celltypes namely atrial myocytes and vascular endothelial cells from the coronary microcirculation. PMID:8739228

  16. Expression of the vertebrate Gli proteins in Drosophila reveals a distribution of activator and repressor activities.

    PubMed

    Aza-Blanc, P; Lin, H Y; Ruiz i Altaba, A; Kornberg, T B

    2000-10-01

    The Cubitus interruptus (Ci) and Gli proteins are transcription factors that mediate responses to Hedgehog proteins (Hh) in flies and vertebrates, respectively. During development of the Drosophila wing, Ci transduces the Hh signal and regulates transcription of different target genes at different locations. In vertebrates, the three Gli proteins are expressed in overlapping domains and are partially redundant. To assess how the vertebrate Glis correlate with Drosophila Ci, we expressed each in Drosophila and monitored their behaviors and activities. We found that each Gli has distinct activities that are equivalent to portions of the regulatory arsenal of Ci. Gli2 and Gli1 have activator functions that depend on Hh. Gli2 and Gli3 are proteolyzed to produce a repressor form able to inhibit hh expression. However, while Gli3 repressor activity is regulated by Hh, Gli2 repressor activity is not. These observations suggest that the separate activator and repressor functions of Ci are unevenly partitioned among the three Glis, yielding proteins with related yet distinct properties. PMID:10976059

  17. 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.

  18. 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

  19. 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...

  20. 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...

  1. 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...

  2. CHARACTERISTICS OF FLAP GATES AT THE END OF DRAIN PIPES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Flap gates are commonly used at the end of pipe drains and pump outlets to prevent back flows of water and entry of small animals. Flap gates are relatively inexpensive, with low maintenance costs, but can trap debris in their hinge systems. Many texts refer to studies performed on flap gates at t...

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. Prospective evaluation of outcome measures in free-flap surgery.

    PubMed

    Kelly, John L; Eadie, Patricia A; Orr, David; Al-Rawi, Mogdad; O'Donnell, Margaret; Lawlor, Denis

    2004-08-01

    Free-flap failure is usually caused by venous or arterial thrombosis. In many cases, lack of experience and surgical delay also contribute to flap loss. The authors prospectively analyzed the outcome of 57 free flaps over a 28-month period (January, 1999 to April, 2001). The setting was a university hospital tertiary referral center. Anastomotic technique, ischemia time, choice of anticoagulant, and the grade of surgeon were recorded. The type of flap, medications, and co-morbidities, including preoperative radiotherapy, were also documented. Ten flaps were re-explored (17 percent). There were four cases of complete flap failure (6.7 percent) and five cases of partial failure (8.5 percent). In patients who received perioperative systemic heparin or dextran, there was no evidence of flap failure (p = .08). The mean ischemia time was similar in flaps that failed (95 +/- 29 min) and in those that survived (92 +/- 34 min). Also, the number of anastomoses performed by trainees in flaps that failed (22 percent), was similar to the number in flaps that survived (28 percent). Nine patients received preoperative radiotherapy, and there was complete flap survival in each case. This study reveals that closely supervised anastomoses performed by trainees may have a similar outcome to those performed by more senior surgeons. There was no adverse effect from radiotherapy or increased ischemia time on flap survival. PMID:15356760

  9. Clinical outcomes of suture delay in forehead flap.

    PubMed

    Isik, Daghan; Kiroglu, Faruk; Isik, Yasemin; Goktas, Ugur; Atik, Bekir

    2012-01-01

    The delay phenomenon is a surgical procedure performed to raise a wider skin flap and to improve the survival of skin flaps. Surgery, chemicals, sutures, and lasers can be used for the delay procedure. In this study, delayed forehead flaps created by suturing were used for coverage of nasal skin defects in eleven patients. In 7 patients, the cross-paramedian forehead flap was used to increase the extent of flap lengthening. In the first session, suture delay was performed on both sides of the forehead flap margin. In the second session, the flap was elevated and sutured to its new position, 7 to 10 days after the initial surgery. All flaps were completely viable, and patient satisfaction was optimal in all cases. The positive effect of surgical delay on flap survival has been shown in experimental and clinical studies. However, experimentally, suture delay or chemical delay procedures have been shown to be beneficial in flap survival only. Suture delay seems to be an inexpensive, effective, easily performed, atraumatic, and safe technique, especially among patients with systemic diseases such as diabetes or cardiovascular diseases, smoking patients who may lose the flap, or patients who need very wide and long flaps. PMID:22337378

  10. 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...

  11. 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...

  12. 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

  13. Peptide biosensors for the electrochemical measurement of protein kinase activity.

    PubMed

    Kerman, Kagan; Song, Haifeng; Duncan, James S; Litchfield, David W; Kraatz, Heinz-Bernhard

    2008-12-15

    The kinase activities are elucidated using the novel redox-active cosubstrate adenosine 5'-[gamma-ferrocene] triphosphate (Fc-ATP), which enables the kinase-catalyzed transfer of a redox active gamma-phosphate-Fc to a hydroxyamino acid. In this report, a versatile electrochemical biosensor is developed for monitoring the activity and inhibition of a serine/threonine kinase, casein kinase 2 (CK2), and protein tyrosine kinases, Abl1-T315I and HER2, in buffered solutions and in cell lysates. The method is based on the labeling of a specific phosphorylation event with Fc, followed by electrochemical detection. The electrochemical response obtained from the "ferrocenylated" peptides enables monitoring the activity of the kinase and its substrate, as well as the inhibition of small molecule inhibitors on protein phosphorylation. Kinetic information was extracted from the electrochemical measurements for the determination of K(m) and V(m) values, which were in agreement with those previously reported. Kinase reactions were also performed in the presence of well-defined inhibitors of CK2, 4,5,6,7-tetrabromo-2-azabenzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, and E-3-(2,3,4,5-tetrabromophenyl)acrylic acid as well as the nonspecific kinase inhibitors, staurosporine and N-benzoylstaurosporine. On the basis of the dependency of the Fc signal on inhibitor concentration, K(i) of the inhibitors was estimated, which were also in agreement with the literature values. The performance of the biosensor was optimized including the kinase reaction, incubation with Fc-ATP, and the small molecule inhibitors. Peptide modified electrochemical biosensors are promising candidates for cost-effective in vitro kinase activity and inhibitor screening assays. PMID:18989981

  14. Juvenile GM2 gangliosidosis (AMB variant): inability to activate hexosaminidase A by activator protein.

    PubMed Central

    Inui, K; Grebner, E E; Jackson, L G; Wenger, D A

    1983-01-01

    Two sibling from a consanguineous Puerto Rican marriage were found to have a juvenile-onset type of lipidosis first noted at age 2 1/2 by expressing difficulties with motor function and developmental delay. They continued to deteriorate, showing muscle atrophy, spasticity, and loss of speech, and death occurred at ages 7 and 8. Examination of the brains from these patients revealed that the concentration of GM2 ganglioside was about 56% of the total gangliosides. Hexosaminidase and percent hexosaminidase A (HEX A) and other lysosomal enzymes were normal in cultured skin fibroblasts, liver, and brain. The concentration of the activator protein required for the enzymatic hydrolysis of GM2 ganglioside was in high normal levels in the brain of the patient available. However, the HEX A from the patient's brain and liver as well as from skin fibroblast lysates could not be activated to hydrolyze GM2 ganglioside by the activator protein from a control or himself. The HEX A from a control could be activated by the activator protein from controls or this patient. These patients appear to have a defect in HEX A, which does not affect it heat stability, electrophoretic migration, and activity toward fluorogenic substrates, but may affect the binding of the activator protein required for GM2 ganglioside hydrolysis. We propose to call these patients the AMB variant of GM2 gangliosidosis to denote the mutation in HEX A but with normal levels of HEX A and B with synthetic substrates. This is to distinguish these patients from those missing the activator protein and normal HEX A and B levels. Images Fig. 1 Fig. 2 PMID:6224417

  15. Collective Flow Enhancement by Tandem Flapping Wings.

    PubMed

    Gravish, Nick; Peters, Jacob M; Combes, Stacey A; Wood, Robert J

    2015-10-30

    We examine the fluid-mechanical interactions that occur between arrays of flapping wings when operating in close proximity at a moderate Reynolds number (Re≈100-1000). Pairs of flapping wings are oscillated sinusoidally at frequency f, amplitude θ_{M}, phase offset ϕ, and wing separation distance D^{*}, and outflow speed v^{*} is measured. At a fixed separation distance, v^{*} is sensitive to both f and ϕ, and we observe both constructive and destructive interference in airspeed. v^{*} is maximized at an optimum phase offset, ϕ_{max}, which varies with wing separation distance, D^{*}. We propose a model of collective flow interactions between flapping wings based on vortex advection, which reproduces our experimental data. PMID:26565499

  16. Collective Flow Enhancement by Tandem Flapping Wings

    NASA Astrophysics Data System (ADS)

    Gravish, Nick; Peters, Jacob M.; Combes, Stacey A.; Wood, Robert J.

    2015-10-01

    We examine the fluid-mechanical interactions that occur between arrays of flapping wings when operating in close proximity at a moderate Reynolds number (Re ≈100 - 1000 ). Pairs of flapping wings are oscillated sinusoidally at frequency f , amplitude θM, phase offset ϕ , and wing separation distance D*, and outflow speed v* is measured. At a fixed separation distance, v* is sensitive to both f and ϕ , and we observe both constructive and destructive interference in airspeed. v* is maximized at an optimum phase offset, ϕmax, which varies with wing separation distance, D*. We propose a model of collective flow interactions between flapping wings based on vortex advection, which reproduces our experimental data.

  17. 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.

  18. Damping in flapping flight and its implications for manoeuvring, scaling and evolution.

    PubMed

    Hedrick, Tyson L

    2011-12-15

    Flying animals exhibit remarkable degrees of both stability and manoeuvrability. Our understanding of these capabilities has recently been improved by the identification of a source of passive damping specific to flapping flight. Examining how this damping effect scales among different species and how it affects active manoeuvres as well as recovery from perturbations provides general insights into the flight of insects, birds and bats. These new damping models offer a means to predict manoeuvrability and stability for a wide variety of flying animals using prior reports of the morphology and flapping motions of these species. Furthermore, the presence of passive damping is likely to have facilitated the evolution of powered flight in animals by providing a stability benefit associated with flapping. PMID:22116750

  19. 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. PMID:26028769

  20. Recombinant Human Peptidoglycan Recognition Proteins Reveal Antichlamydial Activity.

    PubMed

    Bobrovsky, Pavel; Manuvera, Valentin; Polina, Nadezhda; Podgorny, Oleg; Prusakov, Kirill; Govorun, Vadim; Lazarev, Vassili

    2016-07-01

    Peptidoglycan recognition proteins (PGLYRPs) are innate immune components that recognize the peptidoglycan and lipopolysaccharides of bacteria and exhibit antibacterial activity. Recently, the obligate intracellular parasite Chlamydia trachomatis was shown to have peptidoglycan. However, the antichlamydial activity of PGLYRPs has not yet been demonstrated. The aim of our study was to test whether PGLYRPs exhibit antibacterial activity against C. trachomatis Thus, we cloned the regions containing the human Pglyrp1, Pglyrp2, Pglyrp3, and Pglyrp4 genes for subsequent expression in human cell lines. We obtained stable HeLa cell lines that secrete recombinant human PGLYRPs into culture medium. We also generated purified recombinant PGLYRP1, -2, and -4 and confirmed their activities against Gram-positive (Bacillus subtilis) and Gram-negative (Escherichia coli) bacteria. Furthermore, we examined the activities of recombinant PGLYRPs against C. trachomatis and determined their MICs. We also observed a decrease in the infectious ability of chlamydial elementary bodies in the next generation after a single exposure to PGLYRPs. Finally, we demonstrated that PGLYRPs attach to C. trachomatis elementary bodies and activate the expression of the chlamydial two-component stress response system. Thus, PGLYRPs inhibit the development of chlamydial infection. PMID:27160295

  1. The unfolded protein response selectively targets active smoothened mutants.

    PubMed

    Marada, Suresh; Stewart, Daniel P; Bodeen, William J; Han, Young-Goo; Ogden, Stacey K

    2013-06-01

    The Hedgehog signaling pathway, an essential regulator of developmental patterning, has been implicated in playing causative and survival roles in a range of human cancers. The signal-transducing component of the pathway, Smoothened, has revealed itself to be an efficacious therapeutic target in combating oncogenic signaling. However, therapeutic challenges remain in cases where tumors acquire resistance to Smoothened antagonists, and also in cases where signaling is driven by active Smoothened mutants that exhibit reduced sensitivity to these compounds. We previously demonstrated that active Smoothened mutants are subjected to prolonged endoplasmic reticulum (ER) retention, likely due to their mutations triggering conformation shifts that are detected by ER quality control. We attempted to exploit this biology and demonstrate that deregulated Hedgehog signaling driven by active Smoothened mutants is specifically attenuated by ER stressors that induce the unfolded protein response (UPR). Upon UPR induction, active Smoothened mutants are targeted by ER-associated degradation, resulting in attenuation of inappropriate pathway activity. Accordingly, we found that the UPR agonist thapsigargin attenuated mutant Smoothened-induced phenotypes in vivo in Drosophila melanogaster. Wild-type Smoothened and physiological Hedgehog patterning were not affected, suggesting that UPR modulation may provide a novel therapeutic window to be evaluated for targeting active Smoothened mutants in disease. PMID:23572559

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

    PubMed Central

    Zourelidou, Melina; Absmanner, Birgit; Weller, Benjamin; Barbosa, Inês CR; 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

    2014-01-01

    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. DOI: http://dx.doi.org/10.7554/eLife.02860.001 PMID:24948515

  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

    2014-01-01

    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. PMID:24948515

  4. Ribosomal protein S14 negatively regulates c-Myc activity.

    PubMed

    Zhou, Xiang; Hao, Qian; Liao, Jun-Ming; Liao, Peng; Lu, Hua

    2013-07-26

    The ribosomal gene RPS14 is associated with the cancer-prone 5q-syndrome, which is caused by an interstitial deletion of the long arm of human chromosome 5. Previously, we found that ribosomal protein S14 (RPS14) binds to and inactivates MDM2, consequently leading to p53-dependent cell-cycle arrest and growth inhibition. However, it remains elusive whether RPS14 regulates cell proliferation in a p53-independent manner. Here, we show that RPS14 interacts with the Myc homology box II (MBII) and the C-terminal basic helix-loop-helix leucine zipper (bHLH-LZ) domains of the oncoprotein c-Myc. Further, RPS14 inhibited c-Myc transcriptional activity by preventing the recruitment of c-Myc and its cofactor, TRRAP, to the target gene promoters, as thus suppressing c-Myc-induced cell proliferation. Also, siRNA-mediated RPS14 depletion elevated c-Myc transcriptional activity determined by its target gene, Nucleolin, expression. Interestingly, RPS14 depletion also resulted in the induction of c-Myc mRNA and subsequent protein levels. Consistent with this, RPS14 promoted c-Myc mRNA turnover through an Argonaute 2 (Ago2)- and microRNA-mediated pathway. Taken together, our study demonstrates that RPS14 negates c-Myc functions by directly inhibiting its transcriptional activity and mediating its mRNA degradation via miRNA. PMID:23775087

  5. Coq7p relevant residues for protein activity and stability.

    PubMed

    Busso, Cleverson; Ferreira-Júnior, José Ribamar; Paulela, Janaina A; Bleicher, Lucas; Demasi, Marilene; Barros, Mario H

    2015-12-01

    Coenzyme Q (Q) is an isoprenylated benzoquinone electron carrier required for electronic transport in the mitochondrial respiratory chain, shuttling electrons from complexes I and II to complex III. Q synthesis requires proteins termed Coq (Coq1-Coq11). Coq7p is part of the multimeric complex involved in Q synthesis catalyzing the hydroxylation of demethoxy-Q6 (DMQ6), the last monooxygenase step in Q synthesis with a catalytic center containing a carboxylate-bridged di-iron at the active site of the enzyme. Here we indicate a group of Coq7p residues that modulate protein activity: D53, R57, V111 and S114. R57, V111 and S114 are very conserved residues; V111 and S114 are present in separated communities of amino acid correlation analysis. The coq7 double mutant V111G/S114A and S114E show respiratory deficiency at non permissive temperature, DMQ6 accumulation and lower content of Q6. Therefore we conclude that phosphomimetic S114E inhibit Coq7p activity, and propose that S114 phosphorylation is required to move a non-structured loop of 25 amino acids between helix 2 and 3, and that affects the di-iron coordination in Coq7p catalytic center. PMID:26497406

  6. 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. PMID:26631595

  7. Mitogen activated protein kinase at the nuclear pore complex

    PubMed Central

    Faustino, Randolph S; Maddaford, Thane G; Pierce, Grant N

    2011-01-01

    Abstract Mitogen activated protein (MAP) kinases control eukaryotic proliferation, and import of kinases into the nucleus through the nuclear pore complex (NPC) can influence gene expression to affect cellular growth, cell viability and homeostatic function. The NPC is a critical regulatory checkpoint for nucleocytoplasmic traffic that regulates gene expression and cell growth, and MAP kinases may be physically associated with the NPC to modulate transport. In the present study, highly enriched NPC fractions were isolated and investigated for associated kinases and/or activity. Endogenous kinase activity was identified within the NPC fraction, which phosphorylated a 30 kD nuclear pore protein. Phosphomodification of this nucleoporin, here termed Nup30, was inhibited by apigenin and PD-98059, two MAP kinase antagonists as well as with SB-202190, a pharmacological blocker of p38. Furthermore, high throughput profiling of enriched NPCs revealed constitutive presence of all members of the MAP kinase family, extracellular regulated kinases (ERK), p38 and Jun N-terminal kinase. The NPC thus contains a spectrum of associated MAP kinases that suggests an intimate role for ERK and p38 in regulation of nuclear pore function. PMID:20497490

  8. Genome activation by raspberry bushy dwarf virus coat protein.

    PubMed

    Macfarlane, Stuart A; McGavin, Wendy J

    2009-03-01

    Two sets of infectious cDNA clones of raspberry bushy dwarf virus (RBDV) have been constructed, enabling either the synthesis of infectious RNA transcripts or the delivery of infectious binary plasmid DNA by infiltration of Agrobacterium tumefaciens. In whole plants and in protoplasts, inoculation of RBDV RNA1 and RNA2 transcripts led to a low level of infection, which was greatly increased by the addition of RNA3, a subgenomic RNA coding for the RBDV coat protein (CP). Agroinfiltration of RNA1 and RNA2 constructs did not produce a detectable infection but, again, inclusion of a construct encoding the CP led to high levels of infection. Thus, RBDV replication is greatly stimulated by the presence of the CP, a mechanism that also operates with ilarviruses and alfalfa mosaic virus, where it is referred to as genome activation. Mutation to remove amino acids from the N terminus of the CP showed that the first 15 RBDV CP residues are not required for genome activation. Other experiments, in which overlapping regions at the CP N terminus were fused to the monomeric red fluorescent protein, showed that sequences downstream of the first 48 aa are not absolutely required for genome activation. PMID:19218221

  9. Small glutamine-rich protein/viral protein U–binding protein is a novel cochaperone that affects heat shock protein 70 activity

    PubMed Central

    Angeletti, Peter C.; Walker, Doriann; Panganiban, Antonito T.

    2002-01-01

    Molecular chaperone complexes containing heat shock protein (Hsp) 70 and Hsp90 are regulated by cochaperones, including a subclass of regulators, such as Hsp70 interacting protein (Hip), C-terminus of Hsp70 interacting protein (CHIP), and Hsp70-Hsp90 organizing factor (Hop), that contain tetratricopeptide repeats (TPRs), where Hsp70 refers to Hsp70 and its nearly identical constitutive counterpart, Hsc70, together. These proteins interact with the Hsp70 to regulate adenosine triphosphatase (ATPase) and folding activities or to generate the chaperone complex. Here we provide evidence that small glutamine-rich protein/viral protein U–binding protein (SGT/UBP) is a cochaperone that negatively regulates Hsp70. By “Far-Western” and pull-down assays, SGT/UBP was shown to interact directly with Hsp70 and weakly with Hsp90. The interaction of SGT/UBP with both these protein chaperones was mapped to 3 TPRs in SGT/UBP (amino acids 95–195) that are flanked by charged residues. Moreover, SGT/UBP caused an approximately 30% reduction in both the intrinsic ATPase activity of Hsc70 and the ability of Hsc70 to refold denatured luciferase in vitro. This negative effect of SGT/UBP on Hsc70 is similar in magnitude to that observed for the cochaperone CHIP. A role for SGT/UBP in protein folding is also supported by evidence that a yeast strain containing a deletion in the yeast homolog to SGT/UBP (ΔSGT/UBP) displays a 50-fold reduction in recovery from heat shock compared with the wild type parent. Together, these results are consistent with a regulatory role for SGT/UBP in the chaperone complex. PMID:12482202

  10. Ahnak protein activates protein kinase C (PKC) through dissociation of the PKC-protein phosphatase 2A complex.

    PubMed

    Lee, In Hye; Lim, Hee Jung; Yoon, Suhyeon; Seong, Je Kyung; Bae, Duk Soo; Rhee, Sue Goo; Bae, Yun Soo

    2008-03-01

    We have previously reported that central repeated units (CRUs) of Ahnak act as a scaffolding protein networking phospholipase Cgamma and protein kinase C (PKC). Here, we demonstrate that an Ahnak derivative consisting of four central repeated units binds and activates PKC-alpha in a phosphatidylserine/1,2-dioleoyl-sn-glycerol-independent manner. Moreover, NIH3T3 cells expressing the 4 CRUs of Ahnak showed enhanced c-Raf, MEK, and Erk phosphorylation in response to phorbol 12-myristate 13-acetate (PMA) compared with parental cells. To evaluate the effect of loss-of-function of Ahnak in cell signaling, we investigated PKC activation and Raf phosphorylation in embryonic fibroblast cells (MEFs) of the Ahnak knock-out (Ahnak(-/-)) mouse. Membrane translocation of PKC-alpha and phosphorylation of Raf in response to PMA or platelet-derived growth factor were decreased in Ahnak null MEF cells compared with wild type MEFs. Several lines of evidence suggest that PKC-alpha activity is regulated through association with protein phosphatase 2A (PP2A). A co-immunoprecipitation assay indicated that the association of PKC-alpha with PP2A was disrupted in NIH3T3 cells expressing 4 CRUs of Ahnak in response to PMA. Consistently, Ahnak null MEF cells stimulated by PMA showed enhanced PKC-PP2A complex formation, and add-back expression of Ahnak into Ahnak null MEF cells abolished the PKC-PP2A complex formation in response to PMA. These data indicate that Ahnak potentiates PKC activation through inhibiting the interaction of PKC with PP2A. PMID:18174170

  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. 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...

  13. 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...

  14. 50 CFR Figure 16 to Part 223 - Escape Opening and Flap Dimensions for the Double Cover Flap TED

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 10 2012-10-01 2012-10-01 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...

  15. 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...

  16. 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...

  17. 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

  18. Flap-edge aeroacoustic measurements and predictions

    NASA Astrophysics Data System (ADS)

    Brooks, Thomas F.; Humphreys, William M.

    2003-03-01

    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 the prediction models capture much of the physics. 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. The complexity of the directivity results demonstrate the strong role of edge source geometry and frequency in

  19. The trapezius osteomusculocutaneous flap in dogs.

    PubMed

    Philibert, D; Fowler, J D

    1993-01-01

    A pedicled osteomusculocutaneous flap, composed of the cervical part of the trapezius muscle with its overlying skin and the central spine and body of the scapula, was elevated on the prescapular branch of the superficial cervical vascular pedicle in four dogs. The flap was replaced in an orthotopic location. Bone viability was evaluated using histology, fluorescence bone labeling, and angiography. Bone from the scapular spines had a high percentage of viable osteocytes, positive fluorescence, and vessels were outlined in the angiographic study. Bone from the body of the scapula was not viable based on similar criteria. PMID:8116199

  20. 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

  1. 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

  2. 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

  3. Impacts of Activation of the Mitogen-Activated Protein Kinase Pathway in Pancreatic Cancer

    PubMed Central

    Furukawa, Toru

    2015-01-01

    Pancreatic cancer is characterized by constitutive activation of the mitogen-activated protein kinase (MAPK) pathway. Mutations of KRAS or BRAF and epigenetic abrogation of DUSP6 contribute synergistically to the constitutive activation of MAPK. Active MAPK induces the expression of a variety of genes that are thought to play roles in malignant phenotypes of pancreatic cancer. By blocking the functions of such induced genes, it is possible to attenuate the malignant phenotypes. The development of drugs targeting genes downstream of MAPK may provide a novel therapeutic option for pancreatic cancer. PMID:25699241

  4. 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

  5. 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

  6. 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

  7. 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

  8. [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

  9. [The gracilis muscle as musculocutaneous flap. Evaluation of 20 cases].

    PubMed

    Gholam, D; Trevidic, P; Kleimann, P; Hautefeuille, P; Nicoletis, C

    1991-01-01

    The use of gracilis as muscular or myocutaneous flap is very well-known. The authors report 20 cases of gracilis flap including 13 reconstructions of the vaginal cavity following extended abdomino-perineal resection. Some technical points concerning the localization of the cutaneous part of the flap and the pedicle dissection are discussed. The use of gracilis flap is still limited in surgical teams following extended abdomino-perineal resection, nevertheless it is a very useful flap because of its low morbidity, the shortening of patient hospitalization and the very satisfying aesthetic result of the neo-vaginal cavity. PMID:1726389

  10. Long term follow up of neurovascular island flaps.

    PubMed

    Henderson, H P; Reid, D A

    1980-06-01

    The results of a ten year mean follow up of twenty Neurovascular Island Flaps and two Radial Nerve Innervated Cross Finger Flaps are presented. Sensory acuity sufficient for tactile gnosis was achieved in nineteen cases. In only one case had sensory acuity deteriorated since operation. Use of the flap was hampered in one patient by a pre-existing neuroma. Complete sensory reorientation occurred in five patients. Sensory misreference persisted more commonly on dominant hands. It was our impression that Porter's Letter Test revealed the patients making most use of their neurovascular island flaps. The place of neurovascular island flaps in the management of the mutilated hand is discussed. PMID:7409615

  11. Mycobacterium tuberculosis Rv1096 protein: gene cloning, protein expression, and peptidoglycan deacetylase activity

    PubMed Central

    2014-01-01

    Background Many bacteria modulate and evade the immune defenses of their hosts through peptidoglycan (PG) deacetylation. The PG deacetylases from Streptococcus pneumonia, Listeria monocytogenes and Lactococcus lactis have been characterized. However, thus far, the PG deacetylase of Mycobacterium tuberculosis has not been identified. Results In this study, we cloned the Rv1096 gene from the M. tuberculosis H37Rv strain and expressed Rv1096 protein in both Escherichia coli and M. smegmatis. The results showed that the purified Rv1096 protein possessed metallo-dependent PG deacetylase activity, which increased in the presence of Co2+. The kinetic parameters of the PG deacetylase towards M. smegmatis PG as a substrate were as follows: Km, 0.910 ± 0.007 mM; Vmax, 0.514 ± 0.038 μMmin-1; and Kcat = 0.099 ± 0.007 (S-1). Additionally, the viability of M. smegmatis in the presence of over-expressed Rv1096 protein was 109-fold higher than that of wild-type M. smegmatis after lysozyme treatment. Additionally, light microscopy and scanning electron microscopy showed that in the presence of over-expressed Rv1096 protein, M. smegmatis kept its regular shape, with an undamaged cell wall and smooth surface. These results indicate that Rv1096 caused deacetylation of cell wall PG, leading to lysozyme resistance in M. smegmatis. Conclusion We have determined that M. tuberculosis Rv1096 is a PG deacetylase. The PG deacetylase activity of Rv1096 contributed to lysozyme resistance in M. smegmatis. Our findings suggest that deacetylation of cell wall PG may be involved in evasion of host immune defenses by M. tuberculosis. PMID:24975018

  12. 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

  13. Mitogen-activated protein kinases in male reproductive function

    PubMed Central

    Li, Michelle W.M.; Mruk, Dolores D.; Cheng, C. Yan

    2009-01-01

    Recent studies have shown that male reproductive function is modulated via the mitogen-activated protein kinase (MAPK) cascade. The MAPK cascade is involved in numerous male reproductive processes, including spermatogenesis, sperm maturation and activation, capacitation and acrosome reaction, before fertilization of the oocyte. In this review, we discuss the latest findings in this rapidly developing field regarding the role of MAPK in male reproduction in animal models and in human spermatozoa in vitro. This research will facilitate the design of future studies in humans, although much work is needed before this information can be used to manage male infertility and environmental toxicant-induced testicular injury in men, such as blood–testis-barrier disruption. PMID:19303360

  14. 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

  15. 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

  16. 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

  17. Cutaneous necrosis in pregnancy secondary to activated protein C resistance in hereditary angioedema.

    PubMed

    Perkins, W; Downie, I; Keefe, M; Chisholm, M

    1995-04-01

    A 26-year-old woman with hereditary angineurotic oedema (HAE) presented at 22 weeks gestation with severe cutaneous necrosis similar to that seen in coumarin skin necrosis. Protein S deficiency secondary to HAE and pregnancy was postulated. Treatment with heparin, C1-inhibitor concentrates, systemic steroids and surgical debridement resulted in a successful outcome for both mother and child. Subsequent investigations revealed normal levels of protein C, antithrombin III, total protein S, free protein S but reduced function protein S activity with evidence of activated protein C resistance. Cutaneous necrosis has not been reported in associated with activated protein C resistance previously and the possible mechanisms are discussed. PMID:7745572

  18. Bryostatins: potent, new activators of protein kinase C

    SciTech Connect

    Smith, L.; Pettit, G.R.; Smith, J.B.

    1986-03-01

    Bryostatins (B) are a class of 17 macrocyclic lactones that have antineoplastic activity in the murine P388 lymphocytic leukemia system. Bryostatin-1 (B-1) is a potent co-mitogen for the Swiss 3T3 line of murine fibroblasts that have been arrested in G/sub 1//G/sub 0/. B-1 and insulin synergistically increase entry into the S phase of the cell cycle measured autoradiographically as % nuclei labeled with (/sup 3/H)thymidine. A prior treatment of the cells with phorbol 13-myristate 12-acetate (PMA) selectively eliminated the mitogenic response to B-1 or PMA. Conversely, a prior treatment of the cells with B-1 eliminated the mitogenic response to PMA or B-1. Five other B are approximately equipotent to B-1, but B-3 is 5 to 10 times less potent than B-1 as a mitogen. B-1 inhibits the binding of (/sup 3/H)phorbol dibutyrate ((/sup 3/H)PDB) at 4/sup 0/C to a high affinity receptor in the cells. B-3 was also less potent than B-1 as an inhibitor of (/sup 3/H)PDB binding. B-3 differs from B-1 in the diacylglycerol-like component of the molecule. In vitro B-1 and PMA are similarly potent activators of protein kinase C from bovine brain. Further comparisons of the relative activities of the various B are needed to define the structural features that are critical for the activation of protein kinase C which may help in the design of tumor promoter antagonists.

  19. Antistaphylococcal activity of bacteriophage derived chimeric protein P128

    PubMed Central

    2012-01-01

    Background Bacterial drug resistance is one of the most significant challenges to human health today. In particular, effective antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) are urgently needed. A causal relationship between nasal commensal S. aureus and infection has been reported. Accordingly, elimination of nasal S. aureus reduces the risk of infection. Enzymes that degrade bacterial cell walls show promise as antibacterial agents. Bacteriophage-encoded bacterial cell wall-degrading enzymes exhibit intrinsic bactericidal activity. P128 is a chimeric protein that combines the lethal activity of the phage tail-associated muralytic enzyme of Phage K and the staphylococcal cell wall targeting-domain (SH3b) of lysostaphin. Here we report results of in vitro studies evaluating the susceptibility of staphylococcal strains to this novel protein. Results Using the broth microdilution method adapted for lysostaphin, we found that P128 is effective against S. aureus clinical strains including MRSA, methicillin-sensitive S. aureus (MSSA), and a mupirocin-resistant S. aureus. Minimum bactericidal concentrations and minimum inhibitory concentrations of P128 (1-64 μg/mL) were similar across the 32 S. aureus strains tested, demonstrating its bactericidal nature. In time-kill assays, P128 reduced colony-forming units by 99.99% within 1 h and inhibited growth up to 24 h. In an assay simulating topical application of P128 to skin or other biological surfaces, P128 hydrogel was efficacious when layered on cells seeded on solid media. P128 hydrogel was lethal to Staphylococci recovered from nares of healthy people and treated without any processing or culturing steps, indicating its in situ efficacy. This methodology used for in vitro assessment of P128 as an agent for eradicating nasal carriage is unique. Conclusions The novel chimeric protein P128 is a staphylococcal cell wall-degrading enzyme under development for clearance of S. aureus nasal

  20. 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

  1. Negative activation enthalpies in the kinetics of protein folding.

    PubMed

    Oliveberg, M; Tan, Y J; Fersht, A R

    1995-09-12

    Although the rates of chemical reactions become faster with increasing temperature, the converse may be observed with protein-folding reactions. The rate constant for folding initially increases with temperature, goes through a maximum, and then decreases. The activation enthalpy is thus highly temperature dependent because of a large change in specific heat (delta Cp). Such a delta Cp term is usually presumed to be a consequence of a large decrease in exposure of hydrophobic surfaces to water as the reaction proceeds from the denatured state to the transition state for folding: the hydrophobic side chains are surrounded by "icebergs" of water that melt with increasing temperature, thus making a large contribution to the Cp of the denatured state and a smaller one to the more compact transition state. The rate could also be affected by temperature-induced changes in the conformational population of the ground state: the heat required for the progressive melting of residual structure in the denatured state will contribute to delta Cp. By examining two proteins with different refolding mechanisms, we are able to find both of these two processes; barley chymotrypsin inhibitor 2, which refolds from a highly unfolded state, fits well to a hydrophobic interaction model with a constant delta Cp of activation, whereas barnase, which refolds from a more structured denatured state, deviates from this ideal behavior. PMID:7568045

  2. 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

  3. 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. PMID:21639993

  4. Protein Kinase Cδ mediates the activation of Protein Kinase D2 in Platelets

    PubMed Central

    Bhavanasi, Dheeraj; Kim, Soochong; Goldfinger, Lawrence E.; Kunapuli, Satya P.

    2011-01-01

    Protein Kinase D (PKD) is a subfamily of serine/threonine specific family of kinases, comprised of PKD1, PKD2 and PKD3 (PKCμ, PKD2 and PKCν in humans). It is known that PKCs activate PKD, but the relative expression of isoforms of PKD or the specific PKC isoform/s responsible for its activation in platelets is not known. This study is aimed at investigating the pathway involved in activation of PKD in platelets. We show that PKD2 is the major isoform of PKD that is expressed in human as well as murine platelets but not PKD1 or PKD3. PKD2 activation induced by AYPGKF was abolished with a Gq inhibitor YM-254890, but was not affected by Y-27632, a RhoA/p160ROCK inhibitor, indicating that PKD2 activation is Gq-, but not G12/13-mediated Rho-kinase dependent. Calcium-mediated signals are also required for activation of PKD2 as dimethyl BAPTA inhibited its phosphorylation. GF109203X, a pan PKC inhibitor abolished PKD2 phosphorylation but Go6976, a classical PKC inhibitor had no effect suggesting that novel PKC isoforms are involved in PKD2 activation. Importantly, Rottlerin, a non-selective PKCδ inhibitor, inhibited AYPGKF-induced PKD2 activation in human platelets. Similarly, AYPGKF- and Convulxin-induced PKD2 phosphorylation was dramatically inhibited in PKCδ-deficient platelets, but not in PKCθ– or PKCε–deficient murine platelets compared to that of wild type platelets. Hence, we conclude that PKD2 is a common signaling target downstream of various agonist receptors in platelets and Gq-mediated signals along with calcium and novel PKC isoforms, in particular, PKCδ activate PKD2 in platelets. PMID:21736870

  5. Propeller thoracodorsal artery perforator flap for breast reconstruction

    PubMed Central

    Angrigiani, Claudio; Escudero, Ezequiel; Artero, Guillermo; Gercovich, Gustavo; Deza, Ernesto Gil

    2014-01-01

    Background The thoracodorsal artery perforator (TDAP) flap has been described for breast reconstruction. This flap requires intramuscular dissection of the pedicle. A modification of the conventional TDAP surgical technique for breast reconstruction is described, utilizing instead a propeller TDAP flap. The authors present their clinical experience with the propeller TDAP flap in breast reconstruction alone or in combination with expanders or permanent implants. Methods From January 2009 to February 2013, sixteen patients had breast reconstruction utilizing a propeller TDAP flap. Retrospective analysis of patient characteristics, clinical indications, procedure and outcomes were performed. The follow-up period ranged from 4 to 48 months. Results Sixteen patients had breast reconstruction using a TDAP flap with or without simultaneous insertion of an expander or implant. All flaps survived, while two cases required minimal resection due to distal flap necrosis, healing by second intention. There were not donor-site seromas, while minimal wound dehiscence was detected in two cases. Conclusions The propeller TDAP flap appears to be safe and effective for breast reconstruction, resulting in minimal donor site morbidity. The use of this propeller flap emerges as a true alternative to the traditional TDAP flap. PMID:25207210

  6. Rectus abdominis myocutaneous flap for primary vaginal reconstruction.

    PubMed

    Carlson, J W; Soisson, A P; Fowler, J M; Carter, J R; Twiggs, L B; Carson, L F

    1993-12-01

    Reconstructive procedures are being performed with increasing frequency in conjunction with pelvic exenterations and other radical gynecologic surgeries. The most common reconstructive procedures include continent urinary diversion, rectosigmoid anastomosis, and vaginal reconstruction. Historically, the gracilis myocutaneous flap has been the procedure of choice for vaginal reconstruction. However, the gracilis myocutaneous flap has a history of partial to severe necrosis, a propensity to prolapse, and leaves ipsilateral donor scars on the thigh. In contrast, neovaginal reconstruction using a relatively new procedure, the distally based rectus abdominis myocutaneous flaps, has the advantage of using a large, single flap that can be incorporated into the primary incision. This flap is mobilized on a long vascular pedicle, the rectus muscle. In relation to the underlying rectus muscle, the orientation of the cutaneous portion of this flap may be customized to accommodate the pelvic defect or the surgeon's preference. Depending on their primary orientation, they are referred to as either a vertical or transverse rectus abdominis myocutaneous flap. The versatility and reliability of the rectus flap is demonstrated here through the presentation of a small pilot series of seven patients. The technique was used for vaginal reconstruction, primarily in conjunction with pelvic exenteration. The flaps were mobilized from the supraumbilical area and had a flap viability of 100% for the 2 years that they have been followed. There were no postoperative incisional or flap infections. There was one infraumbilical fascial dehiscence. The advantages of primary pelvic reconstruction along with the description of the operative techniques are presented. PMID:8112640

  7. The keystone flap: expanding the dermatologic surgeon's armamentarium.

    PubMed

    Hessam, Schapoor; Sand, Michael; Bechara, Falk G

    2015-01-01

    First described in 2003, the keystone flap has become an established plastic and reconstructive surgery technique for the closure of soft tissue defects following the excision of skin tumors. Complex reconstruction procedures may thus be avoided [3]. In dermatosurgery, however, the keystone flap remains largely unknown.Compared to subcutaneous pedicle flaps, the keystone flap technique warrants better vascular supply by additional-ly preserving musculocutaneous and fasciocutaneous perforator vessels [4]. Moreover, the flap is hyperemic compared to the surrounding skin [5]. Adequate perfusion and its specific design result in high safety of the flap. According to both published data and the authors’ experience, the keystone flap technique is associated with a low complication rate [1, 6].In summary, the keystone flap method yields good aesthetic and functional results by preserving shape and cont-our, avoiding differences in skin coloration and preserving sensitivity (Figure 1b). The flap is particularly well suited for deep defects with exposed bones or tendons, especially on the extremities or the trunk. Alternative closing options such as secondary intention healing, primary closure, or local flaps appear less adequate in these cases. In addition, skin grafting without long-term wound conditioning–for example negative pressure wound therapy–or expensive der-mal replacement products are not very promising. Thus, the keystone flap provides the dermatosurgeon with an effective alternative to reconstruction techniques. PMID:25640508

  8. 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

  9. Delineation of LASIK Flaps with Prednisolone Acetate Eyedrops

    PubMed Central

    Fahd, Daoud C; Fahed, Sharbel D

    2014-01-01

    We describe the use and safety of prednisolone acetate eyedrops at the end of laser in situ keratomileusis (LASIK) to aid proper positioning of the corneal flap. The LASIK flap is created using the preferred technique. Following laser ablation and flap repositioning, one drop of prednisolone acetate is instilled on the eye. This delineates the flap “gutters” and allows perfect flap positioning and centration. We used this technique in 425 eyes undergoing LASIK for correction of spherocylindrical refractive errors. Flap margins were adequately delineated intraoperatively. The only complication related to the use of the steroid suspension was crystal deposition under the flap in one case which resolved completely in 48 hours. PMID:24982743

  10. Aerodynamic characteristics of NACA 4412 airfoil sction with flap

    NASA Astrophysics Data System (ADS)

    Ockfen, Alex E.; Matveev, Konstantin I.

    2009-09-01

    Wing-in-Ground vehicles and aerodynamically assisted boats take advantage of increased lift and reduced drag of wing sections in the ground proximity. At relatively low speeds or heavy payloads of these craft, a flap at the wing trailing-ground-effect flow id numerically investigated in this study. The computational method consists of a steady-state, incompressible, finite volume method utilizing the Spalart-Allmaras turbulence model. Grid generation and solution of the Navier-Stokes equations are completed flow with a flap, as well as ground-effect motion without a flap. Aerodynamic forces are plain flap. Changes in the flow introduced with the flap addition are also discussed. Overall, the use of a flap on wings with small attack angles is found to be beneficial for small flap deflections up to 5% of the chord, where the contribution of lift augmentation exceeds the drag increase, yielding an augmented lift-to-drag ratio

  11. The double opposing myomucosal cheek flap in hard palate reconstruction.

    PubMed

    Pabiszczak, Maciej; Banaszewski, Jacek; Pastusiak, Tomasz; Buczkowska, Agata; Wierzbicka, Małgorzata

    2015-01-01

    Limited defects in the oral cavity can be treated with local and pedicled cheek flaps. It allows to preserve the functions of the resected organ. Large defects in the midline of the hard palate can be reconstructed with double opposing myomucosal cheek flaps. The aim of this study was to discuss the methodology of the flap harvest and to show our experiences of treatment in a group of 15 patients with oral cavity cancer. In 1 patient the double opposing myomucosal cheek flap was harvested due to the wider local defect. The small size of the flap with ability to use the double opposing cheek flap in more extended defects as well as short duration of the surgery procedure can lead to reduced risk of postoperative complications. Finally, cheek flaps form an effective method of treatment of defects in the oral cavity. PMID:26388355

  12. The comparative study on the flapping motion of Venusian magnetotail

    NASA Astrophysics Data System (ADS)

    Rong, Zhaojin; Barabash, Stas; Wieser Stenberg, Gabriella; Futaana, Yoshifumi; Wan, Weixing; Wei, Yong; Wang, Xiaodong; Chai, Lihui; Zhong, Jun

    2016-04-01

    With a newly developed technique and magnetic field measurements obtained by the magnetometer on Venus Express, we study the flapping motion of the Venusian magnetotail. We find that the flapping motion generally comprises contributions both from a nonpropagating steady flapping and a propagating kink-like flapping. The flapping motion tilts the current sheet normal significantly in the plane perpendicular to the Venus-Sun line. The kink-like flapping waves traveling along solar wind electric field or its antidirection can be found in either magnetotail hemisphere where solar wind electric field pointing toward/away. The traveling behaviors suggest that the locations of the triggers for kink-like flappings are near the boundaries between magnetotail current sheet and magnetosheath, not near the central region of magnetotail as is for the Earth's magnetotail.

  13. The split radial forearm flap for lower leg defects.

    PubMed

    van Kampen, Robert J; Corten, Eveline M L; Schellekens, Pascal P A

    2014-11-01

    The conventional free radial forearm flap is a very reliable, long-pedicled flap with thin, pliable skin. These properties make it an excellent choice for high-risk reconstructions or defects requiring only a thin cover. The split radial forearm flap allows primary closure of the donor site and has a large variability in shape and size. In this report, the cutaneous perforators of the radial artery were investigated in fresh cadavers and we present our clinical experience with the split radial artery flap in 9 patients with lower leg defects. Sufficient perforators exist to safely divide the flap proximally and distally into segments. In all clinical cases, the donor site could be closed primarily. All flaps remained viable and 8 of 9 patients obtained an esthetically pleasing result. The split radial forearm free flap is an elegant option for reconstruction of small- to moderate-sized soft tissue defects in the lower extremity. PMID:23657049

  14. 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.

  15. TiAl Scramjet Inlet Flap Subelement Designed and Fabricated

    NASA Technical Reports Server (NTRS)

    Draper, Susan L.

    2004-01-01

    Next-generation launch vehicles are being designed with turbine-based combined cycle (TBCC) propulsion systems having very aggressive thrust/weight targets and long lives. Achievement of these goals requires advanced materials in a wide spectrum of components. TiAl has been identified as a potential backstructure material for maintainable composite panel heat exchangers (HEX) in the inlet, combustor, and nozzle section of a TBCC propulsion system. Weight reduction is the primary objective of this technology. Design tradeoff studies have assessed that a TiAl structure, utilizing a high-strength, hightemperature TiAl alloy called Gamma MET PX,1 reduce weight by 41 to 48 percent in comparison to the baseline Inconel 718 configuration for the TBCC propulsion system inlet, combustor, and nozzle. A collaborative effort between the NASA Glenn Research Center, Pratt & Whitney, Engineering Evaluation & Design, PLANSEE AG (Austria), and the Austrian Space Agency was undertaken to design, manufacture, and validate a Gamma-MET PX TiAl structure for scramjet applications. The TiAl inlet flap was designed with segmented flaps to improve manufacturability, to better control thermal distortion and thermal stresses, and to allow for maintainable HEX segments. The design philosophy was to avoid excessively complicated shapes, to minimize the number of stress concentrations, to keep the part sizes reasonable to match processing capabilities, and to avoid risky processes such as welding. The conceptual design used a standard HEX approach with a double-pass coolant concept for centrally located manifolds. The flowpath side was actively cooled, and an insulation package was placed on the external side to save weight. The inlet flap was analyzed structurally, and local high-stress regions were addressed with local reinforcements.

  16. 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.

  17. Tubularized Penile-Flap Urethroplasty Using a Fasciocutaneous Random Pedicled Flap for Recurrent Anterior Urethral Stricture

    PubMed Central

    Lee, Byung Kwon

    2012-01-01

    This report describes the use of a tubularized random flap for the curative treatment of recurrent anterior urethral stricture. Under the condition of pendulous lithotomy and suprapubic cystostomy, the urethral stricture was removed via a midline ventral penile incision followed by elevation of the flap and insertion of an 18-Fr catheter. Subcutaneous buried interrupted sutures were used to reapproximate the waterproof tubularized neourethra and to coapt with the neourethra and each stump of the urethra, first proximally and then distally. The defect of the penile shaft was covered by advancement of the surrounding scrotal flap. The indwelling catheter was maintained for 21 days. A 9 month postoperative cystoscopy showed no flap necrosis, no mechanical stricture, and no hair growth on the lumen of the neourethra. The patient showed no voiding discomfort 6 months after the operation. The advantages of this procedure are the lack of need for microsurgery, shortening of admission, the use of only spinal anesthesia (no general anesthesia), and a relatively short operative time. The tubularized unilateral penile fasciocutaneous flap should be considered an option for initial flap urethroplasty as a curative technique. PMID:22783537

  18. Histochemical localization of palmitoyl protein thioesterase-1 activity.

    PubMed

    Dearborn, Joshua T; Ramachandran, Subramania; Shyng, Charles; Lu, Jui-Yun; Thornton, Jonah; Hofmann, Sandra L; Sands, Mark S

    2016-02-01

    Infantile neuronal ceroid lipofuscinosis (INCL, Infantile Batten disease) is an invariably fatal neurodegenerative pediatric disorder caused by an inherited mutation in the PPT1 gene. Patients with INCL lack the lysosomal enzyme palmitoyl protein thioesterase-1 (PPT1, EC 3.1.2.22), resulting in intracellular accumulation of autofluorescent storage material and subsequent neuropathology. The Ppt1(-/-) mouse is deficient in PPT1 activity and represents a useful animal model of INCL that recapitulates most of the clinical and pathological aspects of the disease. Preclinical therapeutic experiments performed in the INCL mouse include CNS-directed gene therapy and recombinant enzyme replacement therapy; both seek to re-establish therapeutic levels of the deficient enzyme. We present a novel method for the histochemical localization of PPT1 activity in the Ppt1(-/-) mouse. By utilizing the substrate CUS-9235, tissues known to be positive for PPT1 activity turn varying intensities of blue. Presented here are histochemistry data showing the staining pattern in Ppt1(-/-), wild type, and Ppt1(-/-) mice treated with enzyme replacement therapy or AAV2/9-PPT1-mediated gene therapy. Results are paired with quantitative biochemistry data that confirm the ability of CUS-9235 to detect and localize PPT1 activity. This new method complements the current tools for the study of INCL and evaluation of effective therapies. PMID:26597320

  19. The "open book" flap: a heterodigital cross-finger skin flap and adipofascial flap for coverage of a circumferential soft tissue defect of a digit.

    PubMed

    Tadiparthi, S; Akali, A; Felberg, L

    2009-02-01

    A case of circumferential digital skin loss with exposed tendons from the proximal phalanx to the distal interphalangeal joint is presented. This was treated with a two-layer heterodigital cross-finger ("open book") flap from the adjacent digit, utilising a skin-only cross-finger flap to cover the palmar defect and an adipofascial flap to cover the dorsal defect. PMID:19129359

  20. 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