Structure of a thermophilic F1-ATPase inhibited by an ε-subunit: deeper insight into the ε-inhibition mechanism.

PubMed

Shirakihara, Yasuo; Shiratori, Aya; Tanikawa, Hiromi; Nakasako, Masayoshi; Yoshida, Masasuke; Suzuki, Toshiharu

2015-08-01

F1-ATPase (F1) is the catalytic sector in F(o)F1-ATP synthase that is responsible for ATP production in living cells. In catalysis, its three catalytic β-subunits undergo nucleotide occupancy-dependent and concerted open-close conformational changes that are accompanied by rotation of the γ-subunit. Bacterial and chloroplast F1 are inhibited by their own ε-subunit. In the ε-inhibited Escherichia coli F1 structure, the ε-subunit stabilizes the overall conformation (half-closed, closed, open) of the β-subunits by inserting its C-terminal helix into the α3β3 cavity. The structure of ε-inhibited thermophilic F1 is similar to that of E. coli F1, showing a similar conformation of the ε-subunit, but the thermophilic ε-subunit stabilizes another unique overall conformation (open, closed, open) of the β-subunits. The ε-C-terminal helix 2 and hook are conserved between the two structures in interactions with target residues and in their positions. Rest of the ε-C-terminal domains are in quite different conformations and positions, and have different modes of interaction with targets. This region is thought to serve ε-inhibition differently. For inhibition, the ε-subunit contacts the second catches of some of the β- and α-subunits, the N- and C-terminal helices, and some of the Rossmann fold segments. Those contacts, as a whole, lead to positioning of those β- and α- second catches in ε-inhibition-specific positions, and prevent rotation of the γ-subunit. Some of the structural features are observed even in IF1 inhibition in mitochondrial F1. © 2015 FEBS.

Structure of ratcheted ribosomes with tRNAs in hybrid states

PubMed Central

Julián, Patricia; Konevega, Andrey L.; Scheres, Sjors H. W.; Lázaro, Melisa; Gil, David; Wintermeyer, Wolfgang; Rodnina, Marina V.; Valle, Mikel

2008-01-01

During protein synthesis, tRNAs and mRNA move through the ribosome between aminoacyl (A), peptidyl (P), and exit (E) sites of the ribosome in a process called translocation. Translocation is accompanied by the displacement of the tRNAs on the large ribosomal subunit toward the hybrid A/P and P/E states and by a rotational movement (ratchet) of the ribosomal subunits relative to one another. So far, the structure of the ratcheted state has been observed only when translation factors were bound to the ribosome. Using cryo-electron microscopy and classification, we show here that ribosomes can spontaneously adopt a ratcheted conformation with tRNAs in their hybrid states. The peptidyl-tRNA molecule in the A/P state, which is visualized here, is not distorted compared with the A/A state except for slight adjustments of its acceptor end, suggesting that the displacement of the A-site tRNA on the 50S subunit is passive and is induced by the 30S subunit rotation. Simultaneous subunit ratchet and formation of the tRNA hybrid states precede and may promote the subsequent rapid and coordinated tRNA translocation on the 30S subunit catalyzed by elongation factor G. PMID:18971332

A Viral Packaging Motor Varies Its DNA Rotation and Step Size to Preserve Subunit Coordination as the Capsid Fills

PubMed Central

Tafoya, Sara; Aathavan, K.; Schnitzbauer, Joerg; Grimes, Shelley; Jardine, Paul J.; Bustamante, Carlos

2014-01-01

SUMMARY Multimeric, ring-shaped molecular motors rely on the coordinated action of their subunits to perform crucial biological functions. During these tasks, motors often change their operation in response to regulatory signals. Here, we investigate a viral packaging machine as it fills the capsid with DNA and encounters increasing internal pressure. We find that the motor rotates the DNA during packaging and that the rotation per basepair increases with filling. This change accompanies a reduction in the motor’s step size. We propose that these adjustments preserve motor coordination by allowing one subunit to make periodic, specific, and regulatory contacts with the DNA. At high filling, we also observe the down-regulation of the ATP-binding rate and the emergence of long-lived pauses, suggesting a throttling-down mechanism employed by the motor near the completion of packaging. This study illustrates how a biological motor adjusts its operation in response to changing conditions, while remaining highly coordinated. PMID:24766813

[No interrupted surgical defects of the white upper lip: repair by a combined advancement and rotation flap in the lip subunit].

PubMed

Guillot, P

2013-01-01

A solid understanding of anatomy, basic surgical principles, and tissue movement is essential when undertaking the reconstruction of facial cutaneous surgical defects. Aesthetic facial reconstruction requires understanding ability to use the tissue adjacent to the defect to create a reconstruction that preserves the function of the area and the cosmetic facial units and subunits. The closure of non interrupted white upper lip defects by using a combined advancement and rotation flap is preferred for defects not overtaking 2.5 cm in diameter.

Discovery of a new motion mechanism of biomotors similar to the earth revolving around the sun without rotation

PubMed Central

Guo, Peixuan; Schwartz, Chad; Haak, Jeannie; Zhao, Zhengyi

2013-01-01

Biomotors have been classified into linear and rotational motors. For 35 years, it has been popularly believed that viral dsDNA-packaging apparatuses are pentameric rotation motors. Recently, a third class of hexameric motor has been found in bacteriophage phi29 that utilizes a mechanism of revolution without rotation, friction, coiling, or torque. This review addresses how packaging motors control dsDNA one-way traffic; how four electropositive layers in the channel interact with the electronegative phosphate backbone to generate four steps in translocating one dsDNA helix; how motors resolve the mismatch between 10.5 bases and 12 connector subunits per cycle of revolution; and how ATP regulates sequential action of motor ATPase. Since motors with all number of subunits can utilize the revolution mechanism, this finding helps resolve puzzles and debates concerning the oligomeric nature of packaging motors in many phage systems. This revolution mechanism helps to solve the undesirable dsDNA supercoiling issue involved in rotation. PMID:24074575

Subunit mass analysis for monitoring antibody oxidation.

PubMed

Sokolowska, Izabela; Mo, Jingjie; Dong, Jia; Lewis, Michael J; Hu, Ping

2017-04-01

Methionine oxidation is a common posttranslational modification (PTM) of monoclonal antibodies (mAbs). Oxidation can reduce the in-vivo half-life, efficacy and stability of the product. Peptide mapping is commonly used to monitor the levels of oxidation, but this is a relatively time-consuming method. A high-throughput, automated subunit mass analysis method was developed to monitor antibody methionine oxidation. In this method, samples were treated with IdeS, EndoS and dithiothreitol to generate three individual IgG subunits (light chain, Fd' and single chain Fc). These subunits were analyzed by reversed phase-ultra performance liquid chromatography coupled with an online quadrupole time-of-flight mass spectrometer and the levels of oxidation on each subunit were quantitated based on the deconvoluted mass spectra using the UNIFI software. The oxidation results obtained by subunit mass analysis correlated well with the results obtained by peptide mapping. Method qualification demonstrated that this subunit method had excellent repeatability and intermediate precision. In addition, UNIFI software used in this application allows automated data acquisition and processing, which makes this method suitable for high-throughput process monitoring and product characterization. Finally, subunit mass analysis revealed the different patterns of Fc methionine oxidation induced by chemical and photo stress, which makes it attractive for investigating the root cause of oxidation.

The Phylogenetic Signature Underlying ATP Synthase c-Ring Compliance

DOE PAGES

Pandini, Alessandro; Kleinjung, Jens; Taylor, Willie R.; ...

2015-09-01

The proton-driven ATP synthase (F OF 1) is comprised of two rotary, stepping motors (F O and F 1) coupled by an elastic power transmission. The elastic compliance resides in the rotor module that includes the membrane-embedded FO c-ring. Proton transport by FO is firmly coupled to the rotation of the c-ring relative to other F O subunits (ab 2). It drives ATP synthesis. We used a computational method to investigate the contribution of the c-ring to the total elastic compliance. We performed principal component analysis of conformational ensembles built using distance constraints from the bovine mitochondrial c-ring x-ray structure.more » Angular rotary twist, the dominant ring motion, was estimated to show that the c-ring accounted in part for the measured compliance. Ring rotation was entrained to rotation of the external helix within each hairpin-shaped c-subunit in the ring. Ensembles of monomer and dimers extracted from complete c-rings showed that the coupling between collective ring and the individual subunit motions was independent of the size of the c-ring, which varies between organisms. Molecular determinants were identified by covariance analysis of residue coevolution and structural-alphabet-based local dynamics correlations. The residue coevolution gave a readout of subunit architecture. The dynamic couplings revealed that the hinge for both ring and subunit helix rotations was constructed from the proton-binding site and the adjacent glycine motif (IB-GGGG) in the midmembrane plane. IB-GGGG motifs were linked by long-range couplings across the ring, while intrasubunit couplings connected the motif to the conserved cytoplasmic loop and adjacent segments. The correlation with principal collective motions shows that the couplings underlie both ring rotary and bending motions. Noncontact couplings between IB-GGGG motifs matched the coevolution signal as well as contact couplings. The residue coevolution reflects the physiological importance of the dynamics that may link proton transfer to ring compliance.« less

Self-Assembled Molecular Gear: A 4:1 Complex of Rh(III)Cl Tetraarylporphyrin and Tetra(p-pyridyl)cavitand.

PubMed

Nakamura, Munechika; Kishimoto, Kazuki; Kobori, Yasuhiro; Abe, Tomoka; Yoza, Kenji; Kobayashi, Kenji

2016-09-28

The components of a 4:1 mixture of Rh(III)Cl tetrakis(4-methylphenyl)porphyrin 1 and a bowl-shaped tetra(4-pyridyl)cavitand 4 self-assemble into a 4:1 complex 14•4 via Rh-pyridyl axial coordination bonds. The single-crystal X-ray diffraction analysis and variable-temperature (VT) (1)H NMR study of 14•4 indicated that 14•4 behaves as a quadruple interlocking gear with an inner space, wherein (i) four subunits-1 are gear wheels and four p-pyridyl groups in subunit-4 are axes of gear wheels, (ii) one subunit-1 and two adjacent subunits-1 interlock with one another cooperatively, and (iii) four subunits-1 in 14•4 rotate quickly at 298 K on the NMR time scale. Together, the extremely strong porphyrin-Rh-pyridyl axial coordination bond, the rigidity of the methylene-bridge cavitand as a scaffold of the pyridyl axes, and the cruciform arrangement of the interdigitating p-tolyl groups as the teeth moiety of the gear wheels in the assembling 14-unit make 14•4 function as a quadruple interlocking gear in solution. The gear function of 14•4 was also supported by the rotation behaviors of other 4:1 complexes: 24•4 and 34•4 obtained from Rh(III)Cl tetrakis[4-(4-methylphenyl)phenyl]porphyrin 2 or Rh(III)Cl tetrakis(3,5-dialkoxyphenyl)porphyrin 3 and 4 also served as quadruple interlocking gears, whereas 14•5 obtained from 1 and tetrakis[4-(4-pyridyl)phenyl]cavitand 5 did not behave as a gear. The results of activation parameters (ΔH(⧧), ΔS(⧧), and ΔG(⧧)) obtained from Eyring plots based on line-shape analysis of the VT (1)H NMR spectra of 14•4, 24•4, and 34•4 also support the interlocking rotation (geared coupled rotation) mechanism.

The Phylogenetic Signature Underlying ATP Synthase c-Ring Compliance

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

Pandini, Alessandro; Kleinjung, Jens; Taylor, Willie R.

The proton-driven ATP synthase (F OF 1) is comprised of two rotary, stepping motors (F O and F 1) coupled by an elastic power transmission. The elastic compliance resides in the rotor module that includes the membrane-embedded FO c-ring. Proton transport by FO is firmly coupled to the rotation of the c-ring relative to other F O subunits (ab 2). It drives ATP synthesis. We used a computational method to investigate the contribution of the c-ring to the total elastic compliance. We performed principal component analysis of conformational ensembles built using distance constraints from the bovine mitochondrial c-ring x-ray structure.more » Angular rotary twist, the dominant ring motion, was estimated to show that the c-ring accounted in part for the measured compliance. Ring rotation was entrained to rotation of the external helix within each hairpin-shaped c-subunit in the ring. Ensembles of monomer and dimers extracted from complete c-rings showed that the coupling between collective ring and the individual subunit motions was independent of the size of the c-ring, which varies between organisms. Molecular determinants were identified by covariance analysis of residue coevolution and structural-alphabet-based local dynamics correlations. The residue coevolution gave a readout of subunit architecture. The dynamic couplings revealed that the hinge for both ring and subunit helix rotations was constructed from the proton-binding site and the adjacent glycine motif (IB-GGGG) in the midmembrane plane. IB-GGGG motifs were linked by long-range couplings across the ring, while intrasubunit couplings connected the motif to the conserved cytoplasmic loop and adjacent segments. The correlation with principal collective motions shows that the couplings underlie both ring rotary and bending motions. Noncontact couplings between IB-GGGG motifs matched the coevolution signal as well as contact couplings. The residue coevolution reflects the physiological importance of the dynamics that may link proton transfer to ring compliance.« less

Interacting cytoplasmic loops of subunits a and c of Escherichia coli F1F0 ATP synthase gate H+ transport to the cytoplasm.

PubMed

Steed, P Ryan; Kraft, Kaitlin A; Fillingame, Robert H

2014-11-25

H(+)-transporting F1F0 ATP synthase catalyzes the synthesis of ATP via coupled rotary motors within F0 and F1. H(+) transport at the subunit a-c interface in transmembranous F0 drives rotation of a cylindrical c10 oligomer within the membrane, which is coupled to rotation of subunit γ within the α3β3 sector of F1 to mechanically drive ATP synthesis. F1F0 functions in a reversible manner, with ATP hydrolysis driving H(+) transport. ATP-driven H(+) transport in a select group of cysteine mutants in subunits a and c is inhibited after chelation of Ag(+) and/or Cd(+2) with the substituted sulfhydryl groups. The H(+) transport pathway mapped via these Ag(+)(Cd(+2))-sensitive Cys extends from the transmembrane helices (TMHs) of subunits a and c into cytoplasmic loops connecting the TMHs, suggesting these loop regions could be involved in gating H(+) release to the cytoplasm. Here, using select loop-region Cys from the single cytoplasmic loop of subunit c and multiple cytoplasmic loops of subunit a, we show that Cd(+2) directly inhibits passive H(+) transport mediated by F0 reconstituted in liposomes. Further, in extensions of previous studies, we show that the regions mediating passive H(+) transport can be cross-linked to each other. We conclude that the loop-regions in subunits a and c that are implicated in H(+) transport likely interact in a single structural domain, which then functions in gating H(+) release to the cytoplasm.

Atomic model for the membrane-embedded VO motor of a eukaryotic V-ATPase.

PubMed

Mazhab-Jafari, Mohammad T; Rohou, Alexis; Schmidt, Carla; Bueler, Stephanie A; Benlekbir, Samir; Robinson, Carol V; Rubinstein, John L

2016-11-03

Vacuolar-type ATPases (V-ATPases) are ATP-powered proton pumps involved in processes such as endocytosis, lysosomal degradation, secondary transport, TOR signalling, and osteoclast and kidney function. ATP hydrolysis in the soluble catalytic V 1 region drives proton translocation through the membrane-embedded V O region via rotation of a rotor subcomplex. Variability in the structure of the intact enzyme has prevented construction of an atomic model for the membrane-embedded motor of any rotary ATPase. We induced dissociation and auto-inhibition of the V 1 and V O regions of the V-ATPase by starving the yeast Saccharomyces cerevisiae, allowing us to obtain a ~3.9-Å resolution electron cryomicroscopy map of the V O complex and build atomic models for the majority of its subunits. The analysis reveals the structures of subunits ac 8 c'c″de and a protein that we identify and propose to be a new subunit (subunit f). A large cavity between subunit a and the c-ring creates a cytoplasmic half-channel for protons. The c-ring has an asymmetric distribution of proton-carrying Glu residues, with the Glu residue of subunit c″ interacting with Arg735 of subunit a. The structure suggests sequential protonation and deprotonation of the c-ring, with ATP-hydrolysis-driven rotation causing protonation of a Glu residue at the cytoplasmic half-channel and subsequent deprotonation of a Glu residue at a luminal half-channel.

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

PubMed Central

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

2010-01-01

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

Evidence for rotation of V1-ATPase

PubMed Central

Imamura, Hiromi; Nakano, Masahiro; Noji, Hiroyuki; Muneyuki, Eiro; Ohkuma, Shoji; Yoshida, Masasuke; Yokoyama, Ken

2003-01-01

VoV1-ATPase is responsible for acidification of eukaryotic intracellular compartments and ATP synthesis of Archaea and some eubacteria. From the similarity to FoF1-ATP synthase, VoV1-ATPase has been assumed to be a rotary motor, but to date there are no experimental data to support this. Here we visualized the rotation of single molecules of V1-ATPase, a catalytic subcomplex of VoV1-ATPase. V1-ATPase from Thermus thermophilus was immobilized onto a glass surface, and a bead was attached to the D or F subunit through the biotin-streptavidin linkage. In both cases we observed ATP-dependent rotations of beads, the direction of which was always counterclockwise viewed from the membrane side. Given that three ATP molecules are hydrolyzed per one revolution, rates of rotation agree consistently with rates of ATP hydrolysis at saturating ATP concentrations. This study provides experimental evidence that VoV1-ATPase is a rotary motor and that both D and F subunits constitute a rotor shaft. PMID:12598655

Asymmetry in the F1-ATPase and Its Implications for the Rotational Cycle

PubMed Central

Sun, Sean X.; Wang, Hongyun; Oster, George

2004-01-01

ATP synthase uses a rotary mechanism to carry out its cellular function of manufacturing ATP. The centralγ-shaft rotates inside a hexameric cylinder composed of alternating α- and β-subunits. When operating in the hydrolysis direction under high frictional loads and low ATP concentrations, a coordinated mechanochemical cycle in the three catalytic sites of the β-subunits rotates the γ-shaft in three 120° steps. At low frictional loads, the 120° steps alternate with three ATP-independent substeps separated by ∼30°. We present a quantitative model that accounts for these substeps and show that the observed pauses are due to 1), the asymmetry of the F1 hexamer that produces a propeller-like motion of the power-stroke and 2), the relatively tight binding of ADP to the catalytic sites. PMID:14990467

Anatomy of F1-ATPase powered rotation.

PubMed

Martin, James L; Ishmukhametov, Robert; Hornung, Tassilo; Ahmad, Zulfiqar; Frasch, Wayne D

2014-03-11

F1-ATPase, the catalytic complex of the ATP synthase, is a molecular motor that can consume ATP to drive rotation of the γ-subunit inside the ring of three αβ-subunit heterodimers in 120° power strokes. To elucidate the mechanism of ATPase-powered rotation, we determined the angular velocity as a function of rotational position from single-molecule data collected at 200,000 frames per second with unprecedented signal-to-noise. Power stroke rotation is more complex than previously understood. This paper reports the unexpected discovery that a series of angular accelerations and decelerations occur during the power stroke. The decreases in angular velocity that occurred with the lower-affinity substrate ITP, which could not be explained by an increase in substrate-binding dwells, provides direct evidence that rotation depends on substrate binding affinity. The presence of elevated ADP concentrations not only increased dwells at 35° from the catalytic dwell consistent with competitive product inhibition but also decreased the angular velocity from 85° to 120°, indicating that ADP can remain bound to the catalytic site where product release occurs for the duration of the power stroke. The angular velocity profile also supports a model in which rotation is powered by Van der Waals repulsive forces during the final 85° of rotation, consistent with a transition from F1 structures 2HLD1 and 1H8E (Protein Data Bank).

Anatomy of F1-ATPase powered rotation

PubMed Central

Martin, James L.; Ishmukhametov, Robert; Hornung, Tassilo; Ahmad, Zulfiqar; Frasch, Wayne D.

2014-01-01

F1-ATPase, the catalytic complex of the ATP synthase, is a molecular motor that can consume ATP to drive rotation of the γ-subunit inside the ring of three αβ-subunit heterodimers in 120° power strokes. To elucidate the mechanism of ATPase-powered rotation, we determined the angular velocity as a function of rotational position from single-molecule data collected at 200,000 frames per second with unprecedented signal-to-noise. Power stroke rotation is more complex than previously understood. This paper reports the unexpected discovery that a series of angular accelerations and decelerations occur during the power stroke. The decreases in angular velocity that occurred with the lower-affinity substrate ITP, which could not be explained by an increase in substrate-binding dwells, provides direct evidence that rotation depends on substrate binding affinity. The presence of elevated ADP concentrations not only increased dwells at 35° from the catalytic dwell consistent with competitive product inhibition but also decreased the angular velocity from 85° to 120°, indicating that ADP can remain bound to the catalytic site where product release occurs for the duration of the power stroke. The angular velocity profile also supports a model in which rotation is powered by Van der Waals repulsive forces during the final 85° of rotation, consistent with a transition from F1 structures 2HLD1 and 1H8E (Protein Data Bank). PMID:24567403

Wire-Guide Manipulator For Automated Welding

NASA Technical Reports Server (NTRS)

Morris, Tim; White, Kevin; Gordon, Steve; Emerich, Dave; Richardson, Dave; Faulkner, Mike; Stafford, Dave; Mccutcheon, Kim; Neal, Ken; Milly, Pete

1994-01-01

Compact motor drive positions guide for welding filler wire. Drive part of automated wire feeder in partly or fully automated welding system. Drive unit contains three parallel subunits. Rotations of lead screws in three subunits coordinated to obtain desired motions in three degrees of freedom. Suitable for both variable-polarity plasma arc welding and gas/tungsten arc welding.

Structure and conformational states of the bovine mitochondrial ATP synthase by cryo-EM.

PubMed

Zhou, Anna; Rohou, Alexis; Schep, Daniel G; Bason, John V; Montgomery, Martin G; Walker, John E; Grigorieff, Nikolaus; Rubinstein, John L

2015-10-06

Adenosine triphosphate (ATP), the chemical energy currency of biology, is synthesized in eukaryotic cells primarily by the mitochondrial ATP synthase. ATP synthases operate by a rotary catalytic mechanism where proton translocation through the membrane-inserted FO region is coupled to ATP synthesis in the catalytic F1 region via rotation of a central rotor subcomplex. We report here single particle electron cryomicroscopy (cryo-EM) analysis of the bovine mitochondrial ATP synthase. Combining cryo-EM data with bioinformatic analysis allowed us to determine the fold of the a subunit, suggesting a proton translocation path through the FO region that involves both the a and b subunits. 3D classification of images revealed seven distinct states of the enzyme that show different modes of bending and twisting in the intact ATP synthase. Rotational fluctuations of the c8-ring within the FO region support a Brownian ratchet mechanism for proton-translocation-driven rotation in ATP synthases.

Role of the DELSEED Loop in Torque Transmission of F1-ATPase

PubMed Central

Tanigawara, Mizue; Tabata, Kazuhito V.; Ito, Yuko; Ito, Jotaro; Watanabe, Rikiya; Ueno, Hiroshi; Ikeguchi, Mitsunori; Noji, Hiroyuki

2012-01-01

F1-ATPase is an ATP-driven rotary motor that generates torque at the interface between the catalytic β-subunits and the rotor γ-subunit. The β-subunit inwardly rotates the C-terminal domain upon nucleotide binding/dissociation; hence, the region of the C-terminal domain that is in direct contact with γ—termed the DELSEED loop—is thought to play a critical role in torque transmission. We substituted all the DELSEED loop residues with alanine to diminish specific DELSEED loop-γ interactions and with glycine to disrupt the loop structure. All the mutants rotated unidirectionally with kinetic parameters comparable to those of the wild-type F1, suggesting that the specific interactions between DELSEED loop and γ is not involved in cooperative interplays between the catalytic β-subunits. Glycine substitution mutants generated half the torque of the wild-type F1, whereas the alanine mutant generated comparable torque. Fluctuation analyses of the glycine/alanine mutants revealed that the γ-subunit was less tightly held in the α3β3-stator ring of the glycine mutant than in the wild-type F1 and the alanine mutant. Molecular dynamics simulation showed that the DELSEED loop was disordered by the glycine substitution, whereas it formed an α-helix in the alanine mutant. Our results emphasize the importance of loop rigidity for efficient torque transmissions. PMID:23009846

Molecular mechanics of 30S subunit head rotation.

PubMed

Mohan, Srividya; Donohue, John Paul; Noller, Harry F

2014-09-16

During ribosomal translocation, a process central to the elongation phase of protein synthesis, movement of mRNA and tRNAs requires large-scale rotation of the head domain of the small (30S) subunit of the ribosome. It has generally been accepted that the head rotates by pivoting around the neck helix (h28) of 16S rRNA, its sole covalent connection to the body domain. Surprisingly, we observe that the calculated axis of rotation does not coincide with the neck. Instead, comparative structure analysis across 55 ribosome structures shows that 30S head movement results from flexing at two hinge points lying within conserved elements of 16S rRNA. Hinge 1, although located within the neck, moves by straightening of the kinked helix h28 at the point of contact with the mRNA. Hinge 2 lies within a three-way helix junction that extends to the body through a second, noncovalent connection; its movement results from flexing between helices h34 and h35 in a plane orthogonal to the movement of hinge 1. Concerted movement at these two hinges accounts for the observed magnitudes of head rotation. Our findings also explain the mode of action of spectinomycin, an antibiotic that blocks translocation by binding to hinge 2.

Molecular mechanics of 30S subunit head rotation

PubMed Central

Mohan, Srividya; Donohue, John Paul; Noller, Harry F.

2014-01-01

During ribosomal translocation, a process central to the elongation phase of protein synthesis, movement of mRNA and tRNAs requires large-scale rotation of the head domain of the small (30S) subunit of the ribosome. It has generally been accepted that the head rotates by pivoting around the neck helix (h28) of 16S rRNA, its sole covalent connection to the body domain. Surprisingly, we observe that the calculated axis of rotation does not coincide with the neck. Instead, comparative structure analysis across 55 ribosome structures shows that 30S head movement results from flexing at two hinge points lying within conserved elements of 16S rRNA. Hinge 1, although located within the neck, moves by straightening of the kinked helix h28 at the point of contact with the mRNA. Hinge 2 lies within a three-way helix junction that extends to the body through a second, noncovalent connection; its movement results from flexing between helices h34 and h35 in a plane orthogonal to the movement of hinge 1. Concerted movement at these two hinges accounts for the observed magnitudes of head rotation. Our findings also explain the mode of action of spectinomycin, an antibiotic that blocks translocation by binding to hinge 2. PMID:25187561

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

PubMed

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

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

Structure and conformational states of the bovine mitochondrial ATP synthase by cryo-EM

PubMed Central

Zhou, Anna; Rohou, Alexis; Schep, Daniel G; Bason, John V; Montgomery, Martin G; Walker, John E; Grigorieff, Nikolaus; Rubinstein, John L

2015-01-01

Adenosine triphosphate (ATP), the chemical energy currency of biology, is synthesized in eukaryotic cells primarily by the mitochondrial ATP synthase. ATP synthases operate by a rotary catalytic mechanism where proton translocation through the membrane-inserted FO region is coupled to ATP synthesis in the catalytic F1 region via rotation of a central rotor subcomplex. We report here single particle electron cryomicroscopy (cryo-EM) analysis of the bovine mitochondrial ATP synthase. Combining cryo-EM data with bioinformatic analysis allowed us to determine the fold of the a subunit, suggesting a proton translocation path through the FO region that involves both the a and b subunits. 3D classification of images revealed seven distinct states of the enzyme that show different modes of bending and twisting in the intact ATP synthase. Rotational fluctuations of the c8-ring within the FO region support a Brownian ratchet mechanism for proton-translocation-driven rotation in ATP synthases. DOI: http://dx.doi.org/10.7554/eLife.10180.001 PMID:26439008

Escherichia coli F1Fo-ATP synthase with a b/δ fusion protein allows analysis of the function of the individual b subunits.

PubMed

Gajadeera, Chathurada S; Weber, Joachim

2013-09-13

The "stator stalk" of F1Fo-ATP synthase is essential for rotational catalysis as it connects the nonrotating portions of the enzyme. In Escherichia coli, the stator stalk consists of two (identical) b subunits and the δ subunit. In mycobacteria, one of the b subunits and the δ subunit are replaced by a b/δ fusion protein; the remaining b subunit is of the shorter b' type. In the present study, it is shown that it is possible to generate a functional E. coli ATP synthase containing a b/δ fusion protein. This construct allowed the analysis of the roles of the individual b subunits. The full-length b subunit (which in this case is covalently linked to δ in the fusion protein) is responsible for connecting the stalk to the catalytic F1 subcomplex. It is not required for interaction with the membrane-embedded Fo subcomplex, as its transmembrane helix can be removed. Attachment to Fo is the function of the other b subunit which in turn has only a minor (if any at all) role in binding to δ. Also in E. coli the second b subunit can be shortened to a b' type.

Structural characterization of ribosome recruitment and translocation by type IV IRES.

PubMed

Murray, Jason; Savva, Christos G; Shin, Byung-Sik; Dever, Thomas E; Ramakrishnan, V; Fernández, Israel S

2016-05-09

Viral mRNA sequences with a type IV IRES are able to initiate translation without any host initiation factors. Initial recruitment of the small ribosomal subunit as well as two translocation steps before the first peptidyl transfer are essential for the initiation of translation by these mRNAs. Using electron cryomicroscopy (cryo-EM) we have structurally characterized at high resolution how the Cricket Paralysis Virus Internal Ribosomal Entry Site (CrPV-IRES) binds the small ribosomal subunit (40S) and the translocation intermediate stabilized by elongation factor 2 (eEF2). The CrPV-IRES restricts tvhe otherwise flexible 40S head to a conformation compatible with binding the large ribosomal subunit (60S). Once the 60S is recruited, the binary CrPV-IRES/80S complex oscillates between canonical and rotated states (Fernández et al., 2014; Koh et al., 2014), as seen for pre-translocation complexes with tRNAs. Elongation factor eEF2 with a GTP analog stabilizes the ribosome-IRES complex in a rotated state with an extra ~3 degrees of rotation. Key residues in domain IV of eEF2 interact with pseudoknot I (PKI) of the CrPV-IRES stabilizing it in a conformation reminiscent of a hybrid tRNA state. The structure explains how diphthamide, a eukaryotic and archaeal specific post-translational modification of a histidine residue of eEF2, is involved in translocation.

Spontaneous quaternary and tertiary T-R transitions of human hemoglobin in molecular dynamics simulation.

PubMed

Hub, Jochen S; Kubitzki, Marcus B; de Groot, Bert L

2010-05-06

We present molecular dynamics simulations of unliganded human hemoglobin (Hb) A under physiological conditions, starting from the R, R2, and T state. The simulations were carried out with protonated and deprotonated HC3 histidines His(beta)146, and they sum up to a total length of 5.6 micros. We observe spontaneous and reproducible T-->R quaternary transitions of the Hb tetramer and tertiary transitions of the alpha and beta subunits, as detected from principal component projections, from an RMSD measure, and from rigid body rotation analysis. The simulations reveal a marked asymmetry between the alpha and beta subunits. Using the mutual information as correlation measure, we find that the beta subunits are substantially more strongly linked to the quaternary transition than the alpha subunits. In addition, the tertiary populations of the alpha and beta subunits differ substantially, with the beta subunits showing a tendency towards R, and the alpha subunits showing a tendency towards T. Based on the simulation results, we present a transition pathway for coupled quaternary and tertiary transitions between the R and T conformations of Hb.

Spontaneous Quaternary and Tertiary T-R Transitions of Human Hemoglobin in Molecular Dynamics Simulation

PubMed Central

de Groot, Bert L.

2010-01-01

We present molecular dynamics simulations of unliganded human hemoglobin (Hb) A under physiological conditions, starting from the R, R2, and T state. The simulations were carried out with protonated and deprotonated HC3 histidines His(β)146, and they sum up to a total length of 5.6µs. We observe spontaneous and reproducible T→R quaternary transitions of the Hb tetramer and tertiary transitions of the α and β subunits, as detected from principal component projections, from an RMSD measure, and from rigid body rotation analysis. The simulations reveal a marked asymmetry between the α and β subunits. Using the mutual information as correlation measure, we find that the β subunits are substantially more strongly linked to the quaternary transition than the α subunits. In addition, the tertiary populations of the α and β subunits differ substantially, with the β subunits showing a tendency towards R, and the α subunits showing a tendency towards T. Based on the simulation results, we present a transition pathway for coupled quaternary and tertiary transitions between the R and T conformations of Hb. PMID:20463873

Revolution rather than rotation of AAA+ hexameric phi29 nanomotor for viral dsDNA packaging without coiling☆

PubMed Central

Schwartz, Chad; De Donatis, Gian Marco; Zhang, Hui; Fang, Huaming; Guo, Peixuan

2013-01-01

It has long been believed that the DNA-packaging motor of dsDNA viruses utilizes a rotation mechanism. Here we report a revolution rather than rotation mechanism for the bacteriophage phi29 DNA packaging motor. The phi29 motor contains six copies of the ATPase (Schwartz et al., this issue); ATP binding to one ATPase subunit stimulates the ATPase to adopt a conformation with a high affinity for dsDNA. ATP hydrolysis induces a new conformation with a lower affinity, thus transferring the dsDNA to an adjacent subunit by a power stroke. DNA revolves unidirectionally along the hexameric channel wall of the ATPase, but neither the dsDNA nor the ATPase itself rotates along its own axis. One ATP is hydrolyzed in each transitional step, and six ATPs are consumed for one helical turn of 360°. Transition of the same dsDNA chain along the channel wall, but at a location 60° different from the last contact, urges dsDNA to move forward 1.75 base pairs each step (10.5 bp per turn/6ATP=1.75 bp per ATP). Each connector subunit tilts with a left-handed orientation at a 30° angle in relation to its vertical axis that runs anti-parallel to the right-handed dsDNA helix, facilitating the one-way traffic of dsDNA. The connector channel has been shown to cause four steps of transition due to four positively charged lysine rings that make direct contact with the negatively charged DNA phosphate backbone. Translocation of dsDNA into the procapsid by revolution avoids the difficulties during rotation that are associated with DNA supercoiling. Since the revolution mechanism can apply to any stoichiometry, this motor mechanism might reconcile the stoichiometry discrepancy in many phage systems where the ATPase has been found as a tetramer, hexamer, or nonamer. PMID:23763768

Structural characterization of ribosome recruitment and translocation by type IV IRES

PubMed Central

Murray, Jason; Savva, Christos G; Shin, Byung-Sik; Dever, Thomas E; Ramakrishnan, V; Fernández, Israel S

2016-01-01

Viral mRNA sequences with a type IV IRES are able to initiate translation without any host initiation factors. Initial recruitment of the small ribosomal subunit as well as two translocation steps before the first peptidyl transfer are essential for the initiation of translation by these mRNAs. Using electron cryomicroscopy (cryo-EM) we have structurally characterized at high resolution how the Cricket Paralysis Virus Internal Ribosomal Entry Site (CrPV-IRES) binds the small ribosomal subunit (40S) and the translocation intermediate stabilized by elongation factor 2 (eEF2). The CrPV-IRES restricts the otherwise flexible 40S head to a conformation compatible with binding the large ribosomal subunit (60S). Once the 60S is recruited, the binary CrPV-IRES/80S complex oscillates between canonical and rotated states (Fernández et al., 2014; Koh et al., 2014), as seen for pre-translocation complexes with tRNAs. Elongation factor eEF2 with a GTP analog stabilizes the ribosome-IRES complex in a rotated state with an extra ~3 degrees of rotation. Key residues in domain IV of eEF2 interact with pseudoknot I (PKI) of the CrPV-IRES stabilizing it in a conformation reminiscent of a hybrid tRNA state. The structure explains how diphthamide, a eukaryotic and archaeal specific post-translational modification of a histidine residue of eEF2, is involved in translocation. DOI: http://dx.doi.org/10.7554/eLife.13567.001 PMID:27159451

Allosteric mechanism controls traffic in the chaperone/usher pathway.

PubMed

Di Yu, Xiao; Dubnovitsky, Anatoly; Pudney, Alex F; Macintyre, Sheila; Knight, Stefan D; Zavialov, Anton V

2012-11-07

Many virulence organelles of Gram-negative bacterial pathogens are assembled via the chaperone/usher pathway. The chaperone transports organelle subunits across the periplasm to the outer membrane usher, where they are released and incorporated into growing fibers. Here, we elucidate the mechanism of the usher-targeting step in assembly of the Yersinia pestis F1 capsule at the atomic level. The usher interacts almost exclusively with the chaperone in the chaperone:subunit complex. In free chaperone, a pair of conserved proline residues at the beginning of the subunit-binding loop form a "proline lock" that occludes the usher-binding surface and blocks usher binding. Binding of the subunit to the chaperone rotates the proline lock away from the usher-binding surface, allowing the chaperone-subunit complex to bind to the usher. We show that the proline lock exists in other chaperone/usher systems and represents a general allosteric mechanism for selective targeting of chaperone:subunit complexes to the usher and for release and recycling of the free chaperone. Copyright © 2012 Elsevier Ltd. All rights reserved.

Eukaryotic RNA polymerase subunit RPB8 is a new relative of the OB family.

PubMed

Krapp, S; Kelly, G; Reischl, J; Weinzierl, R O; Matthews, S

1998-02-01

RNA polymerase II subunit RPB8 is an essential subunit that is highly conserved throughout eukaryotic evolution and is present in all three types of nuclear RNA polymerases. We report the first high resolution structural insight into eukaryotic RNA polymerase architecture with the solution structure of RPB8 from Saccharomyces cerevisiae. It consists of an eight stranded, antiparallel beta-barrel, four short helical regions and a large, unstructured omega-loop. The strands are connected in classic Greek-key fashion. The overall topology is unusual and contains a striking C2 rotational symmetry. Furthermore, it is most likely a novel associate of the oligonucleotide/oligosaccharide (OB) binding protein class.

The structure of the protein phosphatase 2A PR65/A subunit reveals the conformation of its 15 tandemly repeated HEAT motifs.

PubMed

Groves, M R; Hanlon, N; Turowski, P; Hemmings, B A; Barford, D

1999-01-08

The PR65/A subunit of protein phosphatase 2A serves as a scaffolding molecule to coordinate the assembly of the catalytic subunit and a variable regulatory B subunit, generating functionally diverse heterotrimers. Mutations of the beta isoform of PR65 are associated with lung and colon tumors. The crystal structure of the PR65/Aalpha subunit, at 2.3 A resolution, reveals the conformation of its 15 tandemly repeated HEAT sequences, degenerate motifs of approximately 39 amino acids present in a variety of proteins, including huntingtin and importin beta. Individual motifs are composed of a pair of antiparallel alpha helices that assemble in a mainly linear, repetitive fashion to form an elongated molecule characterized by a double layer of alpha helices. Left-handed rotations at three interrepeat interfaces generate a novel left-hand superhelical conformation. The protein interaction interface is formed from the intrarepeat turns that are aligned to form a continuous ridge.

Ensemble cryo-EM uncovers inchworm-like translocation of a viral IRES through the ribosome

PubMed Central

Abeyrathne, Priyanka D; Koh, Cha San; Grant, Timothy; Grigorieff, Nikolaus; Korostelev, Andrei A

2016-01-01

Internal ribosome entry sites (IRESs) mediate cap-independent translation of viral mRNAs. Using electron cryo-microscopy of a single specimen, we present five ribosome structures formed with the Taura syndrome virus IRES and translocase eEF2•GTP bound with sordarin. The structures suggest a trajectory of IRES translocation, required for translation initiation, and provide an unprecedented view of eEF2 dynamics. The IRES rearranges from extended to bent to extended conformations. This inchworm-like movement is coupled with ribosomal inter-subunit rotation and 40S head swivel. eEF2, attached to the 60S subunit, slides along the rotating 40S subunit to enter the A site. Its diphthamide-bearing tip at domain IV separates the tRNA-mRNA-like pseudoknot I (PKI) of the IRES from the decoding center. This unlocks 40S domains, facilitating head swivel and biasing IRES translocation via hitherto-elusive intermediates with PKI captured between the A and P sites. The structures suggest missing links in our understanding of tRNA translocation. DOI: http://dx.doi.org/10.7554/eLife.14874.001 PMID:27159452

Structure and mechanism of the ATP synthase membrane motor inferred from quantitative integrative modeling.

PubMed

Leone, Vanessa; Faraldo-Gómez, José D

2016-12-01

Two subunits within the transmembrane domain of the ATP synthase-the c-ring and subunit a-energize the production of 90% of cellular ATP by transducing an electrochemical gradient of H + or Na + into rotational motion. The nature of this turbine-like energy conversion mechanism has been elusive for decades, owing to the lack of definitive structural information on subunit a or its c-ring interface. In a recent breakthrough, several structures of this complex were resolved by cryo-electron microscopy (cryo-EM), but the modest resolution of the data has led to divergent interpretations. Moreover, the unexpected architecture of the complex has cast doubts on a wealth of earlier biochemical analyses conducted to probe this structure. Here, we use quantitative molecular-modeling methods to derive a structure of the a-c complex that is not only objectively consistent with the cryo-EM data, but also with correlated mutation analyses of both subunits and with prior cross-linking and cysteine accessibility measurements. This systematic, integrative approach reveals unambiguously the topology of subunit a and its relationship with the c-ring. Mapping of known Cd 2+ block sites and conserved protonatable residues onto the structure delineates two noncontiguous pathways across the complex, connecting two adjacent proton-binding sites in the c-ring to the space on either side of the membrane. The location of these binding sites and of a strictly conserved arginine on subunit a, which serves to prevent protons from hopping between them, explains the directionality of the rotary mechanism and its strict coupling to the proton-motive force. Additionally, mapping of mutations conferring resistance to oligomycin unexpectedly reveals that this prototypical inhibitor may bind to two distinct sites at the a-c interface, explaining its ability to block the mechanism of the enzyme irrespective of the direction of rotation of the c-ring. In summary, this study is a stepping stone toward establishing the mechanism of the ATP synthase at the atomic level.

Cooperative macromolecular device revealed by meta-analysis of static and time-resolved structures

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

Ren, Zhong; Šrajer, Vukica; Knapp, James E.

2013-04-08

Here we present a meta-analysis of a large collection of static structures of a protein in the Protein Data Bank in order to extract the progression of structural events during protein function. We apply this strategy to the homodimeric hemoglobin HbI from Scapharca inaequivalvis. We derive a simple dynamic model describing how binding of the first ligand in one of the two chemically identical subunits facilitates a second binding event in the other partner subunit. The results of our ultrafast time-resolved crystallographic studies support this model. We demonstrate that HbI functions like a homodimeric mechanical device, such as pliers ormore » scissors. Ligand-induced motion originating in one subunit is transmitted to the other via conserved pivot points, where the E and F' helices from two partner subunits are 'bolted' together to form a stable dimer interface permitting slight relative rotation but preventing sliding.« less

Lightweight Deployable Mirrors with Tensegrity Supports

NASA Technical Reports Server (NTRS)

Zeiders, Glenn W.; Bradford, Larry J.; Cleve, Richard C.

2004-01-01

The upper part of Figure 1 shows a small-scale prototype of a developmental class of lightweight, deployable structures that would support panels in precise alignments. In this case, the panel is hexagonal and supports disks that represent segments of a primary mirror of a large telescope. The lower part of Figure 1 shows a complete conceptual structure containing multiple hexagonal panels that hold mirror segments. The structures of this class are of the tensegrity type, which was invented five decades ago by artist Kenneth Snelson. A tensegrity structure consists of momentfree compression members (struts) and tension members (cables). The structures of this particular developmental class are intended primarily as means to erect large segmented primary mirrors of astronomical telescopes or large radio antennas in outer space. Other classes of tensegrity structures could also be designed for terrestrial use as towers, masts, and supports for general structural panels. An important product of the present development effort is the engineering practice of building a lightweight, deployable structure as an assembly of tensegrity modules like the one shown in Figure 2. This module comprises two octahedral tensegrity subunits that are mirror images of each other joined at their plane of mirror symmetry. In this case, the plane of mirror symmetry is both the upper plane of the lower subunit and the lower plane of the upper subunit, and is delineated by the midheight triangle in Figure 2. In the configuration assumed by the module to balance static forces under mild loading, the upper and lower planes of each sub-unit are rotated about 30 , relative to each other, about the long (vertical) axis of the structure. Larger structures can be assembled by joining multiple modules like this one at their sides or ends. When the module is compressed axially (vertically), the first-order effect is an increase in the rotation angle, but by virtue of the mirror arrangement, the net first-order rotation between the uppermost and lowermost planes is zero. The need to have zero net rotation between these planes under all loading conditions in a typical practical structure is what prompts the use of the mirror configuration. Force and moment loadings other than simple axial compression produce only second-order deformations through strains in the struts and cables.

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

PubMed

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

2007-03-01

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

Protein kinase A catalytic subunit primed for action: Time-lapse crystallography of Michaelis complex formation

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

Das, Amit; Gerlits, Oksana O.; Parks, Jerry M.

The catalytic subunit of the cyclic AMP-dependent protein kinase A (PKAc) catalyzes the transfer of the γ-phosphate of bound Mg 2ATP to a serine or threonine residue of a protein substrate. Here, time-lapse X-ray crystallography was used to capture a series of complexes of PKAc with an oligopeptide substrate and unreacted Mg 2ATP, including the Michaelis complex, that reveal important geometric rearrangements in and near the active site preceding the phosphoryl transfer reaction. Contrary to the prevailing view, Mg 2+ binds first to the M1 site as a complex with ATP and is followed by Mg 2+ binding to themore » M2 site. Furthermore, the target serine hydroxyl of the peptide substrate rotates away from the active site toward the bulk solvent, which breaks the hydrogen bond with D166. In conclusion, the serine hydroxyl of the substrate rotates back toward D166 to form the Michaelis complex with the active site primed for phosphoryl transfer.« less

Protein kinase A catalytic subunit primed for action: Time-lapse crystallography of Michaelis complex formation

DOE PAGES

Das, Amit; Gerlits, Oksana O.; Parks, Jerry M.; ...

2015-11-12

The catalytic subunit of the cyclic AMP-dependent protein kinase A (PKAc) catalyzes the transfer of the γ-phosphate of bound Mg 2ATP to a serine or threonine residue of a protein substrate. Here, time-lapse X-ray crystallography was used to capture a series of complexes of PKAc with an oligopeptide substrate and unreacted Mg 2ATP, including the Michaelis complex, that reveal important geometric rearrangements in and near the active site preceding the phosphoryl transfer reaction. Contrary to the prevailing view, Mg 2+ binds first to the M1 site as a complex with ATP and is followed by Mg 2+ binding to themore » M2 site. Furthermore, the target serine hydroxyl of the peptide substrate rotates away from the active site toward the bulk solvent, which breaks the hydrogen bond with D166. In conclusion, the serine hydroxyl of the substrate rotates back toward D166 to form the Michaelis complex with the active site primed for phosphoryl transfer.« less

Interacting helical surfaces of the transmembrane segments of subunits a and c' of the yeast V-ATPase defined by disulfide-mediated cross-linking.

PubMed

Kawasaki-Nishi, Shoko; Nishi, Tsuyoshi; Forgac, Michael

2003-10-24

Proton translocation by the vacuolar (H+)-ATPase (or V-ATPase) has been shown by mutagenesis to be dependent upon charged residues present within transmembrane segments of subunit a as well as the three proteolipid subunits (c, c', and c"). Interaction between R735 in TM7 of subunit a and the glutamic acid residue in the middle of TM4 of subunits c and c' or TM2 of subunit c" has been proposed to be essential for proton release to the luminal compartment. In order to determine whether the helical face of TM7 of subunit a containing R735 is capable of interacting with the helical face of TM4 of subunit c' containing the essential glutamic acid residue (Glu-145), cysteine-mediated cross-linking between these subunits in yeast has been performed. Cys-less forms of subunits a and c' as well as forms containing unique cysteine residues were constructed, introduced together into a strain disrupted in both endogenous subunits, and tested for growth at neutral pH, for assembly competence and for cross-linking in the presence of cupric-phenanthroline by SDS-PAGE and Western blot analysis. Four different cysteine mutants of subunit a were each tested pairwise with ten different unique cysteine mutants of subunit c'. Strong cross-linking was observed for the pairs aS728C/c'I142C, aA731C/c'E145C, aA738C/c'F143C, aA738C/c'L147C, and aL739C/c'L147C. Partial cross-linking was observed for an additional 13 of 40 pairs analyzed. When arrayed on a helical wheel diagram, the results suggest that the helical face of TM7 of subunit a containing Arg-735 interacts with the helical face of TM4 of subunit c' centered on Val-146 and bounded by Glu-145 and Leu-147. The results are consistent with a possible rotational flexibility of one or both of these transmembrane segments as well as some flexibility of movement perpendicular to the membrane.

Towards elucidation of the mechanism of biological nanomotors

NASA Astrophysics Data System (ADS)

Zhao, Zhengyi

Biological functions such as cell mitosis, bacterial binary fission, DNA replication or repair, homologous recombination, Holliday junction resolution, viral genome packaging, and cell entry all involve biomotor-driven DNA translocation. In the past, the ubiquitous biological nanomotors were classified into two categories: linear and rotation motors. In 2013, we discovered a third type of biomotor, revolving motor without rotation. The revolving motion is further found to be widespread among many biological systems. In addition, the detailed sequential action mechanism of the ATPase ring in the phi29 dsDNA packaging motor has been elucidated: ATP binding induces a conformational entropy alternation of ATPase to a high affinity toward dsDNA; ATP hydrolysis triggers another conformational entropy change in ATPase to a low DNA affinity, by which the dsDNA substrate is pushed toward an adjacent ATPase subunit. The subunit communication is regulated by an arginine finger that extends from one ATPase subunit to the adjacent unit, resulting in an asymmetrical hexameric organization. Continuation of this process promotes the movement and revolving of the dsDNA within the hexameric ATPase ring. Coordination of all the motor components facilitate the motion direction control of the viral DNA packaging motors, and make it unusually powerful and effective. KEYWORDS: Phi29 dsDNA Packaging Motor, Bio-nanomotor, RNA Nanotechnology, DNA Translocase, One-Way Revolving, ASCE Superfamily, AAA+ Superfamily.

Essential arginine in subunit a and aspartate in subunit c of FoF1 ATP synthase: effect of repositioning within helix 4 of subunit a and helix 2 of subunit c.

PubMed

Langemeyer, Lars; Engelbrecht, Siegfried

2007-07-01

FoF1 ATP synthase couples proton flow through the integral membrane portion Fo (ab2c10) to ATP-synthesis in the extrinsic F1-part ((alphabeta)3gammadeltaepsilon) (Escherichia coli nomenclature and stoichiometry). Coupling occurs by mechanical rotation of subunits c10gammaepsilon relative to (alphabeta)3deltaab2. Two residues were found to be essential for proton flow through ab2c10, namely Arg210 in subunit a (aR210) and Asp61 in subunits c (cD61). Their deletion abolishes proton flow, but "horizontal" repositioning, by anchoring them in adjacent transmembrane helices, restores function. Here, we investigated the effects of "vertical" repositioning aR210, cD61, or both by one helical turn towards the N- or C-termini of their original helices. Other than in the horizontal the vertical displacement changes the positions of the side chains within the depth of the membrane. Mutant aR210A/aN214R appeared to be short-circuited in that it supported proton conduction only through EF1-depleted EFo, but not in EFoEF1, nor ATP-driven proton pumping. Mutant cD61N/cM65D grew on succinate, retained the ability to synthesize ATP and supported passive proton conduction but apparently not ATP hydrolysis-driven proton pumping.

Controlled clockwise and anticlockwise rotational switching of a molecular motor.

PubMed

Perera, U G E; Ample, F; Kersell, H; Zhang, Y; Vives, G; Echeverria, J; Grisolia, M; Rapenne, G; Joachim, C; Hla, S-W

2013-01-01

The design of artificial molecular machines often takes inspiration from macroscopic machines. However, the parallels between the two systems are often only superficial, because most molecular machines are governed by quantum processes. Previously, rotary molecular motors powered by light and chemical energy have been developed. In electrically driven motors, tunnelling electrons from the tip of a scanning tunnelling microscope have been used to drive the rotation of a simple rotor in a single direction and to move a four-wheeled molecule across a surface. Here, we show that a stand-alone molecular motor adsorbed on a gold surface can be made to rotate in a clockwise or anticlockwise direction by selective inelastic electron tunnelling through different subunits of the motor. Our motor is composed of a tripodal stator for vertical positioning, a five-arm rotor for controlled rotations, and a ruthenium atomic ball bearing connecting the static and rotational parts. The directional rotation arises from sawtooth-like rotational potentials, which are solely determined by the internal molecular structure and are independent of the surface adsorption site.

Functional conservation of RNA polymerase II in fission and budding yeasts.

PubMed

Shpakovski, G V; Gadal, O; Labarre-Mariotte, S; Lebedenko, E N; Miklos, I; Sakurai, H; Proshkin, S A; Van Mullem, V; Ishihama, A; Thuriaux, P

2000-02-04

The complementary DNAs of the 12 subunits of fission yeast (Schizosaccharomyces pombe) RNA polymerase II were expressed from strong promoters in Saccharomyces cerevisiae and tested for heterospecific complementation by monitoring their ability to replace in vivo the null mutants of the corresponding host genes. Rpb1 and Rpb2, the two largest subunits and Rpb8, a small subunit shared by all three polymerases, failed to support growth in S. cerevisiae. The remaining nine subunits were all proficient for heterospecific complementation and led in most cases to a wild-type level of growth. The two alpha-like subunits (Rpb3 and Rpb11), however, did not support growth at high (37 degrees C) or low (25 degrees C) temperatures. In the case of Rpb3, growth was restored by increasing the gene dosage of the host Rpb11 or Rpb10 subunits, confirming previous evidence of a close genetic interaction between these three subunits. Copyright 2000 Academic Press.

Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria.

PubMed

Formosa, Luke E; Hofer, Annette; Tischner, Christin; Wenz, Tina; Ryan, Michael T

2016-01-01

In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.

Mitochondrial Genome Integrity Mutations Uncouple the Yeast Saccharomyces cerevisiae ATP Synthase*║

PubMed Central

Wang, Yamin; Singh, Usha; Mueller, David M.

2013-01-01

The mitochondrial ATP synthase is a molecular motor, which couples the flow of rotons with phosphorylation of ADP. Rotation of the central stalk within the core of ATP synthase effects conformational changes in the active sites driving the synthesis of ATP. Mitochondrial genome integrity (mgi) mutations have been previously identified in the α-, β-, and γ-subunits of ATP synthase in yeast Kluyveromyces lactis and trypanosome Trypanosoma brucei. These mutations reverse the lethality of the loss of mitochondrial DNA in petite negative strains. Introduction of the homologous mutations in Saccharomyces cerevisiae results in yeast strains that lose mitochondrial DNA at a high rate and accompanied decreases in the coupling of the ATP synthase. The structure of yeast F1-ATPase reveals that the mgi residues cluster around the γ-subunit and selectively around the collar region of F1. These results indicate that residues within the mgi complementation group are necessary for efficient coupling of ATP synthase, possibly acting as a support to fix the axis of rotation of the central stalk. PMID:17244612

Kinetics of acrylodan-labelled cAMP-dependent protein kinase catalytic subunit denaturation.

PubMed

Kivi, Rait; Loog, Mart; Jemth, Per; Järv, Jaak

2013-10-01

Fluorescence spectroscopy was used to study denaturation of cAMP-dependent protein kinase catalytic subunit labeled with an acrylodan moiety. The dye was covalently bound to a cystein residue introduced into the enzyme by replacement of arginine in position 326 in the native sequence, located near the enzyme active center. This labeling had no effect on catalytic activity of the enzyme, but provided possibility to monitor changes in protein structure through measuring the fluorescence spectrum of the dye, which is sensitive to changes in its environment. This method was used to monitor denaturation of the protein kinase catalytic subunit and study the kinetics of this process as well as influence of specific ligands on stability of the protein. Stabilization of the enzyme structure was observed in the presence of adenosine triphosphate, peptide substrate RRYSV and inhibitor peptide PKI[5-24].

None of the Rotor Residues of F1-ATPase Are Essential for Torque Generation

PubMed Central

Chiwata, Ryohei; Kohori, Ayako; Kawakami, Tomonari; Shiroguchi, Katsuyuki; Furuike, Shou; Adachi, Kengo; Sutoh, Kazuo; Yoshida, Masasuke; Kinosita, Kazuhiko

2014-01-01

F1-ATPase is a powerful rotary molecular motor that can rotate an object several hundred times as large as the motor itself against the viscous friction of water. Forced reverse rotation has been shown to lead to ATP synthesis, implying that the mechanical work against the motor’s high torque can be converted into the chemical energy of ATP. The minimal composition of the motor protein is α3β3γ subunits, where the central rotor subunit γ turns inside a stator cylinder made of alternately arranged α3β3 subunits using the energy derived from ATP hydrolysis. The rotor consists of an axle, a coiled coil of the amino- and carboxyl-terminal α-helices of γ, which deeply penetrates the stator cylinder, and a globular protrusion that juts out from the stator. Previous work has shown that, for a thermophilic F1, significant portions of the axle can be truncated and the motor still rotates a submicron sized bead duplex, indicating generation of up to half the wild-type (WT) torque. Here, we inquire if any specific interactions between the stator and the rest of the rotor are needed for the generation of a sizable torque. We truncated the protruding portion of the rotor and replaced part of the remaining axle residues such that every residue of the rotor has been deleted or replaced in this or previous truncation mutants. This protrusionless construct showed an unloaded rotary speed about a quarter of the WT, and generated one-third to one-half of the WT torque. No residue-specific interactions are needed for this much performance. F1 is so designed that the basic rotor-stator interactions for torque generation and control of catalysis rely solely upon the shape and size of the rotor at very low resolution. Additional tailored interactions augment the torque to allow ATP synthesis under physiological conditions. PMID:24853745

Stoichiometry for α-bungarotoxin block of α7 acetylcholine receptors

NASA Astrophysics Data System (ADS)

Dacosta, Corrie J. B.; Free, Chris R.; Sine, Steven M.

2015-08-01

α-Bungarotoxin (α-Btx) binds to the five agonist binding sites on the homopentameric α7-acetylcholine receptor, yet the number of bound α-Btx molecules required to prevent agonist-induced channel opening remains unknown. To determine the stoichiometry for α-Btx blockade, we generate receptors comprised of wild-type and α-Btx-resistant subunits, tag one of the subunit types with conductance mutations to report subunit stoichiometry, and following incubation with α-Btx, monitor opening of individual receptor channels with defined subunit stoichiometry. We find that a single α-Btx-sensitive subunit confers nearly maximal suppression of channel opening, despite four binding sites remaining unoccupied by α-Btx and accessible to the agonist. Given structural evidence that α-Btx locks the agonist binding site in an inactive conformation, we conclude that the dominant mechanism of antagonism is non-competitive, originating from conformational arrest of the binding sites, and that the five α7 subunits are interdependent and maintain conformational symmetry in the open channel state.

Free-Energy Landscape of Reverse tRNA Translocation through the Ribosome Analyzed by Electron Microscopy Density Maps and Molecular Dynamics Simulations

PubMed Central

Ishida, Hisashi; Matsumoto, Atsushi

2014-01-01

To understand the mechanism of reverse tRNA translocation in the ribosome, all-atom molecular dynamics simulations of the ribosome-tRNAs-mRNA-EFG complex were performed. The complex at the post-translocational state was directed towards the translocational and pre-translocational states by fitting the complex into cryo-EM density maps. Between a series of the fitting simulations, umbrella sampling simulations were performed to obtain the free-energy landscape. Multistep structural changes, such as a ratchet-like motion and rotation of the head of the small subunit were observed. The free-energy landscape showed that there were two main free-energy barriers: one between the post-translocational and intermediate states, and the other between the pre-translocational and intermediate states. The former corresponded to a clockwise rotation, which was coupled to the movement of P-tRNA over the P/E-gate made of G1338, A1339 and A790 in the small subunit. The latter corresponded to an anticlockwise rotation of the head, which was coupled to the location of the two tRNAs in the hybrid state. This indicates that the coupled motion of the head rotation and tRNA translocation plays an important role in opening and closing of the P/E-gate during the ratchet-like movement in the ribosome. Conformational change of EF-G was interpreted to be the result of the combination of the external motion by L12 around an axis passing near the sarcin-ricin loop, and internal hinge-bending motion. These motions contributed to the movement of domain IV of EF-G to maintain its interaction with A/P-tRNA. PMID:24999999

Free-energy landscape of reverse tRNA translocation through the ribosome analyzed by electron microscopy density maps and molecular dynamics simulations.

PubMed

Ishida, Hisashi; Matsumoto, Atsushi

2014-01-01

To understand the mechanism of reverse tRNA translocation in the ribosome, all-atom molecular dynamics simulations of the ribosome-tRNAs-mRNA-EFG complex were performed. The complex at the post-translocational state was directed towards the translocational and pre-translocational states by fitting the complex into cryo-EM density maps. Between a series of the fitting simulations, umbrella sampling simulations were performed to obtain the free-energy landscape. Multistep structural changes, such as a ratchet-like motion and rotation of the head of the small subunit were observed. The free-energy landscape showed that there were two main free-energy barriers: one between the post-translocational and intermediate states, and the other between the pre-translocational and intermediate states. The former corresponded to a clockwise rotation, which was coupled to the movement of P-tRNA over the P/E-gate made of G1338, A1339 and A790 in the small subunit. The latter corresponded to an anticlockwise rotation of the head, which was coupled to the location of the two tRNAs in the hybrid state. This indicates that the coupled motion of the head rotation and tRNA translocation plays an important role in opening and closing of the P/E-gate during the ratchet-like movement in the ribosome. Conformational change of EF-G was interpreted to be the result of the combination of the external motion by L12 around an axis passing near the sarcin-ricin loop, and internal hinge-bending motion. These motions contributed to the movement of domain IV of EF-G to maintain its interaction with A/P-tRNA.

Correlation between the conformational states of F1-ATPase as determined from its crystal structure and single-molecule rotation

PubMed Central

Okuno, Daichi; Fujisawa, Ryo; Iino, Ryota; Hirono-Hara, Yoko; Imamura, Hiromi; Noji, Hiroyuki

2008-01-01

F1-ATPase is a rotary molecular motor driven by ATP hydrolysis that rotates the γ-subunit against the α3β3 ring. The crystal structures of F1, which provide the structural basis for the catalysis mechanism, have shown essentially 1 stable conformational state. In contrast, single-molecule studies have revealed that F1 has 2 stable conformational states: ATP-binding dwell state and catalytic dwell state. Although structural and single-molecule studies are crucial for the understanding of the molecular mechanism of F1, it remains unclear as to which catalytic state the crystal structure represents. To address this issue, we introduced cysteine residues at βE391 and γR84 of F1 from thermophilic Bacillus PS3. In the crystal structures of the mitochondrial F1, the corresponding residues in the ADP-bound β (βDP) and γ were in direct contact. The βE190D mutation was additionally introduced into the β to slow ATP hydrolysis. By incorporating a single copy of the mutant β-subunit, the chimera F1, α3β2β(E190D/E391C)γ(R84C), was prepared. In single-molecule rotation assay, chimera F1 showed a catalytic dwell pause in every turn because of the slowed ATP hydrolysis of β(E190D/E391C). When the mutant β and γ were cross-linked through a disulfide bond between βE391C and γR84C, F1 paused the rotation at the catalytic dwell angle of β(E190D/E391C), indicating that the crystal structure represents the catalytic dwell state and that βDP is the catalytically active form. The former point was again confirmed in experiments where F1 rotation was inhibited by adenosine-5′-(β,γ-imino)-triphosphate and/or azide, the most commonly used inhibitors for the crystallization of F1. PMID:19075235

Crystallization of the c14-rotor of the chloroplast ATP synthase reveals that it contains pigments

PubMed Central

Varco-Merth, Benjamin; Fromme, Raimund; Wang, Meitian; Fromme, Petra

2012-01-01

The ATP synthase is one of the most important enzymes on earth as it couples the transmembrane electrochemical potential of protons to the synthesis of ATP from ADP and inorganic phosphate, providing the main ATP source of almost all higher life on earth. During ATP synthesis, stepwise protonation of a conserved carboxylate on each protein subunit of an oligomeric ring of 10–15 c-subunits is commonly thought to drive rotation of the rotor moiety (c10–14γε) relative to stator moiety (α3β3δab2). Here we report the isolation and crystallization of the c14-ring of subunit c from the spinach chloroplast enzyme diffracting as far as 2.8 Å. Though ATP synthase was not previously known to contain any pigments, the crystals of the c-subunit possessed a strong yellow color. The pigment analysis revealed that they contain 1 chlorophyll and 2 carotenoids, thereby showing for the first time that the chloroplast ATP synthase contains cofactors, leading to the question of the possible roles of the functions of the pigments in the chloroplast ATP synthase. PMID:18515064

Errors of car wheels rotation rate measurement using roller follower on test benches

NASA Astrophysics Data System (ADS)

Potapov, A. S.; Svirbutovich, O. A.; Krivtsov, S. N.

2018-03-01

The article deals with rotation rate measurement errors, which depend on the motor vehicle rate, on the roller, test benches. Monitoring of the vehicle performance under operating conditions is performed on roller test benches. Roller test benches are not flawless. They have some drawbacks affecting the accuracy of vehicle performance monitoring. Increase in basic velocity of the vehicle requires increase in accuracy of wheel rotation rate monitoring. It determines the degree of accuracy of mode identification for a wheel of the tested vehicle. To ensure measurement accuracy for rotation velocity of rollers is not an issue. The problem arises when measuring rotation velocity of a car wheel. The higher the rotation velocity of the wheel is, the lower the accuracy of measurement is. At present, wheel rotation frequency monitoring on roller test benches is carried out by following-up systems. Their sensors are rollers following wheel rotation. The rollers of the system are not kinematically linked to supporting rollers of the test bench. The roller follower is forced against the wheels of the tested vehicle by means of a spring-lever mechanism. Experience of the test bench equipment operation has shown that measurement accuracy is satisfactory at small rates of vehicles diagnosed on roller test benches. With a rising diagnostics rate, rotation velocity measurement errors occur in both braking and pulling modes because a roller spins about a tire tread. The paper shows oscillograms of changes in wheel rotation velocity and rotation velocity measurement system’s signals when testing a vehicle on roller test benches at specified rates.

Study on the non-contact FBG vibration sensor and its application

NASA Astrophysics Data System (ADS)

Li, Tianliang; Tan, Yuegang; Zhou, Zude; Cai, Li; Liu, Sai; He, Zhongting; Zheng, Kai

2015-06-01

A non-contact vibration sensor based on the fiber Bragg grating (FBG) sensor has been presented, and it is used to monitor the vibration of rotating shaft. In the paper, we describe the principle of the sensor and make some experimental analyses. The analysis results show that the sensitivity and linearity of the sensor are -1.5 pm/μm and 4.11% within a measuring range of 2 mm-2.6 mm, respectively. When it is used to monitor the vibration of the rotating shaft, the analysis signals of vibration of the rotating shaft and the critical speed of rotation obtained are the same as that obtained from the eddy current sensor. It verifies that the sensor can be used for the non-contact measurement of vibration of the rotating shaft system and for fault monitoring and diagnosis of rotating machinery.

Detachment strength of human osteoblasts cultured on hydroxyapatite with various surface roughness. Contribution of integrin subunits.

PubMed

Kokkinos, Petros A; Koutsoukos, Petros G; Deligianni, Despina D

2012-06-01

Hydroxyapatite (HA) has been widely used as a bone substitute in dental, maxillofacial and orthopaedic surgery and as osteoconductive bone substitute or precoating of pedicle screws and cages in spine surgery. The aim of the present study was to investigate the osteoblastic adhesion strength on HA substrata with different surface topography and biochemistry (pre-adsorption of fibronectin) after blocking of specific integrin subunits with monoclonal antibodies. Stoichiometric HA was prepared by precipitation followed by ageing and characterized by SEM, EDX, powder XRD, Raman spectroscopy, TGA, and specific surface area analysis. Human bone marrow derived osteoblasts were cultured on HA disc-shaped substrata which were sintered and polished resulting in two surface roughness grades. For attachment evaluation, cells were incubated with monoclonal antibodies and seeded for 2 h on the substrata. Cell detachment strength was determined using a rotating disc device. Cell detachment strength was surface roughness, fibronectin preadsorption and intergin subunit sensitive.

Crystallization of isoelectrically homogeneous cholera toxin

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

Spangler, B.D.; Westbrook, E.M.

1989-02-07

Past difficulty in growing good crystals of cholera toxin has prevented the study of the crystal structure of this important protein. The authors have determined that failure of cholera toxin to crystallize well has been due to its heterogeneity. They have now succeeded in overcoming the problem by isolating a single isoelectric variant of this oligomeric protein (one A subunit and five B subunits). Cholera toxin purified by their procedure readily forms large single crystals. The crystal form has been described previously. They have recorded data from native crystals of cholera toxin to 3.0-{angstrom} resolution with our electronic area detectors.more » With these data, they have found the orientation of a 5-fold symmetry axis within these crystals, perpendicular to the screw dyad of the crystal. They are now determining the crystal structure of cholera toxin by a combination of multiple heavy-atom isomorphous replacement and density modification techniques, making use of rotational 5-fold averaging of the B subunits.« less

Revealing an outward-facing open conformational state in a CLC Cl –/H + exchange transporter

DOE PAGES

Khantwal, Chandra M.; Abraham, Sherwin J.; Han, Wei; ...

2016-01-22

CLC secondary active transporters exchange Cl - for H + . Crystal structures have suggested that the conformational change from occluded to outward-facing states is unusually simple, involving only the rotation of a conserved glutamate (Glu ex ) upon its protonation. Using 19 F NMR, we show that as [H + ] is increased to protonate Glu ex and enrich the outward-facing state, a residue ~20 Å away from Glu ex , near the subunit interface, moves from buried to solvent-exposed. Consistent with functional relevance of this motion, constriction via inter-subunit cross-linking reduces transport. Molecular dynamics simulations indicate that themore » cross-link dampens extracellular gate-opening motions. In support of this model, mutations that decrease steric contact between Helix N (part of the extracellular gate) and Helix P (at the subunit interface) remove the inhibitory effect of the cross-link. Together, these results demonstrate the formation of a previously uncharacterized 'outward-facing open' state, and highlight the relevance of global structural changes in CLC function.« less

The Large Ring Laser G for Continuous Earth Rotation Monitoring

NASA Astrophysics Data System (ADS)

Schreiber, K. U.; Klügel, T.; Velikoseltsev, A.; Schlüter, W.; Stedman, G. E.; Wells, J.-P. R.

2009-09-01

Ring Laser gyroscopes exploit the Sagnac effect and measure rotations absolute. They do not require an external reference frame and therefore provide an independent method to monitor Earth rotation. Large-scale versions of these gyroscopes promise to eventually provide a similar high resolution for the measurement of the variations in the Earth rotation rate as the established methods based on VLBI and GNSS. This would open the door to a continuous monitoring of LOD (Length of Day) and polar motion, which is not yet available today. Another advantage is the access to the sub-daily frequency regime of Earth rotation. The ring laser “G” (Grossring), located at the Geodetic Observatory Wettzell (Germany) is the most advanced realization of such a large gyroscope. This paper outlines the current sensor design and properties.

Mechanism of the αβ Conformational Change in F1-ATPase after ATP Hydrolysis: Free-Energy Simulations

PubMed Central

Ito, Yuko; Ikeguchi, Mitsunori

2015-01-01

One of the motive forces for F1-ATPase rotation is the conformational change of the catalytically active β subunit due to closing and opening motions caused by ATP binding and hydrolysis, respectively. The closing motion is accomplished in two steps: the hydrogen-bond network around ATP changes and then the entire structure changes via B-helix sliding, as shown in our previous study. Here, we investigated the opening motion induced by ATP hydrolysis using all-atom free-energy simulations, combining the nudged elastic band method and umbrella sampling molecular-dynamics simulations. Because hydrolysis requires residues in the α subunit, the simulations were performed with the αβ dimer. The results indicate that the large-scale opening motion is also achieved by the B-helix sliding (in the reverse direction). However, the sliding mechanism is different from that of ATP binding because sliding is triggered by separation of the hydrolysis products ADP and Pi. We also addressed several important issues: 1), the timing of the product Pi release; 2), the unresolved half-closed β structure; and 3), the ADP release mechanism. These issues are fundamental for motor function; thus, the rotational mechanism of the entire F1-ATPase is also elucidated through this αβ study. During the conformational change, conserved residues among the ATPase proteins play important roles, suggesting that the obtained mechanism may be shared with other ATPase proteins. When combined with our previous studies, these results provide a comprehensive view of the β-subunit conformational change that drives the ATPase. PMID:25564855

Observation of the noncovalent assembly and disassembly pathways of the chaperone complex MtGimC by mass spectrometry

PubMed Central

Fändrich, Marcus; Tito, Mark A.; Leroux, Michel R.; Rostom, Adam A.; Hartl, F. Ulrich; Dobson, Christopher M.; Robinson, Carol V.

2000-01-01

We have analyzed a newly described archaeal GimC/prefoldin homologue, termed MtGimC, by using nanoflow electrospray coupled with time-of-flight MS. The molecular weight of the complex from Methanobacterium thermoautotrophicum corresponds to a well-defined hexamer of two α subunits and four β subunits. Dissociation of the complex within the gas phase reveals a quaternary arrangement of two central subunits, both α, and four peripheral β subunits. By constructing a thermally controlled nanoflow device, we have monitored the thermal stability of the complex by MS. The results of these experiments demonstrate that a significant proportion of the MtGimC hexamer remains intact under low-salt conditions at elevated temperatures. This finding is supported by data from CD spectroscopy, which show that at physiological salt concentrations, the complex remains stable at temperatures above 65°C. Mass spectrometric methods were developed to monitor in real time the assembly of the MtGimC hexamer from its component subunits. By using this methodology, the mass spectra recorded throughout the time course of the experiment showed the absence of any significantly populated intermediates, demonstrating that the assembly process is highly cooperative. Taken together, these data show that the complex is stable under the elevated temperatures that are appropriate for its hyperthermophile host and demonstrate that the assembly pathway leads exclusively to the hexamer, which is likely to be a structural unit in vivo. PMID:11087821

The cAMP-induced G protein subunits dissociation monitored in live Dictyostelium cells by BRET reveals two activation rates, a positive effect of caffeine and potential role of microtubules.

PubMed

Tariqul Islam, A F M; Yue, Haicen; Scavello, Margarethakay; Haldeman, Pearce; Rappel, Wouter-Jan; Charest, Pascale G

2018-08-01

To study the dynamics and mechanisms controlling activation of the heterotrimeric G protein Gα2βγ in Dictyostelium in response to stimulation by the chemoattractant cyclic AMP (cAMP), we monitored the G protein subunit interaction in live cells using bioluminescence resonance energy transfer (BRET). We found that cAMP induces the cAR1-mediated dissociation of the G protein subunits to a similar extent in both undifferentiated and differentiated cells, suggesting that only a small number of cAR1 (as expressed in undifferentiated cells) is necessary to induce the full activation of Gα2βγ. In addition, we found that treating cells with caffeine increases the potency of cAMP-induced Gα2βγ activation; and that disrupting the microtubule network but not F-actin inhibits the cAMP-induced dissociation of Gα2βγ. Thus, microtubules are necessary for efficient cAR1-mediated activation of the heterotrimeric G protein. Finally, kinetics analyses of Gα2βγ subunit dissociation induced by different cAMP concentrations indicate that there are two distinct rates at which the heterotrimeric G protein subunits dissociate when cells are stimulated with cAMP concentrations above 500 nM versus only one rate at lower cAMP concentrations. Quantitative modeling suggests that the kinetics profile of Gα2βγ subunit dissociation results from the presence of both uncoupled and G protein pre-coupled cAR1 that have differential affinities for cAMP and, consequently, induce G protein subunit dissociation through different rates. We suggest that these different signaling kinetic profiles may play an important role in initial chemoattractant gradient sensing. Copyright © 2018 Elsevier Inc. All rights reserved.

Interallelic complementation of mutations in propionic acidemia by microinjection of mutant cDNAs into fibroblasts of affected patients

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

Loyer, M.; Leclerc, D.; Gravel, R.A.

1994-09-01

Propionic acidemia is a rare autosomal recessive disorder resulting from defects of the {alpha} or {beta} subunit of biotin-dependent propionyl-CoA carboxylase (PCC). Mutations are assigned to defects of the PCCA ({alpha} subunit) or PCCB ({beta} subunit) gene through complementation studies after somatic fusion of patient cell lines. About two-thirds of patients with {beta} subunit defects (complementation group pccBC) show interallelic complementation in cell fusion experiments (subgroups pccB and pccC), monitored by the PCC-dependent metabolisms of {sup 14}C-propionate. Most patient cell lines are heteroallelic for two different mutations, leaving ambiguous the identity of the mutation participating in interallelic complementation. To identifymore » the complementing mutations, we have expressed {beta}-subunit cDNAs containing individual mutations by microinjection of the cDNAs in recipient cells from patients with {beta} subunit defects. Correction of the PCC defect was monitored by autoradiography of {sup 14}C-propionate incorporation. In some experiments, cDNAs were co-injected with a plasmid expressing the E. coli lacZ gene as a positive control for successful injection. Two mutations from the pccB subgroup showed complementation when injected into pccC cells; dupKICK140-143 and Pro228Leu. Similarly, two mutations from the pccC subgroup complemented after injection into pccB cells; {Delta}Ile408 and Arg410Trp. No mutation complemented with mutation of the pccBC group which are classified as non-complementing in cell fusion experiments. The results show that the complementing pccB mutations are found in the N-terminal half of the {beta} subunit, while the complementing pccC mutations cluxter at a site in the C-terminal half. The latter site is a candidate for the propionyl-CoA binding site based on sequence identity with a region of transcarboxylase from Propionibacterium shermanii.« less

An unexpected twist in viral capsid maturation

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

Gertsman, Ilya; Gan, Lu; Guttman, Miklos

2009-04-14

Lambda-like double-stranded (ds) DNA bacteriophage undergo massive conformational changes in their capsid shell during the packaging of their viral genomes. Capsid shells are complex organizations of hundreds of protein subunits that assemble into intricate quaternary complexes that ultimately are able to withstand over 50 atm of pressure during genome packaging. The extensive integration between subunits in capsids requires the formation of an intermediate complex, termed a procapsid, from which individual subunits can undergo the necessary refolding and structural rearrangements needed to transition to the more stable capsid. Although various mature capsids have been characterized at atomic resolution, no such procapsidmore » structure is available for a dsDNA virus or bacteriophage. Here we present a procapsid X-ray structure at 3.65 {angstrom} resolution, termed prohead II, of the lambda-like bacteriophage HK97, the mature capsid structure of which was previously solved to 3.44 {angstrom}. A comparison of the two largely different capsid forms has unveiled an unprecedented expansion mechanism that describes the transition. Crystallographic and hydrogen/deuterium exchange data presented here demonstrate that the subunit tertiary structures are significantly different between the two states, with twisting and bending motions occurring in both helical and -sheet regions. We also identified subunit interactions at each three-fold axis of the capsid that are maintained throughout maturation. The interactions sustain capsid integrity during subunit refolding and provide a fixed hinge from which subunits undergo rotational and translational motions during maturation. Previously published calorimetric data of a closely related bacteriophage, P22, showed that capsid maturation was an exothermic process that resulted in a release of 90 kJ mol{sup -1} of energy. We propose that the major tertiary changes presented in this study reveal a structural basis for an exothermic maturation process probably present in many dsDNA bacteriophage and possibly viruses such as herpesvirus, which share the HK97 subunit fold.« less

Geometry of AN Isolated Dimer of Imidazole Characterised by Rotational Spectroscopy and AB Initio Calculations

NASA Astrophysics Data System (ADS)

Mullaney, John C.; Zaleski, Daniel P.; Tew, David Peter; Walker, Nick; Legon, Anthony

2016-06-01

An isolated, gas-phase dimer of imidazole is generated through laser vaporisation of a solid rod containing a 1:1 mixture of imidazole and copper in the presence of an argon buffer gas undergoing supersonic expansion. The complex is characterised through broadband rotational spectroscopy and is shown to have a twisted, hydrogen-bonded geometry. Calculations at the CCSD(T)(F12*)/cc-pVDZ-F12 level of theory confirm this to be the lowest-energy conformer of the imidazole dimer. The distance between the respective centres of mass of the imidazole monomer subunits is determined to be 5.2751(1) Å, and the twist angle γ describing rotation of one monomer with respect to the other about a line connecting the centres of mass of the monomers is determined to be 87.9(4)o. Four out of six intermolecular parameters in the model geometry are precisely determined from the experimental rotational constants and are consistent with results calculated ab initio.

Regulation of Aerobic Energy Metabolism in Podospora anserina by Two Paralogous Genes Encoding Structurally Different c-Subunits of ATP Synthase.

PubMed

Sellem, Carole H; di Rago, Jean-Paul; Lasserre, Jean-Paul; Ackerman, Sharon H; Sainsard-Chanet, Annie

2016-07-01

Most of the ATP in living cells is produced by an F-type ATP synthase. This enzyme uses the energy of a transmembrane electrochemical proton gradient to synthesize ATP from ADP and inorganic phosphate. Proton movements across the membrane domain (FO) of the ATP synthase drive the rotation of a ring of 8-15 c-subunits, which induces conformational changes in the catalytic part (F1) of the enzyme that ultimately promote ATP synthesis. Two paralogous nuclear genes, called Atp9-5 and Atp9-7, encode structurally different c-subunits in the filamentous fungus Podospora anserina. We have in this study identified differences in the expression pattern for the two genes that correlate with the mitotic activity of cells in vegetative mycelia: Atp9-7 is transcriptionally active in non-proliferating (stationary) cells while Atp9-5 is expressed in the cells at the extremity (apex) of filaments that divide and are responsible for mycelium growth. When active, the Atp9-5 gene sustains a much higher rate of c-subunit synthesis than Atp9-7. We further show that the ATP9-7 and ATP9-5 proteins have antagonist effects on the longevity of P. anserina. Finally, we provide evidence that the ATP9-5 protein sustains a higher rate of mitochondrial ATP synthesis and yield in ATP molecules per electron transferred to oxygen than the c-subunit encoded by Atp9-7. These findings reveal that the c-subunit genes play a key role in the modulation of ATP synthase production and activity along the life cycle of P. anserina. Such a degree of sophistication for regulating aerobic energy metabolism has not been described before.

The Role of Structural Dynamics of Actin in Class-Specific Myosin Motility

PubMed Central

Noguchi, Taro Q. P.; Morimatsu, Masatoshi; Iwane, Atsuko H.; Yanagida, Toshio; Uyeda, Taro Q. P.

2015-01-01

The structural dynamics of actin, including the tilting motion between the small and large domains, are essential for proper interactions with actin-binding proteins. Gly146 is situated at the hinge between the two domains, and we previously showed that a G146V mutation leads to severe motility defects in skeletal myosin but has no effect on motility of myosin V. The present study tested the hypothesis that G146V mutation impaired rotation between the two domains, leading to such functional defects. First, our study showed that depolymerization of G146V filaments was slower than that of wild-type filaments. This result is consistent with the distinction of structural states of G146V filaments from those of the wild type, considering the recent report that stabilization of actin filaments involves rotation of the two domains. Next, we measured intramolecular FRET efficiencies between two fluorophores in the two domains with or without skeletal muscle heavy meromyosin or the heavy meromyosin equivalent of myosin V in the presence of ATP. Single-molecule FRET measurements showed that the conformations of actin subunits of control and G146V actin filaments were different in the presence of skeletal muscle heavy meromyosin. This altered conformation of G146V subunits may lead to motility defects in myosin II. In contrast, distributions of FRET efficiencies of control and G146V subunits were similar in the presence of myosin V, consistent with the lack of motility defects in G146V actin with myosin V. The distribution of FRET efficiencies in the presence of myosin V was different from that in the presence of skeletal muscle heavy meromyosin, implying that the roles of actin conformation in myosin motility depend on the type of myosin. PMID:25945499

Revealing an outward-facing open conformational state in a CLC Cl–/H+ exchange transporter

PubMed Central

Khantwal, Chandra M; Abraham, Sherwin J; Han, Wei; Jiang, Tao; Chavan, Tanmay S; Cheng, Ricky C; Elvington, Shelley M; Liu, Corey W; Mathews, Irimpan I; Stein, Richard A; Mchaourab, Hassane S; Tajkhorshid, Emad; Maduke, Merritt

2016-01-01

CLC secondary active transporters exchange Cl- for H+. Crystal structures have suggested that the conformational change from occluded to outward-facing states is unusually simple, involving only the rotation of a conserved glutamate (Gluex) upon its protonation. Using 19F NMR, we show that as [H+] is increased to protonate Gluex and enrich the outward-facing state, a residue ~20 Å away from Gluex, near the subunit interface, moves from buried to solvent-exposed. Consistent with functional relevance of this motion, constriction via inter-subunit cross-linking reduces transport. Molecular dynamics simulations indicate that the cross-link dampens extracellular gate-opening motions. In support of this model, mutations that decrease steric contact between Helix N (part of the extracellular gate) and Helix P (at the subunit interface) remove the inhibitory effect of the cross-link. Together, these results demonstrate the formation of a previously uncharacterized 'outward-facing open' state, and highlight the relevance of global structural changes in CLC function. DOI: http://dx.doi.org/10.7554/eLife.11189.001 PMID:26799336

Binding of the immunomodulatory drug Bz-423 to mitochondrial FoF1-ATP synthase in living cells by FRET acceptor photobleaching

NASA Astrophysics Data System (ADS)

Starke, Ilka; Johnson, Kathryn M.; Petersen, Jan; Gräber, Peter; Opipari, Anthony W.; Glick, Gary D.; Börsch, Michael

2016-03-01

Bz-423 is a promising new drug for treatment of autoimmune diseases. This small molecule binds to subunit OSCP of the mitochondrial enzyme FoF1-ATP synthase and modulates its catalytic activities. We investigate the binding of Bz-423 to mitochondria in living cells and how subunit rotation in FoF1-ATP synthase, i.e. the mechanochemical mechanism of this enzyme, is affected by Bz-423. Therefore, the enzyme was marked selectively by genetic fusion with the fluorescent protein EGFP to the C terminus of subunit γ. Imaging the threedimensional arrangement of mitochondria in living yeast cells was possible at superresolution using structured illumination microscopy, SIM. We measured uptake and binding of a Cy5-labeled Bz-423 derivative to mitochondrial FoF1-ATP synthase in living yeast cells using FRET acceptor photobleaching microscopy. Our data confirmed the binding of Cy5-labeled Bz-423 to the top of the F1 domain of the enzyme in mitochondria of living Saccharomyces cerevisiae cells.

Structural integration in hypoxia-inducible factors

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

Wu, Dalei; Potluri, Nalini; Lu, Jingping

The hypoxia-inducible factors (HIFs) coordinate cellular adaptations to low oxygen stress by regulating transcriptional programs in erythropoiesis, angiogenesis and metabolism. These programs promote the growth and progression of many tumours, making HIFs attractive anticancer targets. Transcriptionally active HIFs consist of HIF-alpha and ARNT (also called HIF-1 beta) subunits. Here we describe crystal structures for each of mouse HIF-2 alpha-ARNT and HIF-1 alpha-ARNT heterodimers in states that include bound small molecules and their hypoxia response element. A highly integrated quaternary architecture is shared by HIF-2 alpha-ARNT and HIF-1 alpha-ARNT, wherein ARNT spirals around the outside of each HIF-alpha subunit. Five distinctmore » pockets are observed that permit small-molecule binding, including PAS domain encapsulated sites and an interfacial cavity formed through subunit heterodimerization. The DNA-reading head rotates, extends and cooperates with a distal PAS domain to bind hypoxia response elements. HIF-alpha mutations linked to human cancers map to sensitive sites that establish DNA binding and the stability of PAS domains and pockets.« less

Rotation Periods of Open Cluster Stars. IV.

NASA Astrophysics Data System (ADS)

Prosser, Charles F.; Grankin, Konstantin N.

We present the results from a photometric monitoring program of primarily solar-type open cluster stars obtained during 1994 and 1995. Several members of the α Persei cluster have been monitored and the corresponding relation between coronal x-ray activity and rotation period derived. The relation among mid-G/K type members illustrates both the previously noticed downturn in L_X/L_bol at high rotation rates and the sharp decrease in coronal activity at long rotation periods as seen among Pleiades stars. Intensive observation of one slowly rotating G-type member of IC 4665 has enabled a period determination of 8-10 days to be made and illustrates the need for (and limitations of) high quality observations.

Multitiered and Cooperative Surveillance of Mitochondrial Phosphatidylserine Decarboxylase 1.

PubMed

Ogunbona, Oluwaseun B; Onguka, Ouma; Calzada, Elizabeth; Claypool, Steven M

2017-09-01

Phosphatidylserine decarboxylase 1 (Psd1p), an ancient enzyme that converts phosphatidylserine to phosphatidylethanolamine in the inner mitochondrial membrane, must undergo an autocatalytic self-processing event to gain activity. Autocatalysis severs the protein into a large membrane-anchored β subunit that noncovalently associates with the small α subunit on the intermembrane space side of the inner membrane. Here, we determined that a temperature sensitive ( ts ) PSD1 allele is autocatalytically impaired and that its fidelity is closely monitored throughout its life cycle by multiple mitochondrial quality control proteases. Interestingly, the proteases involved in resolving misfolded Psd1 ts vary depending on its autocatalytic status. Specifically, the degradation of a Psd1 ts precursor unable to undergo autocatalysis requires the unprecedented cooperative and sequential actions of two inner membrane proteases, Oma1p and Yme1p. In contrast, upon heat exposure postautocatalysis, Psd1 ts β subunits accumulate in protein aggregates that are resolved by Yme1p acting alone, while the released α subunit is degraded in parallel by an unidentified protease. Importantly, the stability of endogenous Psd1p is also influenced by Yme1p. We conclude that Psd1p, the key enzyme required for the mitochondrial pathway of phosphatidylethanolamine production, is closely monitored at several levels and by multiple mitochondrial quality control mechanisms present in the intermembrane space. Copyright © 2017 American Society for Microbiology.

Central cavity of fructose-1,6-bisphosphatase and the evolution of AMP/fructose 2,6-bisphosphate synergism in eukaryotic organisms.

PubMed

Gao, Yang; Shen, Lu; Honzatko, Richard B

2014-03-21

The effects of AMP and fructose 2,6-bisphosphate (Fru-2,6-P2) on porcine fructose-1,6-bisphosphatase (pFBPase) and Escherichia coli FBPase (eFBPase) differ in three respects. AMP/Fru-2,6-P2 synergism in pFBPase is absent in eFBPase. Fru-2,6-P2 induces a 13° subunit pair rotation in pFBPase but no rotation in eFBPase. Hydrophilic side chains in eFBPase occupy what otherwise would be a central aqueous cavity observed in pFBPase. Explored here is the linkage of AMP/Fru-2,6-P2 synergism to the central cavity and the evolution of synergism in FBPases. The single mutation Ser(45) → His substantially fills the central cavity of pFBPase, and the triple mutation Ser(45) → His, Thr(46) → Arg, and Leu(186) → Tyr replaces porcine with E. coli type side chains. Both single and triple mutations significantly reduce synergism while retaining other wild-type kinetic properties. Similar to the effect of Fru-2,6-P2 on eFBPase, the triple mutant of pFBPase with bound Fru-2,6-P2 exhibits only a 2° subunit pair rotation as opposed to the 13° rotation exhibited by the Fru-2,6-P2 complex of wild-type pFBPase. The side chain at position 45 is small in all available eukaryotic FBPases but large and hydrophilic in bacterial FBPases, similar to eFBPase. Sequence information indicates the likelihood of synergism in the FBPase from Leptospira interrogans (lFBPase), and indeed recombinant lFBPase exhibits AMP/Fru-2,6-P2 synergism. Unexpectedly, however, AMP also enhances Fru-6-P binding to lFBPase. Taken together, these observations suggest the evolution of AMP/Fru-2,6-P2 synergism in eukaryotic FBPases from an ancestral FBPase having a central aqueous cavity and exhibiting synergistic feedback inhibition by AMP and Fru-6-P.

Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

DOE PAGES

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

2015-10-09

SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis. Here in this study, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanismmore » that is strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.« less

Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

DOE PAGES

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

2015-10-09

SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis . Here, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanism that ismore » strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.« less

Analysis and modeling of heat-labile enterotoxins of Escherichia coli suggests a novel space with insights into receptor preference.

PubMed

Krishna Raja, M; Ghosh, Asit Ranjan; Vino, S; Sajitha Lulu, S

2015-01-01

Features of heat-labile enterotoxins of Escherichia coli which make them fit to use as novel receptors for antidiarrheals are not completely explored. Data-set of 14 different serovars of enterotoxigenic Escherichia coli producing heat-labile toxins were taken from NCBI Genbank database and used in the study. Sequence analysis showed mutations in different subunits and also at their interface residues. As these toxins lack crystallography structures, homology modeling using Modeller 9.11 led to the structural approximation for the E. coli producing heat-labile toxins. Interaction of modeled toxin subunits with proanthocyanidin, an antidiarrheal showed several strong hydrogen bonding interactions at the cost of minimized energy. The hits were subsequently characterized by molecular dynamics simulation studies to monitor their binding stabilities. This study looks into novel space where the ligand can choose the receptor preference not as a whole but as an individual subunit. Mutation at interface residues and interaction among subunits along with the binding of ligand to individual subunits would help to design a non-toxic labile toxin and also to improve the therapeutics.

Realistic simulations of the coupling between the protomotive force and the mechanical rotation of the F0-ATPase

PubMed Central

Mukherjee, Shayantani; Warshel, Arieh

2012-01-01

The molecular origin of the action of the F0 proton gradient-driven rotor presents a major puzzle despite significant structural advances. Although important conceptual models have provided guidelines of how such systems should work, it has been challenging to generate a structure-based molecular model using physical principles that will consistently lead to the unidirectional proton-driven rotational motion during ATP synthesis. This work uses a coarse-grained (CG) model to simulate the energetics of the F0-ATPase system in the combined space defined by the rotational coordinate and the proton transport (PTR) from the periplasmic side (P) to the cytoplasmic side (N). The model establishes the molecular origin of the rotation, showing that this effect is due to asymmetry in the energetics of the proton path rather than only the asymmetry of the interaction of the Asp on the c-ring helices and Arg on the subunit-a. The simulation provides a clear conceptual background for further exploration of the electrostatic basis of proton-driven mechanochemical systems. PMID:22927379

Structure and tunneling dynamics in a model system of peptide co-solvents: rotational spectroscopy of the 2,2,2-trifluoroethanol⋯water complex.

PubMed

Thomas, Javix; Xu, Yunjie

2014-06-21

The hydrogen-bonding topology and tunneling dynamics of the binary adduct, 2,2,2-trifluoroethanol (TFE)⋯water, were investigated using chirped pulse and cavity based Fourier transform microwave spectroscopy with the aid of high level ab initio calculations. Rotational spectra of the most stable binary TFE⋯water conformer and five of its deuterium isotopologues were assigned. A strong preference for the insertion binding topology where water is inserted into the existing intramolecular hydrogen-bonded ring of TFE was observed. Tunneling splittings were detected in all of the measured rotational transitions of TFE⋯water. Based on the relative intensity of the two tunneling components and additional isotopic data, the splitting can be unambiguously attributed to the tunneling motion of the water subunit, i.e., the interchange of the bonded and nonbonded hydrogen atoms of water. The absence of any other splitting in the rotational transitions of all isotopologues observed indicates that the tunneling between g+ and g- TFE is quenched in the TFE⋯H2O complex.

Radiation area monitor device and method

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

Vencelj, Matjaz; Stowe, Ashley C.; Petrovic, Toni

A radiation area monitor device/method, utilizing: a radiation sensor; a rotating radiation shield disposed about the radiation sensor, wherein the rotating radiation shield defines one or more ports that are transparent to radiation; and a processor operable for analyzing and storing a radiation fingerprint acquired by the radiation sensor as the rotating radiation shield is rotated about the radiation sensor. Optionally, the radiation sensor includes a gamma and/or neutron radiation sensor. The device/method selectively operates in: a first supervised mode during which a baseline radiation fingerprint is acquired by the radiation sensor as the rotating radiation shield is rotated aboutmore » the radiation sensor; and a second unsupervised mode during which a subsequent radiation fingerprint is acquired by the radiation sensor as the rotating radiation shield is rotated about the radiation sensor, wherein the subsequent radiation fingerprint is compared to the baseline radiation fingerprint and, if a predetermined difference threshold is exceeded, an alert is issued.« less

Distribution and Demographics of Marine Mammals in SOCAL through Photo-Identification, Genetics, and Satellite Telemetry: A Summary of Surveys Conducted 1 July 2011 - 15 June 2012

DTIC Science & Technology

2012-08-01

may have population sub-units with higher than expected residency to the Southern California Bight. Beaked whales particularly show this higher than... killer whale , satellite tags, photo-identification, biopsy sampling, acoustic monitoring, SOAR, Southern California Bight. 16. SECURITY...population sub-units with higher than expected residency to the Southern California Bight, and to SOAR in particular in the case of beaked whales . These

Advances in Rotational Seismic Measurements

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

Pierson, Robert; Laughlin, Darren; Brune, Robert

2016-10-19

Rotational motion is increasingly understood to be a significant part of seismic wave motion. Rotations can be important in earthquake strong motion and in Induced Seismicity Monitoring. Rotational seismic data can also enable shear selectivity and improve wavefield sampling for vertical geophones in 3D surveys, among other applications. However, sensor technology has been a limiting factor to date. The US Department of Energy (DOE) and Applied Technology Associates (ATA) are funding a multi-year project that is now entering Phase 2 to develop and deploy a new generation of rotational sensors for validation of rotational seismic applications. Initial focus is onmore » induced seismicity monitoring, particularly for Enhanced Geothermal Systems (EGS) with fracturing. The sensors employ Magnetohydrodynamic (MHD) principles with broadband response, improved noise floors, robustness, and repeatability. This paper presents a summary of Phase 1 results and Phase 2 status.« less

Chicken embryo fibroblasts exposed to weak, time-varying magnetic fields share cell proliferation, adenosine deaminase activity, and membrane characteristics of transformed cells

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

Parola, A.H.; Porat, N.; Kiesow, L.A.

1993-01-01

Chicken embryo fibroblasts (CEF) exposed to a sinusoidally varying magnetic field (SVMF) (100 Hz, 700 microT, for 24 h) showed a remarkable rise of segmental rotational relaxation rate of adenosine deaminase (ADA, EC 3.5.4.4) as determined by multifrequency phase fluorometry. Pyrene-labeled, small subunit ADA was applied to cultured (normal) CEF, which have available and abundant ADA complexing protein (ADCP) on their plasma membranes. Sine-wave-modulated fluorometry of the pyrene yielded a profile of phase angle vs. modulation frequency. In SVMF-treated cells and in Rous-sarcoma-virus (RSV) transformed cells the differential phase values at low modulation frequencies of the excitation are remarkably reduced.more » This effect is magnetic rather than thermal, because the temperature was carefully controlled and monitored; nevertheless to further check this matter we studied CEF, infected by the RSV-Ts68 temperature-sensitive mutant (36 degrees C transformed, 41 degrees C revertant). When grown at 36 degrees C in the SVMF, cells did not show the slightest trend towards reversion, as would be expected had there been local heating. Concomitant with the increased segmental rotational relaxation rate of ADA, there was a decrease in fluorescence lifetime and a slight, yet significant, increase in membrane lipid microfluidity. These biophysical observations prompted us to examine the effect of SVMF on cell proliferation and ADA activity (a malignancy marker): higher rates of cell proliferation and reduced specific activity of ADA were observed.« less

Rotation periods of open-cluster stars, 3

NASA Technical Reports Server (NTRS)

Prosser, Charles F.; Shetrone, Matthew D.; Dasgupta, Amil; Backman, Dana E.; Laaksonen, Bentley D.; Baker, Shawn W.; Marschall, Laurence A.; Whitney, Barbara A.; Kuijken, Konrad; Stauffer, John R.

1995-01-01

We present the results from a photometric monitoring program of 15 open cluster stars and one weak-lined T Tauri star during late 1993/early 1994. Several show rotators which are members of the Alpha Persei, Pleiades, and Hyades open clusters have been monitored and period estimates derived. Using all available Pleiades stars with photometric periods together with current X-ray flux measurements, we illustrate the X-ray activity/rotation relation among Pleiades late-G/K dwarfs. The data show a clear break in the rotation-activity relation around P approximately 6-7 days -- in general accordance with previous results using more heterogeneous samples of G/K stars.

Overexpressing wild-type γ2 subunits rescued the seizure phenotype in Gabrg2+/Q390X Dravet syndrome mice.

PubMed

Huang, Xuan; Zhou, Chengwen; Tian, Mengnan; Kang, Jing-Qiong; Shen, Wangzhen; Verdier, Kelienne; Pimenta, Aurea; MacDonald, Robert L

2017-08-01

The mutant γ-aminobutyric acid type A (GABA A ) receptor γ2(Q390X) subunit (Q351X in the mature peptide) has been associated with the epileptic encephalopathy, Dravet syndrome, and the epilepsy syndrome genetic epilepsy with febrile seizures plus (GEFS+). The mutation generates a premature stop codon that results in translation of a stable truncated and misfolded γ2 subunit that accumulates in neurons, forms intracellular aggregates, disrupts incorporation of γ2 subunits into GABA A receptors, and affects trafficking of partnering α and β subunits. Heterozygous Gabrg2 +/Q390X knock-in (KI) mice had reduced cortical inhibition, spike wave discharges on electroencephalography (EEG), a lower seizure threshold to the convulsant drug pentylenetetrazol (PTZ), and spontaneous generalized tonic-clonic seizures. In this proof-of-principal study, we attempted to rescue these deficits in KI mice using a γ2 subunit gene (GABRG2) replacement therapy. We introduced the GABRG2 allele by crossing Gabrg2 +/Q390X KI mice with bacterial artificial chromosome (BAC) transgenic mice overexpressing HA (hemagglutinin)-tagged human γ2 HA subunits, and compared GABA A receptor subunit expression by Western blot and immunohistochemical staining, seizure threshold by monitoring mouse behavior after PTZ-injection, and thalamocortical inhibition and network oscillation by slice recording. Compared to KI mice, adult mice carrying both mutant allele and transgene had increased wild-type γ2 and partnering α1 and β2/3 subunits, increased miniature inhibitory postsynaptic current (mIPSC) amplitudes recorded from layer VI cortical neurons, reduced thalamocortical network oscillations, and higher PTZ seizure threshold. Based on these results we suggest that seizures in a genetic epilepsy syndrome caused by epilepsy mutant γ2(Q390X) subunits with dominant negative effects could be rescued potentially by overexpression of wild-type γ2 subunits. Wiley Periodicals, Inc. © 2017 International League Against Epilepsy.

Mechanism of One-Way Traffic of Hexameric Phi29 DNA Packaging Motor with Four Electropositive Relaying Layers Facilitating Antiparallel Revolution

PubMed Central

2013-01-01

The importance of nanomotors in nanotechnology is akin to that of mechanical engines to daily life. The AAA+ superfamily is a class of nanomotors performing various functions. Their hexagonal arrangement facilitates bottom-up assembly for stable structures. The bacteriophage phi29 DNA translocation motor contains three coaxial rings: a dodecamer channel, a hexameric ATPase ring, and a hexameric pRNA ring. The viral DNA packaging motor has been believed to be a rotational machine. However, we discovered a revolution mechanism without rotation. By analogy, the earth revolves around the sun while rotating on its own axis. One-way traffic of dsDNA translocation is facilitated by five factors: (1) ATPase changes its conformation to revolve dsDNA within a hexameric channel in one direction; (2) the 30° tilt of the channel subunits causes an antiparallel arrangement between two helices of dsDNA and channel wall to advance one-way translocation; (3) unidirectional flow property of the internal channel loops serves as a ratchet valve to prevent reversal; (4) 5′–3′ single-direction movement of one DNA strand along the channel wall ensures single direction; and (5) four electropositive layers interact with one strand of the electronegative dsDNA phosphate backbone, resulting in four relaying transitional pauses during translocation. The discovery of a riding system along one strand provides a motion nanosystem for cargo transportation and a tool for studying force generation without coiling, friction, and torque. The revolution of dsDNA among 12 subunits offers a series of recognition sites on the DNA backbone to provide additional spatial variables for nucleotide discrimination for sensing applications. PMID:23510192

Phosphate release coupled to rotary motion of F1-ATPase

PubMed Central

Okazaki, Kei-ichi; Hummer, Gerhard

2013-01-01

F1-ATPase, the catalytic domain of ATP synthase, synthesizes most of the ATP in living organisms. Running in reverse powered by ATP hydrolysis, this hexameric ring-shaped molecular motor formed by three αβ-dimers creates torque on its central γ-subunit. This reverse operation enables detailed explorations of the mechanochemical coupling mechanisms in experiment and simulation. Here, we use molecular dynamics simulations to construct a first atomistic conformation of the intermediate state following the 40° substep of rotary motion, and to study the timing and molecular mechanism of inorganic phosphate (Pi) release coupled to the rotation. In response to torque-driven rotation of the γ-subunit in the hydrolysis direction, the nucleotide-free αβE interface forming the “empty” E site loosens and singly charged Pi readily escapes to the P loop. By contrast, the interface stays closed with doubly charged Pi. The γ-rotation tightens the ATP-bound αβTP interface, as required for hydrolysis. The calculated rate for the outward release of doubly charged Pi from the αβE interface 120° after ATP hydrolysis closely matches the ∼1-ms functional timescale. Conversely, Pi release from the ADP-bound αβDP interface postulated in earlier models would occur through a kinetically infeasible inward-directed pathway. Our simulations help reconcile conflicting interpretations of single-molecule experiments and crystallographic studies by clarifying the timing of Pi exit, its pathway and kinetics, associated changes in Pi protonation, and changes of the F1-ATPase structure in the 40° substep. Important elements of the molecular mechanism of Pi release emerging from our simulations appear to be conserved in myosin despite the different functional motions. PMID:24062450

Dissociating anxiolytic and sedative effects of GABAAergic drugs using temperature and locomotor responses to acute stress

PubMed Central

Klanker, Marianne; Groenink, Lucianne; Korte, S. Mechiel; Cook, James M.; Van Linn, Michael L.; Hopkins, Seth C.; Olivier, Berend

2009-01-01

Rationale The stress-induced hyperthermia (SIH) model is an anxiety model that uses the transient rise in body temperature in response to acute stress. Benzodiazepines produce anxiolytic as well as sedative side effects through nonselective binding to GABAA receptor subunits. The GABAA receptor α1 subunit is associated with sedation, whereas the GABAA receptor α2 and α3 subunits are involved in anxiolytic effects. Objectives We therefore examined the effects of (non) subunit-selective GABAA receptor agonists on temperature and locomotor responses to novel cage stress. Results Using telemetric monitoring of temperature and locomotor activity, we found that nonsubunit-selective GABAA receptor agonist diazepam as well as the α3 subunit-selective receptor agonist TP003 dose-dependently attenuated SIH and locomotor responses. Administration of GABAA receptor α1-selective agonist zolpidem resulted in profound hypothermia and locomotor sedation. The GABAA receptor α1-selective antagonist βCCt antagonized the hypothermia, but did not reverse the SIH response attenuation caused by diazepam and zolpidem. These results suggest an important regulating role for the α1 subunit in thermoregulation and sedation. Ligands of extrasynaptic GABAA receptors such as alcohol and nonbenzodiazepine THIP attenuated the SIH response only at high doses. Conclusions The present study confirms a putative role for the GABAA receptor α1 subunit in hypothermia and sedation and supports a role for α2/3 subunit GABAA receptor agonists in anxiety processes. In conclusion, we show that home cage temperature and locomotor responses to novel home cage stress provide an excellent tool to assess both anxiolytic and sedative effects of various (subunit-selective) GABAAergic compounds. PMID:19169673

Rotation of artificial rotor axles in rotary molecular motors

PubMed Central

Baba, Mihori; Iwamoto, Kousuke; Ueno, Hiroshi; Hara, Mayu; Nakanishi, Atsuko; Kishikawa, Jun-ichi; Noji, Hiroyuki; Yokoyama, Ken

2016-01-01

F1- and V1-ATPase are rotary molecular motors that convert chemical energy released upon ATP hydrolysis into torque to rotate a central rotor axle against the surrounding catalytic stator cylinder with high efficiency. How conformational change occurring in the stator is coupled to the rotary motion of the axle is the key unknown in the mechanism of rotary motors. Here, we generated chimeric motor proteins by inserting an exogenous rod protein, FliJ, into the stator ring of F1 or of V1 and tested the rotation properties of these chimeric motors. Both motors showed unidirectional and continuous rotation, despite no obvious homology in amino acid sequence between FliJ and the intrinsic rotor subunit of F1 or V1. These results showed that any residue-specific interactions between the stator and rotor are not a prerequisite for unidirectional rotation of both F1 and V1. The torque of chimeric motors estimated from viscous friction of the rotation probe against medium revealed that whereas the F1-FliJ chimera generates only 10% of WT F1, the V1-FliJ chimera generates torque comparable to that of V1 with the native axle protein that is structurally more similar to FliJ than the native rotor of F1. This suggests that the gross structural mismatch hinders smooth rotation of FliJ accompanied with the stator ring of F1. PMID:27647891

Fo-driven Rotation in the ATP Synthase Direction against the Force of F1 ATPase in the FoF1 ATP Synthase*

PubMed Central

Martin, James; Hudson, Jennifer; Hornung, Tassilo; Frasch, Wayne D.

2015-01-01

Living organisms rely on the FoF1 ATP synthase to maintain the non-equilibrium chemical gradient of ATP to ADP and phosphate that provides the primary energy source for cellular processes. How the Fo motor uses a transmembrane electrochemical ion gradient to create clockwise torque that overcomes F1 ATPase-driven counterclockwise torque at high ATP is a major unresolved question. Using single FoF1 molecules embedded in lipid bilayer nanodiscs, we now report the observation of Fo-dependent rotation of the c10 ring in the ATP synthase (clockwise) direction against the counterclockwise force of ATPase-driven rotation that occurs upon formation of a leash with Fo stator subunit a. Mutational studies indicate that the leash is important for ATP synthase activity and support a mechanism in which residues aGlu-196 and cArg-50 participate in the cytoplasmic proton half-channel to promote leash formation. PMID:25713065

Advantages of isotopic depletion of proteins for hydrogen/deuterium exchange experiments monitored by mass spectrometry.

PubMed

Bou-Assaf, George M; Chamoun, Jean E; Emmett, Mark R; Fajer, Piotr G; Marshall, Alan G

2010-04-15

Solution-phase hydrogen/deuterium exchange (HDX) monitored by mass spectrometry is an excellent tool to study protein-protein interactions and conformational changes in biological systems, especially when traditional methods such as X-ray crystallography or nuclear magnetic resonance are not feasible. Peak overlap among the dozens of proteolytic fragments (including those from autolysis of the protease) can be severe, due to high protein molecular weight(s) and the broad isotopic distributions due to multiple deuterations of many peptides. In addition, different subunits of a protein complex can yield isomeric proteolytic fragments. Here, we show that depletion of (13)C and/or (15)N for one or more protein subunits of a complex can greatly simplify the mass spectra, increase the signal-to-noise ratio of the depleted fragment ions, and remove ambiguity in assignment of the m/z values to the correct isomeric peptides. Specifically, it becomes possible to monitor the exchange progress for two isobaric fragments originating from two or more different subunits within the complex, without having to resort to tandem mass spectrometry techniques that can lead to deuterium scrambling in the gas phase. Finally, because the isotopic distribution for a small to medium-size peptide is essentially just the monoisotopic species ((12)C(c)(1)H(h)(14)N(n)(16)O(o)(32)S(s)), it is not necessary to deconvolve the natural abundance distribution for each partially deuterated peptide during HDX data reduction.

Crystal structure of release factor RF3 trapped in the GTP state on a rotated conformation of the ribosome

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

Zhou, Jie; Lancaster, Laura; Trakhanov, Sergei

2012-03-26

The class II release factor RF3 is a GTPase related to elongation factor EF-G, which catalyzes release of class I release factors RF1 and RF2 from the ribosome after termination of protein synthesis. The 3.3 {angstrom} crystal structure of the RF3 {center_dot} GDPNP {center_dot} ribosome complex provides a high-resolution description of interactions and structural rearrangements that occur when binding of this translational GTPase induces large-scale rotational movements in the ribosome. RF3 induces a 7{sup o} rotation of the body and 14{sup o} rotation of the head of the 30S ribosomal subunit, and itself undergoes inter- and intradomain conformational rearrangements. Wemore » suggest that ordering of critical elements of switch loop I and the P loop, which help to form the GTPase catalytic site, are caused by interactions between the G domain of RF3 and the sarcin-ricin loop of 23S rRNA. The rotational movements in the ribosome induced by RF3, and its distinctly different binding orientation to the sarcin-ricin loop of 23S rRNA, raise interesting implications for the mechanism of action of EF-G in translocation.« less

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

PubMed Central

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

2013-01-01

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

Reverse engineering the cooperative machinery of human hemoglobin.

PubMed

Ren, Zhong

2013-01-01

Hemoglobin transports molecular oxygen from the lungs to all human tissues for cellular respiration. Its α2β2 tetrameric assembly undergoes cooperative binding and releasing of oxygen for superior efficiency and responsiveness. Over past decades, hundreds of hemoglobin structures were determined under a wide range of conditions for investigation of molecular mechanism of cooperativity. Based on a joint analysis of hemoglobin structures in the Protein Data Bank (Ren, companion article), here I present a reverse engineering approach to elucidate how two subunits within each dimer reciprocate identical motions that achieves intradimer cooperativity, how ligand-induced structural signals from two subunits are integrated to drive quaternary rotation, and how the structural environment at the oxygen binding sites alter their binding affinity. This mechanical model reveals the intricate design that achieves the cooperative mechanism and has previously been masked by inconsistent structural fluctuations. A number of competing theories on hemoglobin cooperativity and broader protein allostery are reconciled and unified.

Ribosome rearrangements at the onset of translational bypassing

PubMed Central

Agirrezabala, Xabier; Samatova, Ekaterina; Klimova, Mariia; Zamora, Miguel; Gil-Carton, David; Rodnina, Marina V.; Valle, Mikel

2017-01-01

Bypassing is a recoding event that leads to the translation of two distal open reading frames into a single polypeptide chain. We present the structure of a translating ribosome stalled at the bypassing take-off site of gene 60 of bacteriophage T4. The nascent peptide in the exit tunnel anchors the P-site peptidyl-tRNAGly to the ribosome and locks an inactive conformation of the peptidyl transferase center (PTC). The mRNA forms a short dynamic hairpin in the decoding site. The ribosomal subunits adopt a rolling conformation in which the rotation of the small subunit around its long axis causes the opening of the A-site region. Together, PTC conformation and mRNA structure safeguard against premature termination and read-through of the stop codon and reconfigure the ribosome to a state poised for take-off and sliding along the noncoding mRNA gap. PMID:28630923

Microwave Spectrum and Structure of the Methane-Propane Complex

NASA Astrophysics Data System (ADS)

Peterson, Karen I.; Lin, Wei; Arsenault, Eric A.; Choi, Yoon Jeong; Novick, Stewart E.

2017-06-01

Methane is exceptional in its solid-phase orientational disorder that persists down to 24 K. Only below that temperature does the structure become partially ordered, and full crystallinity requires even lower temperatures and high pressures. Not surprisingly, methane appears to freely rotate in most van der Waals complexes, although two notable exceptions are CH_4-HF and CH_4-C_5H_5N. Of interest to us is how alkane interactions affect the methane rotation. Except for CH_4-CH_4, rotationally-resolved spectra of alkane-alkane complexes have not been studied. To fill this void, we present the microwave spectrum of CH_4-C_3H_8 which is the smallest alkane complex with a practical dipole moment. The microwave spectrum of CH_4-C_3H_8 was measured using the Fourier Transform microwave spectrometer at Wesleyan University. In the region between 7100 and 25300 MHz, we observed approximately 70 transitions that could plausibly be attributed to the CH_4-C_3H_8 complex (requiring high power and the proper mixture of gases). Of these, 16 were assigned to the A-state (lowest internal rotor state of methane) and four to the F-state. The A-state transitions were fitted with a Watson Hamiltonian using nine spectroscopic constants of which A = 7553.8144(97) MHz, B = 2483.9183(35) MHz, and C = 2041.8630(21) MHz. The A rotational constant is only 1.5 MHz higher than that of Ar-C_3H_8 and, since the a-axis of the complex passes approximately through the centers of mass of the subunits, this indicates a similar relative orientation. Thus, we find that the CH_4 is located above the plane of the propane. The center-of-mass separation of the subunits in CH_4-C_3H_8 is calculated to be 3.993 Å, 0.16 Å longer than the Ar-C_3H_8 distance of 3.825 Å, a reasonable difference considering the larger van der Waals radius of CH_4. The four F-state lines, which were about twice as strong as the A-state lines, could be fitted to A, B, and C rotational constants, and further analysis is in progress.

Functions and ATP-binding responses of the twelve histidine residues in the TF1-ATPase beta subunit.

PubMed

Tozawa, K; Yagi, H; Hisamatsu, K; Ozawa, K; Yoshida, M; Akutsu, H

2001-10-01

The C2 proton signals of all (twelve) histidine residues of the TF1 beta subunit in the 1H-NMR spectrum have been identified and assigned by means of pH change experiments and site-directed substitution of histidines by glutamines. pH and ligand titration experiments were carried out for these signals. Furthermore, the ATPase activity of the reconstituted alpha3beta3gamma complex was examined for the twelve mutant beta subunits. Two of three conserved histidines, namely, His-119 and 324, were found to be important for expression of the ATPase activity. The former fixes the N-terminal domain to the central domain. His-324 is involved in the formation of the interface essential for the alpha3beta3gamma complex assembly. The other conserved residue, His-363, showed a very low pK(a), suggesting that it is involved in the tertiary structure formation. On the binding of a nucleotide, only the signals of His-173, 179, 200, and 324 shifted. These histidines are located in the hinge region, and its proximity, of the beta subunit. This observation provided further support for the conformational change of the beta monomer from the open to the closed form on the binding of a nucleotide proposed by us [Yagi et al. (1999) Biophys. J. 77, 2175-2183]. This conformational change should be one of the essential driving forces in the rotation of the alpha3beta3gamma complex.

Low resolution crystal structure of Arenicola erythrocruorin: influence of coiled coils on the architecture of a megadalton respiratory protein.

PubMed

Royer, William E; Omartian, Michael N; Knapp, James E

2007-01-05

Annelid erythrocruorins are extracellular respiratory complexes assembled from 180 subunits into hexagonal bilayers. Cryo-electron microscopic experiments have identified two different architectural classes. In one, designated type I, the vertices of the two hexagonal layers are partially staggered, with one hexagonal layer rotated by about 16 degrees relative to the other layer, whereas in the other class, termed type II, the vertices are essentially eclipsed. We report here the first crystal structure of a type II erythrocruorin, that from Arenicola marina, at 6.2 A resolution. The structure reveals the presence of long continuous triple-stranded coiled-coil "spokes" projecting towards the molecular center from each one-twelfth unit; interdigitation of these spokes provides the only contacts between the two hexagonal layers of the complex. This arrangement contrasts with that of a type I erythrocruorin from Lumbricus terrestris in which the spokes are broken into two triple-stranded coiled coils with a disjointed connection. The disjointed connection allows formation of a more compact structure in the type I architecture, with the two hexagonal layers closer together and additional extensive contacts between the layers. Comparison of sequences of the coiled-coil regions of various linker subunits shows that the linker subunits from type II erythrocruorins possess continuous heptad repeats, whereas a sequence gap places these repeats out of register in the type I linker subunits, consistent with a disjointed coiled-coil arrangement.

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

PubMed

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

2010-08-19

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

Rotational coherent anti-stokes Raman spectroscopy measurements in a rotating cavity with axial throughflow of cooling air: oxygen concentration measurements.

PubMed

Black, J D; Long, C A

1992-07-20

In a rotating cavity rig, which models cooling air flow in the spaces between disks of a gas turbine compressor, the buildup of oxygen concentration after the cooling gas was changed from nitrogen to air was monitored using rotational coherent anti-Stokes Raman spectroscopy (CARS). From this information an estimate of the fraction of the throughflow entering the rotating cavity was obtained. This demonstrates that rotational CARS can be applied as a nonintrusive concentration-measurement technique in a rotating engineering test rig.

Structural Characterisation of Proteins from the Peroxiredoxin Family

DTIC Science & Technology

2014-01-01

SECURITY CLASSIFICATION OF: The oligomerisation of protein subunits is an area of much research interest, in particular the relationship to protein...or decision, unless so designated by other documentation. 9. SPONSORING/MONITORING AGENCY NAME(S) AND ADDRESS (ES) U.S. Army Research Office P.O...Box 12211 Research Triangle Park, NC 27709-2211 peroxiredoxin, tecton, supramolecular assembly, TEM REPORT DOCUMENTATION PAGE 11. SPONSOR/MONITOR’S

Permanent magnet DC motor control by using arduino and motor drive module BTS7960

NASA Astrophysics Data System (ADS)

Syukriyadin, S.; Syahrizal, S.; Mansur, G.; Ramadhan, H. P.

2018-05-01

This study proposes a control system for permanent magnet DC (PMDC) motor. PMDC drive control system has two critical parameters: control and monitoring. Control system includes rotation speed control and direction of rotation of motor using motor drive module BTS7960. The PWM signal has a fixed frequency of waves with varying duty cycles (between 0% and 100%), so the motor rotation can be regulated gradually using a potentiometer already programmed on the Arduino Uno board. The motor rotation direction setting uses the H-bridge circuit method using a 3-way switch to set the direction of forward-reverse rotation of the motor. The monitoring system includes measurements of rotational speed, current, and voltage. Motor rotation speed can be adjusted from the armature voltage settings through the duty cycle PWM setting so that the motor speed can be increased or decreased by the desired duty cycle. From the unload PMDC motor test results it has also been shown that the torque of the motor is relatively constant when there is a change in speed from low rpm to high rpm or vice versa.

Determination Method of Bridge Rotation Angle Response Using MEMS IMU.

PubMed

Sekiya, Hidehiko; Kinomoto, Takeshi; Miki, Chitoshi

2016-11-09

To implement steel bridge maintenance, especially that related to fatigue damage, it is important to monitor bridge deformations under traffic conditions. Bridges deform and rotate differently under traffic load conditions because their structures differ in terms of length and flexibility. Such monitoring enables the identification of the cause of stress concentrations that cause fatigue damage and the proposal of appropriate countermeasures. However, although bridge deformation monitoring requires observations of bridge angle response as well as the bridge displacement response, measuring the rotation angle response of a bridge subject to traffic loads is difficult. Theoretically, the rotation angle response can be calculated by integrating the angular velocity, but for field measurements of actual in-service bridges, estimating the necessary boundary conditions would be difficult due to traffic-induced vibration. To solve the problem, this paper proposes a method for determining the rotation angle response of an in-service bridge from its angular velocity, as measured by a inertial measurement unit (IMU). To verify our proposed method, field measurements were conducted using nine micro-electrical mechanical systems (MEMS) IMUs and two contact displacement gauges. The results showed that our proposed method provided high accuracy when compared to the reference responses calculated by the contact displacement gauges.

Central Cavity of Fructose-1,6-bisphosphatase and the Evolution of AMP/Fructose 2,6-bisphosphate Synergism in Eukaryotic Organisms*

PubMed Central

Gao, Yang; Shen, Lu; Honzatko, Richard B.

2014-01-01

The effects of AMP and fructose 2,6-bisphosphate (Fru-2,6-P2) on porcine fructose-1,6-bisphosphatase (pFBPase) and Escherichia coli FBPase (eFBPase) differ in three respects. AMP/Fru-2,6-P2 synergism in pFBPase is absent in eFBPase. Fru-2,6-P2 induces a 13° subunit pair rotation in pFBPase but no rotation in eFBPase. Hydrophilic side chains in eFBPase occupy what otherwise would be a central aqueous cavity observed in pFBPase. Explored here is the linkage of AMP/Fru-2,6-P2 synergism to the central cavity and the evolution of synergism in FBPases. The single mutation Ser45 → His substantially fills the central cavity of pFBPase, and the triple mutation Ser45 → His, Thr46 → Arg, and Leu186 → Tyr replaces porcine with E. coli type side chains. Both single and triple mutations significantly reduce synergism while retaining other wild-type kinetic properties. Similar to the effect of Fru-2,6-P2 on eFBPase, the triple mutant of pFBPase with bound Fru-2,6-P2 exhibits only a 2° subunit pair rotation as opposed to the 13° rotation exhibited by the Fru-2,6-P2 complex of wild-type pFBPase. The side chain at position 45 is small in all available eukaryotic FBPases but large and hydrophilic in bacterial FBPases, similar to eFBPase. Sequence information indicates the likelihood of synergism in the FBPase from Leptospira interrogans (lFBPase), and indeed recombinant lFBPase exhibits AMP/Fru-2,6-P2 synergism. Unexpectedly, however, AMP also enhances Fru-6-P binding to lFBPase. Taken together, these observations suggest the evolution of AMP/Fru-2,6-P2 synergism in eukaryotic FBPases from an ancestral FBPase having a central aqueous cavity and exhibiting synergistic feedback inhibition by AMP and Fru-6-P. PMID:24436333

A human-specific, truncated α7 nicotinic receptor subunit assembles with full-length α7 and forms functional receptors with different stoichiometries.

PubMed

Lasala, Matías; Corradi, Jeremías; Bruzzone, Ariana; Esandi, María Del Carmen; Bouzat, Cecilia

2018-05-21

The cholinergic α7 nicotinic receptor gene, CHRNA7, encodes a subunit that forms the homopentameric α7 receptor, involved in learning and memory. In humans, exons 5-10 in CHRNA7 are duplicated and fused to the FAM7A genetic element, giving rise to the hybrid gene CHRFAM7A. Its product, dupα7, is a truncated subunit lacking part of the N-terminal extracellular ligand-binding domain and is associated with neurological disorders, including schizophrenia, and immunomodulation.We combined dupα7 expression on mammalian cells with patch clamp recordings to understand its functional role. Transfected cells expressed dupα7 protein, but they exhibited neither surface binding of the α7 antagonist α-bungarotoxin nor responses to acetylcholine (ACh) or to an allosteric agonist that binds to the conserved transmembrane region. To determine if dupα7 assembles with α7, we generated receptors comprising α7 and dupα7 subunits, one of which was tagged with conductance substitutions that report subunit stoichiometry and monitored ACh-elicited channel openings elicited by ACh in the presence of a positive allosteric α7 modulator. We found that α7 and dupα7 subunits co-assemble into functional heteromeric receptors, that at least two α7 subunits are required for channel opening, and that dupα7's presence in the pentameric arrangement does not affect the duration of the potentiated events compare with that of α7. Using an α7 subunit mutant, we found that activation of (α7)2(dupα7)3 receptors occurs through ACh binding at the α7/α7 interfacial binding site. Our study contributes to the understanding of the modulation of α7 function by the human specific, duplicated subunit, associated with human disorders. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

Diesel emission reduction using internal exhaust gas recirculation

DOEpatents

He, Xin [Denver, CO; Durrett, Russell P [Bloomfield Hills, MI

2012-01-24

A method for controlling combustion in a direct-injection diesel engine includes monitoring a crankshaft rotational position of a cylinder of the engine, monitoring an engine load, determining an intake stroke within the cylinder based upon the crankshaft rotational position, and when the engine load is less than a threshold engine load, opening an exhaust valve for the cylinder during a portion of the intake stroke.

Temperature-dependent instability of the cTnI subunit in NIST SRM2921 characterized by tryptic peptide mapping.

PubMed

van der Burgt, Yuri E M; Cobbaert, Christa M; Dalebout, Hans; Smit, Nico; Deelder, André M

2012-08-01

In this study temperature-dependent instability of the cTnI subunit of the three-protein complex NIST SRM2921 was demonstrated using a mass spectrometric tryptic peptide mapping approach. The results were compared to the cTnI subunit obtained as a protein standard from Calbiochem with identical amino acid sequence. Both the three-protein complex from NIST as well as the cTnI subunit were incubated at elevated temperatures and then evaluated with respect to the primary sequence. The corresponding peptide maps were analyzed using LC-MS/MS. From a Mascot database search in combination with "semiTrypsin" tolerance it was found that two peptide backbone cleavages had occurred in subunit cTnI in NIST SRM2921 material upon incubation at 37°C, namely between amino acids at 148/149 and 194/195. The Calbiochem standard did not show increased levels of "unexpected" peptides in tryptic peptide maps. One of the two peptide backbone cleavages could also be monitored using a "single-step" MALDI-MS approach, i.e. without the need for peptide separation. The amount of degradation appeared rather constant in replicate temperature-instability experiments. However, for accurate quantification internal labelled standards are needed. Copyright © 2012 Elsevier B.V. All rights reserved.

Noise test system of rotating machinery in nuclear power station based on microphone array

NASA Astrophysics Data System (ADS)

Chang, Xincai; Guan, Jishi; Qi, Liangcai

2017-12-01

Rotating machinery plays an important role in all walks of life. Once the equipment fails, equipment maintenance and shutdown will cause great social harm and economic losses. Equipment safety operations at nuclear power stations have always been of top priority. It is prone to noise when the equipment is out of order or aging. Failure to find or develop equipment at the initial stage of equipment failure or ageing will pose a serious threat to the safety of the plant’s equipment. In this paper, sound imaging diagnosis technology is applied as a supplementary method to the condition monitoring and diagnosis system of rotating machinery in nuclear power stations. It provides a powerful guarantee for the condition monitoring and fault diagnosis of rotating machinery in nuclear power stations.

The Involvement of Ser1898 of the Human L-Type Calcium Channel in Evoked Secretion

PubMed Central

Bachnoff, Niv; Cohen-Kutner, Moshe; Atlas, Daphne

2011-01-01

A PKA consensus phosphorylation site S1928 at the α 11.2 subunit of the rabbit cardiac L-type channel, CaV1.2, is involved in the regulation of CaV1.2 kinetics and affects catecholamine secretion. This mutation does not alter basal CaV1.2 current properties or regulation of CaV1.2 current by PKA and the beta-adrenergic receptor, but abolishes CaV1.2 phosphorylation by PKA. Here, we test the contribution of the corresponding PKA phosphorylation site of the human α 11.2 subunit S1898, to the regulation of catecholamine secretion in bovine chromaffin cells. Chromaffin cells were infected with a Semliki-Forest viral vector containing either the human wt or a mutated S1898A α 11.2 subunit. Both subunits harbor a T1036Y mutation conferring nifedipine insensitivity. Secretion evoked by depolarization in the presence of nifedipine was monitored by amperometry. Depolarization-triggered secretion in cells infected with either the wt α 11.2 or α 11.2/S1898A mutated subunit was elevated to a similar extent by forskolin. Forskolin, known to directly activate adenylyl-cyclase, increased the rate of secretion in a manner that is largely independent of the presence of S1898. Our results are consistent with the involvement of additional PKA regulatory site(s) at the C-tail of α 11.2, the pore forming subunit of CaV1.2. PMID:22216029

P. aeruginosa PilT Structures with and without Nucleotide Reveal a Dynamic Type IV Pilus Retraction Motor

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

Misic, Ana M.; Satyshur, Kenneth A.; Forest, Katrina T.

Type IV pili are bacterial extracellular filaments that can be retracted to create force and motility. Retraction is accomplished by the motor protein PilT. Crystal structures of Pseudomonas aeruginosa PilT with and without bound {beta},{gamma}-methyleneadenosine-5{prime}-triphosphate have been solved at 2.6 {angstrom} and 3.1 {angstrom} resolution, respectively, revealing an interlocking hexamer formed by the action of a crystallographic 2-fold symmetry operator on three subunits in the asymmetric unit and held together by extensive ionic interactions. The roles of two invariant carboxylates, Asp Box motif Glu163 and Walker B motif Glu204, have been assigned to Mg{sup 2+} binding and catalysis, respectively. Themore » nucleotide ligands in each of the subunits in the asymmetric unit of the {beta},{gamma}-methyleneadenosine-5{prime}-triphosphate-bound PilT are not equally well ordered. Similarly, the three subunits in the asymmetric unit of both structures exhibit differing relative conformations of the two domains. The 12{sup o} and 20{sup o} domain rotations indicate motions that occur during the ATP-coupled mechanism of the disassembly of pili into membrane-localized pilin monomers. Integrating these observations, we propose a three-state 'Ready, Active, Release' model for the action of PilT.« less

Feasibility of vibration monitoring of small rotating machines for the environmental control and life support systems (ECLSS) of the NASA advanced space craft

NASA Technical Reports Server (NTRS)

Milner, G. Martin; Black, Mike; Hovenga, Mike; Mcclure, Paul; Miller, Patrice

1988-01-01

The application of vibration monitoring to the rotating machinery typical of ECLSS components in advanced NASA spacecraft was studied. It is found that the weighted summation of the accelerometer power spectrum is the most successful detection scheme for a majority of problem types. Other detection schemes studied included high-frequency demodulation, cepstrum, clustering, and amplitude processing.

FOSREM - Fibre-Optic System for Rotational Events&Phenomena Monitoring

NASA Astrophysics Data System (ADS)

Jaroszewicz, Leszek; Krajewski, Zbigniew; Kurzych, Anna; Kowalski, Jerzy; Teisseyre, Krzysztof

2016-04-01

We present the construction and tests of fiber-optic rotational seismometer named FOSREM (Fibre-Optic System for Rotational Events&Phenomena Monitoring). This presented device is designed for detection and monitoring the one-axis rotational motions, brought about to ground or human-made structures both by seismic events and the creep processes. The presented system works by measuring Sagnac effect and generally consists of two basic elements: optical sensor and electronic part. The optical sensor is based on so-called the minimum configuration of FOG (Fibre-Optic Gyroscope) where the Sagnac effect produces a phase shift between two counter-propagating light beams proportional to the measured rotation speed. The main advantage of the sensor of this type is its complete insensitivity to linear motions and a direct measurement of rotational speed. It may work even when tilted, moreover, used in continuous mode it may record the tilt. The electronic system, involving specific electronic solutions, calculates and records rotational events data by realizing synchronous in a digital form by using 32 bit DSP (Digital Signal Processing). Storage data and system control are realised over the internet by using connection between FOSREM and GSM/GPS. The most significant attribute of our system is possibility to measure rotation in wide range both amplitude up to 10 rad/s and frequency up to 328.12 Hz. Application of the wideband, low coherence and high power superluminescent diode with long fibre loop and suitable low losses optical elements assures the theoretical sensitivity of the system equal to 2·10-8 rad/s/Sqrt(Hz). Moreover, the FOSREM is fully remote controlled as well as is suited for continuous, autonomous work in very long period of time (weeks, months, even years), so it is useful for systematic seismological investigation at any place. Possible applications of this system include seismic monitoring in observatories, buildings, mines and even on glaciers and in their vicinity. In geodetic, geomorphological and glaciological survey, joint measurement of tilt and seismic phenomena using a set of three FOSREM devices oriented in perpendicular planes would enable to collect very important information.

Characterization of inhibin forms and their measurement by an inhibin alpha-subunit ELISA in serum from postmenopausal women with ovarian cancer.

PubMed

Robertson, D M; Stephenson, T; Pruysers, E; McCloud, P; Tsigos, A; Groome, N; Mamers, P; Burger, H G

2002-02-01

The aim of this study was to characterize the molecular wt forms of inhibins A and B and its free alpha-subunit present in serum from women with ovarian cancer as a basis for developing improved monoclonal antibody-based inhibin assays for monitoring ovarian cancer. Three new inhibin alpha-subunit (alphaC) ELISAs were developed using monoclonal antibodies directed to three nonoverlapping peptide regions of the alphaC region of the inhibin alpha-subunit. To characterize serum inhibin molecular wt forms present in women with ovarian cancer, existing inhibin immunoassays (inhibin A, inhibin B, and pro-alphaC) and the new alphaC ELISAs were applied to sera from women with granulosa cell tumors and mucinous carcinomas previously fractionated using a combined immunoaffinity chromatography, preparative SDS-PAGE, and electroelution procedure. The distribution and molecular size of dimeric inhibins and alpha-subunit detected were consistent with known mol wt forms of inhibins A and B and inhibin alpha-subunit and their precursor forms present in serum and follicular fluid from healthy women. The alphaC ELISAs recognized all known forms of inhibin and the free inhibin alpha-subunit, although differences between alphaC ELISAs were observed in their ability to detect high mol wt forms. To assess which of the alphaC ELISAs was preferred in application to ovarian cancer, the alphaC ELISAs were applied to serum from a range of normal postmenopausal women (n = 61) and postmenopausal women (n = 152) with ovarian (serous, mucinous, endometrioid, clear cell carcinomas, and granulosa cell tumors) and nonovarian (breast and colon) cancers. Despite differences in their ability to detect high mol wt forms of inhibin, the alphaC ELISAs showed similar sensitivity (i.e. proportion of cancer patients correctly detected) and specificity (proportion of controls correctly detected) indexes in the detection of mucinous carcinomas (84% and 95%) and granulosa cell tumors (100% and 95%) compared with earlier inhibin RIA or polyclonal antibody-based immunofluorometric assays. A combination of the alphaC ELISAs with the CA125 assay, an ovarian tumor marker that has a high sensitivity and specificity for other ovarian cancers (serous, clear cell, and endometrioid), resulted in an increase in sensitivity/specificity indexes (95% and 95%) for the all ovarian cancer group. These new monoclonal antibody-based inhibin alphaC ELISAs now provide practical and sensitive assays suitable for evaluation as diagnostic tests for monitoring ovarian cancers.

Controlling Sample Rotation in Acoustic Levitation

NASA Technical Reports Server (NTRS)

Barmatz, M. B.; Stoneburner, J. D.

1985-01-01

Rotation of acoustically levitated object stopped or controlled according to phase-shift monitoring and control concept. Principle applies to square-cross-section levitation chamber with two perpendicular acoustic drivers operating at same frequency. Phase difference between X and Y acoustic excitation measured at one corner by measuring variation of acoustic amplitude sensed by microphone. Phase of driver adjusted to value that produces no rotation or controlled rotation of levitated object.

Determination Method of Bridge Rotation Angle Response Using MEMS IMU

PubMed Central

Sekiya, Hidehiko; Kinomoto, Takeshi; Miki, Chitoshi

2016-01-01

To implement steel bridge maintenance, especially that related to fatigue damage, it is important to monitor bridge deformations under traffic conditions. Bridges deform and rotate differently under traffic load conditions because their structures differ in terms of length and flexibility. Such monitoring enables the identification of the cause of stress concentrations that cause fatigue damage and the proposal of appropriate countermeasures. However, although bridge deformation monitoring requires observations of bridge angle response as well as the bridge displacement response, measuring the rotation angle response of a bridge subject to traffic loads is difficult. Theoretically, the rotation angle response can be calculated by integrating the angular velocity, but for field measurements of actual in-service bridges, estimating the necessary boundary conditions would be difficult due to traffic-induced vibration. To solve the problem, this paper proposes a method for determining the rotation angle response of an in-service bridge from its angular velocity, as measured by a inertial measurement unit (IMU). To verify our proposed method, field measurements were conducted using nine micro-electrical mechanical systems (MEMS) IMUs and two contact displacement gauges. The results showed that our proposed method provided high accuracy when compared to the reference responses calculated by the contact displacement gauges. PMID:27834871

Fluorescence Polarization and Fluctuation Analysis Monitors Subunit Proximity, Stoichiometry, and Protein Complex Hydrodynamics

PubMed Central

Nguyen, Tuan A.; Sarkar, Pabak; Veetil, Jithesh V.; Koushik, Srinagesh V.; Vogel, Steven S.

2012-01-01

Förster resonance energy transfer (FRET) microscopy is frequently used to study protein interactions and conformational changes in living cells. The utility of FRET is limited by false positive and negative signals. To overcome these limitations we have developed Fluorescence Polarization and Fluctuation Analysis (FPFA), a hybrid single-molecule based method combining time-resolved fluorescence anisotropy (homo-FRET) and fluorescence correlation spectroscopy. Using FPFA, homo-FRET (a 1–10 nm proximity gauge), brightness (a measure of the number of fluorescent subunits in a complex), and correlation time (an attribute sensitive to the mass and shape of a protein complex) can be simultaneously measured. These measurements together rigorously constrain the interpretation of FRET signals. Venus based control-constructs were used to validate FPFA. The utility of FPFA was demonstrated by measuring in living cells the number of subunits in the α-isoform of Venus-tagged calcium-calmodulin dependent protein kinase-II (CaMKIIα) holoenzyme. Brightness analysis revealed that the holoenzyme has, on average, 11.9±1.2 subunit, but values ranged from 10–14 in individual cells. Homo-FRET analysis simultaneously detected that catalytic domains were arranged as dimers in the dodecameric holoenzyme, and this paired organization was confirmed by quantitative hetero-FRET analysis. In freshly prepared cell homogenates FPFA detected only 10.2±1.3 subunits in the holoenzyme with values ranging from 9–12. Despite the reduction in subunit number, catalytic domains were still arranged as pairs in homogenates. Thus, FPFA suggests that while the absolute number of subunits in an auto-inhibited holoenzyme might vary from cell to cell, the organization of catalytic domains into pairs is preserved. PMID:22666486

Scanning sky monitor (SSM) onboard AstroSat

NASA Astrophysics Data System (ADS)

Ramadevi, M. C.; Seetha, S.; Bhattacharya, Dipankar; Ravishankar, B. T.; Sitaramamurthy, N.; Meena, G.; Sharma, M. Ramakrishna; Kulkarni, Ravi; Babu, V. Chandra; Kumar; Singh, Brajpal; Jain, Anand; Yadav, Reena; Vaishali, S.; Ashoka, B. N.; Agarwal, Anil; Balaji, K.; Nagesh, G.; Kumar, Manoj; Gaan, Dhruti Ranjan; Kulshresta, Prashanth; Agarwal, Pankaj; Sebastian, Mathew; Rajarajan, A.; Radhika, D.; Nandi, Anuj; Girish, V.; Agarwal, Vivek Kumar; Kushwaha, Ankur; Iyer, Nirmal Kumar

2017-10-01

Scanning Sky Monitor (SSM) onboard AstroSat is an Xray sky monitor in the soft X-ray band designed with a large field of view to detect and locate transient X-ray sources and alert the astronomical community about interesting phenomena in the X-ray sky. SSM comprises position sensitive proportional counters with 1D coded mask for imaging. There are three detector units mounted on a platform capable of rotation which helps covering about 50% of the sky in one full rotation. This paper discusses the elaborate details of the instrument and few immediate results from the instrument after launch.

Monitoring of wind turbines

DOEpatents

White, Jonathan R.; Adams, Douglas E.; Paquette, Josh

2017-07-25

Method and apparatus for determining the deflection or curvature of a rotating blade, such as a wind turbine blade or a helicopter blade. Also, methods and apparatus for establishing an inertial reference system on a rotating blade.

Spatio-temporal brain activity related to rotation method during a mental rotation task of three-dimensional objects: an MEG study.

PubMed

Kawamichi, Hiroaki; Kikuchi, Yoshiaki; Ueno, Shoogo

2007-09-01

During mental rotation tasks, subjects perform mental simulation to solve tasks. However, detailed neural mechanisms underlying mental rotation of three-dimensional (3D) objects, particularly, whether higher motor areas related to mental simulation are activated, remain unknown. We hypothesized that environmental monitoring-a process based on environmental information and is included in motor execution-is as a key factor affecting the utilization of higher motor areas. Therefore, using magnetoencephalography (MEG), we measured spatio-temporal brain activities during two types (two-dimensional (2D) and 3D rotation tasks) of mental rotation of 3D objects. Only the 3D rotation tasks required subjects to mentally rotate objects in a depth plane with visualization of hidden parts of the visual stimuli by acquiring and retrieving 3D information. In cases showing significant differences in the averaged activities at 100-ms intervals between the two rotations, the activities were located in the right dorsal premotor (PMd) at approximately 500 ms. In these cases, averaged activities during 3D rotation were greater than those during 2D rotation, implying that the right PMd activities are related to environmental monitoring. During 3D rotation, higher activities were observed from 200 to 300 ms in the left PMd and from 400 to 700 ms in the right PMd. It is considered that the left PMd is related to primary motor control, whereas the right PMd plays a supplementary role during mental simulation. Further, during 3D rotation, late higher activities related to mental simulation are observed in the right superior parietal lobule (SPL), which is connected to PMd.

Torque generation mechanism of ATP synthase

NASA Astrophysics Data System (ADS)

Miller, John; Maric, Sladjana; Scoppa, M.; Cheung, M.

2010-03-01

ATP synthase is a rotary motor that produces adenosine triphosphate (ATP), the chemical currency of life. Our proposed electric field driven torque (EFT) model of FoF1-ATP synthase describes how torque, which scales with the number of c-ring proton binding sites, is generated by the proton motive force (pmf) across the mitochondrial inner membrane. When Fo is coupled to F1, the model predicts a critical pmf to drive ATP production. In order to fully understand how the electric field resulting from the pmf drives the c-ring to rotate, it is important to examine the charge distributions in the protonated c-ring and a-subunit containing the proton channels. Our calculations use a self-consistent field approach based on a refinement of reported structural data. The results reveal changes in pKa for key residues on the a-subunit and c-ring, as well as titration curves and protonation state energy diagrams. Health implications will be briefly discussed.

The chloroplast ATP synthase features the characteristic redox regulation machinery.

PubMed

Hisabori, Toru; Sunamura, Ei-Ichiro; Kim, Yusung; Konno, Hiroki

2013-11-20

Regulation of the activity of the chloroplast ATP synthase is largely accomplished by the chloroplast thioredoxin system, the main redox regulation system in chloroplasts, which is directly coupled to the photosynthetic reaction. We review the current understanding of the redox regulation system of the chloroplast ATP synthase. The thioredoxin-targeted portion of the ATP synthase consists of two cysteines located on the central axis subunit γ. The redox state of these two cysteines is under the influence of chloroplast thioredoxin, which directly controls rotation during catalysis by inducing a conformational change in this subunit. The molecular mechanism of redox regulation of the chloroplast ATP synthase has recently been determined. Regulation of the activity of the chloroplast ATP synthase is critical in driving efficiency into the ATP synthesis reaction in chloroplasts. The molecular architecture of the chloroplast ATP synthase, which confers redox regulatory properties requires further investigation, in light of the molecular structure of the enzyme complex as well as the physiological significance of the regulation system.

High-Resolution Structure and Mechanism of an F/V-Hybrid Rotor Ring in a Na+-coupled ATP Synthase

PubMed Central

Matthies, Doreen; Zhou, Wenchang; Klyszejko, Adriana L.; Anselmi, Claudio; Yildiz, Özkan; Brandt, Karsten; Müller, Volker; Faraldo-Gómez, José D.; Meier, Thomas

2014-01-01

All rotary ATPases catalyze the interconversion of ATP and ADP-Pi through a mechanism that is coupled to the transmembrane flow of H+ or Na+. Physiologically, however, F/A-type enzymes specialize in ATP synthesis driven by downhill ion diffusion, while eukaryotic V-type ATPases function as ion pumps. To begin to rationalize the molecular basis for this functional differentiation, we solved the crystal structure of the Na+-driven membrane rotor of the Acetobacterium woodii ATP synthase, at 2.1 Å resolution. Unlike known structures, this rotor ring is a 9:1 heteromer of F- and V-type c-subunits, and therefore features a hybrid configuration of ion-binding sites along its circumference. Molecular and kinetic simulations are used to dissect the mechanisms of Na+ recognition and rotation of this c-ring, and to explain the functional implications of the V-type c-subunit. These structural and mechanistic insights indicate an evolutionary path between synthases and pumps involving adaptations in the rotor ring. PMID:25381992

Structural Basis for Catalytic Activation of a Serine Recombinase

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

Keenholtz, Ross A.; Rowland, Sally-J.; Boocock, Martin R.

2014-10-02

Sin resolvase is a site-specific serine recombinase that is normally controlled by a complex regulatory mechanism. A single mutation, Q115R, allows the enzyme to bypass the entire regulatory apparatus, such that no accessory proteins or DNA sites are required. Here, we present a 1.86 {angstrom} crystal structure of the Sin Q115R catalytic domain, in a tetrameric arrangement stabilized by an interaction between Arg115 residues on neighboring subunits. The subunits have undergone significant conformational changes from the inactive dimeric state previously reported. The structure provides a new high-resolution view of a serine recombinase active site that is apparently fully assembled, suggestingmore » roles for the conserved active site residues. The structure also suggests how the dimer-tetramer transition is coupled to assembly of the active site. The tetramer is captured in a different rotational substate than that seen in previous hyperactive serine recombinase structures, and unbroken crossover site DNA can be readily modeled into its active sites.« less

Cloning, expression, purification, crystallization and preliminary X-ray studies of argininosuccinate lyase (Rv1659) from Mycobacterium tuberculosis

PubMed Central

Paul, A.; Mishra, A.; Surolia, A.; Vijayan, M.

2013-01-01

The last enzyme in the arginine-biosynthesis pathway, argininosuccinate lyase, from Mycobacterium tuberculosis has been cloned, expressed, purified and crystallized, and preliminary X-ray studies have been carried out on the crystals. The His-tagged tetrameric enzyme with a subunit molecular weight of 50.9 kDa crystallized with two tetramers in the asymmetric unit of the orthorhombic unit cell, space group P212121. Molecular-replacement calculations and self-rotation calculations confirmed the space group and the tetrameric nature of the molecule. PMID:24316845

Millimeter Wave Spectrum of the Weakly Bound Complex CH2═CHCN·H2O: Structure, Dynamics, and Implications for Astronomical Search.

PubMed

Calabrese, Camilla; Vigorito, Annalisa; Maris, Assimo; Mariotti, Sergio; Fathi, Pantea; Geppert, Wolf D; Melandri, Sonia

2015-12-03

The weakly bound 1:1 complex between acrylonitrile (CH2═CHCN) and water has been characterized spectroscopically in the millimeter wave range (59.6-74.4 GHz) using a Free Jet Absorption Millimeter Wave spectrometer. Precise values of the rotational and quartic centrifugal distortion constants have been obtained from the measured frequencies of the normal and isotopically substituted water moiety (DOH, DOD, H(18)OH). Structural parameters have been estimated from the rotational constants and their differences among isotopologues: the complex has a planar structure with the two subunits held together by a O-H···N (2.331(3) Å) and a C-H···O (2.508(4) Å) interaction. The ab initio intermolecular binding energy, obtained at the counterpoise corrected MP2/aug-cc-pVTZ level of calculation, is De = 24.4 kJ mol(-1).

Propulsion Health Monitoring of a Turbine Engine Disk Using Spin Test Data

NASA Technical Reports Server (NTRS)

Abdul-Aziz, Ali; Woike, Mark R.; Oza, Nikunj; Matthews, Bryan; Baaklini, George Y.

2010-01-01

This paper considers data collected from an experimental study using high frequency capacitive sensor technology to capture blade tip clearance and tip timing measurements in a rotating turbine engine-like-disk-to predict the disk faults and assess its structural integrity. The experimental results collected at a range of rotational speeds from tests conducted at the NASA Glenn Research Center s Rotordynamics Laboratory are evaluated using multiple data-driven anomaly detection techniques to identify abnormalities in the disk. Further, this study presents a select evaluation of an online health monitoring scheme of a rotating disk using high caliber sensors and test the capability of the in-house spin system.

Monitoring Bridge Dynamic Responses Using Fiber Bragg Grating Tiltmeters.

PubMed

Xiao, Feng; Chen, Gang S; Hulsey, J Leroy

2017-10-20

In bridge health monitoring, tiltmeters have been used for measuring rotation and curvature; however, their application in dynamic parameter identification has been lacking. This study installed fiber Bragg grating (FBG) tiltmeters on the bearings of a bridge and monitored the dynamic rotational angle. The dynamic features, including natural frequencies and mode shapes, have been identified successfully. The innovation presented in this paper is the first-time use of FBG tiltmeter readings to identify the natural frequencies of a long-span steel girder bridge. The identified results have been verified using a bridge finite element model. This paper introduces a new method for the dynamic monitoring of a bridge using FBG tiltmeters. Limitations and future research directions are also discussed in the conclusion.

Identification of marker proteins for the adulteration of meat products with soybean proteins by multidimensional liquid chromatography-tandem mass spectrometry.

PubMed

Leitner, Alexander; Castro-Rubio, Florentina; Marina, Maria Luisa; Lindner, Wolfgang

2006-09-01

Soybean proteins are frequently added to processed meat products for economic reasons and to improve their functional properties. Monitoring of the addition of soybean protein to meat products is of high interest due to the existence of regulations forbidding or limiting the amount of soybean proteins that can be added during the processing of meat products. We have used chromatographic prefractionation on the protein level by perfusion liquid chromatography to isolate peaks of interest from extracts of soybean protein isolate (SPI) and of meat products containing SPI. After enzymatic digestion using trypsin, the collected fractions were analyzed by nanoflow liquid chromatography-tandem mass spectrometry. Several variants and subunits of the major seed proteins, glycinin and beta-conglycinin, were identified in SPI, along with two other proteins. In soybean-protein-containing meat samples, different glycinin A subunits could be identified from the peak discriminating between samples with and without soybean proteins added. Among those, glycinin G4 subunit A4 was consistently found in all samples. Consequently, this protein (subunit) can be used as a target for new analytical techniques in the course of identifying the addition of soybean protein to meat products.

System for monitoring the growth of crystalline films on stationary substrates

DOEpatents

Sheldon, P.

1996-12-31

A system for monitoring the growth of crystalline films on stationary or rotating substrates includes a combination of some or all of the elements including a photodiode sensor for detecting the intensity of incoming light and converting it to a measurable current, a lens for focusing the RHEED pattern emanating from the phosphor screen onto the photodiode, an interference filter for filtering out light other than that which emanates from the phosphor screen, a current amplifier for amplifying and convening the current produced by the photodiode into a voltage, a computer for receiving the amplified photodiode current for RHEED data analysis, and a graphite impregnated triaxial cable for improving the signal to noise ratio obtained while sampling a stationary or rotating substrate. A rotating stage for supporting the substrate with diametrically positioned electron beam apertures and an optically encoded shaft can also be used to accommodate rotation of the substrate during measurement. 16 figs.

System for monitoring the growth of crystalline films on stationary substrates

DOEpatents

Sheldon, P.

1995-10-10

A system for monitoring the growth of crystalline films on stationary or rotating substrates includes a combination of some or all of the elements including a photodiode sensor for detecting the intensity of incoming light and converting it to a measurable current, a lens for focusing the RHEED pattern emanating from the phosphor screen onto the photodiode, an interference filter for filtering out light other than that which emanates from the phosphor screen, a current amplifier for amplifying and converting the current produced by the photodiode into a voltage, a computer for receiving the amplified photodiode current for RHEED data analysis, and a graphite impregnated triaxial cable for improving the signal-to-noise ratio obtained while sampling a stationary or rotating substrate. A rotating stage for supporting the substrate with diametrically positioned electron beam apertures and an optically encoded shaft can also be used to accommodate rotation of the substrate during measurement. 16 figs.

System for monitoring the growth of crystalline films on stationary substrates

DOEpatents

Sheldon, Peter

1995-01-01

A system for monitoring the growth of crystalline films on stationary or rotating substrates includes a combination of some or all of the elements including a photodiode sensor for detecting the intensity of incoming light and converting it to a measurable current, a lens for focusing the RHEED pattern emanating from the phosphor screen onto the photodiode, an interference filter for filtering out light other than that which emanates from the phosphor screen, a current amplifier for amplifying and converting the current produced by the photodiode into a voltage, a computer for receiving the amplified photodiode current for RHEED data analysis, and a graphite impregnated triax cable for improving the signal to noise ratio obtained while sampling a stationary or rotating substrate. A rotating stage for supporting the substrate with diametrically positioned electron beam apertures and an optically encoded shaft can also be used to accommodate rotation of the substrate during measurement.

System for monitoring the growth of crystalline films on stationary substrates

DOEpatents

Sheldon, Peter

1996-01-01

A system for monitoring the growth of crystalline films on stationary or rotating substrates includes a combination of some or all of the elements including a photodiode sensor for detecting the intensity of incoming light and converting it to a measurable current, a lens for focusing the RHEED pattern emanating from the phosphor screen onto the photodiode, an interference filter for filtering out light other than that which emanates from the phosphor screen, a current amplifier for amplifying and convening the current produced by the photodiode into a voltage, a computer for receiving the amplified photodiode current for RHEED data analysis, and a graphite impregnated triax cable for improving the signal to noise ratio obtained while sampling a stationary or rotating substrate. A rotating stage for supporting the substrate with diametrically positioned electron beam apertures and an optically encoded shaft can also be used to accommodate rotation of the substrate during measurement.

Direct Observation of Cooperative Protein Structural Dynamics of Homodimeric Hemoglobin from 100 ps to 10 ms with Pump-Probe X-ray Solution Scattering

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

Kim, Kyung Hwan; Muniyappan, Srinivasan; Oang, Key Young

2012-05-29

Proteins serve as molecular machines in performing their biological functions, but the detailed structural transitions are difficult to observe in their native aqueous environments in real time. For example, despite extensive studies, the solution-phase structures of the intermediates along the allosteric pathways for the transitions between the relaxed (R) and tense (T) forms have been elusive. In this work, we employed picosecond X-ray solution scattering and novel structural analysis to track the details of the structural dynamics of wild-type homodimeric hemoglobin (HbI) from the clam Scapharca inaequivalvis and its F97Y mutant over a wide time range from 100 ps tomore » 56.2 ms. From kinetic analysis of the measured time-resolved X-ray solution scattering data, we identified three structurally distinct intermediates (I 1, I 2, and I 3) and their kinetic pathways common for both the wild type and the mutant. The data revealed that the singly liganded and unliganded forms of each intermediate share the same structure, providing direct evidence that the ligand photolysis of only a single subunit induces the same structural change as the complete photolysis of both subunits does. In addition, by applying novel structural analysis to the scattering data, we elucidated the detailed structural changes in the protein, including changes in the heme-heme distance, the quaternary rotation angle of subunits, and interfacial water gain/loss. The earliest, R-like I 1 intermediate is generated within 100 ps and transforms to the R-like I 2 intermediate with a time constant of 3.2 ± 0.2 ns. Subsequently, the late, T-like I 3 intermediate is formed via subunit rotation, a decrease in the heme-heme distance, and substantial gain of interfacial water and exhibits ligation-dependent formation kinetics with time constants of 730 ± 120 ns for the fully photolyzed form and 5.6 ± 0.8 μs for the partially photolyzed form. For the mutant, the overall kinetics are accelerated, and the formation of the T-like I 3 intermediate involves interfacial water loss (instead of water entry) and lacks the contraction of the heme-heme distance, thus underscoring the dramatic effect of the F97Y mutation. The ability to keep track of the detailed movements of the protein in aqueous solution in real time provides new insights into the protein structural dynamics.« less

All-around viewing display system for group activity on life review therapy

NASA Astrophysics Data System (ADS)

Sakamoto, Kunio; Okumura, Mitsuru

2009-10-01

This paper describes 360 degree viewing display system that can be viewed from any direction. A conventional monitor display is viewed from one direction, i.e., the display has narrow viewing angle and observers cannot view the screen from the opposite side. To solve this problem, we developed the 360 degree viewing display for collaborative tasks on the round table. This developed 360 degree viewing system has a liquid crystal display screen and a 360 degree rotating table by motor. The principle is very simple. The screen of a monitor only rotates at a uniform speed, but the optical techniques are also utilized. Moreover, we have developed a floating 360 degree viewing display that can be viewed from any direction. This new viewing system has a display screen, a rotating table and dual parabolic mirrors. In order to float the only image screen above the table, the rotating mechanism works in the parabolic mirrors. Because the dual parabolic mirrors generate a "mirage" image over the upper mirror, observers can view a floating 2D image on the virtual screen in front of them. Then the observer can view a monitor screen at any position surrounding the round table.

Rotating Desk for Collaboration by Two Computer Programmers

NASA Technical Reports Server (NTRS)

Riley, John Thomas

2005-01-01

A special-purpose desk has been designed to facilitate collaboration by two computer programmers sharing one desktop computer or computer terminal. The impetus for the design is a trend toward what is known in the software industry as extreme programming an approach intended to ensure high quality without sacrificing the quantity of computer code produced. Programmers working in pairs is a major feature of extreme programming. The present desk design minimizes the stress of the collaborative work environment. It supports both quality and work flow by making it unnecessary for programmers to get in each other s way. The desk (see figure) includes a rotating platform that supports a computer video monitor, keyboard, and mouse. The desk enables one programmer to work on the keyboard for any amount of time and then the other programmer to take over without breaking the train of thought. The rotating platform is supported by a turntable bearing that, in turn, is supported by a weighted base. The platform contains weights to improve its balance. The base includes a stand for a computer, and is shaped and dimensioned to provide adequate foot clearance for both users. The platform includes an adjustable stand for the monitor, a surface for the keyboard and mouse, and spaces for work papers, drinks, and snacks. The heights of the monitor, keyboard, and mouse are set to minimize stress. The platform can be rotated through an angle of 40 to give either user a straight-on view of the monitor and full access to the keyboard and mouse. Magnetic latches keep the platform preferentially at either of the two extremes of rotation. To switch between users, one simply grabs the edge of the platform and pulls it around. The magnetic latch is easily released, allowing the platform to rotate freely to the position of the other user

Propulsion health monitoring of a turbine engine disk using spin test data

NASA Astrophysics Data System (ADS)

Abdul-Aziz, Ali; Woike, Mark; Oza, Nikunj; Matthews, Bryan; Baakilini, George

2010-03-01

On line detection techniques to monitor the health of rotating engine components are becoming increasingly attractive options to aircraft engine companies in order to increase safety of operation and lower maintenance costs. Health monitoring remains a challenging feature to easily implement, especially, in the presence of scattered loading conditions, crack size, component geometry and materials properties. The current trend, however, is to utilize noninvasive types of health monitoring or nondestructive techniques to detect hidden flaws and mini cracks before any catastrophic event occurs. These techniques go further to evaluate materials' discontinuities and other anomalies that have grown to the level of critical defects which can lead to failure. Generally, health monitoring is highly dependent on sensor systems that are capable of performing in various engine environmental conditions and able to transmit a signal upon a predetermined crack length, while acting in a neutral form upon the overall performance of the engine system. Efforts are under way at NASA Glenn Research Center through support of the Intelligent Vehicle Health Management Project (IVHM) to develop and implement such sensor technology for a wide variety of applications. These efforts are focused on developing high temperature, wireless, low cost and durable products. Therefore, in an effort to address the technical issues concerning health monitoring of a rotor disk, this paper considers data collected from an experimental study using high frequency capacitive sensor technology to capture blade tip clearance and tip timing measurements in a rotating engine-like-disk-to predict the disk faults and assess its structural integrity. The experimental results collected at a range of rotational speeds from tests conducted at the NASA Glenn Research Center's Rotordynamics Laboratory will be evaluated using multiple data-driven anomaly detection techniques to identify anomalies in the disk. This study is expected to present a select evaluation of online health monitoring of a rotating disk using these high caliber sensors and test the capability of the in-house spin system.

Ligand-Induced Conformational Change in the α7 Nicotinic Receptor Ligand Binding Domain

PubMed Central

Henchman, Richard H.; Wang, Hai-Long; Sine, Steven M.; Taylor, Palmer; McCammon, J. Andrew

2005-01-01

Molecular dynamics simulations of a homology model of the ligand binding domain of the α7 nicotinic receptor are conducted with a range of bound ligands to induce different conformational states. Four simulations of 15 ns each are run with no ligand, antagonist d-tubocurarine (dTC), agonist acetylcholine (ACh), and agonist ACh with potentiator Ca2+, to give insight into the conformations of the active and inactive states of the receptor and suggest the mechanism for conformational change. The main structural factor distinguishing the active and inactive states is that a more open, symmetric arrangement of the five subunits arises for the two agonist simulations, whereas a more closed and asymmetric arrangement results for the apo and dTC cases. Most of the difference arises in the lower portion of the ligand binding domain near its connection to the adjacent transmembrane domain. The transfer of the more open state to the transmembrane domain could then promote ion flow through the channel. Variation in how subunits pack together with no ligand bound appears to give rise to asymmetry in the apo case. The presence of dTC expands the receptor but induces rotations in alternate directions in adjacent subunits that lead to an asymmetric arrangement as in the apo case. Ca2+ appears to promote a slightly greater expansion in the subunits than ACh alone by stabilizing the C-loop and ACh positions. Although the simulations are unlikely to be long enough to view the full conformational changes between open and closed states, a collection of different motions at a range of length scales are observed that are likely to participate in the conformational change. PMID:15665135

STRUCTURE OF THE VIBRIO CHOLERAE TYPE IVB PILUS AND STABILITY COMPARISON WITH THE NEISSERIA GONORRHOEAE TYPE IVA PILUS

PubMed Central

Li, Juliana; Egelman, Edward H.; Craig, Lisa

2012-01-01

Type IV pili are multifunctional filaments displayed on many bacterial pathogens. Members of the Type IVa pilus subclass are found on a diverse group of human pathogens whereas Type IVb pili are almost exclusively on enteric bacteria. The Type IVa and IVb subclasses are distinguished by differences in the pilin subunits, including the fold of the globular domain. To understand the implications of the distinct pilin folds we compared the stabilities of pilin subunits and pilus filaments for the Type IVa GC pilus from Neisseria gonorrhoeae and the Type IVb toxin-coregulated pilus (TCP) from Vibrio cholerae. We show that while recombinant TCP pilin is more stable than GC pilin, the GC pili are more resistant than TCP to proteolysis, heat and chemical denaturation, remaining intact in 8 M urea. To understand these differences we determined the TCP structure by electron microscopy and 3D image reconstruction. TCP have a similar architecture to GC pili, with subunits arranged in a right-handed one-start helix and related by an 8.4 Å axial rise and a 96.8° azimuthal rotation. However, the TCP subunits are not as tightly packed as GC pilins and the distinct Type IVb pilin fold exposes a segment of the α-helical core of TCP. Hydrophobic interactions dominate for both pilus subtypes, but base-stacking by aromatic residues conserved among the Type IVa pilins may contribute to GC pilus stability. The extraordinary stability of GC pili may represent an adaptation of the Type IVa pili to harsh environments and the need to retract against external forces. PMID:22361030

Asymmetry in the function and dynamics of the cytosolic group II chaperonin CCT/TRiC

PubMed Central

Yamamoto, Yohei Y.; Uno, Yuko; Sha, Eiryo; Ikegami, Kentaro; Ishii, Noriyuki; Dohmae, Naoshi; Sekiguchi, Hiroshi; Sasaki, Yuji C.

2017-01-01

The eukaryotic group II chaperonin, the chaperonin-containing t-complex polypeptide 1 (CCT), plays an important role in cytosolic proteostasis. It has been estimated that as much as 10% of cytosolic proteins interact with CCT during their folding process. CCT is composed of 8 different paralogous subunits. Due to its complicated structure, molecular and biochemical investigations of CCT have been difficult. In this study, we constructed an expression system for CCT from a thermophilic fungus, Chaetomium thermophilum (CtCCT), by using E. coli as a host. As expected, we obtained recombinant CtCCT with a relatively high yield, and it exhibited fairly high thermal stability. We showed the advantages of the overproduction system by characterizing CtCCT variants containing ATPase-deficient subunits. For diffracted X-ray tracking experiment, we removed all surface exposed cysteine residues, and added cysteine residues at the tip of helical protrusions of selected two subunits. Gold nanocrystals were attached onto CtCCTs via gold-thiol bonds and applied for the analysis by diffracted X-ray tracking. Irrespective of the locations of cysteines, it was shown that ATP binding induces tilting motion followed by rotational motion in the CtCCT molecule, like the archaeal group II chaperonins. When gold nanocrystals were attached onto two subunits in the high ATPase activity hemisphere, the CtCCT complex exhibited a fairly rapid response to the motion. In contrast, the response of CtCCT, which had gold nanocrystals attached to the low-activity hemisphere, was slow. These results clearly support the possibility that ATP-dependent conformational change starts with the high-affinity hemisphere and progresses to the low-affinity hemisphere. PMID:28463997

Asymmetry in the function and dynamics of the cytosolic group II chaperonin CCT/TRiC.

PubMed

Yamamoto, Yohei Y; Uno, Yuko; Sha, Eiryo; Ikegami, Kentaro; Ishii, Noriyuki; Dohmae, Naoshi; Sekiguchi, Hiroshi; Sasaki, Yuji C; Yohda, Masafumi

2017-01-01

The eukaryotic group II chaperonin, the chaperonin-containing t-complex polypeptide 1 (CCT), plays an important role in cytosolic proteostasis. It has been estimated that as much as 10% of cytosolic proteins interact with CCT during their folding process. CCT is composed of 8 different paralogous subunits. Due to its complicated structure, molecular and biochemical investigations of CCT have been difficult. In this study, we constructed an expression system for CCT from a thermophilic fungus, Chaetomium thermophilum (CtCCT), by using E. coli as a host. As expected, we obtained recombinant CtCCT with a relatively high yield, and it exhibited fairly high thermal stability. We showed the advantages of the overproduction system by characterizing CtCCT variants containing ATPase-deficient subunits. For diffracted X-ray tracking experiment, we removed all surface exposed cysteine residues, and added cysteine residues at the tip of helical protrusions of selected two subunits. Gold nanocrystals were attached onto CtCCTs via gold-thiol bonds and applied for the analysis by diffracted X-ray tracking. Irrespective of the locations of cysteines, it was shown that ATP binding induces tilting motion followed by rotational motion in the CtCCT molecule, like the archaeal group II chaperonins. When gold nanocrystals were attached onto two subunits in the high ATPase activity hemisphere, the CtCCT complex exhibited a fairly rapid response to the motion. In contrast, the response of CtCCT, which had gold nanocrystals attached to the low-activity hemisphere, was slow. These results clearly support the possibility that ATP-dependent conformational change starts with the high-affinity hemisphere and progresses to the low-affinity hemisphere.

A conserved phenylalanine as a relay between the α5 helix and the GDP binding region of heterotrimeric Gi protein α subunit.

PubMed

Kaya, Ali I; Lokits, Alyssa D; Gilbert, James A; Iverson, Tina M; Meiler, Jens; Hamm, Heidi E

2014-08-29

G protein activation by G protein-coupled receptors is one of the critical steps for many cellular signal transduction pathways. Previously, we and other groups reported that the α5 helix in the G protein α subunit plays a major role during this activation process. However, the precise signaling pathway between the α5 helix and the guanosine diphosphate (GDP) binding pocket remains elusive. Here, using structural, biochemical, and computational techniques, we probed different residues around the α5 helix for their role in signaling. Our data showed that perturbing the Phe-336 residue disturbs hydrophobic interactions with the β2-β3 strands and α1 helix, leading to high basal nucleotide exchange. However, mutations in β strands β5 and β6 do not perturb G protein activation. We have highlighted critical residues that leverage Phe-336 as a relay. Conformational changes are transmitted starting from Phe-336 via β2-β3/α1 to Switch I and the phosphate binding loop, decreasing the stability of the GDP binding pocket and triggering nucleotide release. When the α1 and α5 helices were cross-linked, inhibiting the receptor-mediated displacement of the C-terminal α5 helix, mutation of Phe-336 still leads to high basal exchange rates. This suggests that unlike receptor-mediated activation, helix 5 rotation and translocation are not necessary for GDP release from the α subunit. Rather, destabilization of the backdoor region of the Gα subunit is sufficient for triggering the activation process. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Analysis and Quantitation of Glycated Hemoglobin by Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry

NASA Astrophysics Data System (ADS)

Hattan, Stephen J.; Parker, Kenneth C.; Vestal, Marvin L.; Yang, Jane Y.; Herold, David A.; Duncan, Mark W.

2016-03-01

Measurement of glycated hemoglobin is widely used for the diagnosis and monitoring of diabetes mellitus. Matrix assisted laser desorption/ionization (MALDI) time of flight (TOF) mass spectrometry (MS) analysis of patient samples is used to demonstrate a method for quantitation of total glycation on the β-subunit of hemoglobin. The approach is accurate and calibrated with commercially available reference materials. Measurements were linear (R2 > 0.99) across the clinically relevant range of 4% to 20% glycation with coefficients of variation of ≤ 2.5%. Additional and independent measurements of glycation of the α-subunit of hemoglobin are used to validate β-subunit glycation measurements and distinguish hemoglobin variants. Results obtained by MALDI-TOF MS were compared with those obtained in a clinical laboratory using validated HPLC methodology. MALDI-TOF MS sample preparation was minimal and analysis times were rapid making the method an attractive alternative to methodologies currently in practice.

Disulfide bond rearrangement during formation of the chorionic gonadotropin beta-subunit cystine knot in vivo.

PubMed

Wilken, Jason A; Bedows, Elliott

2004-05-04

The intracellular kinetic folding pathway of the human chorionic gonadotropin beta-subunit (hCG-beta) reveals the presence of a disulfide between Cys residues 38-57 that is not detected by X-ray analysis of secreted hCG-beta. This led us to propose that disulfide rearrangement is an essential feature of cystine knot formation during CG-beta folding. To test this, we used disulfide bond formation to monitor progression of intracellular folding intermediates of a previously uncharacterized protein, the CG-beta subunit of cynomolgous macaque (Macaca fascicularis). Like its human counterpart hCG-beta with which it shares 81% identity, macaque (m)CG-beta is a cystine knot-containing subunit that assembles with an alpha-subunit common to all glycoprotein hormone members of its species to form a biologically active heterodimer, mCG, which, like hCG, is required for pregnancy maintenance. An early mCG-beta folding intermediate, mpbeta1, contained two disulfide bonds, one between Cys34 and Cys88 and the other between Cys38 and Cys57. The subsequent folding intermediate, mpbeta2-early, was represented by an ensemble of folding forms that, in addition to the two disulfides mentioned above, included disulfide linkages between Cys9 and Cys57 and between Cys38 and Cys90. These latter two disulfides are those contained within the beta-subunit cystine knot and reveal that a disulfide exchange occurred during the mpbeta2-early folding step leading to formation of the mCG-beta knot. Thus, while defining the intracellular kinetic protein folding pathway of a monkey homologue of CG-beta, we detected the previously predicted disulfide exchange event crucial for CG-beta cystine knot formation and attainment of CG-beta assembly competence.

Unidirectional rotary motion in a molecular system

NASA Astrophysics Data System (ADS)

Kelly, T. Ross; de Silva, Harshani; Silva, Richard A.

1999-09-01

The conversion of energy into controlled motion plays an important role in both man-made devices and biological systems. The principles of operation of conventional motors are well established, but the molecular processes used by `biological motors' such as muscle fibres, flagella and cilia to convert chemical energy into co-ordinated movement remain poorly understood. Although `brownian ratchets' are known to permit thermally activated motion in one direction only, the concept of channelling random thermal energy into controlled motion has not yet been extended to the molecular level. Here we describe a molecule that uses chemical energy to activate and bias a thermally induced isomerization reaction, and thereby achieve unidirectional intramolecular rotary motion. The motion consists of a 120° rotation around a single bond connecting a three-bladed subunit to the bulky remainder of the molecule, and unidirectional motion is achieved by reversibly introducing a tether between the two units to energetically favour one of the two possible rotation directions. Although our system does not achieve continuous and fast rotation, the design principles that we have used may prove relevant for a better understanding of biological and synthetic molecular motors producing unidirectional rotary motion.

Fourier transformation microwave spectroscopy of the methyl glycolate-H2O complex

NASA Astrophysics Data System (ADS)

Fujitake, Masaharu; Tanaka, Toshihiro; Ohashi, Nobukimi

2018-01-01

The rotational spectrum of one conformer of the methyl glycolate-H2O complex has been measured by means of the pulsed jet Fourier transform microwave spectrometer. The observed a- and b-type transitions exhibit doublet splittings due to the internal rotation of the methyl group. On the other hand, most of the c-type transitions exhibit quartet splittings arising from the methyl internal rotation and the inversion motion between two equivalent conformations. The spectrum was analyzed using parameterized expressions of the Hamiltonian matrix elements derived by applying the tunneling matrix formalism. Based on the results obtained from ab initio calculation, the observed complex of methyl glycolate-H2O was assigned to the most stable conformer of the insertion complex, in which a non-planer seven membered-ring structure is formed by the intermolecular hydrogen bonds between methyl glycolate and H2O subunits. The inversion motion observed in the c-type transitions is therefore a kind of ring-inversion motion between two equivalent conformations. Conformational flexibility, which corresponds to the ring-inversion between two equivalent conformations and to the isomerization between two possible conformers of the insertion complex, was investigated with the help of the ab initio calculation.

Role of the HoxZ subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase from Ralstonia eutropha immobilized on electrodes.

PubMed

Sezer, Murat; Frielingsdorf, Stefan; Millo, Diego; Heidary, Nina; Utesch, Tillman; Mroginski, Maria-Andrea; Friedrich, Bärbel; Hildebrandt, Peter; Zebger, Ingo; Weidinger, Inez M

2011-09-01

The role of the diheme cytochrome b (HoxZ) subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase (MBH) heterotrimer from Ralstonia eutropha H16 has been investigated. The MBH in its native heterotrimeric state was immobilized on electrodes and subjected to spectroscopic and electrochemical analysis. Surface enhanced resonance Raman spectroscopy was used to monitor the redox and coordination state of the HoxZ heme cofactors while concomitant protein film voltammetric measurements gave insights into the catalytic response of the enzyme on the electrode. The entire MBH heterotrimer as well as its isolated HoxZ subunit were immobilized on silver electrodes coated with self-assembled monolayers of ω-functionalized alkylthiols, displaying the preservation of the native heme pocket structure and an electrical communication between HoxZ and the electrode. For the immobilized MBH heterotrimer, catalytic reduction of the HoxZ heme cofactors was observed upon H(2) addition. The catalytic currents of MBH with and without the HoxZ subunit were measured and compared with the heterogeneous electron transfer rates of the isolated HoxZ. On the basis of the spectroscopic and electrochemical results, we conclude that the HoxZ subunit under these artificial conditions is not primarily involved in the electron transfer to the electrode but plays a crucial role in stabilizing the enzyme on the electrode. © 2011 American Chemical Society

Measurement of instantaneous rotational speed using double-sine-varying-density fringe pattern

NASA Astrophysics Data System (ADS)

Zhong, Jianfeng; Zhong, Shuncong; Zhang, Qiukun; Peng, Zhike

2018-03-01

Fast and accurate rotational speed measurement is required both for condition monitoring and faults diagnose of rotating machineries. A vision- and fringe pattern-based rotational speed measurement system was proposed to measure the instantaneous rotational speed (IRS) with high accuracy and reliability. A special double-sine-varying-density fringe pattern (DSVD-FP) was designed and pasted around the shaft surface completely and worked as primary angular sensor. The rotational angle could be correctly obtained from the left and right fringe period densities (FPDs) of the DSVD-FP image sequence recorded by a high-speed camera. The instantaneous angular speed (IAS) between two adjacent frames could be calculated from the real-time rotational angle curves, thus, the IRS also could be obtained accurately and efficiently. Both the measurement principle and system design of the novel method have been presented. The influence factors on the sensing characteristics and measurement accuracy of the novel system, including the spectral centrobaric correction method (SCCM) on the FPD calculation, the noise sources introduce by the image sensor, the exposure time and the vibration of the shaft, were investigated through simulations and experiments. The sampling rate of the high speed camera could be up to 5000 Hz, thus, the measurement becomes very fast and the change in rotational speed was sensed within 0.2 ms. The experimental results for different IRS measurements and characterization of the response property of a servo motor demonstrated the high accuracy and fast measurement of the proposed technique, making it attractive for condition monitoring and faults diagnosis of rotating machineries.

Extracting full-field dynamic strain response of a rotating wind turbine using photogrammetry

NASA Astrophysics Data System (ADS)

Baqersad, Javad; Poozesh, Peyman; Niezrecki, Christopher; Avitabile, Peter

2015-04-01

Health monitoring of wind turbines is typically performed using conventional sensors (e.g. strain-gages and accelerometers) that are usually mounted to the nacelle or gearbox. Although many wind turbines stop operating due to blade failures, there are typically few to no sensor mounted on the blades. Placing sensors on the rotating parts of the structure is a challenge due to the wiring and data transmission constraints. Within the current work, an approach to monitor full-field dynamic response of rotating structures (e.g. wind turbine blades or helicopter rotors) is developed and experimentally verified. A wind turbine rotor was used as the test structure and was mounted to a block and horizontally placed on the ground. A pair of bearings connected to the rotor shaft allowed the turbine to freely spin along the shaft. Several optical targets were mounted to the blades and a pair of high-speed cameras was used to monitor the dynamics of the spinning turbine. Displacements of the targets during rotation were measured using three-dimensional point tracking. The point tracking technique measured both rigid body displacement and flexible deformation of the blades at target locations. While the structure is rotating, only flap displacements of optical targets (displacements out of the rotation plane) were used in strain prediction process. The measured displacements were expanded and applied to the finite element model of the turbine to extract full-field dynamic strain on the structure. The proposed approach enabled the prediction of dynamic response on the outer surface as well as within the inner points of the structure where no other sensor could be easily mounted. In order to validate the proposed approach, the predicted strain was compared to strain measured at four locations on the spinning blades using a wireless strain-gage system.

Theory of single-molecule controlled rotation experiments, predictions, tests, and comparison with stalling experiments in F1-ATPase.

PubMed

Volkán-Kacsó, Sándor; Marcus, Rudolph A

2016-10-25

A recently proposed chemomechanical group transfer theory of rotary biomolecular motors is applied to treat single-molecule controlled rotation experiments. In these experiments, single-molecule fluorescence is used to measure the binding and release rate constants of nucleotides by monitoring the occupancy of binding sites. It is shown how missed events of nucleotide binding and release in these experiments can be corrected using theory, with F 1 -ATP synthase as an example. The missed events are significant when the reverse rate is very fast. Using the theory the actual rate constants in the controlled rotation experiments and the corrections are predicted from independent data, including other single-molecule rotation and ensemble biochemical experiments. The effective torsional elastic constant is found to depend on the binding/releasing nucleotide, and it is smaller for ADP than for ATP. There is a good agreement, with no adjustable parameters, between the theoretical and experimental results of controlled rotation experiments and stalling experiments, for the range of angles where the data overlap. This agreement is perhaps all the more surprising because it occurs even though the binding and release of fluorescent nucleotides is monitored at single-site occupancy concentrations, whereas the stalling and free rotation experiments have multiple-site occupancy.

Student and staff experiences of attendance monitoring in undergraduate obstetrics and gynecology: a cross-sectional survey

PubMed Central

Deane, Richard P; Murphy, Deirdre J

2016-01-01

Background Despite the widespread introduction of active learning strategies to engage students across modern medical curricula, student attendance and attendance monitoring remain a challenging issue for medical educators. In addition, there is little published evidence available to medical educators regarding the use of attendance monitoring systems. The aim of this study was to evaluate the opinions of students and staff about the use of a paper-based student logbook to record student attendance across all clinical and classroom-based learning activities within an undergraduate clinical rotation in obstetrics and gynecology (OBGYN). Methods Each student undertaking the clinical rotation in OBGYN was required to complete a paper-based logbook in a booklet format that listed every clinical and classroom-based activity that the student was expected to attend. A cross-sectional survey evaluating the acceptability, practicality, and effect on access to learning opportunities of using the logbook was undertaken. The survey was conducted among all medical students who completed their OBGYN rotation over a full academic year and staff who taught on the program. Results The response rate was 87% (n=128/147) among students and 80% (n=8/10) among staff. Monitoring attendance was widely acceptable to students (n=107/128, 84%) and staff (n=8/8, 100%). Most students (n=95/128, 74%) and staff (n=7/8, 88%) recommended that attendance should be mandatory during rotations. Almost all staff felt that attendance should contribute toward academic credit (n=7/8, 88%), but students were divided (n=73/128, 57%). Students (n=94/128, 73%) and staff (n=6/8, 75%) reported that the use of the logbook to record attendance with tutor signatures was a satisfactory system, although students questioned the need for recording attendance at every classroom-based activity. Most students felt that the logbook facilitated access to learning experiences during the rotation (n=90/128, 71%). Staff felt that the process of signing logbooks improved their interaction with students (n=6/8, 75%). Conclusion The survey showed that the use of a paper-based logbook to record medical student attendance with tutor signatures across all clinical and classroom-based learning activities was acceptable and practical for students and staff and was felt to facilitate access to learning opportunities. The study provides medical educators with evidence to support monitoring of attendance within clinical rotations. PMID:27099545

Student and staff experiences of attendance monitoring in undergraduate obstetrics and gynecology: a cross-sectional survey.

PubMed

Deane, Richard P; Murphy, Deirdre J

2016-01-01

Despite the widespread introduction of active learning strategies to engage students across modern medical curricula, student attendance and attendance monitoring remain a challenging issue for medical educators. In addition, there is little published evidence available to medical educators regarding the use of attendance monitoring systems. The aim of this study was to evaluate the opinions of students and staff about the use of a paper-based student logbook to record student attendance across all clinical and classroom-based learning activities within an undergraduate clinical rotation in obstetrics and gynecology (OBGYN). Each student undertaking the clinical rotation in OBGYN was required to complete a paper-based logbook in a booklet format that listed every clinical and classroom-based activity that the student was expected to attend. A cross-sectional survey evaluating the acceptability, practicality, and effect on access to learning opportunities of using the logbook was undertaken. The survey was conducted among all medical students who completed their OBGYN rotation over a full academic year and staff who taught on the program. The response rate was 87% (n=128/147) among students and 80% (n=8/10) among staff. Monitoring attendance was widely acceptable to students (n=107/128, 84%) and staff (n=8/8, 100%). Most students (n=95/128, 74%) and staff (n=7/8, 88%) recommended that attendance should be mandatory during rotations. Almost all staff felt that attendance should contribute toward academic credit (n=7/8, 88%), but students were divided (n=73/128, 57%). Students (n=94/128, 73%) and staff (n=6/8, 75%) reported that the use of the logbook to record attendance with tutor signatures was a satisfactory system, although students questioned the need for recording attendance at every classroom-based activity. Most students felt that the logbook facilitated access to learning experiences during the rotation (n=90/128, 71%). Staff felt that the process of signing logbooks improved their interaction with students (n=6/8, 75%). The survey showed that the use of a paper-based logbook to record medical student attendance with tutor signatures across all clinical and classroom-based learning activities was acceptable and practical for students and staff and was felt to facilitate access to learning opportunities. The study provides medical educators with evidence to support monitoring of attendance within clinical rotations.

Monitoring of selected non-ionic surfactants in river water by liquid chromatography-tandem mass spectrometry.

PubMed

Zembrzuska, Joanna; Budnik, Irena; Lukaszewski, Zenon

2016-03-15

Alcohol ethoxylates (AEs) are a significant component of the non-ionic surfactant (NS) flux discharged into surface water. Due to the polydispersity of the majority of NS, they are easily recognizable by their 'fingerprints', i.e. a series of mass peaks which differ by m/z = 44, namely the m/z of a single oxyethylene subunit. Dodecanol ethoxylates (C12EOx) represent AEs from both renewable and petrochemical sources. Therefore, C12Ex are suitable fingerprints of NS in the aquatic environment. The aim of this work was to develop an LC-MS/MS method suitable for AE monitoring in river water. River water samples taken from the River Warta in Poznan (Poland) were extracted with ethyl acetate, evaporated, reconstituted in the mobile phase and processed by the LC - Multistage MS procedure (LC-MS/MS) using optimum multiple reaction monitoring (MRM). The method of multiple standard additions was used for the evaluation of each AE fingerprint concentration. The concentration of C12EOx having 2-9 oxyethylene subunits was determined. Standards for higher C12EOx are not yet available. The developed method offers an LOD of between 1 and 9 ng L(-1), and is suitable for the monitoring of NS fingerprints in river water. The range of C12EO2-9 concentrations determined in the River Warta varied within two orders of magnitude in all cases. The lowest determined concentration was 17 ± 1 ng L(-1), while the highest was 2.6 ± 0.14 μg L(-1). The total concentration of C12EO2-C12EO9 homologues varied between 1.4 and 11.2 μg L(-1). A relatively high concentration of short-chained homologues (2-5 oxyethylene subunits) was observed in the investigated river water. This provides evidence of a biodegradation pathway involving the gradual shortening of the AE oxyethylene chain. Distinct evidence was also obtained of unregulated NS discharges into the river. Copyright © 2016 Elsevier Ltd. All rights reserved.

Chronic MK-801 Application in Adolescence and Early Adulthood: A Spatial Working Memory Deficit in Adult Long-Evans Rats But No Changes in the Hippocampal NMDA Receptor Subunits

PubMed Central

Uttl, Libor; Petrasek, Tomas; Sengul, Hilal; Svojanovska, Marketa; Lobellova, Veronika; Vales, Karel; Radostova, Dominika; Tsenov, Grygoriy; Kubova, Hana; Mikulecka, Anna; Svoboda, Jan; Stuchlik, Ales

2018-01-01

The role of NMDA receptors in learning, memory and hippocampal function has long been recognized. Post-mortem studies have indicated that the expression or subunit composition of the NMDA glutamate receptor subtype might be related to the impaired cognitive functions found in schizophrenia patients. NMDA receptor antagonists have been used to develop animal models of this disorder. There is accumulating evidence showing that not only the acute but also the chronic application of NMDA receptor antagonists may induce schizophrenia-like alterations in behavior and brain functions. However, limited evidence is available regarding the consequences of NMDA receptor blockage during periods of adolescence and early adulthood. This study tested the hypothesis that a 2-week treatment of male Long-Evans and Wistar rats with dizocilpine (MK-801; 0.5 mg/kg daily) starting at postnatal days (PD) 30 and 60 would cause a long-term cognitive deficit and changes in the levels of NMDA receptor subunits. The working memory version of the Morris water maze (MWM) and active place avoidance with reversal on a rotating arena (Carousel) requiring cognitive coordination and flexibility probed cognitive functions and an elevated-plus maze (EPM) was used to measure anxiety-like behavior. The western blot method was used to determine changes in NMDA receptor subunit levels in the hippocampus. Our results showed no significant changes in behaviors in Wistar rats. Slightly elevated anxiety-like behavior was observed in the EPM in Long-Evans rats with the onset of treatment on PD 30. Furthermore, Long-Evans rats treated from PD 60 displayed impaired working memory in the MWM. There were; however, no significant changes in the levels of NMDA receptor subunits because of MK-801 administration. These findings suggest that a 2-week treatment starting on PD 60 in Long-Evans rats leads to long-term changes in working memory, but this deficit is not paralleled by changes in NMDA receptor subunits. These results support the face validity, but not construct validity of this model. We suggest that chronic treatment of adolescent and adult rats does not constitute a plausible animal model of schizophrenia. PMID:29487522

Chronic MK-801 Application in Adolescence and Early Adulthood: A Spatial Working Memory Deficit in Adult Long-Evans Rats But No Changes in the Hippocampal NMDA Receptor Subunits.

PubMed

Uttl, Libor; Petrasek, Tomas; Sengul, Hilal; Svojanovska, Marketa; Lobellova, Veronika; Vales, Karel; Radostova, Dominika; Tsenov, Grygoriy; Kubova, Hana; Mikulecka, Anna; Svoboda, Jan; Stuchlik, Ales

2018-01-01

The role of NMDA receptors in learning, memory and hippocampal function has long been recognized. Post-mortem studies have indicated that the expression or subunit composition of the NMDA glutamate receptor subtype might be related to the impaired cognitive functions found in schizophrenia patients. NMDA receptor antagonists have been used to develop animal models of this disorder. There is accumulating evidence showing that not only the acute but also the chronic application of NMDA receptor antagonists may induce schizophrenia-like alterations in behavior and brain functions. However, limited evidence is available regarding the consequences of NMDA receptor blockage during periods of adolescence and early adulthood. This study tested the hypothesis that a 2-week treatment of male Long-Evans and Wistar rats with dizocilpine (MK-801; 0.5 mg/kg daily) starting at postnatal days (PD) 30 and 60 would cause a long-term cognitive deficit and changes in the levels of NMDA receptor subunits. The working memory version of the Morris water maze (MWM) and active place avoidance with reversal on a rotating arena (Carousel) requiring cognitive coordination and flexibility probed cognitive functions and an elevated-plus maze (EPM) was used to measure anxiety-like behavior. The western blot method was used to determine changes in NMDA receptor subunit levels in the hippocampus. Our results showed no significant changes in behaviors in Wistar rats. Slightly elevated anxiety-like behavior was observed in the EPM in Long-Evans rats with the onset of treatment on PD 30. Furthermore, Long-Evans rats treated from PD 60 displayed impaired working memory in the MWM. There were; however, no significant changes in the levels of NMDA receptor subunits because of MK-801 administration. These findings suggest that a 2-week treatment starting on PD 60 in Long-Evans rats leads to long-term changes in working memory, but this deficit is not paralleled by changes in NMDA receptor subunits. These results support the face validity, but not construct validity of this model. We suggest that chronic treatment of adolescent and adult rats does not constitute a plausible animal model of schizophrenia.

Using NDVI to estimate carbon fluxes from small rotationally grazed pastures

USDA-ARS?s Scientific Manuscript database

Satellite-based Normalized Difference Vegetation Index (NDVI) data have been extensively used for estimating gross primary productivity (GPP) and yield of grazing lands throughout the world. However, the usefulness of satellite-based images for monitoring rotationally-grazed pastures in the northea...

Novel localized heating technique on centrifugal microfluidic disc with wireless temperature monitoring system.

PubMed

Joseph, Karunan; Ibrahim, Fatimah; Cho, Jongman

2015-01-01

Recent advances in the field of centrifugal microfluidic disc suggest the need for electrical interface in the disc to perform active biomedical assays. In this paper, we have demonstrated an active application powered by the energy harvested from the rotation of the centrifugal microfluidic disc. A novel integration of power harvester disc onto centrifugal microfluidic disc to perform localized heating technique is the main idea of our paper. The power harvester disc utilizing electromagnetic induction mechanism generates electrical energy from the rotation of the disc. This contributes to the heat generation by the embedded heater on the localized heating disc. The main characteristic observed in our experiment is the heating pattern in relative to the rotation of the disc. The heating pattern is monitored wirelessly with a digital temperature sensing system also embedded on the disc. Maximum temperature achieved is 82 °C at rotational speed of 2000 RPM. The technique proves to be effective for continuous heating without the need to stop the centrifugal motion of the disc.

Stellar Rotation: New Insight from CoRoT

NASA Astrophysics Data System (ADS)

Catala, C.; Goupil, M. J.; Michel, E.; Baglin, A.; de Medeiros, J. Renan; Gondoin, Ph.

2009-02-01

We present an overview of the new insight provided by the CoRoT satellite on stellar rotation. Thanks to its ultra-high precision, high duty cycle, long photometric monitoring of thousands of stars, CoRoT gives us a powerful tool to study stellar rotational modulation, and therefore to measure stellar rotational periods and to study active structures at the surface of stars. This paper presents preliminary results concerning this type of study. CoRoT will also provide us with an insight of internal stellar rotation via the measurement and exploitation of rotational splittings of oscillation modes. This approach to stellar rotation with CoRoT will require a careful analysis of the oscillation power spectra, which is in progress, but prospects for such measurements are presented.

Allosteric Inhibition via R-state Destabilization in ATP Sulfurylase from Penicillium chrysogenum

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

MacRae, I. J.

2002-01-01

The structure of the cooperative hexameric enzyme ATP sulfurylase from Penicillium chrysogenum bound to its allosteric inhibitor, 3'-phosphoadenosine-5'-phosphosulfate (PAPS), was determined to 2.6 {angstrom} resolution. This structure represents the low substrate-affinity T-state conformation of the enzyme. Comparison with the high substrate-affinity R-state structure reveals that a large rotational rearrangement of domains occurs as a result of the R-to-T transition. The rearrangement is accompanied by the 17 {angstrom} movement of a 10-residue loop out of the active site region, resulting in an open, product release-like structure of the catalytic domain. Binding of PAPS is proposed to induce the allosteric transition bymore » destabilizing an R-state-specific salt linkage between Asp 111 in an N-terminal domain of one subunit and Arg 515 in the allosteric domain of a trans-triad subunit. Disrupting this salt linkage by site-directed mutagenesis induces cooperative inhibition behavior in the absence of an allosteric effector, confirming the role of these two residues.« less

Expression, purification, crystallization and preliminary crystallographic analysis of MxiH, a subunit of the Shigella flexneri type III secretion system needle

PubMed Central

Deane, Janet E.; Cordes, Frank S.; Roversi, Pietro; Johnson, Steven; Kenjale, Roma; Picking, William D.; Picking, Wendy L.; Lea, Susan M.; Blocker, Ariel

2006-01-01

A monodisperse truncation mutant of MxiH, the subunit of the needle from the Shigella flexneri type III secretion system (TTSS), has been overexpressed and purified. Crystals were grown of native and selenomethionine-labelled MxiHCΔ5 and diffraction data were collected to 1.9 Å resolution. The crystals belong to space group C2, with unit-cell parameters a = 183.4, b = 28.1, c = 27.8 Å, β = 96.5°. An anomalous difference Patterson map calculated with the data from the SeMet-labelled crystals revealed a single peak on the Harker section v = 0. Inspection of a uranyl derivative also revealed one peak in the isomorphous difference Patterson map on the Harker section v = 0. Analysis of the self-rotation function indicates the presence of a twofold non-crystallographic symmetry axis approximately along a. The calculated Matthews coefficient is 1.9 Å3 Da−1 for two molecules per asymmetric unit, corresponding to a solvent content of 33%. PMID:16511329

Expression, purification, crystallization and preliminary crystallographic analysis of MxiH, a subunit of the Shigella flexneri type III secretion system needle.

PubMed

Deane, Janet E; Cordes, Frank S; Roversi, Pietro; Johnson, Steven; Kenjale, Roma; Picking, William D; Picking, Wendy L; Lea, Susan M; Blocker, Ariel

2006-03-01

A monodisperse truncation mutant of MxiH, the subunit of the needle from the Shigella flexneri type III secretion system (TTSS), has been overexpressed and purified. Crystals were grown of native and selenomethionine-labelled MxiH(CDelta5) and diffraction data were collected to 1.9 A resolution. The crystals belong to space group C2, with unit-cell parameters a = 183.4, b = 28.1, c = 27.8 A, beta = 96.5 degrees. An anomalous difference Patterson map calculated with the data from the SeMet-labelled crystals revealed a single peak on the Harker section v = 0. Inspection of a uranyl derivative also revealed one peak in the isomorphous difference Patterson map on the Harker section v = 0. Analysis of the self-rotation function indicates the presence of a twofold non-crystallographic symmetry axis approximately along a. The calculated Matthews coefficient is 1.9 A3 Da(-1) for two molecules per asymmetric unit, corresponding to a solvent content of 33%.

Steric interactions lead to collective tilting motion in the ribosome during mRNA-tRNA translocation

NASA Astrophysics Data System (ADS)

Nguyen, Kien; Whitford, Paul C.

2016-02-01

Translocation of mRNA and tRNA through the ribosome is associated with large-scale rearrangements of the head domain in the 30S ribosomal subunit. To elucidate the relationship between 30S head dynamics and mRNA-tRNA displacement, we apply molecular dynamics simulations using an all-atom structure-based model. Here we provide a statistical analysis of 250 spontaneous transitions between the A/P-P/E and P/P-E/E ensembles. Consistent with structural studies, the ribosome samples a chimeric ap/P-pe/E intermediate, where the 30S head is rotated ~18°. It then transiently populates a previously unreported intermediate ensemble, which is characterized by a ~10° tilt of the head. To identify the origins of head tilting, we analyse 781 additional simulations in which specific steric features are perturbed. These calculations show that head tilting may be attributed to specific steric interactions between tRNA and the 30S subunit (PE loop and protein S13). Taken together, this study demonstrates how molecular structure can give rise to large-scale collective rearrangements.

Water molecules in the nucleotide binding cleft of actin: effects on subunit conformation and implications for ATP hydrolysis.

PubMed

Saunders, Marissa G; Voth, Gregory A

2011-10-14

In the monomeric actin crystal structure, the positions of a highly organized network of waters are clearly visible within the active site. However, the recently proposed models of filamentous actin (F-actin) did not extend to including these waters. Since the water network is important for ATP hydrolysis, information about water position is critical to understanding the increased rate of catalysis upon filament formation. Here, we show that waters in the active site are essential for intersubdomain rotational flexibility and that they organize the active-site structure. Including the crystal structure waters during simulation setup allows us to observe distinct changes in the active-site structure upon the flattening of the actin subunit, as proposed in the Oda model for F-actin. We identify changes in both protein position and water position relative to the phosphate tail that suggest a mechanism for accelerating the rate of nucleotide hydrolysis in F-actin by stabilizing charge on the β-phosphate and by facilitating deprotonation of catalytic water. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Chloroplast ATP Synthase Features the Characteristic Redox Regulation Machinery

PubMed Central

Sunamura, Ei-Ichiro; Kim, Yusung; Konno, Hiroki

2013-01-01

Abstract Significance: Regulation of the activity of the chloroplast ATP synthase is largely accomplished by the chloroplast thioredoxin system, the main redox regulation system in chloroplasts, which is directly coupled to the photosynthetic reaction. We review the current understanding of the redox regulation system of the chloroplast ATP synthase. Recent Advances: The thioredoxin-targeted portion of the ATP synthase consists of two cysteines located on the central axis subunit γ. The redox state of these two cysteines is under the influence of chloroplast thioredoxin, which directly controls rotation during catalysis by inducing a conformational change in this subunit. The molecular mechanism of redox regulation of the chloroplast ATP synthase has recently been determined. Critical Issues: Regulation of the activity of the chloroplast ATP synthase is critical in driving efficiency into the ATP synthesis reaction in chloroplasts. Future Directions: The molecular architecture of the chloroplast ATP synthase, which confers redox regulatory properties requires further investigation, in light of the molecular structure of the enzyme complex as well as the physiological significance of the regulation system. Antioxid. Redox Signal. 19, 1846–1854. PMID:23145525

Interaction of the p85 subunit of PI 3-kinase and its N-terminal SH2 domain with a PDGF receptor phosphorylation site: structural features and analysis of conformational changes.

PubMed Central

Panayotou, G; Bax, B; Gout, I; Federwisch, M; Wroblowski, B; Dhand, R; Fry, M J; Blundell, T L; Wollmer, A; Waterfield, M D

1992-01-01

Circular dichroism and fluorescence spectroscopy were used to investigate the structure of the p85 alpha subunit of the PI 3-kinase, a closely related p85 beta protein, and a recombinant SH2 domain-containing fragment of p85 alpha. Significant spectral changes, indicative of a conformational change, were observed on formation of a complex with a 17 residue peptide containing a phosphorylated tyrosine residue. The sequence of this peptide is identical to the sequence surrounding Tyr751 in the kinase-insert region of the platelet-derived growth factor beta-receptor (beta PDGFR). The rotational correlation times measured by fluorescence anisotropy decay indicated that phosphopeptide binding changed the shape of the SH2 domain-containing fragment. The CD and fluorescence spectroscopy data support the secondary structure prediction based on sequence analysis and provide evidence for flexible linker regions between the various domains of the p85 proteins. The significance of these results for SH2 domain-containing proteins is discussed. Images PMID:1330535

Common Mechanisms of DNA translocation motors in Bacteria and Viruses Using One-way Revolution Mechanism without Rotation

PubMed Central

Guo, Peixuan; Zhao, Zhengyi; Haak, Jeannie; Wang, Shaoying; Weitao, Tao

2014-01-01

Biomotors were once classified into two categories: linear motor and rotation motor. For decades, the viral DNA-packaging motor has been popularly believed to be a five-fold rotation motor. Recently, a third type of biomotor with revolution mechanism without rotation has been discovered. By analogy, rotation resembles the Earth rotating on its axis in a complete cycle every 24 hours, while revolution resembles the Earth revolving around the Sun one circle per 365 days (see animations http://nanobio.uky.edu/movie.html). The action of revolution that enables a motor free of coiling and torque has solved many puzzles and debates that have occurred throughout the history of viral DNA packaging motor studies. It also settles the discrepancies concerning the structure, stoichiometry, and functioning of DNA translocation motors. This review uses bacteriophages Phi29, HK97, SPP1, P22, T4, T7 as well as bacterial DNA translocase FtsK and SpoIIIE as examples to elucidate the puzzles. These motors use a ATPase, some of which have been confirmed to be a hexamer, to revolve around the dsDNA sequentially. ATP binding induces conformational change and possibly an entropy alteration in ATPase to a high affinity toward dsDNA; but ATP hydrolysis triggers another entropic and conformational change in ATPase to a low affinity for DNA, by which dsDNA is pushed toward an adjacent ATPase subunit. The rotation and revolution mechanisms can be distinguished by the size of channel: the channels of rotation motors are equal to or smaller than 2 nm, whereas channels of revolution motors are larger than 3 nm. Rotation motors use parallel threads to operate with a right-handed channel, while revolution motors use a left-handed channel to drive the right-handed DNA in an anti-parallel arrangement. Coordination of several vector factors in the same direction makes viral DNA-packaging motors unusually powerful and effective. Revolution mechanism avoids DNA coiling in translocating the lengthy genomic dsDNA helix could be advantage for cell replication such as bacterial binary fission and cell mitosis without the need for topoisomerase or helicase to consume additional energy. PMID:24913057

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

PubMed

Kasai, Taishi; Sowa, Yoshiyuki

2017-01-01

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

Effects of fixture rotation on coating uniformity for high-performance optical filter fabrication

NASA Astrophysics Data System (ADS)

Rubin, Binyamin; George, Jason; Singhal, Riju

2018-04-01

Coating uniformity is critical in fabricating high-performance optical filters by various vacuum deposition methods. Simple and planetary rotation systems with shadow masks are used to achieve the required uniformity [J. B. Oliver and D. Talbot, Appl. Optics 45, 13, 3097 (2006); O. Lyngnes, K. Kraus, A. Ode and T. Erguder, in `Method for Designing Coating Thickness Uniformity Shadow Masks for Deposition Systems with a Planetary Fixture', 2014 Technical Conference Proceedings, Optical Coatings, August 13, 2014, DOI: 10.14332/svc14.proc.1817.]. In this work, we discuss the effect of rotation pattern and speed on thickness uniformity in an ion beam sputter deposition system. Numerical modeling is used to determine statistical distribution of random thickness errors in coating layers. The relationship between thickness tolerance and production yield are simulated theoretically and demonstrated experimentally. Production yields for different optical filters produced in an ion beam deposition system with planetary rotation are presented. Single-wavelength and broadband optical monitoring systems were used for endpoint monitoring during filter deposition. Limitations of thickness tolerances that can be achieved in systems with planetary rotation are shown. Paths for improving production yield in an ion beam deposition system are described.

Hydrogen bonds between the alpha and beta subunits of the F1-ATPase allow communication between the catalytic site and the interface of the beta catch loop and the gamma subunit.

PubMed

Boltz, Kathryn W; Frasch, Wayne D

2006-09-19

F(1)-ATPase mutations in Escherichia coli that changed the strength of hydrogen bonds between the alpha and beta subunits in a location that links the catalytic site to the interface between the beta catch loop and the gamma subunit were examined. Loss of the ability to form the hydrogen bonds involving alphaS337, betaD301, and alphaD335 lowered the k(cat) of ATPase and decreased its susceptibility to Mg(2+)-ADP-AlF(n) inhibition, while mutations that maintain or strengthen these bonds increased the susceptibility to Mg(2+)-ADP-AlF(n) inhibition and lowered the k(cat) of ATPase. These data suggest that hydrogen bonds connecting alphaS337 to betaD301 and betaR323 and connecting alphaD335 to alphaS337 are important to transition state stabilization and catalytic function that may result from the proper alignment of catalytic site residues betaR182 and alphaR376 through the VISIT sequence (alpha344-348). Mutations betaD301E, betaR323K, and alphaR282Q changed the rate-limiting step of the reaction as determined by an isokinetic plot. Hydrophobic mutations of betaR323 decreased the susceptibility to Mg(2+)-ADP-AlF(n)() inhibition and lowered the number of interactions required in the rate-limiting step yet did not affect the k(cat) of ATPase, suggesting that betaR323 is important to transition state formation. The decreased rate of ATP synthase-dependent growth and decreased level of lactate-dependent quenching observed with alphaD335, betaD301, and alphaE283 mutations suggest that these residues may be important to the formation of an alternative set of hydrogen bonds at the interface of the alpha and beta subunits that permits the release of intersubunit bonds upon the binding of ATP, allowing gamma rotation in the escapement mechanism.

Construction and laboratory test of the fiber optic rotational seismograph FOSREM for rotational seismology area of interest

NASA Astrophysics Data System (ADS)

Kurzych, Anna; Jaroszewicz, Leszek R.; Kowalski, Jerzy K.

2017-05-01

A relatively young field of study named Rotational Seismology caused a highly interest in an investigation of rotational movements generated by earthquakes, explosions, and ambient vibrations. It includes a wide range of scientific branches. However, this field needs to apply appropriate rotational sensors which should fulfill restrict technical requirements. The presented in this work system FOSREM (Fibre-Optic System for Rotational Events and Phenomena Monitoring) seems to be a promising rotational sensor for such investigation. FOSREM works by measuring the Sagnac effect and generally consists of two basic elements: optical sensor and electronic part. Regarding to its theoretical sensitivity equals 2·10-8 rad/s/Hz1/2, it enables to measure rotation in a wide range of signal amplitude (10-8 rad/s ÷ 10 rad/s) and frequency (DC ÷ 328.12 Hz). Moreover, FOSREM is mobile and remotely controlled via Internet using a special designed software.

A Novel N-Acetylglutamate Synthase Architecture Revealed by the Crystal Structure of the Bifunctional Enzyme from Maricaulis maris

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

Shi, Dashuang; Li, Yongdong; Cabrera-Luque, Juan

2012-05-24

Novel bifunctional N-acetylglutamate synthase/kinases (NAGS/K) that catalyze the first two steps of arginine biosynthesis and are homologous to vertebrate N-acetylglutamate synthase (NAGS), an essential cofactor-producing enzyme in the urea cycle, were identified in Maricaulis maris and several other bacteria. Arginine is an allosteric inhibitor of NAGS but not NAGK activity. The crystal structure of M. maris NAGS/K (mmNAGS/K) at 2.7 {angstrom} resolution indicates that it is a tetramer, in contrast to the hexameric structure of Neisseria gonorrhoeae NAGS. The quaternary structure of crystalline NAGS/K from Xanthomonas campestris (xcNAGS/K) is similar, and cross-linking experiments indicate that both mmNAGS/K and xcNAGS aremore » tetramers in solution. Each subunit has an amino acid kinase (AAK) domain, which is likely responsible for N-acetylglutamate kinase (NAGK) activity and has a putative arginine binding site, and an N-acetyltransferase (NAT) domain that contains the putative NAGS active site. These structures and sequence comparisons suggest that the linker residue 291 may determine whether arginine acts as an allosteric inhibitor or activator in homologous enzymes in microorganisms and vertebrates. In addition, the angle of rotation between AAK and NAT domains varies among crystal forms and subunits within the tetramer. A rotation of 26{sup o} is sufficient to close the predicted AcCoA binding site, thus reducing enzymatic activity. Since mmNAGS/K has the highest degree of sequence homology to vertebrate NAGS of NAGS and NAGK enzymes whose structures have been determined, the mmNAGS/K structure was used to develop a structural model of human NAGS that is fully consistent with the functional effects of the 14 missense mutations that were identified in NAGS-deficient patients.« less

RoboPol: connection between optical polarization plane rotations and gamma-ray flares in blazars

NASA Astrophysics Data System (ADS)

Blinov, D.; Pavlidou, V.; Papadakis, I.; Kiehlmann, S.; Liodakis, I.; Panopoulou, G. V.; Angelakis, E.; Baloković, M.; Hovatta, T.; King, O. G.; Kus, A.; Kylafis, N.; Mahabal, A.; Maharana, S.; Myserlis, I.; Paleologou, E.; Papamastorakis, I.; Pazderski, E.; Pearson, T. J.; Ramaprakash, A.; Readhead, A. C. S.; Reig, P.; Tassis, K.; Zensus, J. A.

2018-02-01

We use results of our 3 yr polarimetric monitoring programme to investigate the previously suggested connection between rotations of the polarization plane in the optical emission of blazars and their gamma-ray flares in the GeV band. The homogeneous set of 40 rotation events in 24 sources detected by RoboPol is analysed together with the gamma-ray data provided by Fermi-LAT. We confirm that polarization plane rotations are indeed related to the closest gamma-ray flares in blazars and the time lags between these events are consistent with zero. Amplitudes of the rotations are anticorrelated with amplitudes of the gamma-ray flares. This is presumably caused by higher relativistic boosting (higher Doppler factors) in blazars that exhibit smaller amplitude polarization plane rotations. Moreover, the time-scales of rotations and flares are marginally correlated.

40 CFR 63.466 - Monitoring procedures.

Code of Federal Regulations, 2013 CFR

2013-07-01

... corner and record the average wind speed. (ii) Monitor on a weekly basis the room parameters established... standards in § 63.463 shall monitor the hoist speed as described in paragraphs (c)(1) through (c)(4) of this...) Determine the direction of the wind current by slowly rotating a velometer or similar device until the...

40 CFR 63.466 - Monitoring procedures.

Code of Federal Regulations, 2012 CFR

2012-07-01

... corner and record the average wind speed. (ii) Monitor on a weekly basis the room parameters established... standards in § 63.463 shall monitor the hoist speed as described in paragraphs (c)(1) through (c)(4) of this...) Determine the direction of the wind current by slowly rotating a velometer or similar device until the...

40 CFR 63.466 - Monitoring procedures.

Code of Federal Regulations, 2010 CFR

2010-07-01

... corner and record the average wind speed. (ii) Monitor on a weekly basis the room parameters established... standards in § 63.463 shall monitor the hoist speed as described in paragraphs (c)(1) through (c)(4) of this...) Determine the direction of the wind current by slowly rotating a velometer or similar device until the...

40 CFR 63.466 - Monitoring procedures.

Code of Federal Regulations, 2011 CFR

2011-07-01

... corner and record the average wind speed. (ii) Monitor on a weekly basis the room parameters established... standards in § 63.463 shall monitor the hoist speed as described in paragraphs (c)(1) through (c)(4) of this...) Determine the direction of the wind current by slowly rotating a velometer or similar device until the...

40 CFR 63.466 - Monitoring procedures.

Code of Federal Regulations, 2014 CFR

2014-07-01

... corner and record the average wind speed. (ii) Monitor on a weekly basis the room parameters established... standards in § 63.463 shall monitor the hoist speed as described in paragraphs (c)(1) through (c)(4) of this...) Determine the direction of the wind current by slowly rotating a velometer or similar device until the...

Measurement of Rate Constants for Homodimer Subunit Exchange Using Double Electron-Electron Resonance and Paramagnetic Relaxation Enhancements

PubMed Central

Yang, Yunhuang; Ramelot, Theresa A.; Ni, Shuisong; McCarrick, Robert M.; Kennedy, Michael A.

2013-01-01

Here, we report novel methods to measure rate constants for homodimer subunit exchange using double electron-electron resonance (DEER) electron paramagnetic resonance spectroscopy measurements and nuclear magnetic resonance spectroscopy based paramagnetic relaxation enhancement (PRE) measurements. The techniques were demonstrated using the homodimeric protein Dsy0195 from the strictly anaerobic bacterium Desulfitobacterium hafniense Y51. At specific times following mixing site-specific MTSL-labeled Dsy0195 with uniformly 15N-labeled Dsy0195, the extent of exchange was determined either by monitoring the decrease of MTSL-labeled homodimer from the decay of the DEER modulation depth or by quantifying the increase of MTSL-labeled/15N-labeled heterodimer using PREs. Repeated measurements at several time points following mixing enabled determination of the homodimer subunit dissociation rate constant, k−1;, which was 0.037 ± 0.005 min−1 derived from DEER experiments with a corresponding half-life time of 18.7 minutes. These numbers agreed with independent measurements obtained from PRE experiments. These methods can be broadly applied to protein-protein and protein-DNA complex studies. PMID:23180051

Conformational Response of 30S-bound IF3 to A-Site Binders Streptomycin and Kanamycin

PubMed Central

Chulluncuy, Roberto; Espiche, Carlos; Nakamoto, Jose Alberto; Fabbretti, Attilio; Milón, Pohl

2016-01-01

Aminoglycoside antibiotics are widely used to treat infectious diseases. Among them, streptomycin and kanamycin (and derivatives) are of importance to battle multidrug-resistant (MDR) Mycobacterium tuberculosis. Both drugs bind the small ribosomal subunit (30S) and inhibit protein synthesis. Genetic, structural, and biochemical studies indicate that local and long-range conformational rearrangements of the 30S subunit account for this inhibition. Here, we use intramolecular FRET between the C- and N-terminus domains of the flexible IF3 to monitor real-time perturbations of their binding sites on the 30S platform. Steady and pre-steady state binding experiments show that both aminoglycosides bring IF3 domains apart, promoting an elongated state of the factor. Binding of Initiation Factor IF1 triggers closure of IF3 bound to the 30S complex, while both aminoglycosides revert the IF1-dependent conformation. Our results uncover dynamic perturbations across the 30S subunit, from the A-site to the platform, and suggest that both aminoglycosides could interfere with prokaryotic translation initiation by modulating the interaction between IF3 domains with the 30S platform. PMID:27983590

Subunit vaccine candidates against Aeromonas salmonicida in rainbow trout Oncorhynchus mykiss.

PubMed

Marana, Moonika Haahr; Jørgensen, Louise von Gersdorff; Skov, Jakob; Chettri, Jiwan Kumar; Holm Mattsson, Andreas; Dalsgaard, Inger; Kania, Per Walter; Buchmann, Kurt

2017-01-01

Aeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis and a major fish health problem in salmonid aquaculture worldwide. Injection vaccination with commercial mineral oil-adjuvanted bacterin vaccines has been partly successful in preventing the disease but in Danish rainbow trout (Oncorhynchus mykiss, Walbaum) aquaculture furunculosis outbreaks still occur. In this study we tested the efficacy of experimental subunit vaccines against A. salmonicida infection in rainbow trout. We utilized in silico screening of the proteome of A. salmonicida subsp. salmonicida strain A449 and identified potential protective protein antigens that were tested by in vivo challenge trial. A total of 14 proteins were recombinantly expressed in Escherichia coli and prepared in 3 different subunit vaccine combinations to immunize 3 groups of rainbow trout by intraperitoneal (i.p.) injection. The fish were exposed to virulent A. salmonicida 7 weeks after immunization. To assess the efficacy of the subunit vaccines we evaluated the immune response in fish after immunization and challenge infection by measuring the antibody levels and monitoring the survival of fish in different groups. The survival of fish at 3 weeks after challenge infection showed that all 3 groups of fish immunized with 3 different protein combinations exhibited significantly lower mortalities (17-30%) compared to the control groups (48% and 56%). The ELISA results revealed significantly elevated antibody levels in fish against several protein antigens, which in some cases were positively correlated to the survival.

Implications of observed and simulated soil carbon sequestration for management options in corn-based rotations

USDA-ARS?s Scientific Manuscript database

Managing cropping systems to sequester soil organic carbon (SOC) improves soil health and a system’s resiliency to impacts of changing climate. Our objectives were to 1) monitor SOC from a bio-energy cropping study in central Pennsylvania that included a corn-soybean-alfalfa rotation, switchgrass, a...

Implications of observed and simulated soil carbon sequestration for management options in corn-based rotations

USDA-ARS?s Scientific Manuscript database

Managing cropping systems to sequester soil organic carbon (SOC) improves soil health and a system’s resiliency to impacts of changing climate. Our objectives were to 1) monitor SOC from a bio-energy cropping study in central Pennsylvania that included a corn-soybean-alfalfa rotation, switchgrass, ...

Systemic Gene Transfer of a Hexosaminidase Variant Using an scAAV9.47 Vector Corrects GM2 Gangliosidosis in Sandhoff Mice.

PubMed

Osmon, Karlaina J L; Woodley, Evan; Thompson, Patrick; Ong, Katalina; Karumuthil-Melethil, Subha; Keimel, John G; Mark, Brian L; Mahuran, Don; Gray, Steven J; Walia, Jagdeep S

2016-07-01

GM2 gangliosidosis is a group of neurodegenerative diseases caused by β-hexosaminidase A (HexA) enzyme deficiency. There is currently no cure. HexA is composed of two similar, nonidentical subunits, α and β, which must interact with the GM2 activator protein (GM2AP), a substrate-specific cofactor, to hydrolyze GM2 ganglioside. Mutations in either subunit or the activator can result in the accumulation of GM2 ganglioside within neurons throughout the central nervous system. The resulting neuronal cell death induces the primary symptoms of the disease: motor impairment, seizures, and sensory impairments. This study assesses the long-term effects of gene transfer in a Sandhoff (β-subunit knockout) mouse model. The study utilized a modified human β-hexosaminidase α-subunit (μ-subunit) that contains critical sequences from the β-subunit that enables formation of a stable homodimer (HexM) and interaction with GM2AP to hydrolyze GM2 ganglioside. We investigated a self-complementary adeno-associated viral (scAAV) vector expressing HexM, through intravenous injections of the neonatal mice. We monitored one cohort for 8 weeks and another cohort long-term for survival benefit, behavioral, biochemical, and molecular analyses. Untreated Sandhoff disease (SD) control mice reached a humane endpoint at approximately 15 weeks, whereas treated mice had a median survival age of 40 weeks, an approximate 2.5-fold survival advantage. On behavioral tests, the treated mice outperformed their knockout age-matched controls and perform similarly to the heterozygous controls. Through the enzymatic and GM2 ganglioside analyses, we observed a significant decrease in the GM2 ganglioside level, even though the enzyme levels were not significantly increased. Molecular analyses revealed a global distribution of the vector between brain and spinal cord regions. In conclusion, the neonatal delivery of a novel viral vector expressing the human HexM enzyme is effective in ameliorating the SD mouse phenotype for long-term. Our data could have implications not only for treatment of SD but also for Tay-Sachs disease (α-subunit deficiency) and similar brain disorders.

Rotation-libration in a hierarchic supramolecular rotor-stator system: arrhenius activation and retardation by local interaction.

PubMed

Wahl, Markus; Stöhr, Meike; Spillmann, Hannes; Jung, Thomas A; Gade, Lutz H

2007-04-07

Fourfold symmetric zinc-octaethylporphyrin (OEP) has been incorporated in the holes of the hexagonal molecular network generated by thermal dehydrogenation of 4,9-diaminoperylene-quinone-3,10-diimine (DPDI) on a Cu(111) surface and displayed hindered rotation; the reorganization between the potential minima, a rotation-libration, which is characterized by an activation energy of ED=0.17+/-0.03 eV, has been monitored in the STM tunnelling currents as a bi-state "switching".

Integrity monitoring of IGS products

NASA Technical Reports Server (NTRS)

Zumberge, James F.; Plag, H. -P.

2005-01-01

The IGS has successfully produced precise GPS and GLONASS transmitter parameters, coordinates of IGS tracking stations, Earth rotation parameters, and atmospheric parameters. In this paper we discuss the concepts of integrity monitoring, system monitoring, and performance assessment, all in the context of IGS products. We report on a recent survey of IGS product users, and propose an integrity strategy for the IGS.

Health Monitoring of a Rotating Disk Using a Combined Analytical-Experimental Approach

NASA Technical Reports Server (NTRS)

Abdul-Aziz, Ali; Woike, Mark R.; Lekki, John D.; Baaklini, George Y.

2009-01-01

Rotating disks undergo rigorous mechanical loading conditions that make them subject to a variety of failure mechanisms leading to structural deformities and cracking. During operation, periodic loading fluctuations and other related factors cause fractures and hidden internal cracks that can only be detected via noninvasive types of health monitoring and/or nondestructive evaluation. These evaluations go further to inspect material discontinuities and other irregularities that have grown to become critical defects that can lead to failure. Hence, the objectives of this work is to conduct a collective analytical and experimental study to present a well-rounded structural assessment of a rotating disk by means of a health monitoring approach and to appraise the capabilities of an in-house rotor spin system. The analyses utilized the finite element method to analyze the disk with and without an induced crack at different loading levels, such as rotational speeds starting at 3000 up to 10 000 rpm. A parallel experiment was conducted to spin the disk at the desired speeds in an attempt to correlate the experimental findings with the analytical results. The testing involved conducting spin experiments which, covered the rotor in both damaged and undamaged (i.e., notched and unnotched) states. Damaged disks had artificially induced through-thickness flaws represented in the web region ranging from 2.54 to 5.08 cm (1 to 2 in.) in length. This study aims to identify defects that are greater than 1.27 cm (0.5 in.), applying available means of structural health monitoring and nondestructive evaluation, and documenting failure mechanisms experienced by the rotor system under typical turbine engine operating conditions.

Measuring Torque and Temperature in a Rotating Shaft Using Commercial SAW Sensors

PubMed Central

Silva, Diogo; Pereira, António B.; Gégot, François

2017-01-01

Real-time monitoring of torque in a rotating shaft is not easy to implement with technologies such as optic fiber sensors or strain gages. Surface acoustic wave (SAW) sensors are wireless and passive and can be used to monitor strain in moving parts. Commercial solutions (sensors, antennas and interrogation unit) can easily be purchased from some companies; however, they are not customized and may not meet the specificity of the measurements. In order to evaluate the adequacy of commercial off-the-shelf (COTS) solutions, temperature and strain sensors fabricated by SENSeOR (Besançon, France) were mounted on a load cell. The sensors were calibrated using a thermal chamber and a universal testing machine. The load cell was then assembled together with a steel shaft that rotated at different speeds inside an oven. The commercial antennas were replaced with an RF (radio frequency) coupler and the sensors were interrogated with the commercial interrogation unit. The influence of rotation in the accuracy on the measurements, as well as the adequacy of the sensors structure, was evaluated. It can be concluded that SAW sensors can be used to measure temperature or torque in a rotating environment; however, some customization of the components is required in order to overcome the limitations posed by COTS sensing solutions. PMID:28671594

Measuring Torque and Temperature in a Rotating Shaft Using Commercial SAW Sensors.

PubMed

Silva, Diogo; Mendes, Joana C; Pereira, António B; Gégot, François; Alves, Luís N

2017-07-02

Real-time monitoring of torque in a rotating shaft is not easy to implement with technologies such as optic fiber sensors or strain gages. Surface acoustic wave (SAW) sensors are wireless and passive and can be used to monitor strain in moving parts. Commercial solutions (sensors, antennas and interrogation unit) can easily be purchased from some companies; however, they are not customized and may not meet the specificity of the measurements. In order to evaluate the adequacy of commercial off-the-shelf (COTS) solutions, temperature and strain sensors fabricated by SENSeOR (Besançon, France) were mounted on a load cell. The sensors were calibrated using a thermal chamber and a universal testing machine. The load cell was then assembled together with a steel shaft that rotated at different speeds inside an oven. The commercial antennas were replaced with an RF (radio frequency) coupler and the sensors were interrogated with the commercial interrogation unit. The influence of rotation in the accuracy on the measurements, as well as the adequacy of the sensors structure, was evaluated. It can be concluded that SAW sensors can be used to measure temperature or torque in a rotating environment; however, some customization of the components is required in order to overcome the limitations posed by COTS sensing solutions.

Magnetostrictive patch sensor system for battery-less real-time measurement of torsional vibrations of rotating shafts

NASA Astrophysics Data System (ADS)

Lee, Jun Kyu; Seung, Hong Min; Park, Chung Il; Lee, Joo Kyung; Lim, Do Hyeong; Kim, Yoon Young

2018-02-01

Real-time uninterrupted measurement for torsional vibrations of rotating shafts is crucial for permanent health monitoring. So far, strain gauge systems with telemetry units have been used for real-time monitoring. However, they have a critical disadvantage in that shaft operations must be stopped intermittently to replace telemetry unit batteries. To find an alternative method to carry out battery-less real-time measurement for torsional vibrations of rotating shafts, a magnetostrictive patch sensor system was proposed in the present study. Since the proposed sensor does not use any powered telemetry system, no battery is needed and thus there is no need to stop rotating shafts for battery replacement. The proposed sensor consists of magnetostrictive patches and small magnets tightly bonded onto a shaft. A solenoid coil is placed around the shaft to convert magnetostrictive patch deformation by shaft torsional vibration into electric voltage output. For sensor design and characterization, investigations were performed in a laboratory on relatively small-sized stationary solid shaft. A magnetostrictive patch sensor system was then designed and installed on a large rotating propulsion shaft of an LPG carrier ship in operation. Vibration signals were measured using the proposed sensor system and compared to those measured with a telemetry unit-equipped strain gauge system.

Investigating the inter-subunit/subdomain interactions and motions relevant to disease mutations in the N-terminal domain of ryanodine receptors by molecular dynamics simulation.

PubMed

Zheng, Wenjun; Liu, Zheng

2017-09-01

The ryanodine receptors (RyR) are essential to calcium signaling in striated muscles, and numerous disease mutations have been identified in two RyR isoforms, RyR1 in skeletal muscle and RyR2 in cardiac muscle. A deep understanding of the activation/regulation mechanisms of RyRs has been hampered by the shortage of high-resolution structures and dynamic information for this giant tetrameric complex in different functional states. Toward elucidating the molecular mechanisms of disease mutations in RyRs, we performed molecular dynamics simulation of the N-terminal domain (NTD) which is not only the best-resolved structural component of RyRs, but also a hotspot of disease mutations. First, we simulated the tetrameric NTD of wild-type RyR1 and three disease mutants (K155E, R157Q, and R164Q) that perturb the inter-subunit interfaces. Our simulations identified a dynamic network of salt bridges involving charged residues at the inter-subunit/subdomain interfaces and disease-mutation sites. By perturbing this key network, the above three mutations result in greater flexibility with the highest inter-subunit opening probability for R157Q. Next, we simulated the monomeric NTD of RyR2 in the presence or absence of a central Cl - anion which is known to stabilize the interfaces between the three NTD subdomains (A, B, and C). We found that the loss of Cl - restructures the salt-bridge network near the Cl - -binding site, leading to rotations of subdomain A/B relative to subdomain C and enhanced mobility between the subdomains. This finding supports a mechanism for disease mutations in the NTD of RyR2 via perturbation of the Cl - binding. The rich structural and dynamic information gained from this study will guide future mutational and functional studies of the NTD of RyRs. Proteins 2017; 85:1633-1644. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Dynamics of molecules in extreme rotational states

PubMed Central

Yuan, Liwei; Teitelbaum, Samuel W.; Robinson, Allison; Mullin, Amy S.

2011-01-01

We have constructed an optical centrifuge with a pulse energy that is more than 2 orders of magnitude larger than previously reported instruments. This high pulse energy enables us to create large enough number densities of molecules in extreme rotational states to perform high-resolution state-resolved transient IR absorption measurements. Here we report the first studies of energy transfer dynamics involving molecules in extreme rotational states. In these studies, the optical centrifuge drives CO2 molecules into states with J ∼ 220 and we use transient IR probing to monitor the subsequent rotational, translational, and vibrational energy flow dynamics. The results reported here provide the first molecular insights into the relaxation of molecules with rotational energy that is comparable to that of a chemical bond.

Monitoring of rotational period variations in magnetic chemically peculiar stars

NASA Astrophysics Data System (ADS)

Mikulášek, Z.

2016-12-01

A majority part of magnetic chemically peculiar (mCP) stars of the upper main sequence exhibits strictly periodic light, magnetic, radio, and spectral variations that can be fully explained by the model of a rigidly rotating main-sequence star with persistent surface structures and stable global magnetic field frozen into the body of the star. Nevertheless, there is an inhomogeneous group consisting of a few mCP stars whose rotation periods vary on timescales of decades, while the shapes of their phase curves remain nearly unchanged. Alternations in the rotational period variations, proven in the case of some of them, offer new insight on this theoretically unpredicted phenomenon. We present a novel and generally applicable method of period analysis based on the simultaneous exploitation of all available observational data containing phase information. This phenomenological method can monitor gradual changes in the observed instantaneous period very efficiently and reliably. We present up to date results of the period monitoring of V901 Ori, CU Vir, σ Ori E, and BS Cir, known to be mCP stars changing their observed periods and discuss the physics of this unusual behaviour. To compare the period behavior of those stars, we treated their data with an orthogonal polynomial model, which was specifically developed for this purpose. We confirmed period variations in all stars and showed that they reflect real changes in the angular velocity of outer layers of the stars, fastened by their global magnetic fields. However, the nature of the observed rotational instabilities has remained elusive up to now. The discussed group of mCP stars is inhomogeneous to such extent that each of the stars may experience a different cause for its period variations.

Review of the Usefulness of Various Rotational Seismometers with Laboratory Results of Fibre-Optic Ones Tested for Engineering Applications

PubMed Central

Jaroszewicz, Leszek R.; Kurzych, Anna; Krajewski, Zbigniew; Marć, Paweł; Kowalski, Jerzy K.; Bobra, Piotr; Zembaty, Zbigniew; Sakowicz, Bartosz; Jankowski, Robert

2016-01-01

Starting with descriptions of rotational seismology, areas of interest and historical field measurements, the fundamental requirements for rotational seismometers for seismological and engineering application are formulated. On the above basis, a review of all existing rotational seismometers is presented with a description of the principles of their operation as well as possibilities to fulfill formulated requirements. This review includes mechanical, acoustical, electrochemical and optical devices and shows that the last of these types are the most promising. It is shown that optical rotational seismometer based on the ring-laser gyroscope concept is the best for seismological applications, whereas systems based on fiber-optic gyroscopes demonstrate parameters which are also required for engineering applications. Laboratory results of the Fibre-Optic System for Rotational Events & Phenomena Monitoring using a small 1-D shaking table modified to generate rotational excitations are presented. The harmonic and time-history tests demonstrate its usefulness for recording rotational motions with rates up to 0.25 rad/s. PMID:27999299

Regulation of Cardiac ATP-sensitive Potassium Channel Surface Expression by Calcium/Calmodulin-dependent Protein Kinase II*

PubMed Central

Sierra, Ana; Zhu, Zhiyong; Sapay, Nicolas; Sharotri, Vikas; Kline, Crystal F.; Luczak, Elizabeth D.; Subbotina, Ekaterina; Sivaprasadarao, Asipu; Snyder, Peter M.; Mohler, Peter J.; Anderson, Mark E.; Vivaudou, Michel; Zingman, Leonid V.; Hodgson-Zingman, Denice M.

2013-01-01

Cardiac ATP-sensitive potassium (KATP) channels are key sensors and effectors of the metabolic status of cardiomyocytes. Alteration in their expression impacts their effectiveness in maintaining cellular energy homeostasis and resistance to injury. We sought to determine how activation of calcium/calmodulin-dependent protein kinase II (CaMKII), a central regulator of calcium signaling, translates into reduced membrane expression and current capacity of cardiac KATP channels. We used real-time monitoring of KATP channel current density, immunohistochemistry, and biotinylation studies in isolated hearts and cardiomyocytes from wild-type and transgenic mice as well as HEK cells expressing wild-type and mutant KATP channel subunits to track the dynamics of KATP channel surface expression. Results showed that activation of CaMKII triggered dynamin-dependent internalization of KATP channels. This process required phosphorylation of threonine at 180 and 224 and an intact 330YSKF333 endocytosis motif of the KATP channel Kir6.2 pore-forming subunit. A molecular model of the μ2 subunit of the endocytosis adaptor protein, AP2, complexed with Kir6.2 predicted that μ2 docks by interaction with 330YSKF333 and Thr-180 on one and Thr-224 on the adjacent Kir6.2 subunit. Phosphorylation of Thr-180 and Thr-224 would favor interactions with the corresponding arginine- and lysine-rich loops on μ2. We concluded that calcium-dependent activation of CaMKII results in phosphorylation of Kir6.2, which promotes endocytosis of cardiac KATP channel subunits. This mechanism couples the surface expression of cardiac KATP channels with calcium signaling and reveals new targets to improve cardiac energy efficiency and stress resistance. PMID:23223335

Regulating Glucose and pH, and Monitoring Oxygen in a Bioreactor

NASA Technical Reports Server (NTRS)

Anderson, Melody M.; Pellis, Neat R.; Jeevarajan, Antony S.; Taylor, Thomas D.; Xu, Yuanhang; Gao, Frank

2006-01-01

A system that automatically regulates the concentration of glucose or pH in a liquid culture medium that is circulated through a rotating-wall perfused bioreactor is described. Another system monitors the concentration of oxygen in the culture medium.

Turbine Engine Disk Rotor Health Monitoring Assessment Using Spin Tests Data

NASA Technical Reports Server (NTRS)

Abdul-Aziz, Ali; Woike, Mark; Baalini, George; Bodis, James R.

2012-01-01

Detecting rotating engine component malfunctions and structural anomalies is increasingly becoming a crucial key feature that will help boost safety and lower maintenance cost. However, achievement of such technology, which can be referred to as a health monitoring remains somewhat challenging to implement. This is mostly due to presence of scattered loading conditions, crack sizes, component geometry and material properties that hinders the simplicity of imposing such application. Different approaches are being considered to assist in developing other means of health monitoring or nondestructive techniques to detect hidden flaws and mini cracks before any catastrophic events occur. These methods extend further to assess material discontinuities and other defects that have matured to the level where a failure is very likely. This paper is focused on presenting data obtained from spin test experiments of a turbine engine like rotor disk and their correlation to the development of a structural health monitoring and fault detection system. The data collected includes blade tip clearance, blade tip timing measurements and shaft displacements. The experimental results are collected at rotational speeds up to 10,000 Rpm and tests are conducted at the NASA Glenn Research Center s Rotordynamics Laboratory via a high precision spin system. Additionally, this study offers a closer glance at a selective online evaluation of a rotating disk using advanced capacitive, microwave and eddy current sensor technology.

Monitoring of Bridges by a Laser Pointer: Dynamic Measurement of Support Rotations and Elastic Line Displacements: Methodology and First Test

PubMed Central

Artese, Serena; Achilli, Vladimiro; Zinno, Raffaele

2018-01-01

Deck inclination and vertical displacements are among the most important technical parameters to evaluate the health status of a bridge and to verify its bearing capacity. Several methods, both conventional and innovative, are used for structural rotations and displacement monitoring; however, none of these allow, at the same time, precision, automation, static and dynamic monitoring without using high cost instrumentation. The proposed system uses a common laser pointer and image processing. The elastic line inclination is measured by analyzing the single frames of an HD video of the laser beam imprint projected on a flat target. For the image processing, a code was developed in Matlab® that provides instantaneous rotation and displacement of a bridge, charged by a mobile load. An important feature is the synchronization of the load positioning, obtained by a GNSS receiver or by a video. After the calibration procedures, a test was carried out during the movements of a heavy truck maneuvering on a bridge. Data acquisition synchronization allowed us to relate the position of the truck on the deck to inclination and displacements. The inclination of the elastic line at the support was obtained with a precision of 0.01 mrad. The results demonstrate the suitability of the method for dynamic load tests, and the control and monitoring of bridges. PMID:29370082

Detection of cerebral oxyhaemoglobin changes during vestibular Coriolis cross-coupling stimulation using near infrared spectroscopy.

PubMed

Kobayashi, A; Cheung, B

2006-02-13

Near infrared spectroscopy (NIRS) has been successful in monitoring cerebral haemodynamics when the subject is immobilized during surgery, and when there is a drastic depletion of blood from the cerebral cortex during positive acceleration. In this study, we monitored subtle changes of cerebral oxygen level using NIRS during vestibular stimulation. For the control conditions, cerebral oxygen status was monitored in six stationary subjects sitting upright, and while they executed head movements in the pitch axis with eyes opened and eyes closed. The experimental conditions involved the subjects making a head movement which required a 45 degrees pitch-down followed by a return to upright head movements 12 s later during yaw rotation (Coriolis cross coupling) at 10 and 20 rotations per minute (rpm) in a random order. Oxyhaemoglobin (O(2)Hb), deoxyhaemoglobin (HHb) and total haemoglobin levels were recorded every 0.5 s from both the parietal and the occipital lobe simultaneously. A significant rotation effect was observed in total Hb level changes from baseline in both regions. Occipital O(2)Hb increased significantly after the head movement with eyes opened at 20 rpm. Our findings appear to be consistent with previous vestibular studies that significant changes in brain blood flow occur during caloric stimulation. NIRS can be used to monitor discrete cortical blood flow changes resulting from vestibular and other forms of stimulation.

Turbine engine disk rotor health monitoring assessment using spin tests data

NASA Astrophysics Data System (ADS)

Abdul-Aziz, Ali; Woike, Mark; Baaklini, George; Bodis, James R.

2012-04-01

Detecting rotating engine component malfunctions and structural anomalies is increasingly becoming a crucial key feature that will help boost safety and lower maintenance cost. However, achievement of such technology, which can be referred to as a health monitoring remains somewhat challenging to implement. This is mostly due to presence of scattered loading conditions, crack sizes, component geometry and material properties that hinders the simplicity of imposing such application. Different approaches are being considered to assist in developing other means of health monitoring or nondestructive techniques to detect hidden flaws and mini cracks before any catastrophic events occur. These methods extend further to assess material discontinuities and other defects that have matured to the level where a failure is very likely. This paper is focused on presenting data obtained from spin test experiments of a turbine engine like rotor disk and their correlation to the development of a structural health monitoring and fault detection system. The data collected includes blade tip clearance, blade tip timing measurements and shaft displacements. The experimental results are collected at rotational speeds up to 10,000 Rpm and tests are conducted at the NASA Glenn Research Center's Rotordynamics Laboratory via a high precision spin system. Additionally, this study offers a closer glance at a selective online evaluation of a rotating disk using advanced capacitive, microwave and eddy current sensor technology.

Visible Thrombolysis Acceleration of a Nanomachine Powered by Light-Driving F0F1-ATPase Motor

NASA Astrophysics Data System (ADS)

Duan, Xiaoxia; Liu, Lifeng; Jiang, Weijian; Yue, Jiachang

2015-05-01

We report on thrombolysis acceleration of a nanomachine powered by light-driving δ-subunit-free F0F1-ATPase motor. It is composed of a mechanical device, locating device, energy storage device, and propeller. The rotory δ-subunit-free F0F1-ATPase motor acts as a mechanical device, which was obtained by reconstructing an original chromatophore extracted from Rhodospirillum rubrum. We found that the bioactivity of the F0F1-ATPase motor improved greatly after reconstruction. The zeta potential of the nanomachine is about -23.4 mV. Cytotoxicity induced by the nanomachine was measured using cell counting kit (CCK)-8 assay. The A549 cells incubated with different fractional concentrations of the nanomachine within 48 h did not show obvious cytotoxicity. The locating device helps the nanomachine bind to the thrombi. Energy was easily stored by exposing the nanomachine to 600-nm-wavelength irradiation, which promoted activity of the motor. The rotation of the long propeller accelerated thrombolysis of a blood clot in vitro in the presence of urokinase (UK). This result was based on visual inspection and confirmed by a series of tests.

Note: Sub-Kelvin refrigeration with dry-coolers on a rotating system.

PubMed

Oguri, S; Ishitsuka, H; Choi, J; Kawai, M; Tajima, O

2014-08-01

We developed a cryogenic system on a rotating table that achieves sub-Kelvin conditions. The cryogenic system consists of a helium sorption cooler and a pulse tube cooler in a cryostat mounted on a rotating table. Two rotary-joint connectors for electricity and helium gas circulation enable the coolers to be operated and maintained with ease. We performed cool-down tests under a condition of continuous rotation at 20 rpm. We obtained a temperature of 0.23 K with a holding time of more than 24 h, thus complying with catalog specifications. We monitored the system's performance for four weeks; two weeks with and without rotation. A few-percent difference in conditions was observed between these two states. Most applications can tolerate such a slight difference. The technology developed is useful for various scientific applications requiring sub-Kelvin conditions on rotating platforms.

A low frequency rotational energy harvesting system

NASA Astrophysics Data System (ADS)

Febbo, M.; Machado, S. P.; Ramirez, J. M.; Gatti, C. D.

2016-11-01

This paper presents a rotary power scavenging unit comprised of two systems of flexible beams connected by two masses which are joined by means of a spring, considering a PZT (QP16N, Midé Corporation) piezoelectric sheet mounted on one of the beams. The energy harvesting (EH) system is mounted rigidly on a rotating hub. The gravitational force on the masses causes sustained oscillatory motion in the flexible beams as long as there is rotary motion. The intention is to use the EH system in the wireless autonomous monitoring of wind turbines under different wind conditions. Specifically, the development is oriented to monitor the dynamic state of the blades of a wind generator of 30 KW which rotates between 50 and 150 rpm. The paper shows a complete set of experimental results on three devices, modifying the amount of beams in the frame supporting the system. The results show an acceptable sustained voltage generation for the expected range, in the three proposed cases. Therefore, it is possible to use this system for generating energy in a low-frequency rotating environment. As an alternative, the system can be easily adapted to include an array of piezoelectric sheets to each of the beams, to provide more power generation.

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

Altenfeld, Anika; Wohlgemuth, Sabine; Wehenkel, Annemarie

The 800 kDa complex of the human Rod, Zwilch and ZW10 proteins (the RZZ complex) was reconstituted in insect cells, purified, crystallized and subjected to preliminary X-ray diffraction analysis. The spindle-assembly checkpoint (SAC) monitors kinetochore–microtubule attachment during mitosis. In metazoans, the three-subunit Rod–Zwilch–ZW10 (RZZ) complex is a crucial SAC component that interacts with additional SAC-activating and SAC-silencing components, including the Mad1–Mad2 complex and cytoplasmic dynein. The RZZ complex contains two copies of each subunit and has a predicted molecular mass of ∼800 kDa. Given the low abundance of the RZZ complex in natural sources, its recombinant reconstitution was attempted bymore » co-expression of its subunits in insect cells. The RZZ complex was purified to homogeneity and subjected to systematic crystallization attempts. Initial crystals containing the entire RZZ complex were obtained using the sitting-drop method and were subjected to optimization to improve the diffraction resolution limit. The crystals belonged to space group P3{sub 1} (No. 144) or P3{sub 2} (No. 145), with unit-cell parameters a = b = 215.45, c = 458.7 Å, α = β = 90.0, γ = 120.0°.« less

A new class of highly potent, broadly neutralizing antibodies isolated from viremic patients infected with dengue virus.

PubMed

Dejnirattisai, Wanwisa; Wongwiwat, Wiyada; Supasa, Sunpetchuda; Zhang, Xiaokang; Dai, Xinghong; Rouvinski, Alexander; Jumnainsong, Amonrat; Edwards, Carolyn; Quyen, Nguyen Than Ha; Duangchinda, Thaneeya; Grimes, Jonathan M; Tsai, Wen-Yang; Lai, Chih-Yun; Wang, Wei-Kung; Malasit, Prida; Farrar, Jeremy; Simmons, Cameron P; Zhou, Z Hong; Rey, Felix A; Mongkolsapaya, Juthathip; Screaton, Gavin R

2015-02-01

Dengue is a rapidly emerging, mosquito-borne viral infection, with an estimated 400 million infections occurring annually. To gain insight into dengue immunity, we characterized 145 human monoclonal antibodies (mAbs) and identified a previously unknown epitope, the envelope dimer epitope (EDE), that bridges two envelope protein subunits that make up the 90 repeating dimers on the mature virion. The mAbs to EDE were broadly reactive across the dengue serocomplex and fully neutralized virus produced in either insect cells or primary human cells, with 50% neutralization in the low picomolar range. Our results provide a path to a subunit vaccine against dengue virus and have implications for the design and monitoring of future vaccine trials in which the induction of antibody to the EDE should be prioritized.

Development of an oligonucleotide probe for Aureobasidium pullulans based on the small-subunit rRNA gene.

PubMed Central

Li, S; Cullen, D; Hjort, M; Spear, R; Andrews, J H

1996-01-01

Aureobasidium pullulans, a cosmopolitan yeast-like fungus, colonizes leaf surfaces and has potential as a biocontrol agent of pathogens. To assess the feasibility of rRNA as a target for A. pullulans-specific oligonucleotide probes, we compared the nucleotide sequences of the small-subunit rRNA (18S) genes of 12 geographically diverse A. pullulans strains. Extreme sequence conservation was observed. The consensus A. pullulans sequence was compared with other fungal sequences to identify potential probes. A 21-mer probe which hybridized to the 12 A. pullulans strains but not to 98 other fungi, including 82 isolates from the phylloplane, was identified. A 17-mer highly specific for Cladosporium herbarum was also identified. These probes have potential in monitoring and quantifying fungi in leaf surface and other microbial communities. PMID:8633850

Chiral self-recognition: direct spectroscopic detection of the homochiral and heterochiral dimers of propylene oxide in the gas phase.

PubMed

Su, Zheng; Borho, Nicole; Xu, Yunjie

2006-12-27

In this report, we describe rotational spectroscopic and high-level ab initio studies of the 1:1 chiral molecular adduct of propylene oxide dimer. The complexes are bound by weak secondary hydrogen bonds, that is, the O(epoxy)...H-C noncovalent interactions. Six homochiral and six heterochiral conformers were predicted to be the most stable configurations where each monomer acts as a proton acceptor and a donor simultaneously, forming two six- or five-membered intermolecular hydrogen-bonded rings. Rotational spectra of six, that is, three homochiral and heterochiral conformer pairs, out of the eight conformers that were predicted to have sufficiently large permanent electric dipole moments were measured and analyzed. The relative conformational stability order and the signs of the chiral recognition energies of the six conformers were determined experimentally and were compared to the ab initio computational results. The experimental observations and the ab initio calculations suggest that the concerted effort of these weak secondary hydrogen bonds can successfully lock the subunits in a particular orientation and that the overall binding strength is comparable to a classic hydrogen bond.

RoboPol: first season rotations of optical polarization plane in blazars

DOE PAGES

Blinov, D.; Pavlidou, V.; Papadakis, I.; ...

2015-08-26

Here, we present first results on polarization swings in optical emission of blazars obtained by RoboPol, a monitoring programme of an unbiased sample of gamma-ray bright blazars specially designed for effective detection of such events. A possible connection of polarization swing events with periods of high activity in gamma-rays is investigated using the data set obtained during the first season of operation. It was found that the brightest gamma-ray flares tend to be located closer in time to rotation events, which may be an indication of two separate mechanisms responsible for the rotations. Blazars with detected rotations during non-rotating periodsmore » have significantly larger amplitude and faster variations of polarization angle than blazars without rotations. Our simulations show that the full set of observed rotations is not a likely outcome (probability ≤1.5 × 10 -2) of a random walk of the polarization vector simulated by a multicell model. Furthermore, it is highly unlikely (~5 × 10 -5) that none of our rotations is physically connected with an increase in gamma-ray activity.« less

NREL Vehicle Testing and Integration Facility (VTIF): Rotating Shadowband Radiometer (RSR); Golden, Colorado (Data)

DOE Data Explorer

Lustbader, J.; Andreas, A.

2012-04-01

This measurement station at NREL's Vehicle Testing and Integration Facility (VTIF) monitors global horizontal, direct normal, and diffuse horizontal irradiance to define the amount of solar energy that hits this particular location. The solar measurement instrumentation is also accompanied by meteorological monitoring equipment.

Graph-based structural change detection for rotating machinery monitoring

NASA Astrophysics Data System (ADS)

Lu, Guoliang; Liu, Jie; Yan, Peng

2018-01-01

Detection of structural changes is critically important in operational monitoring of a rotating machine. This paper presents a novel framework for this purpose, where a graph model for data modeling is adopted to represent/capture statistical dynamics in machine operations. Meanwhile we develop a numerical method for computing temporal anomalies in the constructed graphs. The martingale-test method is employed for the change detection when making decisions on possible structural changes, where excellent performance is demonstrated outperforming exciting results such as the autoregressive-integrated-moving average (ARIMA) model. Comprehensive experimental results indicate good potentials of the proposed algorithm in various engineering applications. This work is an extension of a recent result (Lu et al., 2017).

Monitoring of Earth Rotation by VLBI

NASA Technical Reports Server (NTRS)

Ma., Chopo; Macmillan, D. S.

2000-01-01

Monitoring Earth rotation with Very Long Baseline Interferometry (VLBI) has unique potential because of direct access to the Celestial Reference System (CRF and Terrestrial Reference System (TRF) and the feasibility of re-analyzing the entire data set. While formal precision of better than 0.045 mas for pole and 0.002 ms for UT 1 has been seen in the best 24-hr data, the accuracy of the Earth Orientation Parameter (EOP) time series as a whole is subject to logistical, operational, analytical and conceptual constraints. The current issues related to the VLBI data set and the CORE program for greater time resolution such as analysis consistency, network jitter and reference frame stability will be discussed.

Polar Flagellar Motility of the Vibrionaceae

PubMed Central

McCarter, Linda L.

2001-01-01

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

Monitoring techniques for the X-29A aircraft's high-speed rotating power takeoff shaft

NASA Technical Reports Server (NTRS)

Voracek, David F.

1990-01-01

The experimental X-29A forward swept-wing aircraft has many unique and critical systems that require constant monitoring during ground or flight operation. One such system is the power takeoff shaft, which is the mechanical link between the engine and the aircraft-mounted accessory drive. The X-29A power takeoff shaft opertes in a range between 0 and 16,810 rpm, is longer than most jet engine power takeoff shafts, and is made of graphite epoxy material. Since the X-29A aircraft operates on a single engine, failure of the shaft during flight could lead to loss of the aircraft. The monitoring techniques and test methods used during power takeoff shaft ground and flight operations are discussed. Test data are presented in two case studies where monitoring and testing of the shaft dynamics proved instrumental in discovering and isolating X-29A power takeoff shaft problems. The first study concerns the installation of an unbalanced shaft. The effect of the unbalance on the shaft vibration data and the procedure used to correct the problem are discussed. The second study deals with the shaft exceeding the established vibration limits during flight. This case study found that the vibration of connected rotating machinery unbalances contributed to the excessive vibration level of the shaft. The procedures used to identify the contributions of other rotating machinery unbalances to the power takeoff shaft unbalance are discussed.

A review on prognostic techniques for non-stationary and non-linear rotating systems

NASA Astrophysics Data System (ADS)

Kan, Man Shan; Tan, Andy C. C.; Mathew, Joseph

2015-10-01

The field of prognostics has attracted significant interest from the research community in recent times. Prognostics enables the prediction of failures in machines resulting in benefits to plant operators such as shorter downtimes, higher operation reliability, reduced operations and maintenance cost, and more effective maintenance and logistics planning. Prognostic systems have been successfully deployed for the monitoring of relatively simple rotating machines. However, machines and associated systems today are increasingly complex. As such, there is an urgent need to develop prognostic techniques for such complex systems operating in the real world. This review paper focuses on prognostic techniques that can be applied to rotating machinery operating under non-linear and non-stationary conditions. The general concept of these techniques, the pros and cons of applying these methods, as well as their applications in the research field are discussed. Finally, the opportunities and challenges in implementing prognostic systems and developing effective techniques for monitoring machines operating under non-stationary and non-linear conditions are also discussed.

Laboratory and field testing of commercial rotational seismometers

USGS Publications Warehouse

Nigbor, R.L.; Evans, J.R.; Hutt, C.R.

2009-01-01

There are a small number of commercially available sensors to measure rotational motion in the frequency and amplitude ranges appropriate for earthquake motions on the ground and in structures. However, the performance of these rotational seismometers has not been rigorously and independently tested and characterized for earthquake monitoring purposes as is done for translational strong- and weak-motion seismometers. Quantities such as sensitivity, frequency response, resolution, and linearity are needed for the understanding of recorded rotational data. To address this need, we, with assistance from colleagues in the United States and Taiwan, have been developing performance test methodologies and equipment for rotational seismometers. In this article the performance testing methodologies are applied to samples of a commonly used commercial rotational seismometer, the eentec model R-1. Several examples were obtained for various test sequences in 2006, 2007, and 2008. Performance testing of these sensors consisted of measuring: (1) sensitivity and frequency response; (2) clip level; (3) self noise and resolution; and (4) cross-axis sensitivity, both rotational and translational. These sensor-specific results will assist in understanding the performance envelope of the R-1 rotational seismometer, and the test methodologies can be applied to other rotational seismometers.

Common mechanisms of DNA translocation motors in bacteria and viruses using one-way revolution mechanism without rotation.

PubMed

Guo, Peixuan; Zhao, Zhengyi; Haak, Jeannie; Wang, Shaoying; Wu, Dong; Meng, Bing; Weitao, Tao

2014-01-01

Biomotors were once described into two categories: linear motor and rotation motor. Recently, a third type of biomotor with revolution mechanism without rotation has been discovered. By analogy, rotation resembles the Earth rotating on its axis in a complete cycle every 24h, while revolution resembles the Earth revolving around the Sun one circle per 365 days (see animations http://nanobio.uky.edu/movie.html). The action of revolution that enables a motor free of coiling and torque has solved many puzzles and debates that have occurred throughout the history of viral DNA packaging motor studies. It also settles the discrepancies concerning the structure, stoichiometry, and functioning of DNA translocation motors. This review uses bacteriophages Phi29, HK97, SPP1, P22, T4, and T7 as well as bacterial DNA translocase FtsK and SpoIIIE or the large eukaryotic dsDNA viruses such as mimivirus and vaccinia virus as examples to elucidate the puzzles. These motors use ATPase, some of which have been confirmed to be a hexamer, to revolve around the dsDNA sequentially. ATP binding induces conformational change and possibly an entropy alteration in ATPase to a high affinity toward dsDNA; but ATP hydrolysis triggers another entropic and conformational change in ATPase to a low affinity for DNA, by which dsDNA is pushed toward an adjacent ATPase subunit. The rotation and revolution mechanisms can be distinguished by the size of channel: the channels of rotation motors are equal to or smaller than 2 nm, that is the size of dsDNA, whereas channels of revolution motors are larger than 3 nm. Rotation motors use parallel threads to operate with a right-handed channel, while revolution motors use a left-handed channel to drive the right-handed DNA in an anti-chiral arrangement. Coordination of several vector factors in the same direction makes viral DNA-packaging motors unusually powerful and effective. Revolution mechanism that avoids DNA coiling in translocating the lengthy genomic dsDNA helix could be advantageous for cell replication such as bacterial binary fission and cell mitosis without the need for topoisomerase or helicase to consume additional energy. Copyright © 2014 Elsevier Inc. All rights reserved.

Evaluation of an operating MOD-OA 200 kW wind turbine blade

NASA Technical Reports Server (NTRS)

Donham, R. E.

1979-01-01

Operating loads and structural damage were monitored during operation of the MOD-OA electric generating system. The turbine was damaged locally between stations 48 and 125 after 2.8 million rotations. Loads due to degraded yaw stiffness and fretting at rib station 48 were identified as primary to this distress. The repaired blades operated an additional 4.8 million rotations without problems.

Quench monitoring and control system and method of operating same

DOEpatents

Ryan, David Thomas; Laskaris, Evangelos Trifon; Huang, Xianrui

2006-05-30

A rotating machine comprising a superconductive coil and a temperature sensor operable to provide a signal representative of superconductive coil temperature. The rotating machine may comprise a control system communicatively coupled to the temperature sensor. The control system may be operable to reduce electric current in the superconductive coil when a signal representative of a defined superconducting coil temperature is received from the temperature sensor.

High intensity, short duration rotational grazing on reclaimed cool season tall fescue/legume pastures: II. Forage production, soil and plant tissue comparisons between grazed and ungrazed pastures

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

Carlson, K.E.; Erickson, W.R.; Bonine, R.C.

The Midway Mine is located 50 miles south of Kansas City, Kansas straddling the border of Kansas and Missouri. The Pittsburg & Midway Coal Mining Co. mined the area until 1989, when the mine was closed and reclaimed. Approximately 3,750 acres were topsoiled and revegetated with a cool season tall fescue/legume pasture. High intensity, short duration rotational grazing has become the preferred management practice on these pastures. This study evaluated soil and vegetation data collected on 1,250 acres of pasture which was grazed by about 550 cow/calf units. Ongoing monitoring programs are evaluating the effects of rotational grazing. Soil testingmore » includes macro-nutrients, micro-nutrients and microbial activity. Plant tissue analyses monitor levels of principal macro-nutrients and micro-nutrients. Vegetation monitoring consists of measuring forage production. Results were contrasted between pregrazing and postgrazing, and grazed and ungrazed pasture. Agronomic data from the grazed versus ungrazed treatments documented the following results: (1) higher levels of plant tissue nitrate, phosphorus, potassium, calcium, magnesium, sodium and sulfur; (2) higher microbial activity; (3) similar levels of soil nitrate, phosphorus, potassium, calcium, magnesium, sodium and sulfur; and (4) increased biomass production.« less

The monitoring of transient regimes on machine tools based on speed, acceleration and active electric power absorbed by motors

NASA Astrophysics Data System (ADS)

Horodinca, M.

2016-08-01

This paper intend to propose some new results related with computer aided monitoring of transient regimes on machine-tools based on the evolution of active electrical power absorbed by the electric motor used to drive the main kinematic chains and the evolution of rotational speed and acceleration of the main shaft. The active power is calculated in numerical format using the evolution of instantaneous voltage and current delivered by electrical power system to the electric motor. The rotational speed and acceleration of the main shaft are calculated based on the signal delivered by a sensor. Three real-time analogic signals are acquired with a very simple computer assisted setup which contains a voltage transformer, a current transformer, an AC generator as rotational speed sensor, a data acquisition system and a personal computer. The data processing and analysis was done using Matlab software. Some different transient regimes were investigated; several important conclusions related with the advantages of this monitoring technique were formulated. Many others features of the experimental setup are also available: to supervise the mechanical loading of machine-tools during cutting processes or for diagnosis of machine-tools condition by active electrical power signal analysis in frequency domain.

Stepping-Motion Motor-Control Subsystem For Testing Bearings

NASA Technical Reports Server (NTRS)

Powers, Charles E.

1992-01-01

Control subsystem closed-loop angular-position-control system causing motor and bearing under test to undergo any of variety of continuous or stepping motions. Also used to test bearing-and-motor assemblies, motors, angular-position sensors including rotating shafts, and like. Monitoring subsystem gathers data used to evaluate performance of bearing or other article under test. Monitoring subsystem described in article, "Monitoring Subsystem For Testing Bearings" (GSC-13432).

Accuracy evaluation of the optical surface monitoring system on EDGE linear accelerator in a phantom study

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

Mancosu, Pietro; Fogliata, Antonella, E-mail: Antonella.Fogliata@humanitas.it; Stravato, Antonella

2016-07-01

Frameless stereotactic radiosurgery (SRS) requires dedicated systems to monitor the patient position during the treatment to avoid target underdosage due to involuntary shift. The optical surface monitoring system (OSMS) is here evaluated in a phantom-based study. The new EDGE linear accelerator from Varian (Varian, Palo Alto, CA) integrates, for cranial lesions, the common cone beam computed tomography (CBCT) and kV-MV portal images to the optical surface monitoring system (OSMS), a device able to detect real-time patient's face movements in all 6 couch axes (vertical, longitudinal, lateral, rotation along the vertical axis, pitch, and roll). We have evaluated the OSMS imagingmore » capability in checking the phantoms' position and monitoring its motion. With this aim, a home-made cranial phantom was developed to evaluate the OSMS accuracy in 4 different experiments: (1) comparison with CBCT in isocenter location, (2) capability to recognize predefined shifts up to 2° or 3 cm, (3) evaluation at different couch angles, (4) ability to properly reconstruct the surface when the linac gantry visually block one of the cameras. The OSMS system showed, with a phantom, to be accurate for positioning in respect to the CBCT imaging system with differences of 0.6 ± 0.3 mm for linear vector displacement, with a maximum rotational inaccuracy of 0.3°. OSMS presented an accuracy of 0.3 mm for displacement up to 1 cm and 1°, and 0.5 mm for larger displacements. Different couch angles (45° and 90°) induced a mean vector uncertainty < 0.4 mm. Coverage of 1 camera produced an uncertainty < 0.5 mm. Translations and rotations of a phantom can be accurately detect with the optical surface detector system.« less

Structural and functional aspects of hetero-oligomers formed by the small heat shock proteins αB-crystallin and HSP27.

PubMed

Aquilina, J Andrew; Shrestha, Sudichhya; Morris, Amie M; Ecroyd, Heath

2013-05-10

αB-crystallin and HSP27 are mammalian intracellular small heat shock proteins. These proteins exchange subunits in a rapid and temperature-dependent manner. This facile subunit exchange suggests that differential expression could be used by the cell to regulate the response to stress. A robust technique defines parameters for the dynamic interaction between the major mammalian small heat shock proteins. Small heat shock proteins (sHSPs) exist as large polydisperse species in which there is constant dynamic subunit exchange between oligomeric and dissociated forms. Their primary role in vivo is to bind destabilized proteins and prevent their misfolding and aggregation. αB-Crystallin (αB) and HSP27 are the two most widely distributed and most studied sHSPs in the human body. They are coexpressed in different tissues, where they are known to associate with each other to form hetero-oligomeric complexes. In this study, we aimed to determine how these two sHSPs interact to form hetero-oligomers in vitro and whether, by doing so, there is an increase in their chaperone activity and stability compared with their homo-oligomeric forms. Our results demonstrate that HSP27 and αB formed polydisperse hetero-oligomers in vitro, which had an average molecular mass that was intermediate of each of the homo-oligomers and which were more thermostable than αB, but less so than HSP27. The hetero-oligomer chaperone function was found to be equivalent to that of αB, with each being significantly better in preventing the amorphous aggregation of α-lactalbumin and the amyloid fibril formation of α-synuclein in comparison with HSP27. Using mass spectrometry to monitor subunit exchange over time, we found that HSP27 and αB exchanged subunits 23% faster than the reported rate for HSP27 and αA and almost twice that for αA and αB. This represents the first quantitative evaluation of αB/HSP27 subunit exchange, and the results are discussed in the broader context of regulation of function and cellular proteostasis.

Six Degrees-of-Freedom Prostate and Lung Tumor Motion Measurements Using Kilovoltage Intrafraction Monitoring

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

Huang, Chen-Yu; Tehrani, Joubin Nasehi; Ng, Jin Aun

2015-02-01

Purpose: Tumor positional uncertainty has been identified as a major issue that deteriorates the efficacy of radiation therapy. Tumor rotational movement, which is not well understood, can result in significant geometric and dosimetric inaccuracies. The objective of this study was to measure 6 degrees-of-freedom (6 DoF) prostate and lung tumor motion, focusing on the more novel rotation, using kilovoltage intrafraction monitoring (KIM). Methods and Materials: Continuous kilovoltage (kV) projections of tumors with gold fiducial markers were acquired during radiation therapy for 267 fractions from 10 prostate cancer patients and immediately before or after radiation therapy for 50 fractions from 3more » lung cancer patients. The 6 DoF motion measurements were determined from the individual 3-dimensional (3D) marker positions, after using methods to reject spurious and smooth noisy data, using an iterative closest point algorithm. Results: There were large variations in the magnitude of the tumor rotation among different fractions and patients. Various rotational patterns were observed. The average prostate rotation angles around the left-right (LR), superior-inferior (SI), and anterior-posterior (AP) axes were 1.0 ± 5.0°, 0.6 ± 3.3°, and 0.3 ± 2.0°, respectively. For 35% of the time, the prostate rotated more than 5° about the LR axis, indicating the need for intrafractional adaptation during radiation delivery. For lung patients, the average LR, SI, and AP rotation angles were 0.8 ± 4.2°, −0.8 ± 4.5°, and 1.7 ± 3.1°, respectively. For about 30% of the time, the lung tumors rotated more than 5° around the SI axis. Respiration-induced rotation was detected in 2 of the 3 lung patients. Conclusions: The prostate and lung tumors were found to undergo rotations of more than 5° for about a third of the time. The lung tumor data represent the first 6 DoF tumor motion measured by kV images. The 6 DoF KIM method can enable rotational and translational adaptive radiation therapy and potentially reduce treatment margins.« less

SU-G-JeP4-14: Assessment of Inter- and Intra-Fractional Motion for Extremity Soft Tissue Sarcoma Patients by Using In-House Real-Time Optical Image-Based Monitoring System

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

Kim, H; Kim, I; Ye, S

Purpose: This study aimed to assess inter- and intra-fractional motion for extremity Soft Tissue Sarcoma (STS) patients, by using in-house real-time optical image-based monitoring system (ROIMS) with infra-red (IR) external markers. Methods: Inter- and intra-fractional motions for five extremity (1 upper, 4 lower) STS patients received postoperative 3D conformal radiotherapy (3D-CRT) were measured by registering the image acquired by ROIMS with the planning CT image (REG-ROIMS). To compare with the X-ray image-based monitoring, pre- and post-treatment cone beam computed tomography (CBCT) scans were performed once per week and registered with planning CT image as well (REG-CBCT). If the CBCT scanmore » is not feasible due to the large couch shift, AP and LR on-board imager (OBI) images were acquired. The comparison was done by calculating mutual information (MI) of those registered images. Results: The standard deviation (SD) of the inter-fractional motion was 2.6 mm LR, 2.8 mm SI, and 2.0 mm AP, and the SD of the intra-fractional motion was 1.4 mm, 2.1 mm, and 1.3 mm in each axis, respectively. The SD of rotational inter-fractional motion was 0.6° pitch, 0.9° yaw, and 0.8° roll and the SD of rotational intra-fractional motion was 0.4° pitch, 0.9° yaw, and 0.7° roll. The derived averaged MI values were 0.83, 0.92 for REG-CBCT without rotation and REG-ROIMS with rotation, respectively. Conclusion: The in-house real-time optical image-based monitoring system was implemented clinically and confirmed the feasibility to assess inter- and intra-fractional motion for extremity STS patients while the daily basis and real-time CBCT scan is not feasible in clinic.« less

Real-Time Condition Monitoring and Fault Diagnosis of Gear Train Systems Using Instantaneous Angular Speed (IAS) Analysis

NASA Astrophysics Data System (ADS)

Sait, Abdulrahman S.

This dissertation presents a reliable technique for monitoring the condition of rotating machinery by applying instantaneous angular speed (IAS) analysis. A new analysis of the effects of changes in the orientation of the line of action and the pressure angle of the resultant force acting on gear tooth profile of spur gear under different levels of tooth damage is utilized. The analysis and experimental work discussed in this dissertation provide a clear understating of the effects of damage on the IAS by analyzing the digital signals output of rotary incremental optical encoder. A comprehensive literature review of state of the knowledge in condition monitoring and fault diagnostics of rotating machinery, including gearbox system is presented. Progress and new developments over the past 30 years in failure detection techniques of rotating machinery including engines, bearings and gearboxes are thoroughly reviewed. This work is limited to the analysis of a gear train system with gear tooth surface faults utilizing angular motion analysis technique. Angular motion data were acquired using an incremental optical encoder. Results are compared to a vibration-based technique. The vibration data were acquired using an accelerometer. The signals were obtained and analyzed in the phase domains using signal averaging to determine the existence and position of faults on the gear train system. Forces between the mating teeth surfaces are analyzed and simulated to validate the influence of the presence of damage on the pressure angle and the IAS. National Instruments hardware is used and NI LabVIEW software code is developed for real-time, online condition monitoring systems and fault detection techniques. The sensitivity of optical encoders to gear fault detection techniques is experimentally investigated by applying IAS analysis under different gear damage levels and different operating conditions. A reliable methodology is developed for selecting appropriate testing/operating conditions of a rotating system to generate an alarm system for damage detection.

Dynamic characterization, monitoring and control of rotating flexible beam-mass structures via piezo-embedded techniques

NASA Technical Reports Server (NTRS)

Lai, Steven H.-Y.

1992-01-01

A variational principle and a finite element discretization technique were used to derive the dynamic equations for a high speed rotating flexible beam-mass system embedded with piezo-electric materials. The dynamic equation thus obtained allows the development of finite element models which accommodate both the original structural element and the piezoelectric element. The solutions of finite element models provide system dynamics needed to design a sensing system. The characterization of gyroscopic effect and damping capacity of smart rotating devices are addressed. Several simulation examples are presented to validate the analytical solution.

Precision Photometric Monitoring of Young Low-mass Stars and Brown Dwarfs: Shedding Light on Rotation, Pulsation, and the Star-disk Connection

NASA Astrophysics Data System (ADS)

Cody, Ann Marie; Hillenbrand, L. A.

2010-01-01

Young star-forming regions are host to a variety of optically variable sources, from accreting and flaring stars to those whose light is modulated by surface spots. In addition, recent theory has suggested that a new type of variability-- pulsation powered by deuterium burning-- may be at work on hour timescales in young brown dwarfs and very low mass stars. Photometric studies of these diverse phenomena are key in probing the underlying physics governing the evolution of few-Myr-old cluster members. High-cadence time series provide insight into not only the stochastic nature of accretion, but also trends in rotation via monitoring of magnetic surface spots. Nevertheless, a complete characterization of variability down to low amplitudes, and particularly amongst very-low-mass (0.01-0.3 M⊙) objects, remains elusive. The lower limit to rotation periods in young clusters is not well established, and mechanisms regulating angular momentum down through the substellar regime are not well understood. To expand the census of variability to very low mass and short timescales, as well as assess the possibility of deuterium-burning pulsation, we have carried out a dedicated monitoring campaign on confirmed low-mass members in several 2-5 Myr clusters. Using meter-class telescopes, our survey achieves sensitivity to periodic variations with photometric amplitudes down to the millimagnitude level on timescales ranging from a fraction of an hour to several weeks. We present results from the 5 Myr Sigma Orionis cluster, including a new compilation of rotation rates and a strong correspondence between variability type and presence of a disk, as indicated by Spitzer/IRAC excesses. In contrast to previous reports of 1-4-hour variability amongst brown dwarfs, we find a dearth of periods under 10 hours. However, we identify a significant positive correlation between rotation period and mass.

USSR Report, Military Affairs.

DTIC Science & Technology

1986-09-02

Monitoring the Fulfillment of Pledges"] [Text] Northern Group of Forces—That night the tank company commanded by Captain I. Kuznetsov was to rehearse a...are commanders who reason something like this: We’ll go into the field, we can learn everything there. In particular, Capt Kuznetsov , it emerged...this. 36 But in the regiment where Capt Kuznetsov serves there are subunits which now know to achieve high results in combat training while using

Structure-Function, Stability, and Chemical Modification of the Cyanobacterial Cytochrome b6f Complex from Nostoc sp. PCC 7120*

PubMed Central

Baniulis, Danas; Yamashita, Eiki; Whitelegge, Julian P.; Zatsman, Anna I.; Hendrich, Michael P.; Hasan, S. Saif; Ryan, Christopher M.; Cramer, William A.

2009-01-01

The crystal structure of the cyanobacterial cytochrome b6f complex has previously been solved to 3.0-Å resolution using the thermophilic Mastigocladus laminosus whose genome has not been sequenced. Several unicellular cyanobacteria, whose genomes have been sequenced and are tractable for mutagenesis, do not yield b6f complex in an intact dimeric state with significant electron transport activity. The genome of Nostoc sp. PCC 7120 has been sequenced and is closer phylogenetically to M. laminosus than are unicellular cyanobacteria. The amino acid sequences of the large core subunits and four small peripheral subunits of Nostoc are 88 and 80% identical to those in the M. laminosus b6f complex. Purified b6f complex from Nostoc has a stable dimeric structure, eight subunits with masses similar to those of M. laminosus, and comparable electron transport activity. The crystal structure of the native b6f complex, determined to a resolution of 3.0Å (PDB id: 2ZT9), is almost identical to that of M. laminosus. Two unique aspects of the Nostoc complex are: (i) a dominant conformation of heme bp that is rotated 180° about the α- and γ-meso carbon axis relative to the orientation in the M. laminosus complex and (ii) acetylation of the Rieske iron-sulfur protein (PetC) at the N terminus, a post-translational modification unprecedented in cyanobacterial membrane and electron transport proteins, and in polypeptides of cytochrome bc complexes from any source. The high spin electronic character of the unique heme cn is similar to that previously found in the b6f complex from other sources. PMID:19189962

Cryo-electron microscopy structures of the SARS-CoV spike glycoprotein reveal a prerequisite conformational state for receptor binding.

PubMed

Gui, Miao; Song, Wenfei; Zhou, Haixia; Xu, Jingwei; Chen, Silian; Xiang, Ye; Wang, Xinquan

2017-01-01

The global outbreak of SARS in 2002-2003 was caused by the infection of a new human coronavirus SARS-CoV. The infection of SARS-CoV is mediated mainly through the viral surface glycoproteins, which consist of S1 and S2 subunits and form trimer spikes on the envelope of the virions. Here we report the ectodomain structures of the SARS-CoV surface spike trimer in different conformational states determined by single-particle cryo-electron microscopy. The conformation 1 determined at 4.3 Å resolution is three-fold symmetric and has all the three receptor-binding C-terminal domain 1 (CTD1s) of the S1 subunits in "down" positions. The binding of the "down" CTD1s to the SARS-CoV receptor ACE2 is not possible due to steric clashes, suggesting that the conformation 1 represents a receptor-binding inactive state. Conformations 2-4 determined at 7.3, 5.7 and 6.8 Å resolutions are all asymmetric, in which one RBD rotates away from the "down" position by different angles to an "up" position. The "up" CTD1 exposes the receptor-binding site for ACE2 engagement, suggesting that the conformations 2-4 represent a receptor-binding active state. This conformational change is also required for the binding of SARS-CoV neutralizing antibodies targeting the CTD1. This phenomenon could be extended to other betacoronaviruses utilizing CTD1 of the S1 subunit for receptor binding, which provides new insights into the intermediate states of coronavirus pre-fusion spike trimer during infection.

The NMR solution structure of Mycobacterium tuberculosis F-ATP synthase subunit ε provides new insight into energy coupling inside the rotary engine.

PubMed

Joon, Shin; Ragunathan, Priya; Sundararaman, Lavanya; Nartey, Wilson; Kundu, Subhashri; Manimekalai, Malathy S S; Bogdanović, Nebojša; Dick, Thomas; Grüber, Gerhard

2018-03-01

Mycobacterium tuberculosis (Mt) F 1 F 0 ATP synthase (α 3 :β 3 :γ:δ:ε:a:b:b':c 9 ) is essential for the viability of growing and nongrowing persister cells of the pathogen. Here, we present the first NMR solution structure of Mtε, revealing an N-terminal β-barrel domain (NTD) and a C-terminal domain (CTD) composed of a helix-loop-helix with helix 1 and -2 being shorter compared to their counterparts in other bacteria. The C-terminal amino acids are oriented toward the NTD, forming a domain-domain interface between the NTD and CTD. The Mtε structure provides a novel mechanistic model of coupling c-ring- and ε rotation via a patch of hydrophobic residues in the NTD and residues of the CTD to the bottom of the catalytic α 3 β 3 -headpiece. To test our model, genome site-directed mutagenesis was employed to introduce amino acid changes in these two parts of the epsilon subunit. Inverted vesicle assays show that these mutations caused an increase in ATP hydrolysis activity and a reduction in ATP synthesis. The structural and enzymatic data are discussed in light of the transition mechanism of a compact and extended state of Mtε, which provides the inhibitory effects of this coupling subunit inside the rotary engine. Finally, the employment of these data with molecular docking shed light into the second binding site of the drug Bedaquiline. Structural data are available in the PDB under the accession number 5YIO. © 2018 Federation of European Biochemical Societies.

Structure-Function, Stability, and Chemical Modification of the Cyanobacterial Cytochrome b[subscript 6]f Complex from Nostoc sp. PCC 7120

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

Baniulis, Danas; Yamashita, Eiki; Whitelegge, Julian P.

2009-06-08

The crystal structure of the cyanobacterial cytochrome b{sub 6}f complex has previously been solved to 3.0-{angstrom} resolution using the thermophilic Mastigocladus laminosus whose genome has not been sequenced. Several unicellular cyanobacteria, whose genomes have been sequenced and are tractable for mutagenesis, do not yield b{sub 6}f complex in an intact dimeric state with significant electron transport activity. The genome of Nostoc sp. PCC 7120 has been sequenced and is closer phylogenetically to M. laminosus than are unicellular cyanobacteria. The amino acid sequences of the large core subunits and four small peripheral subunits of Nostoc are 88 and 80% identical tomore » those in the M. laminosus b{sub 6}f complex. Purified b{sub 6}f complex from Nostoc has a stable dimeric structure, eight subunits with masses similar to those of M. laminosus, and comparable electron transport activity. The crystal structure of the native b{sub 6}f complex, determined to a resolution of 3.0{angstrom} (PDB id: 2ZT9), is almost identical to that of M. laminosus. Two unique aspects of the Nostoc complex are: (i) a dominant conformation of heme b{sub p} that is rotated 180 deg. about the {alpha}- and {gamma}-meso carbon axis relative to the orientation in the M. laminosus complex and (ii) acetylation of the Rieske iron-sulfur protein (PetC) at the N terminus, a post-translational modification unprecedented in cyanobacterial membrane and electron transport proteins, and in polypeptides of cytochrome bc complexes from any source. The high spin electronic character of the unique heme cn is similar to that previously found in the b{sub 6}f complex from other sources.« less

Coherent Raman Imaging of Live Muscle Sarcomeres Assisted by SFG Microscopy.

PubMed

Kim, Hyunmin; Kim, Do-Young; Joo, Kyung-Il; Kim, Jung-Hye; Jeong, Soon Moon; Lee, Eun Seong; Hahm, Jeong-Hoon; Kim, Kyuhyung; Moon, Dae Woon

2017-08-23

In this study, we used spectrally focused coherent anti-Stokes Raman scattering (spCARS) microscopy assisted by sum-frequency generation (SFG) to monitor the variations in the structural morphology and molecular vibrations of a live muscle of Caenorhabditis elegans. The subunits of the muscle sarcomeres, such as the M-line, myosin, dense body, and α-actinin, were alternatively observed using spCARS microscopy for different sample orientations, with the guidance of a myosin positional marker captured by SFG microscopy. Interestingly enough, the beam polarization dependence of the spCARS contrasts for two parallel subunits (dense body and myosin) showed a ~90° phase difference. The chemically sensitive spCARS spectra induced by the time-varying overlap of two pulses allowed (after a robust subtraction of the non-resonant background using a modified Kramers-Krönig transformation method) high-fidelity detection of various genetically modified muscle sarcomeres tuned to the C-H vibration (2800-3100 cm -1 ). Conversely, SFG image mapping assisted by phase-retrieved spCARS spectra also facilitated label-free monitoring of the changes in the muscle content of C. elegans that are associated with aging, based on the hypothesis that the C-H vibrational modes could serve as qualitative chemical markers sensitive to the amount and/or structural modulation of the muscle.

New sensors and techniques for the structural health monitoring of propulsion systems.

PubMed

Woike, Mark; Abdul-Aziz, Ali; Oza, Nikunj; Matthews, Bryan

2013-01-01

The ability to monitor the structural health of the rotating components, especially in the hot sections of turbine engines, is of major interest to aero community in improving engine safety and reliability. The use of instrumentation for these applications remains very challenging. It requires sensors and techniques that are highly accurate, are able to operate in a high temperature environment, and can detect minute changes and hidden flaws before catastrophic events occur. The National Aeronautics and Space Administration (NASA), through the Aviation Safety Program (AVSP), has taken a lead role in the development of new sensor technologies and techniques for the in situ structural health monitoring of gas turbine engines. This paper presents a summary of key results and findings obtained from three different structural health monitoring approaches that have been investigated. This includes evaluating the performance of a novel microwave blade tip clearance sensor; a vibration based crack detection technique using an externally mounted capacitive blade tip clearance sensor; and lastly the results of using data driven anomaly detection algorithms for detecting cracks in a rotating disk.

New Sensors and Techniques for the Structural Health Monitoring of Propulsion Systems

PubMed Central

2013-01-01

The ability to monitor the structural health of the rotating components, especially in the hot sections of turbine engines, is of major interest to aero community in improving engine safety and reliability. The use of instrumentation for these applications remains very challenging. It requires sensors and techniques that are highly accurate, are able to operate in a high temperature environment, and can detect minute changes and hidden flaws before catastrophic events occur. The National Aeronautics and Space Administration (NASA), through the Aviation Safety Program (AVSP), has taken a lead role in the development of new sensor technologies and techniques for the in situ structural health monitoring of gas turbine engines. This paper presents a summary of key results and findings obtained from three different structural health monitoring approaches that have been investigated. This includes evaluating the performance of a novel microwave blade tip clearance sensor; a vibration based crack detection technique using an externally mounted capacitive blade tip clearance sensor; and lastly the results of using data driven anomaly detection algorithms for detecting cracks in a rotating disk. PMID:23935425

A Network of Hydrophobic Residues Impeding Helix αC Rotation Maintains Latency of Kinase Gcn2, Which Phosphorylates the α Subunit of Translation Initiation Factor 2▿

PubMed Central

Gárriz, Andrés; Qiu, Hongfang; Dey, Madhusudan; Seo, Eun-Joo; Dever, Thomas E.; Hinnebusch, Alan G.

2009-01-01

Kinase Gcn2 is activated by amino acid starvation and downregulates translation initiation by phosphorylating the α subunit of translation initiation factor 2 (eIF2α). The Gcn2 kinase domain (KD) is inert and must be activated by tRNA binding to the adjacent regulatory domain. Previous work indicated that Saccharomyces cerevisiae Gcn2 latency results from inflexibility of the hinge connecting the N and C lobes and a partially obstructed ATP-binding site in the KD. Here, we provide strong evidence that a network of hydrophobic interactions centered on Leu-856 also promotes latency by constraining helix αC rotation in the KD in a manner relieved during amino acid starvation by tRNA binding and autophosphorylation of Thr-882 in the activation loop. Thus, we show that mutationally disrupting the hydrophobic network in various ways constitutively activates eIF2α phosphorylation in vivo and bypasses the requirement for a key tRNA binding motif (m2) and Thr-882 in Gcn2. In particular, replacing Leu-856 with any nonhydrophobic residue activates Gcn2, while substitutions with various hydrophobic residues maintain kinase latency. We further provide strong evidence that parallel, back-to-back dimerization of the KD is a step on the Gcn2 activation pathway promoted by tRNA binding and autophosphorylation. Remarkably, mutations that disrupt the L856 hydrophobic network or enhance hinge flexibility eliminate the need for the conserved salt bridge at the parallel dimer interface, implying that KD dimerization facilitates the reorientation of αC and remodeling of the active site for enhanced ATP binding and catalysis. We propose that hinge remodeling, parallel dimerization, and reorientation of αC are mutually reinforcing conformational transitions stimulated by tRNA binding and secured by the ensuing autophosphorylation of T882 for stable kinase activation. PMID:19114556

Unidirectional regulation of the F1FO-ATP synthase nanomotor by the ζ pawl-ratchet inhibitor protein of Paracoccus denitrificans and related α-proteobacteria.

PubMed

Zarco-Zavala, Mariel; Mendoza-Hoffmann, Francisco; García-Trejo, José J

2018-06-07

The ATP synthase is a reversible nanomotor that gyrates its central rotor clockwise (CW) to synthesize ATP and in counter clockwise (CCW) direction to hydrolyse it. In bacteria and mitochondria, two natural inhibitor proteins, namely the ε and IF 1 subunits, prevent the wasteful CCW F 1 F O -ATPase activity by blocking γ rotation at the α DP /β DP /γ interface of the F 1 portion. In Paracoccus denitrificans and related α-proteobacteria, we discovered a different natural F 1 -ATPase inhibitor named ζ. Here we revise the functional and structural data showing that this novel ζ subunit, although being different to ε and IF 1 , it also binds to the α DP /β DP /γ interface of the F 1 of P. denitrificans. ζ shifts its N-terminal inhibitory domain from an intrinsically disordered protein region (IDPr) to an α-helix when inserted in the α DP /β DP /γ interface. We showed for the first time the key role of a natural ATP synthase inhibitor by the distinctive phenotype of a Δζ knockout mutant in P. denitrificans. ζ blocks exclusively the CCW F 1 F O -ATPase rotation without affecting the CW-F 1 F O -ATP synthase turnover, confirming that ζ is important for respiratory bacterial growth by working as an unidirectional pawl-ratchet PdF 1 F O -ATPase inhibitor, thus preventing the wasteful consumption of cellular ATP. In summary, ζ is an useful model that mimics mitochondrial IF 1 but in α-proteobacteria. The structural, functional, and endosymbiotic evolutionary implications of this ζ inhibitor are discussed to shed light on the natural control mechanisms of the three natural inhibitor proteins (ε, ζ, and IF 1 ) of this unique ATP synthase nanomotor, essential for life. Copyright © 2018. Published by Elsevier B.V.

A network of hydrophobic residues impeding helix alphaC rotation maintains latency of kinase Gcn2, which phosphorylates the alpha subunit of translation initiation factor 2.

PubMed

Gárriz, Andrés; Qiu, Hongfang; Dey, Madhusudan; Seo, Eun-Joo; Dever, Thomas E; Hinnebusch, Alan G

2009-03-01

Kinase Gcn2 is activated by amino acid starvation and downregulates translation initiation by phosphorylating the alpha subunit of translation initiation factor 2 (eIF2alpha). The Gcn2 kinase domain (KD) is inert and must be activated by tRNA binding to the adjacent regulatory domain. Previous work indicated that Saccharomyces cerevisiae Gcn2 latency results from inflexibility of the hinge connecting the N and C lobes and a partially obstructed ATP-binding site in the KD. Here, we provide strong evidence that a network of hydrophobic interactions centered on Leu-856 also promotes latency by constraining helix alphaC rotation in the KD in a manner relieved during amino acid starvation by tRNA binding and autophosphorylation of Thr-882 in the activation loop. Thus, we show that mutationally disrupting the hydrophobic network in various ways constitutively activates eIF2alpha phosphorylation in vivo and bypasses the requirement for a key tRNA binding motif (m2) and Thr-882 in Gcn2. In particular, replacing Leu-856 with any nonhydrophobic residue activates Gcn2, while substitutions with various hydrophobic residues maintain kinase latency. We further provide strong evidence that parallel, back-to-back dimerization of the KD is a step on the Gcn2 activation pathway promoted by tRNA binding and autophosphorylation. Remarkably, mutations that disrupt the L856 hydrophobic network or enhance hinge flexibility eliminate the need for the conserved salt bridge at the parallel dimer interface, implying that KD dimerization facilitates the reorientation of alphaC and remodeling of the active site for enhanced ATP binding and catalysis. We propose that hinge remodeling, parallel dimerization, and reorientation of alphaC are mutually reinforcing conformational transitions stimulated by tRNA binding and secured by the ensuing autophosphorylation of T882 for stable kinase activation.

Study on fault diagnosis and load feedback control system of combine harvester

NASA Astrophysics Data System (ADS)

Li, Ying; Wang, Kun

2017-01-01

In order to timely gain working status parameters of operating parts in combine harvester and improve its operating efficiency, fault diagnosis and load feedback control system is designed. In the system, rotation speed sensors were used to gather these signals of forward speed and rotation speeds of intermediate shaft, conveying trough, tangential and longitudinal flow threshing rotors, grain conveying auger. Using C8051 single chip microcomputer (SCM) as processor for main control unit, faults diagnosis and forward speed control were carried through by rotation speed ratio analysis of each channel rotation speed and intermediate shaft rotation speed by use of multi-sensor fused fuzzy control algorithm, and these processing results would be sent to touch screen and display work status of combine harvester. Field trials manifest that fault monitoring and load feedback control system has good man-machine interaction and the fault diagnosis method based on rotation speed ratios has low false alarm rate, and the system can realize automation control of forward speed for combine harvester.

The IUE Mega Campaign: Wind Variability and Rotation in Early-Type Stars

NASA Technical Reports Server (NTRS)

Massa, D.; Fullerton, A. W.; Nichols, J. S.; Owocki, S. P.; Prinja, R. K.; St-Louis, N.; Willis, A. J.; Altner, B.; Bolton, C. T.; Cassinelli, J. P.;

Dynamic Radioactive Source for Evaluating and Demonstrating Time-dependent Performance of Continuous Air Monitors.

PubMed

McLean, Thomas D; Moore, Murray E; Justus, Alan L; Hudston, Jonathan A; Barbé, Benoît

2016-11-01

Evaluation of continuous air monitors in the presence of a plutonium aerosol is time intensive, expensive, and requires a specialized facility. The Radiation Protection Services Group at Los Alamos National Laboratory has designed a Dynamic Radioactive Source, intended to replace plutonium aerosol challenge testing. The Dynamic Radioactive Source is small enough to be inserted into the sampler filter chamber of a typical continuous air monitor. Time-dependent radioactivity is introduced from electroplated sources for real-time testing of a continuous air monitor where a mechanical wristwatch motor rotates a mask above an alpha-emitting electroplated disk source. The mask is attached to the watch's minute hand, and as it rotates, more of the underlying source is revealed. The measured alpha activity increases with time, simulating the arrival of airborne radioactive particulates at the air sampler inlet. The Dynamic Radioactive Source allows the temporal behavior of puff and chronic release conditions to be mimicked without the need for radioactive aerosols. The new system is configurable to different continuous air monitor designs and provides an in-house testing capability (benchtop compatible). It is a repeatable and reusable system and does not contaminate the tested air monitor. Test benefits include direct user control, realistic (plutonium) aerosol spectra, and iterative development of continuous air monitor alarm algorithms. Data obtained using the Dynamic Radioactive Source has been used to elucidate alarm algorithms and to compare the response time of two commercial continuous air monitors.

Dynamic Radioactive Source for Evaluating and Demonstrating Time-dependent Performance of Continuous Air Monitors

DOE PAGES

McLean, Thomas D.; Moore, Murray E.; Justus, Alan L.; ...

2016-01-01

Evaluation of continuous air monitors in the presence of a plutonium aerosol is time intensive, expensive, and requires a specialized facility. The Radiation Protection Services Group at Los Alamos National Laboratory has designed a Dynamic Radioactive Source, intended to replace plutonium aerosol challenge testing. Furthermore, the Dynamic Radioactive Source is small enough to be inserted into the sampler filter chamber of a typical continuous air monitor. Time-dependent radioactivity is introduced from electroplated sources for real-time testing of a continuous air monitor where a mechanical wristwatch motor rotates a mask above an alpha-emitting electroplated disk source. The mask is attached tomore » the watch’s minute hand, and as it rotates, more of the underlying source is revealed. The alpha activity we measured increases with time, simulating the arrival of airborne radioactive particulates at the air sampler inlet. The Dynamic Radioactive Source allows the temporal behavior of puff and chronic release conditions to be mimicked without the need for radioactive aerosols. The new system is configurable to different continuous air monitor designs and provides an in-house testing capability (benchtop compatible). It is a repeatable and reusable system and does not contaminate the tested air monitor. Test benefits include direct user control, realistic (plutonium) aerosol spectra, and iterative development of continuous air monitor alarm algorithms. We also used data obtained using the Dynamic Radioactive Source to elucidate alarm algorithms and to compare the response time of two commercial continuous air monitors.« less

Comparison of Water Years 2004-05 and Historical Water-Quality Data, Upper Gunnison River Basin, Colorado

USGS Publications Warehouse

Spahr, Norman E.; Hartle, David M.; Diaz, Paul

2008-01-01

Population growth and changes in land use have the potential to affect water quality and quantity in the upper Gunnison River Basin. In 1995, the U.S. Geological Survey (USGS), in cooperation with the Bureau of Land Management, City of Gunnison, Colorado River Water Conservation District, Crested Butte South Metropolitan District, Gunnison County, Hinsdale County, Mount Crested Butte Water and Sanitation District, National Park Service, Town of Crested Butte, Upper Gunnison River Water Conservancy District, and Western State College, established a water-quality monitoring program in the upper Gunnison River Basin to characterize current water-quality conditions and to assess the effects of increased urban development and other land-use changes on water quality. The monitoring network has evolved into two groups of stations - stations that are considered long term and stations that are considered rotational. The long-term stations are monitored to assist in defining temporal changes in water quality (how conditions may change over time). The rotational stations are monitored to assist in the spatial definition of water-quality conditions (how conditions differ throughout the basin) and to address local and short-term concerns. Some stations in the rotational group were changed beginning in water year 2007. Annual summaries of the water-quality data from the monitoring network provide a point of reference for discussions regarding water-quality monitoring in the upper Gunnison River Basin. This summary includes data collected during water years 2004 and 2005. The introduction provides a map of the sampling sites, definitions of terms, and a one-page summary of selected water-quality conditions at the network stations. The remainder of the summary is organized around the data collected at individual stations. Data collected during water years 2004 and 2005 are compared to historical data, State water-quality standards, and Federal water-quality guidelines. Data were collected following USGS protocols.

Comparison of 2006-2007 Water Years and Historical Water-Quality Data, Upper Gunnison River Basin, Colorado

USGS Publications Warehouse

Solberg, P.A.; Moore, Bryan; Smits, Dennis

2009-01-01

Population growth and changes in land use have the potential to affect water quality and quantity in the upper Gunnison River basin. In 1995, the U.S. Geological Survey (USGS), in cooperation with the Bureau of Land Management, City of Gunnison, Colorado River Water Conservation District, Crested Butte South Metropolitan District, Gunnison County, Hinsdale County, Mount Crested Butte Water and Sanitation District, National Park Service, Town of Crested Butte, Upper Gunnison River Water Conservancy District, and Western State College established a water-quality monitoring program in the upper Gunnison River basin to characterize current water-quality conditions and to assess the effects of increased urban development and other land-use changes on water quality. The monitoring network has evolved into two groups of stations - stations that are considered long term and stations that are considered rotational. The long-term stations are monitored to assist in defining temporal changes in water quality (how conditions may change over time). The rotational stations are monitored to assist in the spatial definition of water-quality conditions (how conditions differ throughout the basin) and to address local and short-term concerns. Some stations in the rotational group were changed beginning in water year 2007. Annual summaries of the water-quality data from the monitoring network provide a point of reference for discussions regarding water-quality monitoring in the upper Gunnison River basin. This summary includes data collected during water years 2006 and 2007. The introduction provides a map of the sampling sites, definitions of terms, and a one-page summary of selected water-quality conditions at the network stations. The remainder of the summary is organized around the data collected at individual stations. Data collected during water years 2006 and 2007 are compared to historical data, State water-quality standards, and Federal water-quality guidelines. Data were collected following USGS protocols (U.S. Geological Survey, variously dated).

Photometric light curves for ten rapidly rotating stars in Alpha Persei, the Pleiades, and the field

NASA Technical Reports Server (NTRS)

Prosser, Charles F.; Schild, Rudolph E.; Stauffer, John R.; Jones, Burton F.

1993-01-01

We present the results from a photometric monitoring program of ten rapidly rotating stars observed during 1991 using the FLWO 48-in. telescope. Brightness variations for an additional six cluster stars observed with the Lick 40-in. telescope are also given. The periods and light curves for seven Alpha Persei members, two Pleiades members, and one naked T Tauri field star are reported.

Changes in non-pine woody species density, composition, and diversity following herbicide and fertilization application to mid-rotation loblolly pine stands

Treesearch

Hal O. Liechty; Conner Fristoe

2012-01-01

We monitored woody vegetation (dbh>1.0 in) response for up to six years following a herbicide (16 ounces imazapyr /acre), a fertilizer (365 pounds urea and 175 pounds diammonium phosphate/acre ) and a combined fertilizer and herbicide application in four mid-rotation loblolly pine stands located within the Upper Gulf Coastal Plain in Arkansas. Approximately 60-80%...

Balance Function Disorders

NASA Technical Reports Server (NTRS)

1991-01-01

Researchers at the Balance Function Laboratory and Clinic at the Minneapolis (MN) Neuroscience Institute on the Abbot Northwestern Hospital Campus are using a rotational chair (technically a "sinusoidal harmonic acceleration system") originally developed by NASA to investigate vestibular (inner ear) function in weightlessness to diagnose and treat patients with balance function disorders. Manufactured by ICS Medical Corporation, Schaumberg, IL, the chair system turns a patient and monitors his or her responses to rotational stimulation.

Rapid Diffusion of Green Fluorescent Protein in the Mitochondrial Matrix

PubMed Central

Partikian, Arthur; Ölveczky, Bence; Swaminathan, R.; Li, Yuxin; Verkman, A.S.

1998-01-01

Abstract. It is thought that the high protein density in the mitochondrial matrix results in severely restricted solute diffusion and metabolite channeling from one enzyme to another without free aqueous-phase diffusion. To test this hypothesis, we measured the diffusion of green fluorescent protein (GFP) expressed in the mitochondrial matrix of fibroblast, liver, skeletal muscle, and epithelial cell lines. Spot photobleaching of GFP with a 100× objective (0.8-μm spot diam) gave half-times for fluorescence recovery of 15–19 ms with >90% of the GFP mobile. As predicted for aqueous-phase diffusion in a confined compartment, fluorescence recovery was slowed or abolished by increased laser spot size or bleach time, and by paraformaldehyde fixation. Quantitative analysis of bleach data using a mathematical model of matrix diffusion gave GFP diffusion coefficients of 2–3 × 10−7 cm2/s, only three to fourfold less than that for GFP diffusion in water. In contrast, little recovery was found for bleaching of GFP in fusion with subunits of the fatty acid β-oxidation multienzyme complex that are normally present in the matrix. Measurement of the rotation of unconjugated GFP by time-resolved anisotropy gave a rotational correlation time of 23.3 ± 1 ns, similar to that of 20 ns for GFP rotation in water. A rapid rotational correlation time of 325 ps was also found for a small fluorescent probe (BCECF, ∼0.5 kD) in the matrix of isolated liver mitochondria. The rapid and unrestricted diffusion of solutes in the mitochondrial matrix suggests that metabolite channeling may not be required to overcome diffusive barriers. We propose that the clustering of matrix enzymes in membrane-associated complexes might serve to establish a relatively uncrowded aqueous space in which solutes can freely diffuse. PMID:9472034

Wavelet analysis of stellar differential rotation. III. The Sun in white light

NASA Astrophysics Data System (ADS)

Hempelmann, A.

2003-02-01

Future space projects like KEPLER will deliver a vast quantity of high precision light curves of stars. This paper describes a test concerning the observability of rotation and even differential rotation of slowly rotating stars from such data. Two published light curves of solar total irradiance measures are investigated: the Nimbus-7 Earth Radiation Budget (ERB) observations between 1978 and 1993 and the Active Cavity Radiometer Irradiance Monitor I (ACRIM I) measurements between 1980 and 1989. Light curve analysis show that oscillations on time-scales comparable to solar rotation but of a complex pattern are visible. Neither Fourier analysis nor time-frequency Wavelet analysis yield the true rotation period during the more active phases of the solar cycle. The true rotation period dominates only for a short time during solar minimum. In the light of this study even space-born broad band photometry may turn out an inappropriate instrument to study stellar butterfly diagrams of stars rotating as slow as the Sun. However, it was shown in Papers I and II of this series that chromospheric tracers like Lyman alpha , Mg II h+k and CaII H+K are appropriate instruments to perform this task.

Developmental effects of wheel running on hippocampal glutamate receptor expression in young and mature adult rats

PubMed Central

Staples, Miranda C.; Somkuwar, Sucharita S.; Mandyam, Chitra D.

2015-01-01

Recent evidence suggests that the behavioral benefits associated with voluntary wheel running in rodents may be due to modulation of glutamatergic transmission in the hippocampus, a brain region implicated in learning and memory. However, the expression of the n-Methyl-d-Aspartate glutamate receptor subunits (GluNs) in the hippocampus in response to chronic sustained voluntary wheel running has not yet been investigated. Further, the developmental effects during young and mature adulthood on wheel running output and GluN expression in hippocampal subregions has not been determined, and therefore is the main focus of this investigation. Eight-week-old and sixteen-week-old male Wistar rats were housed in home cages with free access to running wheels and running output was monitored for four weeks. Wheel access was terminated and tissue from the dorsal and ventral hippocampi were processed for Western blot analysis of GluN subunit expression. Young adult runners demonstrated an escalation in running output but this behavior was not evident in mature adult runners. In parallel, young adult runners demonstrated a significant increase in total GluN (1 and 2A) subunit expression in the dorsal hippocampus, and an opposing effect in the ventral hippocampus compared to age-matched sedentary controls; these changes in total protein expression were not associated with significant alterations in the phosphorylation of the GluN subunits. In contrast, mature adult runners demonstrated a reduction in total GluN2A expression in the dorsal hippocampus, without producing alterations in the ventral hippocampus compared to age-matched sedentary controls. In conclusion, differential running activity-mediated modulation of GluN subunit expression in the hippocampal subregions was revealed to be associated with developmental effects on running activity, which may contribute to altered hippocampal synaptic activity and behavioral outcomes in young and mature adult subjects. PMID:26220171

Meta-Analyses of Dehalococcoides mccartyi Strain 195 Transcriptomic Profiles Identify a Respiration Rate-Related Gene Expression Transition Point and Interoperon Recruitment of a Key Oxidoreductase Subunit

PubMed Central

Mansfeldt, Cresten B.; Rowe, Annette R.; Heavner, Gretchen L. W.; Zinder, Stephen H.

2014-01-01

A cDNA-microarray was designed and used to monitor the transcriptomic profile of Dehalococcoides mccartyi strain 195 (in a mixed community) respiring various chlorinated organics, including chloroethenes and 2,3-dichlorophenol. The cultures were continuously fed in order to establish steady-state respiration rates and substrate levels. The organization of array data into a clustered heat map revealed two major experimental partitions. This partitioning in the data set was further explored through principal component analysis. The first two principal components separated the experiments into those with slow (1.6 ± 0.6 μM Cl−/h)- and fast (22.9 ± 9.6 μM Cl−/h)-respiring cultures. Additionally, the transcripts with the highest loadings in these principal components were identified, suggesting that those transcripts were responsible for the partitioning of the experiments. By analyzing the transcriptomes (n = 53) across experiments, relationships among transcripts were identified, and hypotheses about the relationships between electron transport chain members were proposed. One hypothesis, that the hydrogenases Hup and Hym and the formate dehydrogenase-like oxidoreductase (DET0186-DET0187) form a complex (as displayed by their tight clustering in the heat map analysis), was explored using a nondenaturing protein separation technique combined with proteomic sequencing. Although these proteins did not migrate as a single complex, DET0112 (an FdhB-like protein encoded in the Hup operon) was found to comigrate with DET0187 rather than with the catalytic Hup subunit DET0110. On closer inspection of the genome annotations of all Dehalococcoides strains, the DET0185-to-DET0187 operon was found to lack a key subunit, an FdhB-like protein. Therefore, on the basis of the transcriptomic, genomic, and proteomic evidence, the place of the missing subunit in the DET0185-to-DET0187 operon is likely filled by recruiting a subunit expressed from the Hup operon (DET0112). PMID:25063656

Energy Capture and Use in Plants and Bacteria. Final Technical Report

DOE R&D Accomplishments Database

Boyer, P. D.

1993-12-31

The project has centered on elucidation of the mechanism of ATP synthase. The metabolic importance of ATP and the complexity of the ATP synthase have made the problem particularly important and challenging. The development of the binding change mechanism depended upon our recognition of features that were novel in bioenergetics and indeed to the field of enzymology. One important feature of mechanism is that the principal way that energy input from transmembrane proton movement is coupled to ATP formation is to drive conformational changes that cause the release of ATP readily formed and tightly bound at a catalytic site. Another is that three equivalent catalytic sites on the enzyme show strong catalytic cooperativity as they proceed sequentially through different conformations. A more speculative features is that this cooperativity and energy coupling involve a rotational movement of minor subunits relative to the catalytic subunits. During this period these studies have extended and clarified aspects of the synthase mechanism. During assessments of interactions of Mg{sup 2+} and ADP with the synthase we recognized unexpectedly that whether ADP and P{sub i}, or their complexes with Mg{sup 2+} served as substrates for ATP formation by photophosphorylation was not known. Our studies showed that MgADP and free P{sub i} act as substrates.

Structure and function of the adhesive type IV pilus of Sulfolobus acidocaldarius

PubMed Central

Henche, Anna-Lena; Ghosh, Abhrajyoti; Yu, Xiong; Jeske, Torsten; Egelman, Edward; Albers, Sonja-Verena

2014-01-01

Archaea display a variety of type IV pili on their surface and employ them in different physiological functions. In the crenarchaeon Sulfolobus acidocaldarius the most abundant surface structure is the aap pilus (archaeal adhesive pilus). The construction of in frame deletions of the aap genes revealed that all the five genes (aapA, aapX, aapE, aapF, aapB) are indispensible for assembly of the pilus and an impact on surface motility and biofilm formation was observed. Our analyses revealed that there exists a regulatory cross-talk between the expression of aap genes and archaella (formerly archaeal flagella) genes during different growth phases. The structure of the aap pilus is entirely different from the known bacterial type IV pili as well as other archaeal type IV pili. An aap pilus displayed 3 stranded helices where there is a rotation per subunit of ~ 138° and a rise per subunit of ~ 5.7 Å. The filaments have a diameter of ~ 110 Å and the resolution was judged to be ~ 9 Å. We concluded that small changes in sequence might be amplified by large changes in higher-order packing. Our finding of an extraordinary stability of aap-pili possibly represents an adaptation to harsh environments that S. acidocaldarius encounters. PMID:23078543

Crystal Structure of the Eukaryotic Origin Recognition Complex

PubMed Central

Bleichert, Franziska; Botchan, Michael R.; Berger, James M.

2015-01-01

Initiation of cellular DNA replication is tightly controlled to sustain genomic integrity. In eukaryotes, the heterohexameric origin recognition complex (ORC) is essential for coordinating replication onset. The 3.5 Å resolution crystal structure of Drosophila ORC reveals that the 270 kDa initiator core complex comprises a two-layered notched ring in which a collar of winged-helix domains from the Orc1-5 subunits sits atop a layer of AAA+ ATPase folds. Although canonical inter-AAA+ domain interactions exist between four of the six ORC subunits, unanticipated features are also evident, including highly interdigitated domain-swapping interactions between the winged-helix folds and AAA+ modules of neighboring protomers, and a quasi-spiral arrangement of DNA binding elements that circumnavigate a ~20 Å wide channel in the center of the complex. Comparative analyses indicate that ORC encircles DNA, using its winged-helix domain face to engage the MCM2-7 complex during replicative helicase loading; however, an observed >90° out-of-plane rotation for the Orc1 AAA+ domain disrupts interactions with catalytic amino acids in Orc4, narrowing and sealing off entry into the central channel. Prima facie, our data indicate that Drosophila ORC can switch between active and autoinhibited conformations, suggesting a novel means for cell cycle and/or developmental control of ORC functions. PMID:25762138

Fluorescence-based assay probing regulator of G protein signaling partner proteins.

PubMed

Huang, Po-Shiun; Yeh, Hsin-Sung; Yi, Hsiu-Ping; Lin, Chain-Jia; Yang, Chii-Shen

2012-04-01

The regulator of G protein signaling (RGS) proteins are one of the essential modulators for the G protein system. Besides regulating G protein signaling by accelerating the GTPase activity of Gα subunits, RGS proteins are implicated in exerting other functions; they are also known to be involved in several diseases. Moreover, the existence of a single RGS protein in plants and its seven-transmembrane domain found in 2003 triggered efforts to unveil detailed structural and functional information of RGS proteins. We present a method for real-time examination of the protein-protein interactions between RGS and Gα subunits. AtRGS1 from plants and RGS4 from mammals were site-directedly labeled with the fluorescent probe Lucifer yellow on engineered cysteine residues and used to interact with different Gα subunits. The physical interactions can be revealed by monitoring the real-time fluorescence changes (8.6% fluorescence increase in mammals and 27.6% in plants); their correlations to functional exertion were shown with a GTPase accelerating activity assay and further confirmed by measurement of K(d). We validate the effectiveness of this method and suggest its application to the exploration of more RGS signaling partner proteins in physiological and pathological studies. Copyright © 2012 Elsevier Inc. All rights reserved.

Heterodimerization controls localization of Duox-DuoxA NADPH oxidases in airway cells.

PubMed

Luxen, Sylvia; Noack, Deborah; Frausto, Monika; Davanture, Suzel; Torbett, Bruce E; Knaus, Ulla G

2009-04-15

Duox NADPH oxidases generate hydrogen peroxide at the air-liquid interface of the respiratory tract and at apical membranes of thyroid follicular cells. Inactivating mutations of Duox2 have been linked to congenital hypothyroidism, and epigenetic silencing of Duox is frequently observed in lung cancer. To study Duox regulation by maturation factors in detail, its association with these factors, differential use of subunits and localization was analyzed in a lung cancer cell line and undifferentiated or polarized lung epithelial cells. We show here that Duox proteins form functional heterodimers with their respective DuoxA subunits, in close analogy to the phagocyte NADPH oxidase. Characterization of novel DuoxA1 isoforms and mispaired Duox-DuoxA complexes revealed that heterodimerization is a prerequisite for reactive oxygen species production. Functional Duox1 and Duox2 localize to the leading edge of migrating cells, augmenting motility and wound healing. DuoxA subunits are responsible for targeting functional oxidases to distinct cellular compartments in lung epithelial cells, including Duox2 expression in ciliated cells in an ex vivo differentiated lung epithelium. As these locations probably define signaling specificity of Duox1 versus Duox2, these findings will facilitate monitoring Duox isoform expression in lung disease, a first step for early screening procedures and rational drug development.

NEXT GENERATION ANALYSIS SOFTWARE FOR COMPONENT EVALUATION - Results of Rotational Seismometer Evaluation

NASA Astrophysics Data System (ADS)

Hart, D. M.; Merchant, B. J.; Abbott, R. E.

2012-12-01

The Component Evaluation project at Sandia National Laboratories supports the Ground-based Nuclear Explosion Monitoring program by performing testing and evaluation of the components that are used in seismic and infrasound monitoring systems. In order to perform this work, Component Evaluation maintains a testing facility called the FACT (Facility for Acceptance, Calibration, and Testing) site, a variety of test bed equipment, and a suite of software tools for analyzing test data. Recently, Component Evaluation has successfully integrated several improvements to its software analysis tools and test bed equipment that have substantially improved our ability to test and evaluate components. The software tool that is used to analyze test data is called TALENT: Test and AnaLysis EvaluatioN Tool. TALENT is designed to be a single, standard interface to all test configuration, metadata, parameters, waveforms, and results that are generated in the course of testing monitoring systems. It provides traceability by capturing everything about a test in a relational database that is required to reproduce the results of that test. TALENT provides a simple, yet powerful, user interface to quickly acquire, process, and analyze waveform test data. The software tool has also been expanded recently to handle sensors whose output is proportional to rotation angle, or rotation rate. As an example of this new processing capability, we show results from testing the new ATA ARS-16 rotational seismometer. The test data was collected at the USGS ASL. Four datasets were processed: 1) 1 Hz with increasing amplitude, 2) 4 Hz with increasing amplitude, 3) 16 Hz with increasing amplitude and 4) twenty-six discrete frequencies between 0.353 Hz to 64 Hz. The results are compared to manufacture-supplied data sheets.

A remote monitoring system for patients with implantable ventricular assist devices with a personal handy phone system.

PubMed

Okamoto, E; Shimanaka, M; Suzuki, S; Baba, K; Mitamura, Y

1999-01-01

The usefulness of a remote monitoring system that uses a personal handy phone for artificial heart implanted patients was investigated. The type of handy phone used in this study was a personal handy phone system (PHS), which is a system developed in Japan that uses the NTT (Nippon Telephone and Telegraph, Inc.) telephone network service. The PHS has several advantages: high-speed data transmission, low power output, little electromagnetic interference with medical devices, and easy locating of patients. In our system, patients have a mobile computer (Toshiba, Libretto 50, Kawasaki, Japan) for data transmission control between an implanted controller and a host computer (NEC, PC-9821V16) in the hospital. Information on the motor rotational angle (8 bits) and motor current (8 bits) of the implanted motor driven heart is fed into the mobile computer from the implanted controller (Hitachi, H8/532, Yokohama, Japan) according to 32-bit command codes from the host computer. Motor current and motor rotational angle data from inside the body are framed together by a control code (frame number and parity) for data error checking and correcting at the receiving site, and the data are sent through the PHS connection to the mobile computer. The host computer calculates pump outflow and arterial pressure from the motor rotational angle and motor current values and displays the data in real-time waveforms. The results of this study showed that accurate data on motor rotational angle and current could be transmitted from the subjects while they were walking or driving a car to the host computer at a data transmission rate of 9600 bps. This system is useful for remote monitoring of patients with an implanted artificial heart.

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

Pierson, Bob; Laughlin, Darren

Under this Department of Energy (DOE) grant, A-Tech Corporation d.b.a. Applied Technology Associates (ATA), seeks to develop a seven-degree-of-freedom (7-DOF) seismic measurement tool for high-temperature geothermal applications. The Rotational-Enabled 7-DOF Seismometer includes a conventional tri-axial accelerometer, a conventional pressure sensor or hydrophone, and a tri-axial rotational sensor. The rotational sensing capability is novel, based upon ATA's innovative research in rotational sensing technologies. The geothermal industry requires tools for high-precision seismic monitoring of crack formation associated with Enhanced Geothermal System (EGS) stimulation activity. Currently, microseismic monitoring is conducted by deploying many seismic tools at different depth levels along a 'string' withinmore » drilled observation wells. Costs per string can be hundreds of thousands of dollars. Processing data from the spatial arrays of linear seismometers allows back-projection of seismic wave states. In contrast, a Rotational-Enabled 7-DOF Seismometer would simultaneously measure p-wave velocity, s-wave velocity, and incident seismic wave direction all from a single point measurement. In addition, the Rotational-Enabled 7-DOF Seismometer will, by its nature, separate p- and s-waves into different data streams, simplifying signal processing and facilitating analysis of seismic source signatures and geological characterization. By adding measurements of three additional degrees-of-freedom at each level and leveraging the information from this new seismic observable, it is likely that an equally accurate picture of subsurface seismic activity could be garnered with fewer levels per hole. The key cost savings would come from better siting of the well due to increased information content and a decrease in the number of confirmation wells drilled, also due to the increase in information per well. Improved seismic tools may also increase knowledge, understanding, and confidence, thus removing some current blocks to feasibility and significantly increasing access to potential geothermal sites. During the Phase 1 effort summarized in this final report, the ATA Team modeled and built two TRL 3 proof-of-concept test units for two competing rotational sensor technologies. The two competing technologies were based on ATA's angular rate and angular displacement measurement technologies; Angular rate: ATA's Magnetohydrodynamic Angular Rate Sensor (Seismic MHD); and Angular displacement: ATA's Low Frequency Improved Torsional Seismometer (LFITS). In order to down-select between these two technologies and formulate a go / no go decision, the ATA Team analyzed and traded scientific performance requirements and market constraints against sensor characteristics and components, acquiring field data where possible to validate the approach and publishing results from these studies of rotational technology capability. Based on the results of Phase 1, the ATA Team finds that the Seismic MHD (SMHD) technology is the best choice for enabling rotational seismometry and significant technical potential exists for micro-seismic monitoring using a downhole 7-DOF device based on the SMHD. Recent technical papers and field data confirm the potential of rotational sensing for seismic mapping, increasing confidence that cost-reduction benefits are achievable for EGS. However, the market for geothermal rotational sensing is small and undeveloped. As a result, this report recommends modifying the Phase 2 plan to focus on prototype development aimed at partnering with early adopters within the geothermal industry and the scientific research community. The highest public benefit will come from development and deployment of a science-grade SMHD rotational seismometer engineered for geothermal downhole conditions and an integrated test tool for downhole measurements at active geothermal test sites.« less

Single molecule measurements of F1-ATPase reveal an interdependence between the power stroke and the dwell duration.

PubMed

Spetzler, David; Ishmukhametov, Robert; Hornung, Tassilo; Day, Lixia Jin; Martin, James; Frasch, Wayne D

2009-08-25

Increases in the power stroke and dwell durations of single molecules of Escherichia coli F(1)-ATPase were measured in response to viscous loads applied to the motor and inhibition of ATP hydrolysis. The load was varied using different sizes of gold nanorods attached to the rotating gamma subunit and/or by increasing the viscosity of the medium using PEG-400, a noncompetitive inhibitor of ATPase activity. Conditions that increase the duration of the power stroke were found to cause 20-fold increases in the length of the dwell. These results suggest that the order of hydrolysis, product release, and substrate binding may change as the result of external load on the motor or inhibition of hydrolysis.

Interferometric angle monitor

NASA Technical Reports Server (NTRS)

Minott, P. O. (Inventor)

1983-01-01

Two mutually coherent light beams formed from a single monochromatic light source were directed to a reflecting surface of a rotatable object. They were reflected into an imaging optical lens having a focal plane optically at infinity. A series of interference fringes were formed in the focal plane which were translated linearly in response to angular rotation of the object. Photodetectors were located adjacent the focal plane to detect the fringe translation and output a signal in response to the translation. The signal was fed to a signal processor which was adapted to count the number of fringes detected and develop a measure of the angular rotation and direction of the object.

Earth rotation, station coordinates and orbit determination from satellite laser ranging

NASA Astrophysics Data System (ADS)

Murata, Masaaki

The Project MERIT, a special program of international colaboration to Monitor Earth Rotation and Intercompare the Techniques of observation and analysis, has come to an end with great success. Its major objective was to evaluate the ultimate potential of space techniques such as VLBI and satellite laser ranging, in contrast with the other conventional techniques, in the determination of rotational dynamics of the earth. The National Aerospace Laboratory (NAL) has officially participated in the project as an associate analysis center for satellite laser technique for the period of the MERIT Main Campaign (September 1983-October 1984). In this paper, the NAL analysis center results are presented.

Systems and Methods for the Electrodeposition of a Nickel-cobalt Alloy

NASA Technical Reports Server (NTRS)

Ogozalek, Nance Jo (Inventor); Wistrand, Richard E. (Inventor)

2013-01-01

Systems and methods for electrodepositing a nickel-cobalt alloy using a rotating cylinder electrode assembly with a plating surface and an electrical contact. The assembly is placed within a plating bath and rotated while running a plating cycle. Nickel-cobalt alloy deposition is selectively controlled by controlling current density distribution and/or cobalt content in the plating bath while running the plating cycle to deposit an alloy of a desired yield strength onto the plating surface in a single plating cycle. In various embodiments, the rotating cylinder may be used as an insitu monitoring method to assist in obtaining the properties desired.

Online monitoring of dynamic tip clearance of turbine blades in high temperature environments

NASA Astrophysics Data System (ADS)

Han, Yu; Zhong, Chong; Zhu, Xiaoliang; Zhe, Jiang

2018-04-01

Minimized tip clearance reduces the gas leakage over turbine blade tips and improves the thrust and efficiency of turbomachinery. An accurate tip clearance sensor, measuring the dynamic clearances between blade tips and the turbine case, is a critical component for tip clearance control. This paper presents a robust inductive tip clearance sensor capable of monitoring dynamic tip clearances of turbine machines in high-temperature environments and at high rotational speeds. The sensor can also self-sense the temperature at a blade tip in situ such that temperature effect on tip clearance measurement can be estimated and compensated. To evaluate the sensor’s performance, the sensor was tested for measuring the tip clearances of turbine blades under various working temperatures ranging from 700 K to 1300 K and at turbine rotational speeds ranging from 3000 to 10 000 rpm. The blade tip clearance was varied from 50 to 2000 µm. The experiment results proved that the sensor can accurately measure the blade tip clearances with a temporal resolution of 10 µm. The capability of accurately measuring the tip clearances at high temperatures (~1300 K) and high turbine rotation speeds (~30 000 rpm), along with its compact size, makes it promising for online monitoring and active control of blade tip clearances of high-temperature turbomachinery.

In situ control of industrial processes using laser light scattering and optical rotation

NASA Astrophysics Data System (ADS)

Mendoza Sanchez, Patricia Judith; López Echevarria, Daniel; Huerta Ruelas, Jorge Adalberto

2006-02-01

We present results of optical measurements in products or processes usually found in industrial processes, which can be used to control them. Laser light scattering was employed during semiconductor epitaxial growth by molecular beam epitaxy. With this technique, it was possible to determine growth rate, roughness and critical temperatures related to substrate degradation. With the same scattering technique, oil degradation as function of temperature was monitored for different automotive lubricants. Clear differences can be studied between monograde and multigrade oils. Optical rotation measurements as function of temperature were performed in apple juice in a pasteurization process like. Average variations related to optical rotation dependence of sugars were measured and monitored during heating and cooling process, finding a reversible behavior. As opposite behavior, sugar-protein solution was measured in a similar heating and cooling process. Final result showed a non-reversible behavior related to protein denaturation. Potential applications are discussed for metal-mechanic, electronic, food, and pharmaceutical industry. Future improvements in optical systems to make them more portable and easily implemented under typical industry conditions are mentioned.

Symmetrical refolding of protein domains and subunits: example of the dimeric two-domain 3-isopropylmalate dehydrogenases.

PubMed

Gráczer, Eva; Varga, Andrea; Melnik, Bogdan; Semisotnov, Gennady; Závodszky, Péter; Vas, Mária

2009-02-10

The refolding mechanism of the homodimeric two-domain 3-isopropylmalate dehydrogenase (IPMDH) from the organisms adapted to different temperatures, Thermus thermophilus (Tt), Escherichia coli (Ec), and Vibrio sp. I5 (Vib), is described. In all three cases, instead of a self-template mechanism, the high extent of symmetry and cooperativity in folding of subunits and domains have been concluded from the following experimental findings: The complex time course of refolding, monitored by Trp fluorescence, consists of a fast (the rate constant varies as 16.5, 25.0, and 11.7 min-1 in the order of Tt, Ec, and Vib IPMDHs) and a slow (the rate constants are 0.11, 0.80, and 0.23 min-1 for the three different species) first-order process. However, a burst increase of Trp fluorescence anisotropy to the value of the native states indicates that in all three cases the association of the two polypeptide chains occurs at the beginning of refolding. This dimeric species binds the substrate IPM, but the native-like interactions of the tertiary and quaternary structures are only formed during the slow phase of refolding, accompanied by further increase of protein fluorescence and appearance of FRET between Trp side chain(s) and the bound NADH. Joining the contacting arms of each subunit also takes place exclusively during this slow phase. To monitor refolding of each domain within the intact molecule of T. thermophilus IPMDH, Trp's (located in separate domains) were systematically replaced with Phe's. The refolding processes of the mutants were followed by measuring changes in Trp fluorescence and in FRET between the particular Trp and NADH. The high similarity of time courses (both in biphasicity and in their rates) strongly suggests cooperative folding of the domains during formation of the native three-dimensional structure of IPMDH.

Rotational Spectrum of the Methyl Salicylate-Water Complex: the Missing Conformer and the Tunneling Motions

NASA Astrophysics Data System (ADS)

Ghosh, Supriya; Thomas, Javix; Xu, Yunjie; Jäger, Wolfgang

2015-06-01

Methyl salicylate is a naturally occurring organic ester produced by wintergreen and other plants. It is also found in many over-the-counter remedies, such as muscle ache creams. The rotational spectrum of the methyl salicylate monomer was reported previously, where the most stable, dominant conformer was identified. The methyl salicylate-water complex was first studied using fluorescence-detected infrared spectroscopy; only one monohydrate conformer was found in that work. In the present study, we employed both broadband chirped and cavity based Fourier transform microwave spectroscopy to examine the competition between intra- and intermolecular hydrogen-bonding interactions and possible large amplitude motions associated with the methyl group and the water subunit. In contrast to the previous infrared study, two monohydrate conformers were identified, with carbonyl O or hydroxyl O as the hydrogen bond acceptors. Detailed analyses of the observed hyperfine structures will be presented, as well as our efforts to extend the study to larger methyl salicylate hydration clusters. S. Melandri, B. M. Giuliano, A. Maris, L. B. Favero, P. Ottaviani, B. Velino, W. Caminati, J. Phys. Chem. A. 2007, 111, 9076. A. Mitsuzuka, A. Fujii, T. Ebata, N. Mikami, J. Phys. Chem. A 1998, 102, 9779.

Elevation scanning laser/multi-sensor hazard detection system controller and mirror/mast speed control components. [roving vehicle electromechanical devices

NASA Technical Reports Server (NTRS)

Craig, J.; Yerazunis, S. W.

1978-01-01

The electro-mechanical and electronic systems involved with pointing a laser beam from a roving vehicle along a desired vector are described. A rotating 8 sided mirror, driven by a phase-locked dc motor servo system, and monitored by a precision optical shaft encoder is used. This upper assembly is then rotated about an orthogonal axis to allow scanning into all 360 deg around the vehicle. This axis is also driven by a phase locked dc motor servo-system, and monitored with an optical shaft encoder. The electronics are realized in standard TTL integrated circuits with UV-erasable proms used to store desired coordinates of laser fire. Related topics such as the interface to the existing test vehicle are discussed.

Design and performance of daily quality assurance system for carbon ion therapy at NIRS

NASA Astrophysics Data System (ADS)

Saotome, N.; Furukawa, T.; Hara, Y.; Mizushima, K.; Tansho, R.; Saraya, Y.; Shirai, T.; Noda, K.

2017-09-01

At National Institute of Radiological Sciences (NIRS), we have been commissioning a rotating-gantry system for carbon-ion radiotherapy. This rotating gantry can transport heavy ions at 430 MeV/u to an isocenter with irradiation angles of ±180° that can rotate around the patient so that the tumor can be irradiated from any direction. A three-dimensional pencil-beam scanning irradiation system equipped with the rotating gantry enables the optimal use of physical characteristics of carbon ions to provide accurate treatment. To ensure the treatment quality using such a complex system, the calibration of the primary dose monitor, output check, range check, dose rate check, machine safety check, and some mechanical tests should be performed efficiently. For this purpose, we have developed a measurement system dedicated for quality assurance (QA) of this gantry system: the Daily QA system. The system consists of an ionization chamber system and a scintillator system. The ionization chamber system is used for the calibration of the primary dose monitor, output check, and dose rate check, and the scintillator system is used for the range check, isocenter, and gantry angle. The performance of the Daily QA system was verified by a beam test. The stability of the output was within 0.5%, and the range was within 0.5 mm. The coincidence of the coordinates between the patient-positioning system and the irradiation system was verified using the Daily QA system. Our present findings verified that the new Daily QA system for a rotating gantry is capable of verifying the irradiation system with sufficient accuracy.

ROTATING STARS FROM KEPLER OBSERVED WITH GAIA DR1

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

Davenport, James R. A.

2017-01-20

Astrometric data from the recent Gaia Data Release 1 have been matched against the sample of stars from Kepler with known rotation periods. A total of 1299 bright rotating stars were recovered from the subset of Gaia sources with good astrometric solutions, most with temperatures above 5000 K. From these sources, 894 were selected as lying near the main sequence using their absolute G -band magnitudes. These main-sequence stars show a bimodality in their rotation period distribution, centered roughly around a 600 Myr rotation isochrone. This feature matches the bimodal period distribution found in cooler stars with Kepler , butmore » was previously undetected for solar-type stars due to sample contamination by subgiants. A tenuous connection between the rotation period and total proper motion is found, suggesting that the period bimodality is due to the age distribution of stars within ∼300 pc of the Sun, rather than a phase of rapid angular momentum loss. This work emphasizes the unique power for understanding stellar populations that is created by combining temporal monitoring from Kepler with astrometric data from Gaia .« less

Methods for Expanding Rotary Wing Aircraft Health and Usage Monitoring Systems to the Rotating Frame through Real-time Rotor Blade Kinematics Estimation

NASA Astrophysics Data System (ADS)

Allred, Charles Jefferson

Since the advent of Health and Usage Monitoring Systems (HUMS) in the early 1990's, there has been a steady decrease in the number of component failure related helicopter accidents. Additionally, measurable cost benefits due to improved maintenance practices based on HUMS data has led to a desire to expand HUMS from its traditional area of helicopter drive train monitoring. One of the areas of greatest interest for this expansion of HUMS is monitoring of the helicopter rotor head loads. Studies of rotor head load and blade motions have primarily focused on wind tunnel testing with technology which would not be applicable for production helicopter HUMS deployment, or measuring bending along the blade, rather than where it is attached to the rotor head and the location through which all the helicopter loads pass. This dissertation details research into finding methods for real time methods of estimating rotor blade motion which could be applied across helicopter fleets as an expansion of current HUMS technology. First, there is a brief exploration of supporting technologies which will be crucial in enabling the expansion of HUMS from the fuselage of helicopters to the rotor head: wireless data transmission and energy harvesting. A brief overview of the commercially available low power wireless technology selected for this research is presented. The development of a relatively high-powered energy harvester specific to the motion of helicopter rotor blades is presented and two different prototypes of the device are shown. Following the overview of supporting technologies, two novel methods of monitoring rotor blade motion in real time are developed. The first method employs linear displacement sensors embedded in the elastomer layers of a high-capacity laminate bearing of the type commonly used in fully articulated rotors throughout the helicopter industry. The configuration of these displacement sensors allows modeling of the sensing system as a robotic parallel mechanism, similar to a Stewart Platform. A calibration method for this device is developed and the improved orientation estimation results are shown. The second method is not specific to the fully articulated rotor head mounting geometry of the first method. Rather, it utilizes micro-electromechanical (MEMS) accelerometers and gyroscopes configured to measure the centrifugal acceleration and rotation rate induced through rotor head rotation differentially. By measuring these quantities differentially, other accelerations from the fuselage reference frame are removed from the measurement, resulting in acceleration and rate quantities that are impacted only by the angle of the sensors relative to the plane of rotation. By mounting these sensors strategically and symmetrically about the rotor blade root center of rotation, the orientation of the rotor blade can be estimated in real time.

Evaluation of the lubrication mechanism at various rotation speeds and granule filling levels in a container mixer using a thermal effusivity sensor.

PubMed

Uchiyama, Jumpei; Aoki, Shigeru

2015-01-01

To research the detailed mechanism of the lubrication process using the thermal effusivity sensor, the relationships of the lubrication progress with the pattern of powder flow, the rotation speed and the filling level were investigated. The thermal effusivity profile was studied as a function of the number of rotations at various rotation speeds. It was observed that at lower rotation speeds, the profiles of the lubrication progress were almost the same, regardless of the rotation speed. In this region, the highest speed was defined as the critical rotation speed (CRS), which was found to be one of the important factors. The CRS had close relations with avalanche flow in the blender. The first and the second phases were observed in the lubrication process. The first phase was influenced by the CRS and the filling level in the blender. The second phase was influenced by the rotation speed. The mechanism of two-phase process was proposed as a macro progression of the dispersion of the lubricant (first phase) and micro progression of the coating of the powder particles with lubricant (second phase). The accurate monitoring by the thermal effusivity sensor was able to help a better understanding in the lubrication process.

Vision-based measurement for rotational speed by improving Lucas-Kanade template tracking algorithm.

PubMed

Guo, Jie; Zhu, Chang'an; Lu, Siliang; Zhang, Dashan; Zhang, Chunyu

2016-09-01

Rotational angle and speed are important parameters for condition monitoring and fault diagnosis of rotating machineries, and their measurement is useful in precision machining and early warning of faults. In this study, a novel vision-based measurement algorithm is proposed to complete this task. A high-speed camera is first used to capture the video of the rotational object. To extract the rotational angle, the template-based Lucas-Kanade algorithm is introduced to complete motion tracking by aligning the template image in the video sequence. Given the special case of nonplanar surface of the cylinder object, a nonlinear transformation is designed for modeling the rotation tracking. In spite of the unconventional and complex form, the transformation can realize angle extraction concisely with only one parameter. A simulation is then conducted to verify the tracking effect, and a practical tracking strategy is further proposed to track consecutively the video sequence. Based on the proposed algorithm, instantaneous rotational speed (IRS) can be measured accurately and efficiently. Finally, the effectiveness of the proposed algorithm is verified on a brushless direct current motor test rig through the comparison with results obtained by the microphone. Experimental results demonstrate that the proposed algorithm can extract accurately rotational angles and can measure IRS with the advantage of noncontact and effectiveness.

Monitoring arrangement for vented nuclear fuel elements

DOEpatents

Campana, Robert J.

1981-01-01

In a nuclear fuel reactor core, fuel elements are arranged in a closely packed hexagonal configuration, each fuel element having diametrically opposed vents permitting 180.degree. rotation of the fuel elements to counteract bowing. A grid plate engages the fuel elements and forms passages for communicating sets of three, four or six individual vents with respective monitor lines in order to communicate vented radioactive gases from the fuel elements to suitable monitor means in a manner readily permitting detection of leakage in individual fuel elements.

Movable Cameras And Monitors For Viewing Telemanipulator

NASA Technical Reports Server (NTRS)

Diner, Daniel B.; Venema, Steven C.

1993-01-01

Three methods proposed to assist operator viewing telemanipulator on video monitor in control station when video image generated by movable video camera in remote workspace of telemanipulator. Monitors rotated or shifted and/or images in them transformed to adjust coordinate systems of scenes visible to operator according to motions of cameras and/or operator's preferences. Reduces operator's workload and probability of error by obviating need for mental transformations of coordinates during operation. Methods applied in outer space, undersea, in nuclear industry, in surgery, in entertainment, and in manufacturing.

The disaggregation theory of signal transduction revisited: further evidence that G proteins are multimeric and disaggregate to monomers when activated.

PubMed

Jahangeer, S; Rodbell, M

1993-10-01

We have compared the sedimentation rates on sucrose gradients of the heterotrimeric GTP-binding regulatory (G) proteins Gs, G(o), Gi, and Gq extracted from rat brain synaptoneurosomes with Lubrol and digitonin. The individual alpha and beta subunits were monitored with specific antisera. In all cases, both subunits cosedimented, indicating that the subunits are likely complexed as heterotrimers. When extracted with Lubrol all of the G proteins sedimented with rates of about 4.5 S (consistent with heterotrimers) whereas digitonin extracted 60% of the G proteins with peaks at 11 S; 40% pelleted as larger structures. Digitonin-extracted Gi was cross-linked by p-phenylenedimaleimide, yielding structures too large to enter polyacrylamide gels. No cross-linking of Lubrol-extracted Gi occurred. Treatment of the membranes with guanosine 5'-[gamma-thio]triphosphate and Mg2+ yielded digitonin-extracted structures with peak sedimentation values of 8.5 S--i.e., comparable to that of purified G(o) in digitonin and considerably larger than the Lubrol-extracted 2S structures representing the separated alpha and beta gamma subunits formed by the actions of guanosine 5'-[gamma-thio]triphosphate. It is concluded that the multimeric structures of G proteins in brain membranes are at least partially preserved in digitonin and that activation of these structures in membranes yields monomers of G proteins rather than the disaggregated products (alpha and beta gamma complexes) observed in Lubrol. It is proposed that hormones and GTP affect the dynamic interplay between multimeric G proteins and receptors in a fashion analogous to the actions of ATP on the dynamic interactions between myosin and actin filaments. Signal transduction is mediated by activated monomers released from the multimers during the activation process.

The disaggregation theory of signal transduction revisited: further evidence that G proteins are multimeric and disaggregate to monomers when activated.

PubMed Central

Jahangeer, S; Rodbell, M

1993-01-01

We have compared the sedimentation rates on sucrose gradients of the heterotrimeric GTP-binding regulatory (G) proteins Gs, G(o), Gi, and Gq extracted from rat brain synaptoneurosomes with Lubrol and digitonin. The individual alpha and beta subunits were monitored with specific antisera. In all cases, both subunits cosedimented, indicating that the subunits are likely complexed as heterotrimers. When extracted with Lubrol all of the G proteins sedimented with rates of about 4.5 S (consistent with heterotrimers) whereas digitonin extracted 60% of the G proteins with peaks at 11 S; 40% pelleted as larger structures. Digitonin-extracted Gi was cross-linked by p-phenylenedimaleimide, yielding structures too large to enter polyacrylamide gels. No cross-linking of Lubrol-extracted Gi occurred. Treatment of the membranes with guanosine 5'-[gamma-thio]triphosphate and Mg2+ yielded digitonin-extracted structures with peak sedimentation values of 8.5 S--i.e., comparable to that of purified G(o) in digitonin and considerably larger than the Lubrol-extracted 2S structures representing the separated alpha and beta gamma subunits formed by the actions of guanosine 5'-[gamma-thio]triphosphate. It is concluded that the multimeric structures of G proteins in brain membranes are at least partially preserved in digitonin and that activation of these structures in membranes yields monomers of G proteins rather than the disaggregated products (alpha and beta gamma complexes) observed in Lubrol. It is proposed that hormones and GTP affect the dynamic interplay between multimeric G proteins and receptors in a fashion analogous to the actions of ATP on the dynamic interactions between myosin and actin filaments. Signal transduction is mediated by activated monomers released from the multimers during the activation process. Images Fig. 1 Fig. 2 PMID:8415607

Sensorless Estimation and Nonlinear Control of a Rotational Energy Harvester

NASA Astrophysics Data System (ADS)

Nunna, Kameswarie; Toh, Tzern T.; Mitcheson, Paul D.; Astolfi, Alessandro

2013-12-01

It is important to perform sensorless monitoring of parameters in energy harvesting devices in order to determine the operating states of the system. However, physical measurements of these parameters is often a challenging task due to the unavailability of access points. This paper presents, as an example application, the design of a nonlinear observer and a nonlinear feedback controller for a rotational energy harvester. A dynamic model of a rotational energy harvester with its power electronic interface is derived and validated. This model is then used to design a nonlinear observer and a nonlinear feedback controller which yield a sensorless closed-loop system. The observer estimates the mechancial quantities from the measured electrical quantities while the control law sustains power generation across a range of source rotation speeds. The proposed scheme is assessed through simulations and experiments.

Monitoring methanol-induced protein unfolding by fluorescence anisotropy measurements of covalently labelled rhodamine probe*

NASA Astrophysics Data System (ADS)

Soleilhac, Antonin; Bertorelle, Franck; Dugourd, Philippe; Girod, Marion; Antoine, Rodolphe

2017-06-01

We describe the use of an extrinsic fluorophore (rhodamine B isothiocyanate) as a versatile probe to measure rotational motions of proteins. To illustrate the usefulness of this probe, we describe the fluorescence anisotropy values of this fluorophore covalently linked to myoglobin protein measured in aqueous solutions of increased methanol content. Methanol-induced unfolding is revealed by the transition from constrained to free rotation of the covalently attached rhodamine B fluorophore.

Protein brownian rotation at the glass transition temperature of a freeze-concentrated buffer probed by superparamagnetic nanoparticles.

PubMed

Eloi, J-C; Okuda, M; Jones, S E Ward; Schwarzacher, W

2013-06-18

For applications from food science to the freeze-thawing of proteins it is important to understand the often complex freezing behavior of solutions of biomolecules. Here we use a magnetic method to monitor the Brownian rotation of a quasi-spherical cage-shaped protein, apoferritin, approaching the glass transition Tg in a freeze-concentrated buffer (Tris-HCl). The protein incorporates a synthetic magnetic nanoparticle (Co-doped Fe3O4 (magnetite)). We use the magnetic signal from the nanoparticles to monitor the protein orientation. As T decreases toward Tg of the buffer solution the protein's rotational relaxation time increases exponentially, taking values in the range from a few seconds up to thousands of seconds, i.e., orders of magnitude greater than usually accessed, e.g., by NMR. The longest relaxation times measured correspond to estimated viscosities >2 MPa s. As well as being a means to study low-temperature, high-viscosity environments, our method provides evidence that, for the cooling protocol used, the following applies: 1), the concentration of the freeze-concentrated buffer at Tg is independent of its initial concentration; 2), little protein adsorption takes place at the interface between ice and buffer; and 3), the protein is free to rotate even at temperatures as low as 207 K. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Retro- and prospection for mental time travel: Emergence of episodic remembering and mental rotation in 5- to 8-year old children☆

PubMed Central

Perner, Josef; Kloo, Daniela; Rohwer, Michael

2010-01-01

We investigate the common development of children’s ability to “look back in time” (retrospection, episodic remembering) and to “look into the future” (prospection). Experiment 1 with 59 children 5 to 8.5 years old showed mental rotation, as a measure of prospection, explaining specific variance of free recall, as a measure of episodic remembering (retrospection) when controlled for cued recall. Experiment 2 with 31 children from 5 to 6.5 years measured episodic remembering with recall of visually experienced events (seeing which picture was placed inside a box) when controlling for recall of indirectly conveyed events (being informed about the pictures placed inside the box by showing the pictures on a monitor). Quite unexpectedly rotators were markedly worse on indirect items than non-rotators. We speculate that with the ability to rotate children switch from knowledge retrieval to episodic remembering, which maintains success for experienced events but has detrimental effects for indirect information. PMID:20650660

Non-resonant energy harvester with elastic constraints for low rotating frequencies

NASA Astrophysics Data System (ADS)

Machado, Sebastián P.; Febbo, Mariano; Gatti, Claudio D.; Ramirez, José M.

2017-11-01

This paper presents a non-resonant piezoelectric energy harvester (PEH) which is designed to capture energy from low frequency rotational vibration. The proposed device works out of the plane of rotation where the motion of a mass-spring system is transferred to a piezoelectric layer with the intention to generate energy to power wireless structural monitoring systems or sensors. The mechanical structure is formed by two beams with rigid and elastic boundary conditions at the clamped end. On the free boundaries, heavy masses connected by a spring are placed in order to increase voltage generation and diminish the natural frequency. A mathematical framework and the equations governing the energy-harvesting system are presented. Numerical simulations and experimental verifications are performed for different rotation speeds ranging from 0.7 to 2.5 Hz. An output power of 125 μW is obtained for maximum rotating frequency demonstrating that the proposed design can collect enough energy for the suggested application.

Doppler-resolved kinetics of saturation recovery

DOE PAGES

Forthomme, Damien; Hause, Michael L.; Yu, Hua -Gen; ...

2015-04-08

Frequency modulated laser transient absorption has been used to monitor the ground state rotational energy transfer rates of CN radicals in a double-resonance, depletion recovery experiment. When a pulsed laser is used to burn a hole in the equilibrium ground state population of one rotational state without velocity selection, the population recovery rate is found to depend strongly on the Doppler detuning of a narrow-band probe laser. Similar effects should be apparent for any relaxation rate process that competes effectively with velocity randomization. Alternative methods of extracting thermal rate constants in the presence of these non-thermal conditions are evaluated. Totalmore » recovery rate constants, analogous to total removal rate constants in an experiment preparing a single initial rotational level, are in good agreement with quantum scattering calculations, but are slower than previously reported experiments and show qualitatively different rotational state dependence between Ar and He collision partners. As a result, quasi-classical trajectory studies confirm that the differing rotational state dependence is primarily a kinematic effect.« less

Multivariate EMD and full spectrum based condition monitoring for rotating machinery

NASA Astrophysics Data System (ADS)

Zhao, Xiaomin; Patel, Tejas H.; Zuo, Ming J.

2012-02-01

Early assessment of machinery health condition is of paramount importance today. A sensor network with sensors in multiple directions and locations is usually employed for monitoring the condition of rotating machinery. Extraction of health condition information from these sensors for effective fault detection and fault tracking is always challenging. Empirical mode decomposition (EMD) is an advanced signal processing technology that has been widely used for this purpose. Standard EMD has the limitation in that it works only for a single real-valued signal. When dealing with data from multiple sensors and multiple health conditions, standard EMD faces two problems. First, because of the local and self-adaptive nature of standard EMD, the decomposition of signals from different sources may not match in either number or frequency content. Second, it may not be possible to express the joint information between different sensors. The present study proposes a method of extracting fault information by employing multivariate EMD and full spectrum. Multivariate EMD can overcome the limitations of standard EMD when dealing with data from multiple sources. It is used to extract the intrinsic mode functions (IMFs) embedded in raw multivariate signals. A criterion based on mutual information is proposed for selecting a sensitive IMF. A full spectral feature is then extracted from the selected fault-sensitive IMF to capture the joint information between signals measured from two orthogonal directions. The proposed method is first explained using simple simulated data, and then is tested for the condition monitoring of rotating machinery applications. The effectiveness of the proposed method is demonstrated through monitoring damage on the vane trailing edge of an impeller and rotor-stator rub in an experimental rotor rig.

Rotational glenohumeral adaptations are associated with shoulder pathology in professional male handball players.

PubMed

Lubiatowski, Przemyslaw; Kaczmarek, Piotr; Cisowski, Pawel; Breborowicz, Ewa; Grygorowicz, Monika; Dzianach, Marcin; Krupecki, Tomasz; Laver, Lior; Romanowski, Leszek

2018-01-01

Glenohumeral range of motion adaptations may affect throwing athletes and contribute to shoulder injury. The purpose of this study was to evaluate shoulder rotation deficits among elite professional handball players and its correlation to the presence of shoulder pain and morphological changes. Eighty-seven elite professional handball players and 41 healthy non-athlete volunteers participated in the study. Evaluations included measurement of range of internal and external rotation, total arch of motion, identification of shoulder pain and ultrasound scan for diagnosis of rotator cuff tears and internal impingement. Glenohumeral rotational deficits (>20-25°) were found among 11 players group (13%). The throwing shoulders in the players group showed a decrease in internal rotation and an increase in external rotation with significantly larger ranges among players compared to the non-athlete group. Internal rotation deficit >20° was associated with higher incidence of shoulder pain among players. Both internal rotation deficits (>25°) and total arch of motion deficit (>20°) co-existed with higher incidence of internal impingement. Shoulder pain was common (36/97-41%) and was associated with decreased external rotation and total arch of motion. Internal impingement (found in 13/87-15%) correlated with decreased rotation ranges and a greater deficit in total arch of motion, whereas higher gain in external rotation correlated with a partial rotator cuff tear (found in 12/87-14%). Shoulder pathologies and problems commonly affected the group of handball players. Greater glenohumeral rotational deficits in throwing shoulders of handball players correlate with shoulder pain and internal impingement, while increased external rotation with partial rotator cuff tears. Such deficits affect 13% of the athlete population. Major clinical relevance of the study is to monitor handball players' shoulders both clinically and by proper imaging. Evaluation of range of rotation seems to identify shoulders at risk of the pathology. Cross-Sectional study with control group, Level II.

Comparison of 2002 Water Year and Historical Water-Quality Data, Upper Gunnison River Basin, Colorado

USGS Publications Warehouse

Spahr, N.E.

2003-01-01

Introduction: Population growth and changes in land-use practices have the potential to affect water quality and quantity in the upper Gunnison River basin. In 1995, the U.S. Geological Survey (USGS), in cooperation with local sponsors, City of Gunnison, Colorado River Water Conservation District, Crested Butte South Metropolitan District, Gunnison County, Mount Crested Butte Water and Sanitation District, National Park Service, Town of Crested Butte, and Upper Gunnison River Water Conservancy District, established a water-quality monitoring program in the upper Gunnison River basin to characterize current water-quality conditions and to assess the effects of increased urban development and other land-use changes on water quality. The monitoring network has evolved into two groups of stations, stations that are considered as long term and stations that are rotational. The long-term stations are monitored to assist in defining temporal changes in water quality (how conditions have changed over time). The rotational stations are monitored to assist in the spatial definition of water-quality conditions (how conditions differ throughout the basin) and to address local and short term concerns. Another group of stations (rotational group 2) will be chosen and sampled beginning in water year 2004. Annual summaries of the water-quality data from the monitoring network provide a point of reference for discussions regarding water-quality sampling in the upper Gunnison River basin. This summary includes data collected during water year 2002. The introduction provides a map of the sampling locations, definitions of terms, and a one-page summary of selected water-quality conditions at the network stations. The remainder of the summary is organized around the data collected at individual stations. Data collected during water year 2002 are compared to historical data (data collected for this network since 1995), state water-quality standards, and federal water-quality guidelines. Data were collected during water year 2002 following USGS protocols (U.S. Geological Survey, variously dated).

Monitoring and Surveillance of Marine Invasive Species in Californian Waters by DNA Barcoding: Methodological and Analytical Solutions

NASA Astrophysics Data System (ADS)

Campbell, T. L.; Geller, J. B.; Heller, P.; Ruiz, G.; Chang, A.; McCann, L.; Ceballos, L.; Marraffini, M.; Ashton, G.; Larson, K.; Havard, S.; Meagher, K.; Wheelock, M.; Drake, C.; Rhett, G.

2016-02-01

The Ballast Water Management Act, the Marine Invasive Species Act, and the Coastal Ecosystem Protection Act require the California Department of Fish and Wildlife to monitor and evaluate the extent of biological invasions in the state's marine and estuarine waters. This has been performed statewide, using a variety of methodologies. Conventional sample collection and processing is laborious, slow and costly, and may require considerable taxonomic expertise requiring detailed time-consuming microscopic study of multiple specimens. These factors limit the volume of biomass that can be searched for introduced species. New technologies continue to reduce the cost and increase the throughput of genetic analyses, which become efficient alternatives to traditional morphological analysis for identification, monitoring and surveillance of marine invasive species. Using next-generation sequencing of mitochondrial Cytochrome c oxidase subunit I (COI) and nuclear large subunit ribosomal RNA (LSU), we analyzed over 15,000 individual marine invertebrates collected in Californian waters. We have created sequence databases of California native and non-native species to assist in molecular identification and surveillance in North American waters. Metagenetics, the next-generation sequencing of environmental samples with comparison to DNA sequence databases, is a faster and cost-effective alternative to individual sample analysis. We have sequenced from biomass collected from whole settlement plates and plankton in California harbors, and used our introduced species database to create species lists. We can combine these species lists for individual marinas with collected environmental data, such as temperature, salinity, and dissolved oxygen to understand the ecology of marine invasions. Here we discuss high throughput sampling, sequencing, and COASTLINE, our data analysis answer to challenges working with hundreds of millions of sequencing reads from tens of thousands of specimens.

Mass spectrometric real-time monitoring of an enzymatic phosphorylation assay using internal standards and data-handling freeware.

PubMed

Krappmann, Michael; de Boer, Arjen R; Kool, Daniël R W; Irth, Hubertus; Letzel, Thomas

2016-04-30

Continuous-flow reaction detection systems (monitoring enzymatic reactions with mass spectrometry (MS)) lack quantitative values so far. Therefore, two independent internal standards (IS) are implemented in a way that the online system stability can be observed, quantitative conversion values for substrate and product can be obtained and they can be used as mass calibration standards for high MS accuracy. An application previously developed for the MS detection of peptide phosphorylation by cAMP-dependent protein kinase A (PKA) (De Boer et al., Anal. Bioanal. Chem. 2005, 381, 647-655) was transferred to a continuous-flow reaction detection system. This enzymatic reaction, involving enzyme activation as well as the transfer of a phosphate group from ATP to a peptide substrate, was used to prove the compatibility of a quantitative enzymatic assay in a continuous-flow real-time system (connected to MS). Moreover (using internal standards), the critical parameter reaction temperature (including solution density variations depending on temperature) was studied in the continuous-flow mixing system. Furthermore, two substrates (malantide and kemptide), two enzyme types (catalytic subunit of PKA and complete PKA) and one inhibitor were tested to determine system robustness and long-term availability. Even spraying solutions that contained significant amount of MS contaminants (e.g. the polluted catalytic subunit) resulted in quantifiable MS signal intensities. Subsequent recalculations using the internal standards led to results representing the power of this application. The presented methodology and the data evaluation with available Achroma freeware enable the direct coupling of biochemical assays with quantitative MS detection. Monitoring changes such as temperature, reaction time, inhibition, or compound concentrations can be observed quantitatively and thus enzymatic activity can be calculated. Copyright © 2016 John Wiley & Sons, Ltd.

Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

PubMed Central

Xiao, Lihua; Ryan, Una M.; Graczyk, Thaddeus K.; Limor, Josef; Li, Lixia; Kombert, Mark; Junge, Randy; Sulaiman, Irshad M.; Zhou, Ling; Arrowood, Michael J.; Koudela, Břetislav; Modrý, David; Lal, Altaf A.

2004-01-01

The genetic diversity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 samples were analyzed, of which 48 snake samples, 24 lizard samples, and 3 tortoise samples were positive for Cryptosporidium. Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis, Cryptosporidium desert monitor genotype, Cryptosporidium muris, Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis-like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum. Two host-adapted C. serpentis genotypes were found in snakes and lizards. PMID:14766569

The first clinical implementation of a real-time six degree of freedom target tracking system during radiation therapy based on Kilovoltage Intrafraction Monitoring (KIM).

PubMed

Nguyen, Doan Trang; O'Brien, Ricky; Kim, Jung-Ha; Huang, Chen-Yu; Wilton, Lee; Greer, Peter; Legge, Kimberley; Booth, Jeremy T; Poulsen, Per Rugaard; Martin, Jarad; Keall, Paul J

2017-04-01

We present the first clinical implementation of a real-time six-degree of freedom (6DoF) Kilovoltage Intrafraction Monitoring (KIM) system which tracks the cancer target translational and rotational motions during treatment. The method was applied to measure and correct for target motion during stereotactic body radiotherapy (SBRT) for prostate cancer. Patient: A patient with prostate adenocarcinoma undergoing SBRT with 36.25Gy, delivered in 5 fractions was enrolled in the study. 6DoF KIM technology: 2D positions of three implanted gold markers in each of the kV images (125kV, 10mA at 11Hz) were acquired continuously during treatment. The 2D→3D target position estimation was based on a probability distribution function. The 3D→6DoF target rotation was calculated using an iterative closest point algorithm. The accuracy and precision of the KIM method was measured by comparing the real-time results with kV-MV triangulation. Of the five treatment fractions, KIM was utilised successfully in four fractions. The intrafraction prostate motion resulted in three couch shifts in two fractions when the prostate motion exceeded the pre-set action threshold of 2mm for more than 5s. KIM translational accuracy and precision were 0.3±0.6mm, -0.2±0.3mm and 0.2±0.7mm in the Left-Right (LR), Superior-Inferior (SI) and Anterior-Posterior (AP) directions, respectively. The KIM rotational accuracy and precision were 0.8°±2.0°, -0.5°±3.3° and 0.3°±1.6° in the roll, pitch and yaw directions, respectively. This treatment represents, to the best of our knowledge, the first time a cancer patient's tumour position and rotation have been monitored in real-time during treatment. The 6 DoF KIM system has sub-millimetre accuracy and precision in all three translational axes, and less than 1° accuracy and 4° precision in all three rotational axes. Copyright © 2017 Elsevier B.V. All rights reserved.

Optical Strain and Crack-Detection Measurements on a Rotating Disk

NASA Technical Reports Server (NTRS)

Woike, Mark; Abdul-Aziz, Ali; Clem, Michelle; Fralick, Gustave

2013-01-01

The development of techniques for the in-situ measurement and structural health monitoring of the rotating components in gas turbine engines is of major interest to NASA. As part of this on-going effort, several experiments have been undertaken to develop methods for detecting cracks and measuring strain on rotating turbine engine like disks. Previous methods investigated have included the use of blade tip clearance sensors to detect the presence of cracks by monitoring the change in measured blade tip clearance and analyzing the combined disk-rotor system's vibration response. More recently, an experiment utilizing a novel optical Moiré based concept has been conducted on a subscale turbine engine disk to demonstrate a potential strain measurement and crack detection technique. Moiré patterns result from the overlap of two repetitive patterns with slightly different spacing. When this technique is applied to a rotating disk, it has the potential to allow for the detection of very small changes in spacing and radial growth in a rotating disk due to a flaw such as a crack. This investigation was a continuation of previous efforts undertaken in 2011-2012 to validate this optical concept. The initial demonstration attempted on a subscale turbine engine disk was inconclusive due to the minimal radial growth experienced by the disk during operation. For the present experiment a new subscale Aluminum disk was fabricated and improvements were made to the experimental setup to better demonstrate the technique. A circular reference pattern was laser etched onto a subscale engine disk and the disk was operated at speeds up to 12 000 rpm as a means of optically monitoring the Moiré created by the shift in patterns created by the radial growth due the presence of the simulated crack. Testing was first accomplished on a clean defect free disk as a means of acquiring baseline reference data. A notch was then machined in to the disk to simulate a crack and testing was repeated for the purposes of demonstrating the concept. Displacement data was acquired using external blade tip clearance and shaft displacement sensors as a means of confirming the optical data and for validating other sensor based crack detection techniques.

Optical Strain and Crack-Detection Measurements on a Rotating Disk

NASA Technical Reports Server (NTRS)

Woike, Mark; Abdul-Aziz, Ali; Clem, Michelle M.; Fralick, Gustave

2013-01-01

The development of techniques for the in-situ measurement and structural health monitoring of the rotating components in gas turbine engines is of major interest to NASA. As part of this on-going effort, several experiments have been undertaken to develop methods for detecting cracks and measuring strain on rotating turbine engine like disks. Previous methods investigated have included the use of blade tip clearance sensors to detect the presence of cracks by monitoring the change in measured blade tip clearance and analyzing the combined disk-rotor system's vibration response. More recently, an experiment utilizing a novel optical Moiré based concept has been conducted on a subscale turbine engine disk to demonstrate a potential strain measurement and crack detection technique. Moiré patterns result from the overlap of two repetitive patterns with slightly different spacing. When this technique is applied to a rotating disk, it has the potential to allow for the detection of very small changes in spacing and radial growth in a rotating disk due to a flaw such as a crack. This investigation was a continuation of previous efforts undertaken in 2011 to 2012 to validate this optical concept. The initial demonstration attempted on a subscale turbine engine disk was inconclusive due to the minimal radial growth experienced by the disk during operation. For the present experiment a new subscale Aluminum disk was fabricated and improvements were made to the experimental setup to better demonstrate the technique. A circular reference pattern was laser etched onto a subscale engine disk and the disk was operated at speeds up to 12 000 rpm as a means of optically monitoring the Moiré created by the shift in patterns created by the radial growth due the presence of the simulated crack. Testing was first accomplished on a clean defect free disk as a means of acquiring baseline reference data. A notch was then machined in to the disk to simulate a crack and testing was repeated for the purposes of demonstrating the concept. Displacement data was acquired using external blade tip clearance and shaft displacement sensors as a means of confirming the optical data and for validating other sensor based crack detection techniques.

MO-G-BRE-03: Automated Continuous Monitoring of Patient Setup with Second-Check Independent Image Registration

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

Jiang, X; Fox, T; Schreibmann, E

2014-06-15

Purpose: To create a non-supervised quality assurance program to monitor image-based patient setup. The system acts a secondary check by independently computing shifts and rotations and interfaces with Varian's database to verify therapist's work and warn against sub-optimal setups. Methods: Temporary digitally-reconstructed radiographs (DRRs) and OBI radiographic image files created by Varian's treatment console during patient setup are intercepted and used as input in an independent registration module customized for accuracy that determines the optimal rotations and shifts. To deal with the poor quality of OBI images, a histogram equalization of the live images to the DDR counterparts is performedmore » as a pre-processing step. A search for the most sensitive metric was performed by plotting search spaces subject to various translations and convergence analysis was applied to ensure the optimizer finds the global minima. Final system configuration uses the NCC metric with 150 histogram bins and a one plus one optimizer running for 2000 iterations with customized scales for translations and rotations in a multi-stage optimization setup that first corrects and translations and subsequently rotations. Results: The system was installed clinically to monitor and provide almost real-time feedback on patient positioning. On a 2 month-basis uncorrected pitch values were of a mean 0.016° with standard deviation of 1.692°, and couch rotations of − 0.090°± 1.547°. The couch shifts were −0.157°±0.466° cm for the vertical, 0.045°±0.286 laterally and 0.084°± 0.501° longitudinally. Uncorrected pitch angles were the most common source of discrepancies. Large variations in the pitch angles were correlated with patient motion inside the mask. Conclusion: A system for automated quality assurance of therapist's registration was designed and tested in clinical practice. The approach complements the clinical software's automated registration in terms of algorithm configuration and performance and constitutes a practical approach to implement safe and cost-effective radiotherapy.« less

Effects of underestimating the kinematics of trunk rotation on simultaneous reaching movements: predictions of a biomechanical model

PubMed Central

2013-01-01

Background Rotation of the torso while reaching produces torques (e.g., Coriolis torque) that deviate the arm from its planned trajectory. To ensure an accurate reaching movement, the brain may take these perturbing torques into account during movement planning or, alternatively, it may correct hand trajectory during movement execution. Irrespective of the process selected, it is expected that an underestimation of trunk rotation would likely induce inaccurate shoulder and elbow torques, resulting in hand deviation. Nonetheless, it is still undetermined to what extent a small error in the perception of trunk rotations, translating into an inappropriate selection of motor commands, would affect reaching accuracy. Methods To investigate, we adapted a biomechanical model (J Neurophysiol 89: 276-289, 2003) to predict the consequences of underestimating trunk rotations on right hand reaching movements performed during either clockwise or counter clockwise torso rotations. Results The results revealed that regardless of the degree to which the torso rotation was underestimated, the amplitude of hand deviation was much larger for counter clockwise rotations than for clockwise rotations. This was attributed to the fact that the Coriolis and centripetal joint torques were acting in the same direction during counter clockwise rotation yet in opposite directions during clockwise rotations, effectively cancelling each other out. Conclusions These findings suggest that in order to anticipate and compensate for the interaction torques generated during torso rotation while reaching, the brain must have an accurate prediction of torso rotation kinematics. The present study proposes that when designing upper limb prostheses controllers, adding a sensor to monitor trunk kinematics may improve prostheses control and performance. PMID:23758968

Coordinated Control of Three-Dimensional Components of Smooth Pursuit to Rotating and Translating Textures.

PubMed

Edinger, Janick; Pai, Dinesh K; Spering, Miriam

2017-01-01

The neural control of pursuit eye movements to visual textures that simultaneously translate and rotate has largely been neglected. Here we propose that pursuit of such targets-texture pursuit-is a fully three-dimensional task that utilizes all three degrees of freedom of the eye, including torsion. Head-fixed healthy human adults (n = 8) tracked a translating and rotating random dot pattern, shown on a computer monitor, with their eyes. Horizontal, vertical, and torsional eye positions were recorded with a head-mounted eye tracker. The torsional component of pursuit is a function of the rotation of the texture, aligned with its visual properties. We observed distinct behaviors between those trials in which stimulus rotation was in the same direction as that of a rolling ball ("natural") in comparison to those with the opposite rotation ("unnatural"): Natural rotation enhanced and unnatural rotation reversed torsional velocity during pursuit, as compared to torsion triggered by a nonrotating random dot pattern. Natural rotation also triggered pursuit with a higher horizontal velocity gain and fewer and smaller corrective saccades. Furthermore, we show that horizontal corrective saccades are synchronized with torsional corrective saccades, indicating temporal coupling of horizontal and torsional saccade control. Pursuit eye movements have a torsional component that depends on the visual stimulus. Horizontal and torsional eye movements are separated in the motor periphery. Our findings suggest that translational and rotational motion signals might be coordinated in descending pursuit pathways.

Rotation periods of open-cluster stars, 2

NASA Technical Reports Server (NTRS)

Prosser, Charles F.; Shetrone, Matthew D.; Marilli, Ettore; Catalano, Santo; Williams, Scott D.; Backman, Dana E.; Laaksonen, Bentley D.; Adige, Vikram; Marschall, Laurence A.; Stauffer, John R.

1993-01-01

We present the results from a photometric monitoring program of 21 stars observed during 1992 in the Pleiades and Alpha Persei open clusters. Period determinations for 16 stars are given, 13 of which are the first periods reported for these stars. Brightness variations for an additional five cluster stars are also given. One K dwarf member of the alpha Per cluster is observed to have a period of rotation of only 4.39 hr. perhaps the shortest period currently known among BY Draconis variables. The individual photometric measurements have been deposited with the NSSDC. Combining current X-ray flux determinations with known photometric periods, we illustrate the X-ray activity/rotation relation among Pleiades K dwarfs based on available data.

Non-invasive dynamic measurement of helicopter blades

NASA Astrophysics Data System (ADS)

Serafini, J.; Bernardini, G.; Mattioni, L.; Vezzari, V.; Ficuciello, C.

2017-08-01

This paper presents the development and the application on helicopter blades of a measurement system based on FBG strain gauges. Here, the main goal is the structural characterization of the main rotor blades, with the aim of showing the potentialities of such a system in blades quality check applications, as well as in the development of structural health monitoring and rotor state feedback devices. The device has been used in both non-rotating and rotating tests, and does not require the presence of slip rings or optical joint since it is completely allocated in the rotating system. It has been successfully applied to characterize the frequency response of blades lead-lag, flap and torsion deformations, up to 250 Hz.

Palmitoylation of the β4-Subunit Regulates Surface Expression of Large Conductance Calcium-activated Potassium Channel Splice Variants*

PubMed Central

Chen, Lie; Bi, Danlei; Tian, Lijun; McClafferty, Heather; Steeb, Franziska; Ruth, Peter; Knaus, Hans Guenther; Shipston, Michael J.

2013-01-01

Regulatory β-subunits of large conductance calcium- and voltage-activated potassium (BK) channels play an important role in generating functional diversity and control of cell surface expression of the pore forming α-subunits. However, in contrast to α-subunits, the role of reversible post-translational modification of intracellular residues on β-subunit function is largely unknown. Here we demonstrate that the human β4-subunit is S-acylated (palmitoylated) on a juxtamembrane cysteine residue (Cys-193) in the intracellular C terminus of the regulatory β-subunit. β4-Subunit palmitoylation is important for cell surface expression and endoplasmic reticulum (ER) exit of the β4-subunit alone. Importantly, palmitoylated β4-subunits promote the ER exit and surface expression of the pore-forming α-subunit, whereas β4-subunits that cannot be palmitoylated do not increase ER exit or surface expression of α-subunits. Strikingly, however, this palmitoylation- and β4-dependent enhancement of α-subunit surface expression was only observed in α-subunits that contain a putative trafficking motif (… REVEDEC) at the very C terminus of the α-subunit. Engineering this trafficking motif to other C-terminal α-subunit splice variants results in α-subunits with reduced surface expression that can be rescued by palmitoylated, but not depalmitoylated, β4-subunits. Our data reveal a novel mechanism by which palmitoylated β4-subunit controls surface expression of BK channels through masking of a trafficking motif in the C terminus of the α-subunit. As palmitoylation is dynamic, this mechanism would allow precise control of specific splice variants to the cell surface. Our data provide new insights into how complex interplay between the repertoire of post-transcriptional and post-translational mechanisms controls cell surface expression of BK channels. PMID:23504458

Structure of the protein phosphatase 2A holoenzyme.

PubMed

Xu, Yanhui; Xing, Yongna; Chen, Yu; Chao, Yang; Lin, Zheng; Fan, Eugene; Yu, Jong W; Strack, Stefan; Jeffrey, Philip D; Shi, Yigong

2006-12-15

Protein Phosphatase 2A (PP2A) plays an essential role in many aspects of cellular physiology. The PP2A holoenzyme consists of a heterodimeric core enzyme, which comprises a scaffolding subunit and a catalytic subunit, and a variable regulatory subunit. Here we report the crystal structure of the heterotrimeric PP2A holoenzyme involving the regulatory subunit B'/B56/PR61. Surprisingly, the B'/PR61 subunit has a HEAT-like (huntingtin-elongation-A subunit-TOR-like) repeat structure, similar to that of the scaffolding subunit. The regulatory B'/B56/PR61 subunit simultaneously interacts with the catalytic subunit as well as the conserved ridge of the scaffolding subunit. The carboxyterminus of the catalytic subunit recognizes a surface groove at the interface between the B'/B56/PR61 subunit and the scaffolding subunit. Compared to the scaffolding subunit in the PP2A core enzyme, formation of the holoenzyme forces the scaffolding subunit to undergo pronounced conformational rearrangements. This structure reveals significant ramifications for understanding the function and regulation of PP2A.

Age-related similarities and differences in monitoring spatial cognition.

PubMed

Ariel, Robert; Moffat, Scott D

2018-05-01

Spatial cognitive performance is impaired in later adulthood but it is unclear whether the metacognitive processes involved in monitoring spatial cognitive performance are also compromised. Inaccurate monitoring could affect whether people choose to engage in tasks that require spatial thinking and also the strategies they use in spatial domains such as navigation. The current experiment examined potential age differences in monitoring spatial cognitive performance in a variety of spatial domains including visual-spatial working memory, spatial orientation, spatial visualization, navigation, and place learning. Younger and older adults completed a 2D mental rotation test, 3D mental rotation test, paper folding test, spatial memory span test, two virtual navigation tasks, and a cognitive mapping test. Participants also made metacognitive judgments of performance (confidence judgments, judgments of learning, or navigation time estimates) on each trial for all spatial tasks. Preference for allocentric or egocentric navigation strategies was also measured. Overall, performance was poorer and confidence in performance was lower for older adults than younger adults. In most spatial domains, the absolute and relative accuracy of metacognitive judgments was equivalent for both age groups. However, age differences in monitoring accuracy (specifically relative accuracy) emerged in spatial tasks involving navigation. Confidence in navigating for a target location also mediated age differences in allocentric navigation strategy use. These findings suggest that with the possible exception of navigation monitoring, spatial cognition may be spared from age-related decline even though spatial cognition itself is impaired in older age.

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

Jack Preiss

Conversion of the Potato tuber ADP-glucose Pyrophopshorylase Regulatory Subunit into a Catalytic Subunit. ADP-glucose synthesis, a rate-limiting reaction in starch synthesis, is catalyzed by ADP-glucose pyrophosphorylase (ADPGlc PPase). The enzyme in plants is allosterically activated by 3-phosphoglycerate (3PGA) and inhibited by inorganic phosphate (Pi) and is composed of two subunits as a heterotetramer, a2b2. Subunit a is the catalytic subunit and subunit b is designated as the regulatory subunit.The b subunit increases the affinty of the activator for the catalytic subunit. Recent results have shown that the subunits are derived from the same ancestor subunit as the regulatory subunit canmore » be converted to a catalytically subunit via mutation of just two amino acids. Lys44 and Thr54 in the large subunit from potato tuber were converted to the homologous catalytic subunit residues, Arg33 and Lys43. The activity of the large subunit mutants cannot be readily tested with a co-expressed wild-type small (catalytic) subunit because of the intrinsic activity of the latter. We co-expressed the regulatory-subunit mutants with SmallD145N, an inactive S subunit in which the catalytic Asp145 was mutated. The activity of the small (catalytic) subunit was reduced more than three orders of magnitude. Coexpression of the L subunit double mutant LargeK44R/T54K with SmallD145N generated an enzyme with considerable activity, 10% and 18% of the wildtype enzyme, in the ADP-glucose synthetic and pyrophosphorolytic direction, respectively. Replacement of those two residues in the small subunit by the homologous amino acids in the L subunits (mutations R33K and K43T) decreased the activity one and two orders of magnitude. The wild-type enzyme and SmallD145NLargeK44R/T54K had very similar kinetic properties indicating that the substrate site has been conserved. The fact that only two mutations in the L subunit restored enzyme activity is very strong evidence that the large subunit is derived from the catalytic ancestor. Previous results showed that Asp145 in the small subunit of the wild-type is essential for catalysis, whereas the homologous Asp160 in the Large WT subunit is not. However, in this study, mutation D160N or D160E in the LK44R/T54K subunit abolished the activity, which shows the ancestral essential role of this residue and confirms that the catalysis of SmallD145NLarge K44R/T54K occurs in the L(b) subunit. A phylogenetic tree of the ADP-Glc PPases present in photosynthetic eukaryotes also sheds information about the origin of the subunits. The tree showed that plant Small and Large subunits can be divided into two and four distinct groups, respectively. The two main groups of S subunits are from dicot and monocot plants, whereas Large subunit groups correlate better with their documented tissue expression. The first Large-subunit group is generally expressed in photosynthetic tissues and comprises Large subunits from dicots and monocots. Group II displays a broader expression pattern, whereas groups III and IV are expressed in storage organs (roots, stems, tubers, seeds). Subunits from group III are only from dicot plants, whereas group IV are seed-specific subunits from monocots. These last two groups stem from the same branch of the phylogenetic tree and split before monocot and dicot separation. Thus few as two mutations turned the L subunit from Solanum tuberosum catalytic, showing that L and S subunits share a common catalytic ancestor, rather than a non-catalytic one. The L subunit evolved to have a regulatory role, lost catalytic residues more than 130 million years ago before monocots and dicots diverged, and preserved, possibly as a byproduct, the active site domain.« less

Protease-resistant modified human β-hexosaminidase B ameliorates symptoms in GM2 gangliosidosis model

PubMed Central

Mizutani, Yasumichi; Sugiyama, Eiji; Tasaki, Chikako; Tsuji, Daisuke; Maita, Nobuo; Hirokawa, Takatsugu; Asanuma, Daisuke; Kamiya, Mako; Sato, Kohei; Setou, Mitsutoshi; Urano, Yasuteru; Togawa, Tadayasu; Otaka, Akira; Sakuraba, Hitoshi

2016-01-01

GM2 gangliosidoses, including Tay-Sachs and Sandhoff diseases, are neurodegenerative lysosomal storage diseases that are caused by deficiency of β-hexosaminidase A, which comprises an αβ heterodimer. There are no effective treatments for these diseases; however, various strategies aimed at restoring β-hexosaminidase A have been explored. Here, we produced a modified human hexosaminidase subunit β (HexB), which we have termed mod2B, composed of homodimeric β subunits that contain amino acid sequences from the α subunit that confer GM2 ganglioside–degrading activity and protease resistance. We also developed fluorescent probes that allow visualization of endocytosis of mod2B via mannose 6-phosphate receptors and delivery of mod2B to lysosomes in GM2 gangliosidosis models. In addition, we applied imaging mass spectrometry to monitor efficacy of this approach in Sandhoff disease model mice. Following i.c.v. administration, mod2B was widely distributed and reduced accumulation of GM2, asialo-GM2, and bis(monoacylglycero)phosphate in brain regions including the hypothalamus, hippocampus, and cerebellum. Moreover, mod2B administration markedly improved motor dysfunction and a prolonged lifespan in Sandhoff disease mice. Together, the results of our study indicate that mod2B has potential for intracerebrospinal fluid enzyme replacement therapy and should be further explored as a gene therapy for GM2 gangliosidoses. PMID:27018595

Protease-resistant modified human β-hexosaminidase B ameliorates symptoms in GM2 gangliosidosis model.

PubMed

Kitakaze, Keisuke; Mizutani, Yasumichi; Sugiyama, Eiji; Tasaki, Chikako; Tsuji, Daisuke; Maita, Nobuo; Hirokawa, Takatsugu; Asanuma, Daisuke; Kamiya, Mako; Sato, Kohei; Setou, Mitsutoshi; Urano, Yasuteru; Togawa, Tadayasu; Otaka, Akira; Sakuraba, Hitoshi; Itoh, Kohji

2016-05-02

GM2 gangliosidoses, including Tay-Sachs and Sandhoff diseases, are neurodegenerative lysosomal storage diseases that are caused by deficiency of β-hexosaminidase A, which comprises an αβ heterodimer. There are no effective treatments for these diseases; however, various strategies aimed at restoring β-hexosaminidase A have been explored. Here, we produced a modified human hexosaminidase subunit β (HexB), which we have termed mod2B, composed of homodimeric β subunits that contain amino acid sequences from the α subunit that confer GM2 ganglioside-degrading activity and protease resistance. We also developed fluorescent probes that allow visualization of endocytosis of mod2B via mannose 6-phosphate receptors and delivery of mod2B to lysosomes in GM2 gangliosidosis models. In addition, we applied imaging mass spectrometry to monitor efficacy of this approach in Sandhoff disease model mice. Following i.c.v. administration, mod2B was widely distributed and reduced accumulation of GM2, asialo-GM2, and bis(monoacylglycero)phosphate in brain regions including the hypothalamus, hippocampus, and cerebellum. Moreover, mod2B administration markedly improved motor dysfunction and a prolonged lifespan in Sandhoff disease mice. Together, the results of our study indicate that mod2B has potential for intracerebrospinal fluid enzyme replacement therapy and should be further explored as a gene therapy for GM2 gangliosidoses.

Evidence that viral RNAs have evolved for efficient, two-stage packaging.

PubMed

Borodavka, Alexander; Tuma, Roman; Stockley, Peter G

2012-09-25

Genome packaging is an essential step in virus replication and a potential drug target. Single-stranded RNA viruses have been thought to encapsidate their genomes by gradual co-assembly with capsid subunits. In contrast, using a single molecule fluorescence assay to monitor RNA conformation and virus assembly in real time, with two viruses from differing structural families, we have discovered that packaging is a two-stage process. Initially, the genomic RNAs undergo rapid and dramatic (approximately 20-30%) collapse of their solution conformations upon addition of cognate coat proteins. The collapse occurs with a substoichiometric ratio of coat protein subunits and is followed by a gradual increase in particle size, consistent with the recruitment of additional subunits to complete a growing capsid. Equivalently sized nonviral RNAs, including high copy potential in vivo competitor mRNAs, do not collapse. They do support particle assembly, however, but yield many aberrant structures in contrast to viral RNAs that make only capsids of the correct size. The collapse is specific to viral RNA fragments, implying that it depends on a series of specific RNA-protein interactions. For bacteriophage MS2, we have shown that collapse is driven by subsequent protein-protein interactions, consistent with the RNA-protein contacts occurring in defined spatial locations. Conformational collapse appears to be a distinct feature of viral RNA that has evolved to facilitate assembly. Aspects of this process mimic those seen in ribosome assembly.

LOOP IIId of the HCV IRES is essential for the structural rearrangement of the 40S-HCV IRES complex

PubMed Central

Angulo, Jenniffer; Ulryck, Nathalie; Deforges, Jules; Chamond, Nathalie; Lopez-Lastra, Marcelo; Masquida, Benoît; Sargueil, Bruno

2016-01-01

As obligatory intracellular parasites, viruses rely on cellular machines to complete their life cycle, and most importantly they recruit the host ribosomes to translate their mRNA. The Hepatitis C viral mRNA initiates translation by directly binding the 40S ribosomal subunit in such a way that the initiation codon is correctly positioned in the P site of the ribosome. Such a property is likely to be central for many viruses, therefore the description of host-pathogen interaction at the molecular level is instrumental to provide new therapeutic targets. In this study, we monitored the 40S ribosomal subunit and the viral RNA structural rearrangement induced upon the formation of the binary complex. We further took advantage of an IRES viral mutant mRNA deficient for translation to identify the interactions necessary to promote translation. Using a combination of structure probing in solution and molecular modeling we establish a whole atom model which appears to be very similar to the one obtained recently by cryoEM. Our model brings new information on the complex, and most importantly reveals some structural rearrangement within the ribosome. This study suggests that the formation of a ‘kissing complex’ between the viral RNA and the 18S ribosomal RNA locks the 40S ribosomal subunit in a conformation proficient for translation. PMID:26626152

Compilation of 1988 Annual Reports of the Navy ELF (Extremely Low Frequency) Communications System Ecological Monitoring Program. Volume 2

DTIC Science & Technology

1989-08-01

i3, 112-119. SW~right, R.T. & Coffin, R.B. 1984. Measuring microzopl , ’-., grazing on planktonic marine bacteria bv its impact on bacteria...activities are 3 shown for 1987 in Figure 4, along with antenna on and off times. Testing of the antenna was conducted on a 33% time rotation schedule...noticed that different observers differ slightly in their techniques for measuring 3 weights and body parts. Therefore we had all observers rotate among

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

Senesi, Andrew; Lee, Byeongdu

Herein, a general method to calculate the scattering functions of polyhedra, including both regular and semi-regular polyhedra, is presented. These calculations may be achieved by breaking a polyhedron into sets of congruent pieces, thereby reducing computation time by taking advantage of Fourier transforms and inversion symmetry. Each piece belonging to a set or subunit can be generated by either rotation or translation. Further, general strategies to compute truncated, concave and stellated polyhedra are provided. Using this method, the asymptotic behaviors of the polyhedral scattering functions are compared with that of a sphere. It is shown that, for a regular polyhedron,more » the form factor oscillation at highqis correlated with the face-to-face distance. In addition, polydispersity affects the Porod constant. The ideas presented herein will be important for the characterization of nanomaterials using small-angle scattering.« less

Turbine rotor disk health monitoring assessment based on sensor technology and spin tests data.

PubMed

Abdul-Aziz, Ali; Woike, Mark

2013-01-01

The paper focuses on presenting data obtained from spin test experiments of a turbine engine like rotor disk and assessing their correlation to the development of a structural health monitoring and fault detection system. The data were obtained under various operating conditions such as the rotor disk being artificially induced with and without a notch and rotated at a rotational speed of up to 10,000 rpm under balanced and imbalanced state. The data collected included blade tip clearance, blade tip timing measurements, and shaft displacements. Two different sensor technologies were employed in the testing: microwave and capacitive sensors, respectively. The experimental tests were conducted at the NASA Glenn Research Center's Rotordynamics Laboratory using a high precision spin system. Disk flaw observations and related assessments from the collected data for both sensors are reported and discussed.

Ebselen and congeners inhibit NADPH oxidase 2-dependent superoxide generation by interrupting the binding of regulatory subunits.

PubMed

Smith, Susan M E; Min, Jaeki; Ganesh, Thota; Diebold, Becky; Kawahara, Tsukasa; Zhu, Yerun; McCoy, James; Sun, Aiming; Snyder, James P; Fu, Haian; Du, Yuhong; Lewis, Iestyn; Lambeth, J David

2012-06-22

NADPH oxidases (Nox) are a primary source of reactive oxygen species (ROS), which function in normal physiology and, when overproduced, in pathophysiology. Recent studies using mice deficient in Nox2 identify this isoform as a novel target against Nox2-implicated inflammatory diseases. Nox2 activation depends on the binding of the proline-rich domain of its heterodimeric partner p22phox to p47phox. A high-throughput screen that monitored this interaction via fluorescence polarization identified ebselen and several of its analogs as inhibitors. Medicinal chemistry was performed to explore structure-activity relationships and to optimize potency. Ebselen and analogs potently inhibited Nox1 and Nox2 activity but were less effective against other isoforms. Ebselen also blocked translocation of p47phox to neutrophil membranes. Thus, ebselen and its analogs represent a class of compounds that inhibit ROS generation by interrupting the assembly of Nox2-activating regulatory subunits. Copyright © 2012 Elsevier Ltd. All rights reserved.

Bio-layer interferometry for measuring kinetics of protein-protein interactions and allosteric ligand effects.

PubMed

Shah, Naman B; Duncan, Thomas M

2014-02-18

We describe the use of Bio-layer Interferometry to study inhibitory interactions of subunit ε with the catalytic complex of Escherichia coli ATP synthase. Bacterial F-type ATP synthase is the target of a new, FDA-approved antibiotic to combat drug-resistant tuberculosis. Understanding bacteria-specific auto-inhibition of ATP synthase by the C-terminal domain of subunit ε could provide a new means to target the enzyme for discovery of antibacterial drugs. The C-terminal domain of ε undergoes a dramatic conformational change when the enzyme transitions between the active and inactive states, and catalytic-site ligands can influence which of ε's conformations is predominant. The assay measures kinetics of ε's binding/dissociation with the catalytic complex, and indirectly measures the shift of enzyme-bound ε to and from the apparently nondissociable inhibitory conformation. The Bio-layer Interferometry signal is not overly sensitive to solution composition, so it can also be used to monitor allosteric effects of catalytic-site ligands on ε's conformational changes.

The rotational dynamics of Titan from Cassini RADAR images

NASA Astrophysics Data System (ADS)

Meriggiola, Rachele; Iess, Luciano; Stiles, Bryan. W.; Lunine, Jonathan. I.; Mitri, Giuseppe

2016-09-01

Between 2004 and 2009 the RADAR instrument of the Cassini mission provided 31 SAR images of Titan. We tracked the position of 160 surface landmarks as a function of time in order to monitor the rotational dynamics of Titan. We generated and processed RADAR observables using a least squares fit to determine the updated values of the rotational parameters. We provide a new rotational model of Titan, which includes updated values for spin pole location, spin rate, precession and nutation terms. The estimated pole location is compatible with the occupancy of a Cassini state 1. We found a synchronous value of the spin rate (22.57693 deg/day), compatible at a 3-σ level with IAU predictions. The estimated obliquity is equal to 0.31°, incompatible with the assumption of a rigid body with fully-damped pole and a moment of inertia factor of 0.34, as determined by gravity measurements.

Remote detection of rotating machinery with a portable atomic magnetometer.

PubMed

Marmugi, Luca; Gori, Lorenzo; Hussain, Sarah; Deans, Cameron; Renzoni, Ferruccio

2017-01-20

We demonstrate remote detection of rotating machinery, using an atomic magnetometer at room temperature and in an unshielded environment. The system relies on the coupling of the AC magnetic signature of the target with the spin-polarized, precessing atomic vapor of a radio-frequency optical atomic magnetometer. The AC magnetic signatures of rotating equipment or electric motors appear as sidebands in the power spectrum of the atomic sensor, which can be tuned to avoid noisy bands that would otherwise hamper detection. A portable apparatus is implemented and experimentally tested. Proof-of-concept investigations are performed with test targets mimicking possible applications, and the operational conditions for optimum detection are determined. Our instrument provides comparable or better performance than a commercial fluxgate and allows detection of rotating machinery behind a wall. These results demonstrate the potential for ultrasensitive devices for remote industrial and usage monitoring, security, and surveillance.

On fast X-ray rotators with long-term periodicities

NASA Technical Reports Server (NTRS)

Naranan, S.; Elsner, R. F.; Darbro, W.; Ramsey, B. D.; Leahy, D. A.; Weisskopf, M. C.; Williams, A. C.; Hardee, P. E.; Sutherland, P. G.; Grindlay, J. E.

1985-01-01

The support of previous SAS 3 spacecraft observations by new data gathered by the Monitor Proportional Counter aboard the HEAO 2 spacecraft indicates that the pulse period history of the 13.5 sec-pulsing X-ray source LMC X-4 is consistent with standard accretion and torque models only if LMC X-4 is a fast rotator for which the accretion torques nearly cancel. This result leads to a neutron star magnetic field strength estimate of about 1.2 x 10 to the 13th G. Strong evidence is noted for Her X-1's status as a fast rotator, while SMC X-1 is probably an intermediate-to-fast rotator. In the context of slaved disk models for these objects, it is noted that the precession periods expected for the companion stars are significantly longer than the observed 1-2 month time scales; slaved disk models are thereby undermined.

Highly directional transurethral ultrasound applicators with rotational control for MRI-guided prostatic thermal therapy

NASA Astrophysics Data System (ADS)

Ross, Anthony B.; Diederich, Chris J.; Nau, William H.; Gill, Harcharan; Bouley, Donna M.; Daniel, Bruce; Rieke, Viola; Butts, R. Kim; Sommer, Graham

2004-01-01

Transurethral ultrasound applicators with highly directional energy deposition and rotational control were investigated for precise treatment of benign prostatic hyperplasia (BPH) and adenocarcinoma of the prostate (CaP). Two types of catheter-based applicators were fabricated, using either 90° sectored tubular (3.5 mm OD × 10 mm) or planar transducers (3.5 mm × 10 mm). They were constructed to be MRI compatible, minimally invasive and allow for manual rotation of the transducer array within a 10 mm cooling balloon. In vivo evaluations of the applicators were performed in canine prostates (n = 3) using MRI guidance (0.5 T interventional magnet). MR temperature imaging (MRTI) utilizing the proton resonance frequency shift method was used to acquire multiple-slice temperature overlays in real time for monitoring and guiding the thermal treatments. Post-treatment T1-weighted contrast-enhanced imaging and triphenyl tetrazolium chloride stained tissue sections were used to define regions of tissue coagulation. Single sonications with the 90° tubular applicator (9-15 W, 12 min, 8 MHz) produced coagulated zones covering an 80° wedge of the prostate extending from 1-2 mm outside the urethra to the outer boundary of the gland (16 mm radial coagulation). Single sonications with the planar applicator (15-20 W, 10 min, ~8 MHz) generated thermal lesions of ~30° extending to the prostate boundary. Multiple sequential sonications (sweeping) of a planar applicator (12 W with eight rotations of 30° each) demonstrated controllable coagulation of a 270° contiguous section of the prostate extending to the capsule boundary. The feasibility of using highly directional transurethral ultrasound applicators with rotational capabilities to selectively coagulate regions of the prostate while monitoring and controlling the treatments with MRTI was demonstrated in this study.

Fault kinematics and depocenter evolution of oil-bearing, continental successions of the Mina del Carmen Formation (Albian) in the Golfo San Jorge basin, Argentina

NASA Astrophysics Data System (ADS)

Paredes, José Matildo; Plazibat, Silvana; Crovetto, Carolina; Stein, Julián; Cayo, Eric; Schiuma, Ariel

2013-10-01

Up to 10% of the liquid hydrocarbons of the Golfo San Jorge basin come from the Mina del Carmen Formation (Albian), an ash-dominated fluvial succession preserved in a variably integrated channel network that evolved coeval to an extensional tectonic event, poorly analyzed up to date. Fault orientation, throw distribution and kinematics of fault populations affecting the Mina del Carmen Formation were investigated using a 3D seismic dataset in the Cerro Dragón field (Eastern Sector of the Golfo San Jorge basin). Thickness maps of the seismic sub-units that integrate the Mina del Carmen Formation, named MEC-A-MEC-C in ascending order, and mapping of fluvial channels performed applying geophysical tools of visualization were integrated to the kinematical analysis of 20 main normal faults of the field. The study provides examples of changes in fault throw patterns with time, associated with faults of different orientations. The "main synrift phase" is characterized by NE-SW striking (mean Az = 49°), basement-involved normal faults that attains its maximum throw on top of the volcanic basement; this set of faults was active during deposition of the Las Heras Group and Pozo D-129 formation. A "second synrift phase" is recognized by E-W striking normal faults (mean Az = 91°) that nucleated and propagated from the Albian Mina del Carmen Formation. Fault activity was localized during deposition of the MEC-A sub-unit, but generalized during deposition of MEC-B sub-unit, producing centripetal and partially isolated depocenters. Upward decreasing in fault activity is inferred by more gradual thickness variation of MEC-C and the overlying Lower Member of Bajo Barreal Formation, evidencing passive infilling of relief associated to fault boundaries, and conformation of wider depocenters with well integrated networks of channels of larger dimensions but random orientation. Lately, the Mina del Carmen Formation was affected by the downward propagation of E-W to ESE-WNW striking normal faults (mean Az = 98°) formed during the "third rifting phase", which occurs coeval with the deposition of the Upper Member of the Bajo Barreal Formation. The fault characteristics indicate a counterclockwise rotation of the stress field during the deposition of the Chubut Group of the Golfo San Jorge basin, likely associated to the rotation of Southern South America during the fragmentation of the Gondwana paleocontinent. Understanding the evolution of fault-controlled topography in continental basins allow to infer location and orientation of coeval fluvial systems, providing a more reliable scenario for location of producing oil wells.

Simulated microgravity reduces mRNA levels of multidrug resistance genes 4 and 5 in non-metastatic human melanoma cells

NASA Astrophysics Data System (ADS)

Eiermann, Peter; Tsiockas, Wasiliki; Hauslage, Jens; Hemmersbach, Ruth; Gerzer, Rupert; Ivanova, Krassimira

Multidrug resistance proteins (MRP) are members of the ATP-binding cassette transporter superfamily that are able to export a large variety of substances into the extracellular space in-cluding nucleoside and nucleotide base analogs used in antiviral and anticancer therapy. MRP4 and 5 (MRP4/5) particularly transport cyclic nucleotides, e.g. guanosine 3',5'-cyclic monophos-phate (cGMP). The second messenger cGMP, which is synthesized by the catalytic activity of the guanylyl cyclase (GC), plays an import role in vasodilatation, smooth muscle relaxation, and nitric oxide (NO)-induced perturbation of melanocyte-extracellular matrix interactions. In previous studies we have reported that different GC isoforms are responsible for cGMP synthe-sis in melanocytic cells. Normal human melanocytes and non-metastatic melanoma cell lines predominantly express the NO-sensitive soluble GC isoform (sGC), a heterodimeric protein consisting of α and β subunits. Metastatic melanoma cells lack the expression of the β sub-unit and show up-regulated activities of the particulate isoforms. We have further found that long-term exposure to hypergravity (5 g for 24 h) induced an increased cGMP export in normal human melanocytes, and non-metastatic, but not in metastatic human melanoma cells as a re-sult of up-regulated MRP4/5 expression. The aim of the present study is to investigate whether simulated microgravity may also alter the expression of MRP4/5 in non-metastatic melanoma cells. Experiments were performed using a fast-rotating clinostat (60 rpm) with one rotation axis. The non-metastatic 1F6 melanoma cells were exposed to simulated microgravity (up to 1.21x10-2 g) for 24 h. The mRNA analyses were performed by a relative calibrator-normalized and efficiency corrected quantitative polymerase chain reaction (Light Cycler R , Roche). Our data show a reduced expression of approximately 35% for MRP4 and of 50% for MRP5 in simulated microgravity in comparison to 1 g controls. Also, the mRNA levels of sGC α and β were down-regulated by about 31% and 22%, respectively. Thus, the reduced expression of MRP4/5 could be related to the decrease in mRNA levels for the sGC subunits. In addition, the long-term exposure to simulated microgravity did not alter cellular morphology. Taken together, the results of our studies indicate that the expression of MRP4/5 in non-metastatic melanoma cells is inversely regulated by hypergravity and simulated microgravity. Finally, a reduced expression of MRP4 and MRP5 may increase the availability of drugs in cells and influence astronaut medication.

First record of Orthopodomyia pulcripalpis (Rondani, 1872) (Diptera: Culicidae) in Austria.

PubMed

Zittra, Carina; Obwaller, Adelheid G; Wimmer, Victoria; Berer, Dominik; Eigner, Barbara; Fuehrer, Hans-Peter

2017-06-01

During a three-year mosquito monitoring from 2014 to 2016, the strictly ornithophilic, originally Mediterranean species Orthopodomyia pulcripalpis (Rondani, 1872) was collected as single specimen for the first time in Austria in the district of Penzing in Vienna. Morphological species determination was confirmed by analysis of the mitochondrial cytochrome c oxidase subunit I gene. We thus not only confirm the existence of another mosquito species in Austria, but also add a new genus to the Austrian Culicidae taxa list.

Operational manual for MX-290 data-voice PN Mod, MX-291 data-voice PN DEMOD

NASA Technical Reports Server (NTRS)

1971-01-01

This operation manual is also the final report of the program to design, assemble, checkout, and deliver to the customer three MX-290 transmitters and two MX-291 companion receivers. These equipments are designed and assembled to provide for maximum flexibility with respect to making changes in electrical circuits which may be required for future applications. A number of test points for monitoring and troubleshooting are provided along with easy access to subunits.

Multimodality animal rotation imaging system (Mars) for in vivo detection of intraperitoneal tumors.

PubMed

Pizzonia, John; Holmberg, Jennie; Orton, Sean; Alvero, Ayesha; Viteri, Oscar; McLaughlin, William; Feke, Gil; Mor, Gil

2012-01-01

PROBLEM Ovarian cancer stem cells (OCSCs) have been postulated as the potential source of recurrence and chemoresistance. Therefore identification of OvCSC and their complete removal is a pivotal stage for the treatment of ovarian cancer. The objective of the following study was to develop a new in vivo imaging model that allows for the detection and monitoring of OCSCs. METHOD OF STUDY  OCSCs were labeled with X-Sight 761 Nanospheres and injected intra-peritoneally (i.p.) and sub-cutaneously (s.c.) to Athymic nude mice. The Carestream In-Vivo Imaging System FX was used to obtain X-ray and, concurrently, near-infrared fluorescence images. Tumor images in the mouse were observed from different angles by automatic rotation of the mouse. RESULTS  X-Sight 761 Nanospheres labeled almost 100% of the cells. No difference on growth rate was observed between labeled and unlabeled cells. Tumors were observed and monitoring revealed strong signaling up to 21 days. CONCLUSION  We describe the use of near-infrared nanoparticle probes for in vivo imaging of metastatic ovarian cancer models. Visualization of multiple sites around the animals was enhanced with the use of the Carestream Multimodal Animal Rotation System. © 2011 John Wiley & Sons A/S.

Development of a software and hardware system for monitoring the air cleaning process using a cyclone-separator

NASA Astrophysics Data System (ADS)

Nicolaeva, B. K.; Borisov, A. P.; Zlochevskiy, V. L.

2017-08-01

The article is devoted to the development of a hardware-software complex for monitoring and controlling the process of air purification by means of a cyclone-separator. The hardware of this complex is the Arduino platform, to which are connected pressure sensors, air velocities, dustmeters, which allow monitoring of the main parameters of the cyclone-separator. Also, a frequency converter was developed to regulate the rotation speed of an asynchronous motor necessary to correct the flow rate, the control signals of which come with Arduino. The program part of the complex is written in the form of a web application in the programming language JavaScript and inserts into CSS and HTML for the user interface. This program allows you to receive data from sensors, build dependencies in real time and control the speed of rotation of an asynchronous electric drive. The conducted experiment shows that the cleaning efficiency is 95-99.9%, while the airflow at the cyclone inlet is 16-18 m/s, and at the exit 50-70 m/s.

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

Faubel, M.; Weiner, E.R.

Rotational level populations of N/sub 2/ were measured downstream from the skimmer in beams of pure N/sub 2/ and in mixtures of N/sub 2/ with He, Ne, and Ar expanded from room temperature nozzles. The range of p/sub 0/D was from 5 to 50 Torr cm. The formation of dimers and higher condensates of beam species was monitored during the runs. The effect of condensation energy release on rotational populations and parallel temperatures was readily observed. Two different methods for evaluating the rotational population distributions were compared. One method is based on a dipole-excitation model and the other on anmore » excitation matrix obtained empirically. Neither method proved clearly superior. Both methods indicated nonequilibrium rotational populations for all of our room temperature nozzle expansion conditions. Much of the nonequilibrium character appears to be due to the behavior of the K = 2 and K = 4 levels, which may be accounted for in terms of the rotational energy level spacing. In particular, the overpopulation of the K = 4 level is explained by a near-resonant transfer of rotational energy between molecules in the K = 6 and K = 0 states, to give two molecules in the K = 4 state. Rotational and vibrational temperatures were determined for pure N/sub 2/ beams from nozzles heated up to 1700 /sup 0/K. The heated nozzle experiments indicated a 40% increase in the rotational collision number between 300 and 1700 /sup 0/K.« less

Towards the Detection of Explosive Taggants: Microwave and Millimetre-Wave Gas-Phase Spectroscopies of 3-Nitrotoluene.

PubMed

Roucou, Anthony; Kleiner, Isabelle; Goubet, Manuel; Bteich, Sabath; Mouret, Gael; Bocquet, Robin; Hindle, Francis; Meerts, W Leo; Cuisset, Arnaud

2018-05-07

The monitoring of gas-phase mononitrotoluenes is crucial for defence, civil security and environmental interests because they are used as taggant for TNT detection and in the manufacturing of industrial compounds such as dyestuffs. In this study, we have succeeded to measure and analyse at high-resolution a room temperature rotationally resolved millimetre-wave spectrum of meta-nitrotoluene (3-NT). Experimental and theoretical difficulties have been overcome, in particular, those related to the low vapour pressure of 3-NT and to the presence of a CH 3 internal rotation in an almost free rotation regime (V 3 =6.7659(24) cm -1 ). Rotational spectra have been recorded in the microwave and millimetre-wave ranges using a supersonic jet Fourier Transform microwave spectrometer (T rot <10 K) and a millimetre-wave frequency multiplication chain (T=293 K), respectively. Spectral analysis of pure rotation lines in the vibrational ground state and in the first torsional excited state supported by quantum chemistry calculations permits the rotational energy of the molecule, the hyperfine structure due to the 14 N nucleus, and the internal rotation of the methyl group to be characterised. A line list is provided for future in situ detection. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

[The development of the multifunctional automatic rotating bed with process-monitoring].

PubMed

Geng, Hongzhu; Hu, Monong; Cheng, Ping; Dong, Kejiang; Zhang, Jiaxia; Sun, Juefei

2013-04-01

We have developed a new rotating bed for the old and the paralised people. This rotating bed is composed of two bed heads at front and at end, bed boards, guardrails, an electric motor, a reducer, an induction locator and a set of electronic controls. With the preestablished program, the angle between the left/right bed board and the middle board is changed by rotating the left/right board around the rotation axis, and the gravity direction between the human body and the ground is changed by the rotation of the middle board as a whole, so that the middle bed board and the left and right ones will act respectively as supporters of weight of the person who is lying on his back or on his side. In this way, a person can turn over automatically, comfortably and naturally when he/she is asleep. This rotating bed meets the physiological needs of a sleeping person, and people with turning over problems can turn over in a comfortable and natural way by means of biotechnology. It can also improve the quality of sleep and help avoid decubitus. In addition, it can be used to promote the rehabilitation of those who are paralysed by reason of its passive exercising function.

Rotation periods and photometric variability of rapidly rotating ultracool dwarfs

NASA Astrophysics Data System (ADS)

Miles-Páez, P. A.; Pallé, E.; Zapatero Osorio, M. R.

2017-12-01

We used the optical and near-infrared imagers located on the Liverpool, the IAC80, and the William Herschel telescopes to monitor 18 M7-L9.5 dwarfs with the objective of measuring their rotation periods. We achieved accuracies typically in the range ±1.5-28 mmag by means of differential photometry, which allowed us to detect photometric variability at the 2σ level in the 50 per cent of the sample. We also detected periodic modulation with periods in the interval 1.5-4.4 h in 9 out of 18 dwarfs that we attribute to rotation. Our variability detections were combined with data from the literature; we found that 65 ± 18 per cent of M7-L3.5 dwarfs with v sin i ≥ 30 km s-1 exhibit photometric variability with typical amplitudes ≤20 mmag in the I band. For those targets and field ultracool dwarfs with measurements of v sin i and rotation period we derived the expected inclination angle of their rotation axis, and found that those with v sin i ≥ 30 km s-1 are more likely to have inclinations ≳40 deg. In addition, we used these rotation periods and others from the literature to study the likely relationship between rotation and linear polarization in dusty ultracool dwarfs. We found a correlation between short rotation periods and large values of linear polarization at optical and near-infrared wavelengths.

Computation by symmetry operations in a structured model of the brain: Recognition of rotational invariance and time reversal

NASA Astrophysics Data System (ADS)

McGrann, John V.; Shaw, Gordon L.; Shenoy, Krishna V.; Leng, Xiaodan; Mathews, Robert B.

1994-06-01

Symmetries have long been recognized as a vital component of physical and biological systems. What we propose here is that symmetry operations are an important feature of higher brain function and result from the spatial and temporal modularity of the cortex. These symmetry operations arise naturally in the trion model of the cortex. The trion model is a highly structured mathematical realization of the Mountcastle organizational principle [Mountcastle, in The Mindful Brain (MIT, Cambridge, 1978)] in which the cortical column is the basic neural network of the cortex and is comprised of subunit minicolumns, which are idealized as trions with three levels of firing. A columnar network of a small number of trions has a large repertoire of quasistable, periodic spatial-temporal firing magic patterns (MP's), which can be excited. The MP's are related by specific symmetries: Spatial rotation, parity, ``spin'' reversal, and time reversal as well as other ``global'' symmetry operations in this abstract internal language of the brain. These MP's can be readily enhanced (as well as inherent categories of MP's) by only a small change in connection strengths via a Hebb learning rule. Learning introduces small breaking of the symmetries in the connectivities which enables a symmetry in the patterns to be recognized in the Monte Carlo evolution of the MP's. Examples of the recognition of rotational invariance and of a time-reversed pattern are presented. We propose the possibility of building a logic device from the hardware implementation of a higher level architecture of trion cortical columns.

Power Stroke Angular Velocity Profiles of Archaeal A-ATP Synthase Versus Thermophilic and Mesophilic F-ATP Synthase Molecular Motors*

PubMed Central

Sielaff, Hendrik; Martin, James; Singh, Dhirendra; Biuković, Goran; Grüber, Gerhard; Frasch, Wayne D.

2016-01-01

The angular velocities of ATPase-dependent power strokes as a function of the rotational position for the A-type molecular motor A3B3DF, from the Methanosarcina mazei Gö1 A-ATP synthase, and the thermophilic motor α3β3γ, from Geobacillus stearothermophilus (formerly known as Bacillus PS3) F-ATP synthase, are resolved at 5 μs resolution for the first time. Unexpectedly, the angular velocity profile of the A-type was closely similar in the angular positions of accelerations and decelerations to the profiles of the evolutionarily distant F-type motors of thermophilic and mesophilic origins, and they differ only in the magnitude of their velocities. M. mazei A3B3DF power strokes occurred in 120° steps at saturating ATP concentrations like the F-type motors. However, because ATP-binding dwells did not interrupt the 120° steps at limiting ATP, ATP binding to A3B3DF must occur during the catalytic dwell. Elevated concentrations of ADP did not increase dwells occurring 40° after the catalytic dwell. In F-type motors, elevated ADP induces dwells 40° after the catalytic dwell and slows the overall velocity. The similarities in these power stroke profiles are consistent with a common rotational mechanism for A-type and F-type rotary motors, in which the angular velocity is limited by the rotary position at which ATP binding occurs and by the drag imposed on the axle as it rotates within the ring of stator subunits. PMID:27729450

Exchange of rotor components in functioning bacterial flagellar motor

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

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

2010-03-26

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

The molecular motor F-ATP synthase is targeted by the tumoricidal protein HAMLET.

PubMed

Ho, James; Sielaff, Hendrik; Nadeem, Aftab; Svanborg, Catharina; Grüber, Gerhard

2015-05-22

HAMLET (human alpha-lactalbumin made lethal to tumor cells) interacts with multiple tumor cell compartments, affecting cell morphology, metabolism, proteasome function, chromatin structure and viability. This study investigated if these diverse effects of HAMLET might be caused, in part, by a direct effect on the ATP synthase and a resulting reduction in cellular ATP levels. A dose-dependent reduction in cellular ATP levels was detected in A549 lung carcinoma cells, and by confocal microscopy, co-localization of HAMLET with the nucleotide-binding subunits α (non-catalytic) and β (catalytic) of the energy converting F1F0 ATP synthase was detected. As shown by fluorescence correlation spectroscopy, HAMLET binds to the F1 domain of the F1F0 ATP synthase with a dissociation constant (KD) of 20.5μM. Increasing concentrations of the tumoricidal protein HAMLET added to the enzymatically active α3β3γ complex of the F-ATP synthase lowered its ATPase activity, demonstrating that HAMLET binding to the F-ATP synthase effects the catalysis of this molecular motor. Single-molecule analysis was applied to study HAMLET-α3β3γ complex interaction. Whereas the α3β3γ complex of the F-ATP synthase rotated in a counterclockwise direction with a mean rotational rate of 3.8±0.7s(-1), no rotation could be observed in the presence of bound HAMLET. Our findings suggest that direct effects of HAMLET on the F-ATP synthase may inhibit ATP-dependent cellular processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

SU-F-T-640: Feasibility of Using a Commercially Available Surface Guided Radiotherapy System with An Open-Face SRS Immobilization System

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

Chinsky, B; Patel, R; Roeske, J

Purpose: To evaluate the inherent accuracy of using a surface guided radiotherapy system (SGRT) in the setup and monitoring of patients receiving stereotactic radiosurgery with an open-face SRS immobilization system. Methods: An anthropomorphic head phantom was set up using the Qfix Encompass SRS Immobilization System on a Varian Edge with OSMS and Varian TrueBeam with AlignRT. The phantom was positioned at 0° gantry and couch. A reference image was acquired using the SGRT system and an ROI was created over the mask opening. The couch and gantry were rotated to different combinations focusing on clinically used SRS gantry/couch combinations andmore » those blocking the SGRT cameras. Perceived surface deviation by the SGRT system from the reference image was recorded. A Winston-Lutz test was performed on couch angles tested and used to exclude couch walkout. The deviation magnitude was calculated using translational values and rotational raw values were recorded. Results: The maximum couch walkouts were: 0.4mm (Edge) and 0.5mm (TB). Solely rotating the gantry resulted in a median couch deviation of 0.2mm and range of 0.1–0.3mm for both linacs. Only rotating the couch (0° gantry) resulted in median deviations of 0.6mm and 0.5mm with ranges of 0.3–1.0mm and 0.3–0.7mm for the Edge and TB, respectively. Combining gantry and couch rotations, the median deviations were 0.7mm and 0.9mm with ranges of 0.3–1.1mm and 0.2–1.9mm for the Edge and TB, respectively. Including all combinations, rotation, roll, and pitch median deviations ranged from 0.1–0.3° with pitch demonstrating consistently higher values and a maximum deviation of 1.0° (both linacs). Conclusion: SGRT is a reliable monitoring tool, though taking into account system fluctuations, 1mm is too restrictive a site tolerance to use with the Qfix Encompass mask. Gantry rotation has little effect on system fluctuation even with camera blockage, whereas couch rotation has a larger effect.« less

Multi-method automated diagnostics of rotating machines

NASA Astrophysics Data System (ADS)

Kostyukov, A. V.; Boychenko, S. N.; Shchelkanov, A. V.; Burda, E. A.

2017-08-01

The automated machinery diagnostics and monitoring systems utilized within the petrochemical plants are an integral part of the measures taken to ensure safety and, as a consequence, the efficiency of these industrial facilities. Such systems are often limited in their functionality due to the specifics of the diagnostic techniques adopted. As the diagnostic techniques applied in each system are limited, and machinery defects can have different physical nature, it becomes necessary to combine several diagnostics and monitoring systems to control various machinery components. Such an approach is inconvenient, since it requires additional measures to bring the diagnostic results in a single view of the technical condition of production assets. In this case, we mean by a production facility a bonded complex of a process unit, a drive, a power source and lines. A failure of any of these components will cause an outage of the production asset, which is unacceptable. The purpose of the study is to test a combined use of vibration diagnostics and partial discharge techniques within the diagnostic systems of enterprises for automated control of the technical condition of rotating machinery during maintenance and at production facilities. The described solutions allow you to control the condition of mechanical and electrical components of rotating machines. It is shown that the functionality of the diagnostics systems can be expanded with minimal changes in technological chains of repair and operation of rotating machinery. Automation of such systems reduces the influence of the human factor on the quality of repair and diagnostics of the machinery.

Some aspects of cool main sequence star ages derived from stellar rotation (gyrochronology)

NASA Astrophysics Data System (ADS)

Barnes, S. A.; Spada, F.; Weingrill, J.

2016-09-01

Rotation periods for cool stars can be measured with good precision by monitoring starspot light modulation. Observations have shown that the rotation periods of dwarf stars of roughly solar metallicity have such systematic dependencies on stellar age and mass that they can be used to derive reliable ages, a procedure called gyrochronology. We review the method and show illustrative cases, including recent ground- and space-based data. The age uncertainties approach 10 % in the best cases, making them a valuable complement to, and constraint on, asteroseismic or other ages. Edited, updated, and refereed version of a presentation at the WE-Heraeus-Seminar in Bad Honnef, Germany: Reconstructing the Milky Way's History: Spectroscopic Surveys, Asteroseismology and Chemodynamical Models

Effects of Successive Rotation Regimes on Carbon Stocks in Eucalyptus Plantations in Subtropical China Measured over a Full Rotation.

PubMed

Li, Xiaoqiong; Ye, Duo; Liang, Hongwen; Zhu, Hongguang; Qin, Lin; Zhu, Yuling; Wen, Yuanguang

2015-01-01

Plantations play an important role in carbon sequestration and the global carbon cycle. However, there is a dilemma in that most plantations are managed on short rotations, and the carbon sequestration capacities of these short-rotation plantations remain understudied. Eucalyptus has been widely planted in the tropics and subtropics due to its rapid growth, high adaptability, and large economic return. Eucalyptus plantations are primarily planted in successive rotations with a short rotation length of 6~8 years. In order to estimate the carbon-stock potential of eucalyptus plantations over successive rotations, we chose a first rotation (FR) and a second rotation (SR) stand and monitored the carbon stock dynamics over a full rotation from 1998 to 2005. Our results showed that carbon stock in eucalyptus trees (TC) did not significantly differ between rotations, while understory vegetation (UC) and soil organic matter (SOC) stored less carbon in the SR (1.01 vs. 2.76 Mg.ha(-1) and 70.68 vs. 81.08 Mg. ha(-1), respectively) and forest floor carbon (FFC) conversely stored more (2.80 vs. 2.34 Mg. ha(-1)). The lower UC and SOC stocks in the SR stand resulted in 1.13 times lower overall ecosystem carbon stock. Mineral soils and overstory trees were the two dominant carbon pools in eucalyptus plantations, accounting for 73.77%~75.06% and 20.50%~22.39%, respectively, of the ecosystem carbon pool. However, the relative contribution (to the ecosystem pool) of FFC stocks increased 1.38 times and that of UC decreased 2.30 times in the SR versus FR stand. These carbon pool changes over successive rotations were attributed to intensive successive rotation regimes of eucalyptus plantations. Our eight year study suggests that for the sustainable development of short-rotation plantations, a sound silvicultural strategy is required to achieve the best combination of high wood yield and carbon stock potential.

MOJAVE: Monitoring of Jets in Active Galactic Nuclei with VLBA Experiments. VIII. Faraday Rotation in Parsec-scale AGN Jets

NASA Astrophysics Data System (ADS)

Hovatta, Talvikki; Lister, Matthew L.; Aller, Margo F.; Aller, Hugh D.; Homan, Daniel C.; Kovalev, Yuri Y.; Pushkarev, Alexander B.; Savolainen, Tuomas

2012-10-01

We report observations of Faraday rotation measures for a sample of 191 extragalactic radio jets observed within the MOJAVE program. Multifrequency Very Long Baseline Array observations were carried out over 12 epochs in 2006 at four frequencies between 8 and 15 GHz. We detect parsec-scale Faraday rotation measures in 149 sources and find the quasars to have larger rotation measures on average than BL Lac objects. The median core rotation measures are significantly higher than in the jet components. This is especially true for quasars where we detect a significant negative correlation between the magnitude of the rotation measure and the de-projected distance from the core. We perform detailed simulations of the observational errors of total intensity, polarization, and Faraday rotation, and concentrate on the errors of transverse Faraday rotation measure gradients in unresolved jets. Our simulations show that the finite image restoring beam size has a significant effect on the observed rotation measure gradients, and spurious gradients can occur due to noise in the data if the jet is less than two beams wide in polarization. We detect significant transverse rotation measure gradients in four sources (0923+392, 1226+023, 2230+114, and 2251+158). In 1226+023 the rotation measure is for the first time seen to change sign from positive to negative over the transverse cuts, which supports the presence of a helical magnetic field in the jet. In this source we also detect variations in the jet rotation measure over a timescale of three months, which are difficult to explain with external Faraday screens and suggest internal Faraday rotation. By comparing fractional polarization changes in jet components between the four frequency bands to depolarization models, we find that an external purely random Faraday screen viewed through only a few lines of sight can explain most of our polarization observations, but in some sources, such as 1226+023 and 2251+158, internal Faraday rotation is needed.

Effects of Successive Rotation Regimes on Carbon Stocks in Eucalyptus Plantations in Subtropical China Measured over a Full Rotation

PubMed Central

Li, Xiaoqiong; Ye, Duo; Liang, Hongwen; Zhu, Hongguang; Qin, Lin; Zhu, Yuling; Wen, Yuanguang

2015-01-01

Plantations play an important role in carbon sequestration and the global carbon cycle. However, there is a dilemma in that most plantations are managed on short rotations, and the carbon sequestration capacities of these short-rotation plantations remain understudied. Eucalyptus has been widely planted in the tropics and subtropics due to its rapid growth, high adaptability, and large economic return. Eucalyptus plantations are primarily planted in successive rotations with a short rotation length of 6~8 years. In order to estimate the carbon-stock potential of eucalyptus plantations over successive rotations, we chose a first rotation (FR) and a second rotation (SR) stand and monitored the carbon stock dynamics over a full rotation from 1998 to 2005. Our results showed that carbon stock in eucalyptus trees (TC) did not significantly differ between rotations, while understory vegetation (UC) and soil organic matter (SOC) stored less carbon in the SR (1.01 vs. 2.76 Mg.ha-1 and 70.68 vs. 81.08 Mg. ha-1, respectively) and forest floor carbon (FFC) conversely stored more (2.80 vs. 2.34 Mg. ha-1). The lower UC and SOC stocks in the SR stand resulted in 1.13 times lower overall ecosystem carbon stock. Mineral soils and overstory trees were the two dominant carbon pools in eucalyptus plantations, accounting for 73.77%~75.06% and 20.50%~22.39%, respectively, of the ecosystem carbon pool. However, the relative contribution (to the ecosystem pool) of FFC stocks increased 1.38 times and that of UC decreased 2.30 times in the SR versus FR stand. These carbon pool changes over successive rotations were attributed to intensive successive rotation regimes of eucalyptus plantations. Our eight year study suggests that for the sustainable development of short-rotation plantations, a sound silvicultural strategy is required to achieve the best combination of high wood yield and carbon stock potential. PMID:26186367

Purification of R-phycoerythrin from Gracilaria lemaneiformis by centrifugal precipitation chromatography.

PubMed

Gu, Dongyu; Lazo-Portugal, Rodrigo; Fang, Chen; Wang, Zhantong; Ma, Ying; Knight, Martha; Ito, Yoichiro

2018-06-15

Centrifugal precipitation chromatography (CpC) is a powerful chromatographic technique invented in the year 2000 but so far very little applied. The method combines dialysis, counter-current and salting out processes. The separation rotor consists of two identical spiral channels separated by a dialysis membrane (6-8 K MW cut-off) in which the upper channel is eluted with an ammonium sulfate gradient and the lower channel with water, and the mixtures are separated according to their solubility in ammonium sulfate as a chromatographic technique. In the present study, the method was successfully applied for separation and purification of R-phycoerythrin (R-PE), a protein widely used as a fluorescent probe, from the red alga Gracilaria lemaneiformis. The separation was performed with the elution of ammonium sulfate from 50% to 0% in 21.5 h at a flow rate of 0.5 ml/min, while the lower channel was eluted with water at a flow rate of 0.05 ml/min after sample charge, and the column was rotated at 200 rpm. After a single run, the absorbance ratio A 565 /A 280 (a criterion for the purity of R-PE) was increased from 0.5 of the crude to 6.5. The purified R-PE exhibited a typical "three peaks" spectrum with absorbance maximum at 497, 538 and 565 nm. The Native-PAGE showed one single protein band and 20 kDa (subunits α and β) and 30 kDa (subunit γ) can be observed in SDS-PAGE analysis which were consistent with the (αβ) 6 γ subunit composition of R-PE. The results indicated that CpC is an efficient method to obtain protein with the high purity from a complex source. Copyright © 2018 Elsevier B.V. All rights reserved.

A semiempirical study for the ground and excited states of free-base and zinc porphyrin-fullerene dyads

NASA Technical Reports Server (NTRS)

Parusel, A. B.

2000-01-01

The ground and excited states of a covalently linked porphyrin-fullerene dyad in both its free-base and zinc forms (D. Kuciauskas et al., J. Phys. Chem. 100 (1996) 15926) have been investigated by semiempirical methods. The excited-state properties are discussed by investigation of the character of the molecular orbitals. All frontier MOs are mainly localized on either the donor or the acceptor subunit. Thus, the absorption spectra of both systems are best described as the sum of the spectra of the single components. The experimentally observed spectra are well reproduced by the theoretical computations. Both molecules undergo efficient electron transfer in polar but not in apolar solvents. This experimental finding is explained theoretically by explicitly considering solvent effects. The tenth excited state in the gas phase is of charge-separated character where an electron is transferred from the porphyrin donor to the fullerene acceptor subunit. This state is stabilized in energy in polar solvents due to its large formal dipole moment. The stabilization energy for an apolar environment such as benzene is not sufficient to lower this state to become the first excited singlet state. Thus, no electron transfer is observed, in agreement with experiment. In a polar environment such as acetonitrile, the charge-separated state becomes the S, state and electron transfer takes place, as observed experimentally. The flexible single bond connecting both the donor and acceptor subunits allows free rotation by ca. +/- 30 degrees about the optimized ground-state conformation. For the charge-separated state this optimized geometry has a maximum dipole moment. The geometry of the charge-separated state thus does not change relatively to the ground-state conformation. The electron-donating properties of porphyrin are enhanced in the zinc derivative due to a reduced porphyrin HOMO-LUMO energy gap. This yields a lower energy for the charge-separated state compared to the free-base dyad.

A method to track rotational motion for use in single-molecule biophysics.

PubMed

Lipfert, Jan; Kerssemakers, Jacob J W; Rojer, Maylon; Dekker, Nynke H

2011-10-01

The double helical nature of DNA links many cellular processes such as DNA replication, transcription, and repair to rotational motion and the accumulation of torsional strain. Magnetic tweezers (MTs) are a single-molecule technique that enables the application of precisely calibrated stretching forces to nucleic acid tethers and to control their rotational motion. However, conventional magnetic tweezers do not directly monitor rotation or measure torque. Here, we describe a method to directly measure rotational motion of particles in MT. The method relies on attaching small, non-magnetic beads to the magnetic beads to act as fiducial markers for rotational tracking. CCD images of the beads are analyzed with a tracking algorithm specifically designed to minimize crosstalk between translational and rotational motion: first, the in-plane center position of the magnetic bead is determined with a kernel-based tracker, while subsequently the height and rotation angle of the bead are determined via correlation-based algorithms. Evaluation of the tracking algorithm using both simulated images and recorded images of surface-immobilized beads demonstrates a rotational resolution of 0.1°, while maintaining a translational resolution of 1-2 nm. Example traces of the rotational fluctuations exhibited by DNA-tethered beads confined in magnetic potentials of varying stiffness demonstrate the robustness of the method and the potential for simultaneous tracking of multiple beads. Our rotation tracking algorithm enables the extension of MTs to magnetic torque tweezers (MTT) to directly measure the torque in single molecules. In addition, we envision uses of the algorithm in a range of biophysical measurements, including further extensions of MT, tethered particle motion, and optical trapping measurements.

Comparison of 180-degree and 90-degree needle rotation to reduce wound size in PIT-injected juvenile Chinook salmon

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

Bryson, Amanda J.; Woodley, Christa M.; Karls, Rhonda K.

2013-04-30

Animal telemetry, which requires the implantation of passive transponders or active transmitters, is used to monitor and assess fish stock and conservation to gain an understanding of fish movement and behavior. As new telemetry technologies become available, studies of their effects on species of interest are imperative as is development of implantation techniques. In this study, we investigated the effects of bevel rotation (0-, 90-, 180-degree axis rotation) on wound extent, tag loss, and wound healing rates in juvenile Chinook salmon injected with an 8-gauge needle, which is required for implantation of the novel injectable Juvenile Salmon Acoustic Telemetry Systemsmore » (JSATS) acoustic transmitter or large passive integrated transponder (PIT) tags. Although the injection sites were not closed after injection (e.g., with sutures or glue), there were no mortalities, dropped tags, or indications of fungus, ulceration, and/or redness around the wound. On Day 0 and post-implantation Day 7, the 90-degree bevel rotation produced smaller wound extent than the 180-degree bevel rotation. No axis rotation (0-degrees) resulted in the PIT tag frequently misleading or falling out upon injection. The results of this study indicated the 90-degree bevel rotation was the more efficient technique, produced less wound extent. Given the wound extent compared to size of fish, we recommend researchers should consider a 90-degree rotation over the 180-degree rotation in telemetry studies. Highlights •Three degrees of needle rotation were examined for effects in Chinook salmon. •Mortality, tag loss, wound extent, healing, and infection indicators were measured. •There were no mortalities, tag loss, or indications of infection. •The 90-degree needle rotation through Day 7 produced the smallest wound extent.« less

The RINGO2 and DIPOL optical polarization catalogue of blazars

NASA Astrophysics Data System (ADS)

Jermak, H.; Steele, I. A.; Lindfors, E.; Hovatta, T.; Nilsson, K.; Lamb, G. P.; Mundell, C.; Barres de Almeida, U.; Berdyugin, A.; Kadenius, V.; Reinthal, R.; Takalo, L.

2016-11-01

We present ˜2000 polarimetric and ˜3000 photometric observations of 15 γ-ray bright blazars over a period of 936 days (2008-10-11 to 2012-10-26) using data from the Tuorla blazar monitoring program (KVA DIPOL) and Liverpool Telescope (LT) RINGO2 polarimeters (supplemented with data from SkyCamZ (LT) and Fermi-LAT γ-ray data). In 11 out of 15 sources we identify a total of 19 electric vector position angle (EVPA) rotations and 95 flaring episodes. We group the sources into subclasses based on their broad-band spectral characteristics and compare their observed optical and γ-ray properties. We find that (1) the optical magnitude and γ-ray flux are positively correlated, (2) EVPA rotations can occur in any blazar subclass, four sources show rotations that go in one direction and immediately rotate back, (3) we see no difference in the γ-ray flaring rates in the sample; flares can occur during and outside of rotations with no preference for this behaviour, (4) the average degree of polarization (DoP), optical magnitude and γ-ray flux are lower during an EVPA rotation compared with during non-rotation and the distribution of the DoP during EVPA rotations is not drawn from the same parent sample as the distribution outside rotations, (5) the number of observed flaring events and optical polarization rotations are correlated, however we find no strong evidence for a temporal association between individual flares and rotations and (6) the maximum observed DoP increases from ˜10 per cent to ˜30 per cent to ˜40 per cent for subclasses with synchrotron peaks at high, intermediate and low frequencies, respectively.

Prefoldin Subunits Are Protected from Ubiquitin-Proteasome System-mediated Degradation by Forming Complex with Other Constituent Subunits*

PubMed Central

Miyazawa, Makoto; Tashiro, Erika; Kitaura, Hirotake; Maita, Hiroshi; Suto, Hiroo; Iguchi-Ariga, Sanae M. M.; Ariga, Hiroyoshi

2011-01-01

The molecular chaperone prefoldin (PFD) is a complex comprised of six different subunits, PFD1-PFD6, and delivers newly synthesized unfolded proteins to cytosolic chaperonin TRiC/CCT to facilitate the folding of proteins. PFD subunits also have functions different from the function of the PFD complex. We previously identified MM-1α/PFD5 as a novel c-Myc-binding protein and found that MM-1α suppresses transformation activity of c-Myc. However, it remains unclear how cells regulate protein levels of individual subunits and what mechanisms alter the ratio of their activities between subunits and their complex. In this study, we found that knockdown of one subunit decreased protein levels of other subunits and that transfection of five subunits other than MM-1α into cells increased the level of endogenous MM-1α. We also found that treatment of cells with MG132, a proteasome inhibitor, increased the level of transfected/overexpressed MM-1α but not that of endogenous MM-1α, indicating that overexpressed MM-1α, but not endogenous MM-1α, was degraded by the ubiquitin proteasome system (UPS). Experiments using other PFD subunits showed that the UPS degraded a monomer of PFD subunits, though extents of degradation varied among subunits. Furthermore, the level of one subunit was increased after co-transfection with the respective subunit, indicating that there are specific combinations between subunits to be stabilized. These results suggest mutual regulation of protein levels among PFD subunits and show how individual subunits form the PFD complex without degradation. PMID:21478150

Magnetic Actuator Modelling for Rotating Machinery Analysis

NASA Astrophysics Data System (ADS)

Mendes, Ricardo Ugliara; de Castro, Hélio Fiori; Cavalca, Kátia Lucchesi; Ferreira, Luiz Otávio Saraiva

Rotating machines have a wide range of application such as airplanes, factories, laboratories and power plants. Lately, with computer aid design, shafts finite element models including bearings, discs, seals and couplings have been developed, allowing the prediction of the machine behavior. In order to keep confidence during operation, it is necessary to monitor these systems, trying to predict future failures. One of the most applied technique for this purpose is the modal analysis. It consists of applying a perturbation force into the system and then to measure its response. However, there is a difficulty that brings limitations to the excitation of systems with rotating shafts when using impact hammers or shakers, once due to friction, undesired tangential forces and noise can be present in the measurements. Therefore, the study of a non-contact technique of external excitation becomes of high interest. In this sense, the present work deals with the study and development of a finite element model for rotating machines using a magnetic actuator as an external excitation source. This work also brings numerical simulations where the magnetic actuator was used to obtain the frequency response function of the rotating system.

A Novel Permanent Magnetic Angular Acceleration Sensor

PubMed Central

Zhao, Hao; Feng, Hao

2015-01-01

Angular acceleration is an important parameter for status monitoring and fault diagnosis of rotary machinery. Therefore, we developed a novel permanent magnetic angular acceleration sensor, which is without rotation angle limitations and could directly measure the instantaneous angular acceleration of the rotating system. The sensor rotor only needs to be coaxially connected with the rotating system, which enables convenient sensor installation. For the cup structure of the sensor rotor, it has a relatively small rotational inertia. Due to the unique mechanical structure of the sensor, the output signal of the sensor can be directed without a slip ring, which avoids signal weakening effect. In this paper, the operating principle of the sensor is described, and simulated using finite element method. The sensitivity of the sensor is calibrated by torsional pendulum and angle sensor, yielding an experimental result of about 0.88 mV/(rad·s−2). Finally, the angular acceleration of the actual rotating system has been tested, using both a single-phase asynchronous motor and a step motor. Experimental result confirms the operating principle of the sensor and indicates that the sensor has good practicability. PMID:26151217

Normal Mode Analysis on the Relaxation of AN Excited Nitromethane Molecule in Argon Bath

NASA Astrophysics Data System (ADS)

Rivera-Rivera, Luis A.; Wagner, Albert F.

2017-06-01

In our previous work [Rivera-Rivera et al. J. Chem. Phys. 142, 014303 (2015).] classical molecular dynamics simulations followed, in an Ar bath, the relaxation of nitromethane (CH_3NO_2) instantaneously excited by statistically distributing 50 kcal/mol among all its internal degrees of freedom. The 300 K Ar bath was at pressures of 10 to 400 atm. Both rotational and vibrational energies exhibited multi-exponential decay. This study explores mode-specific mechanisms at work in the decay process. With the separation of rotation and vibration developed by Rhee and Kim [J. Chem. Phys. 107, 1394 (1997).], one can show that the vibrational kinetic energy decomposes only into vibrational normal modes while the rotational and Coriolis energies decompose into both vibrational and rotational normal modes. Then the saved CH_3NO_2 positions and momenta can be converted into mode-specific energies whose decay over 1000 ps can be monitored. The results identify vibrational and rotational modes that promote/resist energy lost and drive multi-exponential behavior. In addition to mode-specificity, the results show disruption of IVR with increasing pressure.

Photometric study of HD 155555C in the β Pictoris Association

NASA Astrophysics Data System (ADS)

Messina, Sergio; Millward, Mervyn; Bradstreet, David H.

2015-05-01

We are carrying out a series of photometric monitoring to measure the rotation periods of members in the young β Pictoris Association, as part of the RACE-OC project (Rotation and ACtivity Evolution in Open Clusters). In this paper, we present the results for HD 155555C which is believed to be physically associated to the spectroscopic binary V824 Ara (HD 155555) and thus constituting a triple system. We collected B, V, and R-band photometric data timeseries and discovered from periodogram analysis the rotation period P = 4.43 d. Combined with stellar radius and projected rotational velocity, we find this star almost equator-on with an inclination i ≃ 90 ° . The rotational properties of HD 155555C fit well into the period distribution of other β Pic members, giving further support to the suggested membership to the association and to its physical association to V824 Ara. A comparison with pre-main-sequence isochrones from various models allows us to estimate an age of 20 ± 15 Myr for this triple system.

Radiation area monitor device and method

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

Vencelj, Matjaz; Stowe, Ashley C.; Petrovic, Toni

A radiation area monitor device/method, utilizing: a radiation sensor having a directional radiation sensing capability; a rotation mechanism operable for selectively rotating the radiation sensor such that the directional radiation sensing capability selectively sweeps an area of interest; and a processor operable for analyzing and storing a radiation fingerprint acquired by the radiation sensor as the directional radiation sensing capability selectively sweeps the area of interest. Optionally, the radiation sensor includes a gamma and/or neutron radiation sensor. The device/method selectively operates in: a first supervised mode during which a baseline radiation fingerprint is acquired by the radiation sensor; and amore » second unsupervised mode during which a subsequent radiation fingerprint is acquired by the radiation sensor, wherein the subsequent radiation fingerprint is compared to the baseline radiation fingerprint and, if a predetermined difference threshold is exceeded, an alert is issued.« less

Turbine Rotor Disk Health Monitoring Assessment Based on Sensor Technology and Spin Tests Data

PubMed Central

2013-01-01

The paper focuses on presenting data obtained from spin test experiments of a turbine engine like rotor disk and assessing their correlation to the development of a structural health monitoring and fault detection system. The data were obtained under various operating conditions such as the rotor disk being artificially induced with and without a notch and rotated at a rotational speed of up to 10,000 rpm under balanced and imbalanced state. The data collected included blade tip clearance, blade tip timing measurements, and shaft displacements. Two different sensor technologies were employed in the testing: microwave and capacitive sensors, respectively. The experimental tests were conducted at the NASA Glenn Research Center's Rotordynamics Laboratory using a high precision spin system. Disk flaw observations and related assessments from the collected data for both sensors are reported and discussed. PMID:23844396

Organization of model helical peptides in lipid bilayers: insight into the behavior of single-span protein transmembrane domains.

PubMed Central

Sharpe, Simon; Barber, Kathryn R; Grant, Chris W M; Goodyear, David; Morrow, Michael R

2002-01-01

Selectively deuterated transmembrane peptides comprising alternating leucine-alanine subunits were examined in fluid bilayer membranes by solid-state nuclear magnetic resonance (NMR) spectroscopy in an effort to gain insight into the behavior of membrane proteins. Two groups of peptides were studied: 21-mers having a 17-amino-acid hydrophobic domain calculated to be close in length to the hydrophobic thickness of 1-palmitoyl-2-oleoyl phosphatidylcholine and 26-mers having a 22-amino-acid hydrophobic domain calculated to exceed the membrane hydrophobic thickness. (2)H NMR spectral features similar to ones observed for transmembrane peptides from single-span receptors of higher animal cells were identified which apparently correspond to effectively monomeric peptide. Spectral observations suggested significant distortion of the transmembrane alpha-helix, and/or potential for restriction of rotation about the tilted helix long axis for even simple peptides. Quadrupole splittings arising from the 26-mer were consistent with greater peptide "tilt" than were those of the analogous 21-mer. Quadrupole splittings associated with monomeric peptide were relatively insensitive to concentration and temperature over the range studied, indicating stable average conformations, and a well-ordered rotation axis. At high peptide concentration (6 mol% relative to phospholipid) it appeared that the peptide predicted to be longer than the membrane thickness had a particular tendency toward reversible peptide-peptide interactions occurring on a timescale comparable with or faster than approximately 10(-5) s. This interaction may be direct or lipid-mediated and was manifest as line broadening. Peptide rotational diffusion rates within the membrane, calculated from quadrupolar relaxation times, T(2e), were consistent with such interactions. In the case of the peptide predicted to be equal to the membrane thickness, at low peptide concentration spectral lineshape indicated the additional presence of a population of peptide having rotational motion that was restricted on a timescale of 10(-5) s. PMID:12080125

Role of regulatory subunits and protein kinase inhibitor (PKI) in determining nuclear localization and activity of the catalytic subunit of protein kinase A.

PubMed

Wiley, J C; Wailes, L A; Idzerda, R L; McKnight, G S

1999-03-05

Regulation of protein kinase A by subcellular localization may be critical to target catalytic subunits to specific substrates. We employed epitope-tagged catalytic subunit to correlate subcellular localization and gene-inducing activity in the presence of regulatory subunit or protein kinase inhibitor (PKI). Transiently expressed catalytic subunit distributed throughout the cell and induced gene expression. Co-expression of regulatory subunit or PKI blocked gene induction and prevented nuclear accumulation. A mutant PKI lacking the nuclear export signal blocked gene induction but not nuclear accumulation, demonstrating that nuclear export is not essential to inhibit gene induction. When the catalytic subunit was targeted to the nucleus with a nuclear localization signal, it was not sequestered in the cytoplasm by regulatory subunit, although its activity was completely inhibited. PKI redistributed the nuclear catalytic subunit to the cytoplasm and blocked gene induction, demonstrating that the nuclear export signal of PKI can override a strong nuclear localization signal. With increasing PKI, the export process appeared to saturate, resulting in the return of catalytic subunit to the nucleus. These results demonstrate that both the regulatory subunit and PKI are able to completely inhibit the gene-inducing activity of the catalytic subunit even when the catalytic subunit is forced to concentrate in the nuclear compartment.

The Immediate Effects of Upper Thoracic Translatoric Spinal Manipulation on Cervical Pain and Range of Motion: A Randomized Clinical Trial

PubMed Central

Krauss, John; Creighton, Doug; Ely, Jonathan D.; Podlewska-Ely, Joanna

2008-01-01

This study examined the effect of translatoric spinal manipulation (TSM) on cervical pain and cervical active motion restriction when applied to upper thoracic (T1-T4) segments. Active cervical rotation range of motion was measured re- and post-intervention with a cervical inclinometer (CROM), and cervical pain status was monitored before and after manipulation with a Faces Pain Scale. Study participants included a sample of convenience that included 32 patients referred to physical therapy with complaints of pain in the mid-cervical region and restricted active cervical rotation. Twenty-two patients were randomly assigned to the experimental group and ten were assigned to the control group. Pre- and post-intervention cervical range of motion and pain scale measurements were taken by a physical therapist assistant who was blinded to group assignment. The experimental group received TSM to hypomobile upper thoracic segments. The control group received no intervention. Paired t-tests were used to analyze within-group changes in cervical rotation and pain, and a 2-way repeated-measure ANOVA was used to analyze between-group differences in cervical rotation and pain. Significance was accepted at p = 0.05. Significant changes that exceeded the MDC95 were detected for cervical rotation both within group and between groups with the TSM group demonstrating increased mean (SD) in right rotation of 8.23° (7.41°) and left rotation of 7.09° (5.83°). Pain levels perceived during post-intervention cervical rotation showed significant improvement during right rotation for patients experiencing pain during bilateral rotation only (p=.05). This study supports the hypothesis that spinal manipulation applied to the upper thoracic spine (T1-T4 motion segments) significantly increases cervical rotation ROM and may reduce cervical pain at end range rotation for patients experiencing pain during bilateral cervical rotation. PMID:19119394

Apparatus and method for controlling the rotary airlocks in a coal processing system by reversing the motor current rotating the air lock

DOEpatents

Groombridge, Clifton E.

1996-01-01

An improvement to a coal processing system where hard materials found in the coal may cause jamming of either inflow or outflow rotary airlocks, each driven by a reversible motor. The instantaneous current used by the motor is continually monitored and compared to a predetermined value. If an overcurrent condition occurs, indicating a jamming of the airlock, a controller means starts a "soft" reverse rotation of the motor thereby clearing the jamming. Three patterns of the motor reversal are provided.

Association with β-COP Regulates the Trafficking of the Newly Synthesized Na,K-ATPase*

PubMed Central

Morton, Michael J.; Farr, Glen A.; Hull, Michael; Capendeguy, Oihana; Horisberger, Jean-Daniel; Caplan, Michael J.

2010-01-01

Plasma membrane expression of the Na,K-ATPase requires assembly of its α- and β-subunits. Using a novel labeling technique to identify Na,K-ATPase partner proteins, we detected an interaction between the Na,K-ATPase α-subunit and the coat protein, β-COP, a component of the COP-I complex. When expressed in the absence of the Na,K-ATPase β-subunit, the Na,K-ATPase α-subunit interacts with β-COP, is retained in the endoplasmic reticulum, and is targeted for degradation. In the presence of the Na,K-ATPase β-subunit, the α-subunit does not interact with β-COP and traffics to the plasma membrane. Pulse-chase experiments demonstrate that in cells expressing both the Na,K-ATPase α- and β-subunits, newly synthesized α-subunit associates with β-COP immediately after its synthesis but that this interaction does not constitute an obligate intermediate in the assembly of the α- and β-subunits to form the pump holoenzyme. The interaction with β-COP was reduced by mutating a dibasic motif at Lys54 in the Na,K-ATPase α-subunit. This mutant α-subunit is not retained in the endoplasmic reticulum and reaches the plasma membrane, even in the absence of Na,K-ATPase β-subunit expression. Although the Lys54 α-subunit reaches the cell surface without need for β-subunit assembly, it is only functional as an ion-transporting ATPase in the presence of the β-subunit. PMID:20801885

Amino acid sequence of the human fibronectin receptor

PubMed Central

1987-01-01

The amino acid sequence deduced from cDNA of the human placental fibronectin receptor is reported. The receptor is composed of two subunits: an alpha subunit of 1,008 amino acids which is processed into two polypeptides disulfide bonded to one another, and a beta subunit of 778 amino acids. Each subunit has near its COOH terminus a hydrophobic segment. This and other sequence features suggest a structure for the receptor in which the hydrophobic segments serve as transmembrane domains anchoring each subunit to the membrane and dividing each into a large ectodomain and a short cytoplasmic domain. The alpha subunit ectodomain has five sequence elements homologous to consensus Ca2+- binding sites of several calcium-binding proteins, and the beta subunit contains a fourfold repeat strikingly rich in cysteine. The alpha subunit sequence is 46% homologous to the alpha subunit of the vitronectin receptor. The beta subunit is 44% homologous to the human platelet adhesion receptor subunit IIIa and 47% homologous to a leukocyte adhesion receptor beta subunit. The high degree of homology (85%) of the beta subunit with one of the polypeptides of a chicken adhesion receptor complex referred to as integrin complex strongly suggests that the latter polypeptide is the chicken homologue of the fibronectin receptor beta subunit. These receptor subunit homologies define a superfamily of adhesion receptors. The availability of the entire protein sequence for the fibronectin receptor will facilitate studies on the functions of these receptors. PMID:2958481

Fiber optical sensors for aircraft applications

NASA Astrophysics Data System (ADS)

Pechstedt, Ralf D.

2014-09-01

In this paper selected fiber optical point sensors that are of potential interest for deployment in aircraft are discussed. The operating principles together with recent measurement results are described. Examples include a high-temperature combined pressure and temperature sensor for engine health, hydraulics and landing gear monitoring, an ultra-high sensitive pressure sensor for oil, pneumatic and fluid aero systems applications and a combined acceleration and temperature sensor for condition monitoring of rotating components.

Method and apparatus for monitoring motor operated valve motor output torque and power at valve seating

DOEpatents

Casada, D.A.

1996-01-16

A method and apparatus are provided for monitoring a motor operated valve during the brief period when the valve seats and the torque switch trips to deenergize the valve motor. The method uses voltage measurements on the load side of a deenergizing switch that opens to deenergize the motor to determine, among other things, final motor rotational speed and the decelerating torque at motor deenergization. 14 figs.

Method and apparatus for monitoring motor operated valve motor output torque and power at valve seating

DOEpatents

Casada, Donald A.

1996-01-01

A method and apparatus are provided for monitoring a motor operated valve during the brief period when the valve seats and the torque switch trips to deenergize the valve motor. The method uses voltage measurements on the load side of a deenergizing switch that opens to deenergize the motor to determine, among other things, final motor rotational speed and the decelerating torque at motor deenergization.

An on-line monitoring system for oil-film, pressure and temperature distributions in large-scale hydro-generator bearings

NASA Astrophysics Data System (ADS)

Höbel, M.; Haffner, K.

1999-05-01

Instrumentation that allows the behaviour of a hydro-generator thrust bearing to be monitored during operation is described. The measurement system was developed at the Asea Brown Boveri corporate research centre in Switzerland and was tested under realistic operating conditions at the Harbin Electric Machinery Company bearing-testing facility in the People's Republic of China. Newly developed fibre-optical proximity probes were used for the on-line monitoring of the thin oil film between the static and rotating parts of the bearing. These sensors are based on a back-reflection technique and can be used for various target materials such as Babbitt and Teflon. The monitoring system comprises about 120 temperature sensors, four pressure sensors and five optical oil-film thickness sensors. Temperature sensors are installed at specific static locations, whereas pressure and oil-film sensors are positioned in the runner and generate data during rotation. A special feature of the monitoring equipment is its on-line processing capability. Digital signal processors operating in parallel handle pressure and oil-film thickness data. Important measurement parameters such as the maximum pressure, maximum temperature and minimum oil-film thickness are displayed on-line. Detailed three-dimensional temperature information on one of the load segments can be obtained from subsequent off-line data analysis. The system also calculates two-dimensional plots of the oil-film thickness and pressure for most of the 12 load segments.

Linking structural features from mitochondrial and bacterial F-type ATP synthases to their distinct mechanisms of ATPase inhibition.

PubMed

Krah, Alexander

2015-10-01

ATP synthases are molecular motors, which synthesize ATP, the ubiquitous energy source in all living cells. They use an electrochemical gradient to drive a rotation in the membrane embedded Fo domain, namely the c-ring, causing a conformational change in the soluble F1 domain which leads to the catalytic event. In the opposite fashion, they can also hydrolyse ATP to maintain the ion gradient across the membrane. To prevent wasteful ATP hydrolysis, bacteria and mammals have developed peculiar mechanistic features in addition to a common one, namely MgADP inhibition. Here I discuss the distinct ATPase inhibition mechanism in mitochondrial (IF1) and bacterial (subunits ε and ζ) F-type ATP synthases, based on available structural, biophysical and biochemical data. Copyright © 2015 Elsevier Ltd. All rights reserved.

Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target

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

Mulepati, Sabin; Heroux, Annie; Bailey, Scott

In prokaryotes, RNA derived from type I and type III CRISPR loci direct large ribonucleoprotein complexes to destroy invading bacteriophage and plasmids. In Escherichia coli, this 405-kilodalton complex is called Cascade. We report the crystal structure of Cascade bound to a single-stranded DNA (ssDNA) target at a resolution of 3.03 angstroms. The structure reveals that the CRISPR RNA and target strands do not form a double helix but instead adopt an underwound ribbon-like structure. This noncanonical structure is facilitated by rotation of every sixth nucleotide out of the RNA-DNA hybrid and is stabilized by the highly interlocked organization of proteinmore » subunits. These studies provide insight into both the assembly and the activity of this complex and suggest a mechanism to enforce fidelity of target binding.« less

Crystal structure of a CRISPR RNA-guided surveillance complex bound to a ssDNA target

PubMed Central

Mulepati, Sabin; Héroux, Annie; Bailey, Scott

2015-01-01

In prokaryotes, RNA derived from type I and type III CRISPR loci direct large ribonucleoprotein complexes to destroy invading bacteriophage and plasmids. In Escherichia coli, this 405-kDa complex is called Cascade. Here we report the 3.03Å crystal structure of Cascade bound to a single-stranded DNA target. The structure reveals that the CRISPR RNA and target strands do not form a double helix but instead adopt an underwound ribbon-like structure. This non-canonical structure is facilitated by rotation of every sixth nucleotide out of the RNA-DNA hybrid and is stabilized by the highly interlocked organization of protein subunits. These studies provide insight into both the assembly and the activity of this complex and suggest a mechanism to enforce fidelity of target binding. PMID:25123481

WHY ARE RAPIDLY ROTATING M DWARFS IN THE PLEIADES SO (INFRA)RED? NEW PERIOD MEASUREMENTS CONFIRM ROTATION-DEPENDENT COLOR OFFSETS FROM THE CLUSTER SEQUENCE

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

Covey, Kevin R.; Agüeros, Marcel A.; Liu, Jiyu

2016-05-10

Stellar rotation periods ( P {sub rot}) measured in open clusters have proved to be extremely useful for studying stars’ angular momentum content and rotationally driven magnetic activity, which are both age- and mass-dependent processes. While P {sub rot} measurements have been obtained for hundreds of solar-mass members of the Pleiades, measurements exist for only a few low-mass (<0.5 M {sub ⊙}) members of this key laboratory for stellar evolution theory. To fill this gap, we report P {sub rot} for 132 low-mass Pleiades members (including nearly 100 with M ≤ 0.45 M {sub ⊙}), measured from photometric monitoring ofmore » the cluster conducted by the Palomar Transient Factory in late 2011 and early 2012. These periods extend the portrait of stellar rotation at 125 Myr to the lowest-mass stars and re-establish the Pleiades as a key benchmark for models of the transport and evolution of stellar angular momentum. Combining our new P {sub rot} with precise BVIJHK photometry reported by Stauffer et al. and Kamai et al., we investigate known anomalies in the photometric properties of K and M Pleiades members. We confirm the correlation detected by Kamai et al. between a star's P {sub rot} and position relative to the main sequence in the cluster's color–magnitude diagram. We find that rapid rotators have redder ( V − K ) colors than slower rotators at the same V , indicating that rapid and slow rotators have different binary frequencies and/or photospheric properties. We find no difference in the photometric amplitudes of rapid and slow rotators, indicating that asymmetries in the longitudinal distribution of starspots do not scale grossly with rotation rate.« less

Rotation periods for nearby, mid-to-late M dwarfs estimated from the MEarth Project

NASA Astrophysics Data System (ADS)

Newton, Elisabeth R.; Irwin, Jonathan; Charbonneau, David; Berta-Thompson, Zachory K.; Dittmann, Jason

2015-01-01

Knowledge of M dwarfs' rotation is essential to understanding the generation of their magnetic fields and the mechanism by which they lose angular momentum. It is also important for characterizing the environment of planets that might orbit them. The most direct way to infer rotation periods is from variations in stars' brightnesses as dark spots rotate in and out of view. Most rotation periods estimated prior to this decade are the result of dedicated photometric studies. If care is taken to preserve astrophysical variability and limit systematics, transiting planet surveys generate the high-cadence monitoring required to estimate stellar rotation periods. While targeted surveys of clusters have provided data at young ages, observations of field M dwarfs are required to constrain their late-term evolution. Rotation periods of the smallest stars are also needed: the Kepler mission produced exquisite light curves of several thousand cool dwarfs, but field stars below 0.3 solar masses are not well-represented in the sample. The MEarth Project is a transiting planet survey targeting mid-to-late M dwarfs within 33 parsecs; it provides a unique data set for exploring rotation in a large sample of fully convective stars. We present a catalog of rotation periods for these stars. Our measurements are particularly useful because many of the MEarth targets have parallaxes, multi-wavelength photometry, and optical and near-infrared spectra. We present our methods for estimating rotation periods and quantifying our uncertainties, and discuss our results in the context of other surveys.The MEarth project gratefully acknowledges funding from the David and Lucile Packard Fellowship for Science and Engineering, the National Science Foundation under grants AST-0807690, AST-1109468, and AST-1004488, and the John Templeton Foundation

Rotational Spectrum of Sarin.

PubMed

Walker, A. R. Hight; Suenram, R. D.; Samuels, Alan; Jensen, James; Ellzy, Michael W.; Lochner, J. Michael; Zeroka, Daniel

2001-05-01

As part of an effort to examine the possibility of using molecular-beam Fourier-transform microwave spectroscopy to unambiguously detect and monitor chemical warfare agents, we report the first observation and assignment of the rotational spectrum of the nerve agent Sarin (GB) (Methylphosphonofluoridic acid 1-methyl-ethyl ester, CAS #107-44-8) at frequencies between 10 and 22 GHz. Only one of the two low-energy conformers of this organophosphorus compound (C(4)H(10)FO(2)P) was observed in the rotationally cold (T(rot)<2 K) molecular beam. The experimental asymmetric-rotor ground-state rotational constants of this conformer are A=2874.0710(9) MHz, B=1168.5776(4) MHz, C=1056.3363(4) MHz (Type A standard uncertainties are given, i.e., 1sigma), as obtained from a least-squares analysis of 74 a-, b-, and c-type rotational transitions. Several of the transitions are split into doublets due to the internal rotation of the methyl group attached to the phosphorus. The three-fold-symmetry barrier to internal rotation estimated from these splittings is 677.0(4) cm(-1). Ab initio electronic structure calculations using Hartree-Fock, density functional, and Moller-Plesset perturbation theories have also been made. The structure of the lowest-energy conformer determined from a structural optimization at the MP2/6-311G(**) level of theory is consistent with our experimental findings. Copyright 2001 Academic Press.

Rotational Spectrum of Sarin

NASA Astrophysics Data System (ADS)

Walker, A. R. Hight; Suenram, R. D.; Samuels, Alan; Jensen, James; Ellzy, Michael W.; Lochner, J. Michael; Zeroka, Daniel

2001-05-01

As part of an effort to examine the possibility of using molecular-beam Fourier-transform microwave spectroscopy to unambiguously detect and monitor chemical warfare agents, we report the first observation and assignment of the rotational spectrum of the nerve agent Sarin (GB) (Methylphosphonofluoridic acid 1-methyl-ethyl ester, CAS #107-44-8) at frequencies between 10 and 22 GHz. Only one of the two low-energy conformers of this organophosphorus compound (C4H10FO2P) was observed in the rotationally cold (Trot<2 K) molecular beam. The experimental asymmetric-rotor ground-state rotational constants of this conformer are A=2874.0710(9) MHz, B=1168.5776(4) MHz, C=1056.3363(4) MHz (Type A standard uncertainties are given, i.e., 1σ), as obtained from a least-squares analysis of 74 a-, b-, and c-type rotational transitions. Several of the transitions are split into doublets due to the internal rotation of the methyl group attached to the phosphorus. The three-fold-symmetry barrier to internal rotation estimated from these splittings is 677.0(4) cm-1. Ab initio electronic structure calculations using Hartree-Fock, density functional, and Moller-Plesset perturbation theories have also been made. The structure of the lowest-energy conformer determined from a structural optimization at the MP2/6-311G** level of theory is consistent with our experimental findings.

Properties of human brain sodium channel α-subunits expressed in HEK293 cells and their modulation by carbamazepine, phenytoin and lamotrigine

PubMed Central

Qiao, Xin; Sun, Guangchun; Clare, Jeffrey J; Werkman, Taco R; Wadman, Wytse J

2014-01-01

Background and purpose Voltage-activated Na+ channels contain one distinct α-subunit. In the brain NaV1.1, NaV1.2, NaV1.3 and NaV1.6 are the four most abundantly expressed α-subunits. The antiepileptic drugs (AEDs) carbamazepine, phenytoin and lamotrigine have voltage-gated Na+ channels as their primary therapeutic targets. This study provides a systematic comparison of the biophysical properties of these four α-subunits and characterizes their interaction with carbamazepine, phenytoin and lamotrigine. Experimental approach Na+ currents were recorded in voltage-clamp mode in HEK293 cells stably expressing one of the four α-subunits. Key results NaV1.2 and NaV1.3 subunits have a relatively slow recovery from inactivation, compared with the other subunits and NaV1.1 subunits generate the largest window current. Lamotrigine evokes a larger maximal shift of the steady-state inactivation relationship than carbamazepine or phenytoin. Carbamazepine shows the highest binding rate to the α-subunits. Lamotrigine binding to NaV1.1 subunits is faster than to the other α-subunits. Lamotrigine unbinding from the α-subunits is slower than that of carbamazepine and phenytoin. Conclusions and implications The four Na+ channel α-subunits show subtle differences in their biophysical properties, which, in combination with their (sub)cellular expression patterns in the brain, could contribute to differences in neuronal excitability. We also observed differences in the parameters that characterize AED binding to the Na+ channel subunits. Particularly, lamotrigine binding to the four α-subunits suggests a subunit-specific response. Such differences will have consequences for the clinical efficacy of AEDs. Knowledge of the biophysical and binding parameters could be employed to optimize therapeutic strategies and drug development. PMID:24283699

Electrophysiology and Beyond: Multiple roles of Na+ channel β subunits in development and disease

PubMed Central

Patino, Gustavo A.; Isom, Lori L.

2010-01-01

Voltage-gated Na+ channel (VGSC) β subunits are not “auxiliary.” These multifunctional molecules not only modulate Na+ current (INa), but also function as cell adhesion molecules (CAMs) – playing roles in aggregation, migration, invasion, neurite outgrowth, and axonal fasciculation. β subunits are integral members of VGSC signaling complexes at nodes of Ranvier, axon initial segments, and cardiac intercalated disks, regulating action potential propagation through critical intermolecular and cell-cell communication events. At least in vitro, many β subunit cell adhesive functions occur both in the presence and absence of pore-forming VGSC α subunits, and in vivo β subunits are expressed in excitable as well as non-excitable cells, thus β subunits may play important functional roles on their own, in the absence of α subunits. VGSC β1 subunits are essential for life and appear to be especially important during brain development. Mutations in β subunit genes result in a variety of human neurological and cardiovascular diseases. Moreover, some cancer cells exhibit alterations in β subunit expression during metastasis. In short, these proteins, originally thought of as merely accessory to α subunits, are critical players in their own right in human health and disease. Here we discuss the role of VGSC β subunits in the nervous system. PMID:20600605

The Sulfur Oxygenase Reductase from the Mesophilic Bacterium Halothiobacillus neapolitanus Is a Highly Active Thermozyme

PubMed Central

Veith, Andreas; Botelho, Hugo M.; Kindinger, Florian; Gomes, Cláudio M.

2012-01-01

A biochemical, biophysical, and phylogenetic study of the sulfur oxygenase reductase (SOR) from the mesophilic gammaproteobacterium Halothiobacillus neapolitanus (HnSOR) was performed in order to determine the structural and biochemical properties of the enzyme. SOR proteins from 14 predominantly chemolithoautotrophic bacterial and archaeal species are currently available in public databases. Sequence alignment and phylogenetic analysis showed that they form a coherent protein family. The HnSOR purified from Escherichia coli after heterologous gene expression had a temperature range of activity of 10 to 99°C with an optimum at 80°C (42 U/mg protein). Sulfite, thiosulfate, and hydrogen sulfide were formed at various stoichiometries in a range between pH 5.4 and 11 (optimum pH 8.4). Circular dichroism (CD) spectroscopy and dynamic light scattering showed that the HnSOR adopts secondary and quaternary structures similar to those of the 24-subunit enzyme from the hyperthermophile Acidianus ambivalens (AaSOR). The melting point of the HnSOR was ≈20°C lower than that of the AaSOR, when analyzed with CD-monitored thermal unfolding. Homology modeling showed that the secondary structure elements of single subunits are conserved. Subtle changes in the pores of the outer shell and increased flexibility might contribute to activity at low temperature. We concluded that the thermostability was the result of a rigid protein core together with the stabilizing effect of the 24-subunit hollow sphere. PMID:22139503

New discoveries on the biology and detection of human chorionic gonadotropin

PubMed Central

Cole, Laurence A

2009-01-01

Human chorionic gonadotropin (hCG) is a glycoprotein hormone comprising 2 subunits, alpha and beta joined non covalently. While similar in structure to luteinizing hormone (LH), hCG exists in multiple hormonal and non-endocrine agents, rather than as a single molecule like LH and the other glycoprotein hormones. These are regular hCG, hyperglycosylated hCG and the free beta-subunit of hyperglycosylated hCG. For 88 years regular hCG has been known as a promoter of corpus luteal progesterone production, even though this function only explains 3 weeks of a full gestations production of regular hCG. Research in recent years has explained the full gestational production by demonstration of critical functions in trophoblast differentiation and in fetal nutrition through myometrial spiral artery angiogenesis. While regular hCG is made by fused villous syncytiotrophoblast cells, extravillous invasive cytotrophoblast cells make the variant hyperglycosylated hCG. This variant is an autocrine factor, acting on extravillous invasive cytotrophoblast cells to initiate and control invasion as occurs at implantation of pregnancy and the establishment of hemochorial placentation, and malignancy as occurs in invasive hydatidiform mole and choriocarcinoma. Hyperglycosylated hCG inhibits apoptosis in extravillous invasive cytotrophoblast cells promoting cell invasion, growth and malignancy. Other non-trophoblastic malignancies retro-differentiate and produce a hyperglycosylated free beta-subunit of hCG (hCG free beta). This has been shown to be an autocrine factor antagonizing apoptosis furthering cancer cell growth and malignancy. New applications have been demonstrated for total hCG measurements and detection of the 3 hCG variants in pregnancy detection, monitoring pregnancy outcome, determining risk for Down syndrome fetus, predicting preeclampsia, detecting pituitary hCG, detecting and managing gestational trophoblastic diseases, diagnosing quiescent gestational trophoblastic disease, diagnosing placental site trophoblastic tumor, managing testicular germ cell malignancies, and monitoring other human malignancies. There are very few molecules with such wide and varying functions as regular hCG and its variants, and very few tests with such a wide spectrum of clinical applications as total hCG. PMID:19171054

CIM at GE's factory of the future

NASA Astrophysics Data System (ADS)

Waldman, H.

Functional features of a highly automated aircraft component batch processing factory are described. The system has processing, working, and methodology components. A rotating parts operation installed 20 yr ago features a high density of numerically controlled machines, and is connected to a hierarchical network of data communications and apparatus for moving the rotating parts and tools of engines. Designs produced at one location in the country are sent by telephone link to other sites for development of manufacturing plans, tooling, numerical control programs, and process instructions for the rotating parts. Direct numerical control is implemented at the work stations, which have instructions stored on tape for back-up in case the host computer goes down. Each machine is automatically monitored at 48 points and notice of failure can originate from any point in the system.

Probing the gravitational Faraday rotation using quasar X-ray microlensing.

PubMed

Chen, Bin

2015-11-17

The effect of gravitational Faraday rotation was predicted in the 1950s, but there is currently no practical method for measuring this effect. Measuring this effect is important because it will provide new evidence for correctness of general relativity, in particular, in the strong field limit. We predict that the observed degree and angle of the X-ray polarization of a cosmologically distant quasar microlensed by the random star field in a foreground galaxy or cluster lens vary rapidly and concurrently with flux during caustic-crossing events using the first simulation of quasar X-ray microlensing polarization light curves. Therefore, it is possible to detect gravitational Faraday rotation by monitoring the X-ray polarization of gravitationally microlensed quasars. Detecting this effect will also confirm the strong gravity nature of quasar X-ray emission.

Inherent conformational flexibility of F1-ATPase α-subunit.

PubMed

Hahn-Herrera, Otto; Salcedo, Guillermo; Barril, Xavier; García-Hernández, Enrique

2016-09-01

The core of F1-ATPase consists of three catalytic (β) and three noncatalytic (α) subunits, forming a hexameric ring in alternating positions. A wealth of experimental and theoretical data has provided a detailed picture of the complex role played by catalytic subunits. Although major conformational changes have only been seen in β-subunits, it is clear that α-subunits have to respond to these changes in order to be able to transmit information during the rotary mechanism. However, the conformational behavior of α-subunits has not been explored in detail. Here, we have combined unbiased molecular dynamics (MD) simulations and calorimetrically measured thermodynamic signatures to investigate the conformational flexibility of isolated α-subunits, as a step toward deepening our understanding of its function inside the α3β3 ring. The simulations indicate that the open-to-closed conformational transition of the α-subunit is essentially barrierless, which is ideal to accompany and transmit the movement of the catalytic subunits. Calorimetric measurements of the recombinant α-subunit from Geobacillus kaustophilus indicate that the isolated subunit undergoes no significant conformational changes upon nucleotide binding. Simulations confirm that the nucleotide-free and nucleotide-bound subunits show average conformations similar to that observed in the F1 crystal structure, but they reveal an increased conformational flexibility of the isolated α-subunit upon MgATP binding, which might explain the evolutionary conserved capacity of α-subunits to recognize nucleotides with considerable strength. Furthermore, we elucidate the different dependencies that α- and β-subunits show on Mg(II) for recognizing ATP. Copyright © 2016 Elsevier B.V. All rights reserved.

A Family of G Protein βγ Subunits Translocate Reversibly from the Plasma Membrane to Endomembranes on Receptor Activation*S

PubMed Central

Saini, Deepak Kumar; Kalyanaraman, Vani; Chisari, Mariangela; Gautam, Narasimhan

2008-01-01

The present model of G protein activation by G protein-coupled receptors exclusively localizes their activation and function to the plasma membrane (PM). Observation of the spatiotemporal response of G protein subunits in a living cell to receptor activation showed that 6 of the 12 members of the G protein γ subunit family translocate specifically from the PM to endomembranes. The γ subunits translocate as βγ complexes, whereas the α subunit is retained on the PM. Depending on the γ subunit, translocation occurs predominantly to the Golgi complex or the endoplasmic reticulum. The rate of translocation also varies with the γ subunit type. Different γ subunits, thus, confer distinct spatiotemporal properties to translocation. A striking relationship exists between the amino acid sequences of various γ subunits and their translocation properties. γ subunits with similar translocation properties are more closely related to each other. Consistent with this relationship, introducing residues conserved in translocating subunits into a non-translocating subunit results in a gain of function. Inhibitors of vesicle-mediated trafficking and palmitoylation suggest that translocation is diffusion-mediated and controlled by acylation similar to the shuttling of G protein subunits (Chisari, M., Saini, D. K., Kalyanaraman, V., and Gautam, N. (2007) J. Biol. Chem. 282, 24092–24098). These results suggest that the continual testing of cytosolic surfaces of cell membranes by G protein subunits facilitates an activated cell surface receptor to direct potentially active G protein βγ subunits to intracellular membranes. PMID:17581822

Loss of Complex I activity in the Escherichia coli enzyme results from truncating the C-terminus of subunit K, but not from cross-linking it to subunits N or L.

PubMed

Zhu, Shaotong; Canales, Alejandra; Bedair, Mai; Vik, Steven B

2016-06-01

Complex I is a multi-subunit enzyme of the respiratory chain with seven core subunits in its membrane arm (A, H, J, K, L, M, and N). In the enzyme from Escherichia coli the C-terminal ten amino acids of subunit K lie along the lateral helix of subunit L, and contribute to a junction of subunits K, L and N on the cytoplasmic surface. Using double cysteine mutagenesis, the cross-linking of subunit K (R99C) to either subunit L (K581C) or subunit N (T292C) was attempted. A partial yield of cross-linked product had no effect on the activity of the enzyme, or on proton translocation, suggesting that the C-terminus of subunit K has no dynamic role in function. To further elucidate the role of subunit K genetic deletions were constructed at the C-terminus. Upon the serial deletion of the last 4 residues of the C-terminus of subunit K, various results were obtained. Deletion of one amino acid had little effect on the activity of Complex I, but deletions of 2 or more amino acids led to total loss of enzyme activity and diminished levels of subunits L, M, and N in preparations of membrane vesicles. Together these results suggest that while the C-terminus of subunit K has no dynamic role in energy transduction by Complex I, it is vital for the correct assembly of the enzyme.

Fluorescence excitation-emission matrix spectroscopy for degradation monitoring of machinery lubricants

NASA Astrophysics Data System (ADS)

Sosnovski, Oleg; Suresh, Pooja; Dudelzak, Alexander E.; Green, Benjamin

2018-02-01

Lubrication oil is a vital component of heavy rotating machinery defining the machine's health, operational safety and effectiveness. Recently, the focus has been on developing sensors that provide real-time/online monitoring of oil condition/lubricity. Industrial practices and standards for assessing oil condition involve various analytical methods. Most these techniques are unsuitable for online applications. The paper presents the results of studying degradation of antioxidant additives in machinery lubricants using Fluorescence Excitation-Emission Matrix (EEM) Spectroscopy and Machine Learning techniques. EEM Spectroscopy is capable of rapid and even standoff sensing; it is potentially applicable to real-time online monitoring.

Automated pharmaceutical tablet coating layer evaluation of optical coherence tomography images

NASA Astrophysics Data System (ADS)

Markl, Daniel; Hannesschläger, Günther; Sacher, Stephan; Leitner, Michael; Khinast, Johannes G.; Buchsbaum, Andreas

2015-03-01

Film coating of pharmaceutical tablets is often applied to influence the drug release behaviour. The coating characteristics such as thickness and uniformity are critical quality parameters, which need to be precisely controlled. Optical coherence tomography (OCT) shows not only high potential for off-line quality control of film-coated tablets but also for in-line monitoring of coating processes. However, an in-line quality control tool must be able to determine coating thickness measurements automatically and in real-time. This study proposes an automatic thickness evaluation algorithm for bi-convex tables, which provides about 1000 thickness measurements within 1 s. Beside the segmentation of the coating layer, optical distortions due to refraction of the beam by the air/coating interface are corrected. Moreover, during in-line monitoring the tablets might be in oblique orientation, which needs to be considered in the algorithm design. Experiments were conducted where the tablet was rotated to specified angles. Manual and automatic thickness measurements were compared for varying coating thicknesses, angles of rotations, and beam displacements (i.e. lateral displacement between successive depth scans). The automatic thickness determination algorithm provides highly accurate results up to an angle of rotation of 30°. The computation time was reduced to 0.53 s for 700 thickness measurements by introducing feasibility constraints in the algorithm.

A novel approach of magnetorheological abrasive fluid finishing with swirling-assisted inlet flow

NASA Astrophysics Data System (ADS)

Kheradmand, Saeid; Esmailian, Mojtaba; Fatahy, A.

Abrasive flow machining has been the pioneer of new finishing processes. Rotating workpiece and imposing a magnetic field using magnetorheological working medium are some assisting manipulations to improve surface finishing, because they can increase the forces on the workpiece surface. Similarly, swirling the inlet flow using stationary swirler vanes, as a novel idea, may also increase forces on the surface, and then raise the material removal, with a lower expense and energy consumption compared with the case of workpiece rotation. Thus, in this paper, surface roughness improvement is studied in a pipe with rotating inlet flow of a magnetorheological finishing medium under imposing a magnetic field. The results are compared with the case of rotating workpiece, using 3D numerical simulation. The governing hydrodynamic parameters are investigated in both cases to monitor the flow variations. It is shown that surface roughness is improved by rotating inlet flow. However, it is found that finishing in the entrance length of swirling-assisted inlet flow can be so economical for short length workpieces, compared with the case of rotating workpiece, with very near Ra values. By comparison of the numerical results and published experimental data, current study also shows the ability of the numerical simulation, as an inexpensive and efficient tool, to predict the surface roughness changes in finishing processes.

VizieR Online Data Catalog: Photometric rotation periods of stars in α Per (Prosser+ 1997)

NASA Astrophysics Data System (ADS)

Prosser, C. F.; Grankin, K. N.

2013-07-01

Members of the Alpha Perseus open cluster were monitored and their rotation periods and amplitudes were derived. These are combined with their physical characteristics to estimate rotational velocities. V-band observations of α Per stars were obtained by both C.P. and K.G. Observations by C.P. were obtained with the Whipple Observatory 48-in. telescope on Mt. Hopkins. Observations by K.G. were obtained during photometric conditions at Mt. Maidanak (Tashkent) Observatory, Uzbekistan using a 0.48m telescope. K.G. obtained absolute V (Johnson) photometric magnitudes by observing five standard stars several times each night. All together, rotational period information was obtained for 35 members of the α Per cluster in Aug-Dec 1994 and Oct, Dec 1995. This work was never published in a refereed journal because Charles Prosser was killed in an auto accident in 1998. See http://aas.org/obituaries/charles-franklin-prosser-jr-1963-1998 (1 data file).

In situ synchrotron study of electromigration induced grain rotations in Sn solder joints

NASA Astrophysics Data System (ADS)

Shen, Hao; Zhu, Wenxin; Li, Yao; Tamura, Nobumichi; Chen, Kai

2016-04-01

Here we report an in situ study of the early stage of microstructure evolution induced by electromigration in a Pb-free β-Sn based solder joint by synchrotron polychromatic X-ray microdiffraction. With this technique, crystal orientation evolution is monitored at intragranular levels with high spatial and angular resolution. During the entire experiment, no crystal growth is detected, and rigid grain rotation is observed only in the two grains within the current crowding region, where high density and divergence of electric current occur. Theoretical calculation indicates that the trend of electrical resistance drop still holds under the present conditions in the grain with high electrical resistivity, while the other grain with low resistivity reorients to align its a-axis more parallel with the ones of its neighboring grains. A detailed study of dislocation densities and subgrain boundaries suggests that grain rotation in β-Sn, unlike grain rotation in high melting temperature metals which undergo displacive deformation, is accomplished via diffusional process mainly, due to the high homologous temperature.

Natural History of Rotator Cuff Disease and Implications on Management

PubMed Central

Hsu, Jason

2015-01-01

Degenerative rotator cuff disease is commonly associated with ageing and is often asymptomatic. The factors related to tear progression and pain development are just now being defined through longitudinal natural history studies. The majority of studies that follow conservatively treated painful cuff tears or asymptomatic tears that are monitored at regular intervals show slow progression of tear enlargement and muscle degeneration over time. These studies have highlighted greater risks for disease progression for certain variables, such as the presence of a full-thickness tear and involvement of the anterior aspect supraspinatus tendon. Coupling the knowledge of the natural history of degenerative cuff tear progression with variables associated with greater likelihood of successful tendon healing following surgery will allow better refinement of surgical indications for rotator cuff disease. In addition, natural history studies may better define the risks of nonoperative treatment over time. This article will review pertinent literature regarding degenerative rotator cuff disease with emphasis on variables important to defining appropriate initial treatments and refining surgical indications. PMID:26726288

Solar Cycle Fine Structure and Surface Rotation from Ca II K-Line Time Series Data

NASA Technical Reports Server (NTRS)

Scargle, Jeff; Keil, Steve; Worden, Pete

2011-01-01

Analysis of three and a half decades of data from the NSO/AFRL/Sac Peak K-line monitoring program yields evidence for four components to the variation: (a) the solar cycle, with considerable fine structure and a quasi-periodicity of 122.4 days; (b) a stochastic process, faster than (a) and largely independent of it, (c) a quasi-periodic signal due to rotational modulation, and of course (d) observational errors (shown to be quite small). Correlation and power spectrum analyses elucidate periodic and aperiodic variation of these chromospheric parameters. Time-frequency analysis is especially useful for extracting information about differential rotation, and in particular elucidates the connection between its behavior and fine structure of the solar cycle on approximately one-year time scales. These results further suggest that similar analyses will be useful at detecting and characterizing differential rotation in stars from stellar light-curves such as those being produced at NASA's Kepler observatory.

Differential rotation of stars with multiple transiting planets

NASA Astrophysics Data System (ADS)

Netto, Yuri; Valio, Adriana

2017-10-01

If a star hosts a planet in an orbit such that it eclipses the star periodically, can be estimated the rotation profile of this star. If planets in multiplanetary system occult different stellar areas, spots in more than one latitude of the stellar disc can be detected. The monitored study of theses starspots in different latitudes allow us to infer the rotation profile of the star. We use the model described in Silva (2003) to characterize the starspots of Kepler-210, an active star with two planets. Kepler-210 is a late K star with an estimated age of 350 +/- 50 Myrs, average rotation period of 12.33 days, mass of 0.63 M⊙ and radius of 0.69 R⊙. The planets that eclipses this star have radii of 0.0498 R s and 0.0635 R s with orbital periods of 2.4532 +/- 0.0007 days and 7.9725 +/- 0.0014 days, respectively, where R s is the star radius.

Developmental effects of wheel running on hippocampal glutamate receptor expression in young and mature adult rats.

PubMed

Staples, M C; Somkuwar, S S; Mandyam, C D

2015-10-01

Recent evidence suggests that the behavioral benefits associated with voluntary wheel running in rodents may be due to modulation of glutamatergic transmission in the hippocampus, a brain region implicated in learning and memory. However, the expression of the glutamatergic ionotropic N-methyl-d-aspartate receptor (GluN) in the hippocampus in response to chronic sustained voluntary wheel running has not yet been investigated. Further, the developmental effects during young and mature adulthood on wheel running output and GluN expression in hippocampal subregions has not been determined, and therefore is the main focus of this investigation. Eight-week-old and 16-week-old male Wistar rats were housed in home cages with free access to running wheels and running output was monitored for 4weeks. Wheel access was terminated and tissues from the dorsal and ventral hippocampi were processed for Western blot analysis of GluN subunit expression. Young adult runners demonstrated an escalation in running output but this behavior was not evident in mature adult runners. In parallel, young adult runners demonstrated a significant increase in total GluN (1 and 2A) subunit expression in the dorsal hippocampus (DH), and an opposing effect in the ventral hippocampus (VH) compared to age-matched sedentary controls; these changes in total protein expression were not associated with significant alterations in the phosphorylation of the GluN subunits. In contrast, mature adult runners demonstrated a reduction in total GluN2A expression in the DH, without producing alterations in the VH compared to age-matched sedentary controls. In conclusion, differential running activity-mediated modulation of GluN subunit expression in the hippocampal subregions was revealed to be associated with developmental effects on running activity, which may contribute to altered hippocampal synaptic activity and behavioral outcomes in young and mature adult subjects. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

Deciphering the function of the CNGB1b subunit in olfactory CNG channels.

PubMed

Nache, Vasilica; Wongsamitkul, Nisa; Kusch, Jana; Zimmer, Thomas; Schwede, Frank; Benndorf, Klaus

2016-07-11

Olfactory cyclic nucleotide-gated (CNG) ion channels are key players in the signal transduction cascade of olfactory sensory neurons. The second messengers cAMP and cGMP directly activate these channels, generating a depolarizing receptor potential. Olfactory CNG channels are composed of two CNGA2 subunits and two modulatory subunits, CNGA4, and CNGB1b. So far the exact role of the modulatory subunits for channel activation is not fully understood. By measuring ligand binding and channel activation simultaneously, we show that in functional heterotetrameric channels not only the CNGA2 subunits and the CNGA4 subunit but also the CNGB1b subunit binds cyclic nucleotides and, moreover, also alone translates this signal to open the pore. In addition, we show that the CNGB1b subunit is the most sensitive subunit in a heterotetrameric channel to cyclic nucleotides and that it accelerates deactivation to a similar extent as does the CNGA4 subunit. In conclusion, the CNGB1b subunit participates in ligand-gated activation of olfactory CNG channels and, particularly, contributes to rapid termination of odorant signal in an olfactory sensory neuron.

Sparse filtering with the generalized lp/lq norm and its applications to the condition monitoring of rotating machinery

NASA Astrophysics Data System (ADS)

Jia, Xiaodong; Zhao, Ming; Di, Yuan; Li, Pin; Lee, Jay

2018-03-01

Sparsity is becoming a more and more important topic in the area of machine learning and signal processing recently. One big family of sparse measures in current literature is the generalized lp /lq norm, which is scale invariant and is widely regarded as normalized lp norm. However, the characteristics of the generalized lp /lq norm are still less discussed and its application to the condition monitoring of rotating devices has been still unexplored. In this study, we firstly discuss the characteristics of the generalized lp /lq norm for sparse optimization and then propose a method of sparse filtering with the generalized lp /lq norm for the purpose of impulsive signature enhancement. Further driven by the trend of industrial big data and the need of reducing maintenance cost for industrial equipment, the proposed sparse filter is customized for vibration signal processing and also implemented on bearing and gearbox for the purpose of condition monitoring. Based on the results from the industrial implementations in this paper, the proposed method has been found to be a promising tool for impulsive feature enhancement, and the superiority of the proposed method over previous methods is also demonstrated.

Superstructure for high current applications in superconducting linear accelerators

DOEpatents

Sekutowicz, Jacek [Elbchaussee, DE; Kneisel, Peter [Williamsburg, VA

2008-03-18

A superstructure for accelerating charged particles at relativistic speeds. The superstructure consists of two weakly coupled multi-cell subunits equipped with HOM couplers. A beam pipe connects the subunits and an HOM damper is included at the entrance and the exit of each of the subunits. A coupling device feeds rf power into the subunits. The subunits are constructed of niobium and maintained at cryogenic temperatures. The length of the beam pipe between the subunits is selected to provide synchronism between particles and rf fields in both subunits.

Assembly of the stator in Escherichia coli ATP synthase. Complexation of alpha subunit with other F1 subunits is prerequisite for delta subunit binding to the N-terminal region of alpha

PubMed Central

Senior, Alan E.; Muharemagi, Alma; Wilke-Mounts, Susan

2008-01-01

Alpha subunit of Escherichia coli ATP synthase was expressed with a C-terminal 6-His tag and purified. Pure alpha was monomeric, competent in nucleotide binding, and had normal N-terminal sequence. In F1-subunit dissociation/reassociation experiments it supported full reconstitution of ATPase, and reassociated complexes were able to bind to F1-depleted membranes with restoration of ATP-driven proton pumping. Therefore interaction between the stator delta subunit and the N-terminal residue 1-22 region of alpha occurred normally when pure alpha was complexed with other F1 subunits. On the other hand, three different types of experiment showed that no interaction occurred between pure delta and isolated alpha subunit. Unlike in F1, the N-terminal region of isolated alpha was not susceptible to trypsin cleavage. Therefore, during assembly of ATP synthase, complexation of alpha subunit with other F1 subunits is prerequisite for delta subunit binding to the N-terminal region of alpha. We suggest that the N-terminal 1-22 residues of alpha are sequestered in isolated alpha until released by binding of beta to alpha subunit. This prevents 1/1 delta/alpha complexes from forming, and provides a satisfactory explanation of the stoichiometry of one delta per three alpha seen in the F1 sector of ATP synthase, assuming that steric hindrance prevents binding of more than one delta to the alpha3/beta3 hexagon. The cytoplasmic fragment of the b subunit (bsol) did not bind to isolated alpha. It might also be that complexation of alpha with beta subunits is prerequisite for direct binding of stator b subunit to the F1-sector. PMID:17176112

Topology of subunits of the mammalian cytochrome c oxidase: Relationship to the assembly of the enzyme complex

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

Yu-Zhong Zhang; Ewart, G.; Capaldi, R.A.

The arrangement of three subunits of beef heart cytochrome c oxidase, subunits Va, VIa, and VIII, has been explored by chemical labeling and protease digestion studies. Subunit Va is an extrinsic protein located on the C side of the mitochondrial inner membrane. This subunit was found to label with N-(4-azido-2-nitrophenyl)-2-aminoethane({sup 35}S)sulfonate and sodium methyl 4-({sup 3}H)formylphenyl phosphate in reconstituted vesicles in which 90% of cytochrome c oxidase complexes were oriented with the C domain outermost. Subunit VIa was cleaved by trypsin both in these reconstituted vesicles and in submitochondrial particles, indicating a transmembrane orientation. The epitope for a monoclonal antibodymore » (mAb) to subunit VIa was lost or destroyed when cleavage occurred in reconstituted vesicles. This epitope was localized to the C-terminal part of the subunit by antibody binding to a fusion protein consisting of glutathione S-transferase (G-ST) and the C-terminal amino acids 55-85 of subunit VIa. No antibody binding was obtained with a fusion protein containing G-ST and the N-terminal amino acids 1-55. The mAb reaction orients subunit VIa with its C-terminus in the C-domain. Subunit VIII was cleaved by trypsin in submitochondrial particles but not in reconstituted vesicles. N-Terminal sequencing of the subunit VIII cleavage produce from submitochondrial particles gave the same sequence as the untreated subunit, i.e., ITA, indicating that it is the C-terminus which is cleaved from the M side. Subunits Va and VIII each contain N-terminal extensions or leader sequences in the precursor polypeptides; subunit VIa is made without an N-terminal extension.« less

An Fe-S cluster in the conserved Cys-rich region in the catalytic subunit of FAD-dependent dehydrogenase complexes.

PubMed

Shiota, Masaki; Yamazaki, Tomohiko; Yoshimatsu, Keiichi; Kojima, Katsuhiro; Tsugawa, Wakako; Ferri, Stefano; Sode, Koji

2016-12-01

Several bacterial flavin adenine dinucleotide (FAD)-harboring dehydrogenase complexes comprise three distinct subunits: a catalytic subunit with FAD, a cytochrome c subunit containing three hemes, and a small subunit. Owing to the cytochrome c subunit, these dehydrogenase complexes have the potential to transfer electrons directly to an electrode. Despite various electrochemical applications and engineering studies of FAD-dependent dehydrogenase complexes, the intra/inter-molecular electron transfer pathway has not yet been revealed. In this study, we focused on the conserved Cys-rich region in the catalytic subunits using the catalytic subunit of FAD dependent glucose dehydrogenase complex (FADGDH) as a model, and site-directed mutagenesis and electron paramagnetic resonance (EPR) were performed. By co-expressing a hitch-hiker protein (γ-subunit) and a catalytic subunit (α-subunit), FADGDH γα complexes were prepared, and the properties of the catalytic subunit of both wild type and mutant FADGDHs were investigated. Substitution of the conserved Cys residues with Ser resulted in the loss of dye-mediated glucose dehydrogenase activity. ICP-AEM and EPR analyses of the wild-type FADGDH catalytic subunit revealed the presence of a 3Fe-4S-type iron-sulfur cluster, whereas none of the Ser-substituted mutants showed the EPR spectrum characteristic for this cluster. The results suggested that three Cys residues in the Cys-rich region constitute an iron-sulfur cluster that may play an important role in the electron transfer from FAD (intra-molecular) to the multi-heme cytochrome c subunit (inter-molecular) electron transfer pathway. These features appear to be conserved in the other three-subunit dehydrogenases having an FAD cofactor. Copyright © 2016 Elsevier B.V. All rights reserved.

DNA sequences, recombinant DNA molecules and processes for producing the A and B subunits of cholera toxin and preparations containing so-obtained subunit or subunits

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

Harford, N.; De Wilde, M.

1987-05-19

A recombinant DNA molecule is described comprising at least a portion coding for subunits A and B of cholera toxin, or a fragment or derivative of the portion wherein the fragment or derivative codes for a polypeptide have an activity which can induce an immune response to subunit A; can induce an immune response to subunit A and cause epithelial cell penetration and the enzymatic effect leading to net loss of fluid into the gut lumen; can bind to the membrane receptor for the B subunit of cholera toxin; can induce an immune response to subunit B; can induce anmore » immune response to subunit B and bind to the membrane receptor; or has a combination of the activities.« less

The first transmembrane domain (TM1) of β2-subunit binds to the transmembrane domain S1 of α-subunit in BK potassium channels

PubMed Central

Morera, Francisco J.; Alioua, Abderrahmane; Kundu, Pallob; Salazar, Marcelo; Gonzalez, Carlos; Martinez, Agustin D.; Stefani, Enrico; Toro, Ligia; Latorre, Ramon

2012-01-01

The BK channel is one of the most broadly expressed ion channels in mammals. In many tissues, the BK channel pore-forming α-subunit is associated to an auxiliary β-subunit that modulates the voltage- and Ca2+-dependent activation of the channel. Structural components present in β-subunits that are important for the physical association with the α-subunit are yet unknown. Here, we show through co-immunoprecipitation that the intracellular C-terminus, the second transmembrane domain (TM2) and the extracellular loop of the β2-subunit are dispensable for association with the α-subunit pointing transmembrane domain 1 (TM1) as responsible for the interaction. Indeed, the TOXCAT assay for transmembrane protein–protein interactions demonstrated for the first time that TM1 of the β2-subunit physically binds to the transmembrane S1 domain of the α-subunit. PMID:22710124

Multiple alpha subunits of integrin are involved in cell-mediated responses of the Manduca immune system.

PubMed

Zhuang, Shufei; Kelo, Lisha; Nardi, James B; Kanost, Michael R

2008-01-01

The cell-mediated responses of the insect innate immune system-phagocytosis, nodulation, encapsulation-involve multiple cell adhesion molecules of hemocyte surfaces. A hemocyte-specific (HS) integrin and a member of the immunoglobulin (Ig) superfamily (neuroglian) are involved in the encapsulation response of hemocytes in Manduca sexta. In addition, two new integrin alpha (alpha) subunits have been found on these hemocytes. The alpha2 subunit is mainly expressed in epidermis and Malphigian tubules, whereas the alpha3 subunit is primarily expressed on hemocytes and fat body cells. Of the three known alpha subunits, the alpha1 subunit found in HS integrin is the predominant subunit of hemocytes. Cell adhesion assays indicate that alpha2 belongs to the integrin family with RGD-binding motifs, confirming the phylogenetic analysis of alpha subunits based on the amino-acid sequence alignment of different alpha subunits. Double-stranded RNAs (dsRNAs) targeting each of these three integrin alpha subunits not only specifically decreased transcript expression of each alpha subunit in hemocytes, but also abolished the cell-mediated encapsulation response of hemocytes to foreign surfaces. The individual alpha subunits of M. sexta integrins, like their integrin counterparts in mammalian immune systems, have critical, individual roles in cell-substrate and cell-cell interactions during immune responses.

Mechanism of nucleotide sensing in group II chaperonins.

PubMed

Pereira, Jose H; Ralston, Corie Y; Douglas, Nicholai R; Kumar, Ramya; Lopez, Tom; McAndrew, Ryan P; Knee, Kelly M; King, Jonathan A; Frydman, Judith; Adams, Paul D

2012-02-01

Group II chaperonins mediate protein folding in an ATP-dependent manner in eukaryotes and archaea. The binding of ATP and subsequent hydrolysis promotes the closure of the multi-subunit rings where protein folding occurs. The mechanism by which local changes in the nucleotide-binding site are communicated between individual subunits is unknown. The crystal structure of the archaeal chaperonin from Methanococcus maripaludis in several nucleotides bound states reveals the local conformational changes associated with ATP hydrolysis. Residue Lys-161, which is extremely conserved among group II chaperonins, forms interactions with the γ-phosphate of ATP but shows a different orientation in the presence of ADP. The loss of the ATP γ-phosphate interaction with Lys-161 in the ADP state promotes a significant rearrangement of a loop consisting of residues 160-169. We propose that Lys-161 functions as an ATP sensor and that 160-169 constitutes a nucleotide-sensing loop (NSL) that monitors the presence of the γ-phosphate. Functional analysis using NSL mutants shows a significant decrease in ATPase activity, suggesting that the NSL is involved in timing of the protein folding cycle.

Utility of COX1 phylogenetics to differentiate between locally acquired and imported Plasmodium knowlesi infections in Singapore

PubMed Central

Loh, Jin Phang; Gao, Qiu Han Christine; Lee, Vernon J; Tetteh, Kevin; Drakeley, Chris

2016-01-01

INTRODUCTION Although there have been several phylogenetic studies on Plasmodium knowlesi (P. knowlesi), only cytochrome c oxidase subunit 1 (COX1) gene analysis has shown some geographical differentiation between the isolates of different countries. METHODS Phylogenetic analysis of locally acquired P. knowlesi infections, based on circumsporozoite, small subunit ribosomal ribonucleic acid (SSU rRNA), merozoite surface protein 1 and COX1 gene targets, was performed. The results were compared with the published sequences of regional isolates from Malaysia and Thailand. RESULTS Phylogenetic analysis of the circumsporozoite, SSU rRNA and merozoite surface protein 1 gene sequences for regional P. knowlesi isolates showed no obvious differentiation that could be attributed to their geographical origin. However, COX1 gene analysis showed that it was possible to differentiate between Singapore-acquired P. knowlesi infections and P. knowlesi infections from Peninsular Malaysia and Sarawak, Borneo, Malaysia. CONCLUSION The ability to differentiate between locally acquired P. knowlesi infections and imported P. knowlesi infections has important utility for the monitoring of P. knowlesi malaria control programmes in Singapore. PMID:26805667

Utility of COX1 phylogenetics to differentiate between locally acquired and imported Plasmodium knowlesi infections in Singapore.

PubMed

Loh, Jin Phang; Gao, Qiu Han Christine; Lee, Vernon J; Tetteh, Kevin; Drakeley, Chris

2016-12-01

Although there have been several phylogenetic studies on Plasmodium knowlesi (P. knowlesi), only cytochrome c oxidase subunit 1 (COX1) gene analysis has shown some geographical differentiation between the isolates of different countries. Phylogenetic analysis of locally acquired P. knowlesi infections, based on circumsporozoite, small subunit ribosomal ribonucleic acid (SSU rRNA), merozoite surface protein 1 and COX1 gene targets, was performed. The results were compared with the published sequences of regional isolates from Malaysia and Thailand. Phylogenetic analysis of the circumsporozoite, SSU rRNA and merozoite surface protein 1 gene sequences for regional P. knowlesi isolates showed no obvious differentiation that could be attributed to their geographical origin. However, COX1 gene analysis showed that it was possible to differentiate between Singapore-acquired P. knowlesi infections and P. knowlesi infections from Peninsular Malaysia and Sarawak, Borneo, Malaysia. The ability to differentiate between locally acquired P. knowlesi infections and imported P. knowlesi infections has important utility for the monitoring of P. knowlesi malaria control programmes in Singapore. Copyright: © Singapore Medical Association

Epidemiology of echinocandin resistance in Candida.

PubMed

Grossman, Nina T; Chiller, Tom M; Lockhart, Shawn R

2014-12-01

Echinocandins are the newest antifungal agents approved for use in treating Candida infections in the US. They act by interfering with 1,3-β-D-glucan synthase and therefore disrupt cell wall production and lead to Candida cell death. There is no intrinsic resistance to echinocandins among Candida species, and isolates from historic collections archived before the release of the echinocandins show no resistance. Resistance to the echinocandins remains low among most Candida species and ranges overall from 0-1%. Among isolates of Candida glabrata , the proportion of resistant isolates is higher and has been reported to be as high as 13.5% in at least one hospital. Antifungal resistance is due to specific amino acid mutations in the Fksp subunit(s) of the 1,3-β-D-glucan synthase protein which are localized to one of two hotspots. These mutations are being recognized in isolates from patients who have failed echinocandin therapy, and often lead to a poor outcome. While the future looks bright for the echinocandins against most Candida species, C. glabrata remains a species of concern and resistance rates of C. glabrata to the echinocandins should be monitored closely.

Origin and Evolution of the Sodium -Pumping NADH: Ubiquinone Oxidoreductase

PubMed Central

Reyes-Prieto, Adrian; Barquera, Blanca; Juárez, Oscar

2014-01-01

The sodium -pumping NADH: ubiquinone oxidoreductase (Na+-NQR) is the main ion pump and the primary entry site for electrons into the respiratory chain of many different types of pathogenic bacteria. This enzymatic complex creates a transmembrane gradient of sodium that is used by the cell to sustain ionic homeostasis, nutrient transport, ATP synthesis, flagellum rotation and other essential processes. Comparative genomics data demonstrate that the nqr operon, which encodes all Na+-NQR subunits, is found in a large variety of bacterial lineages with different habitats and metabolic strategies. Here we studied the distribution, origin and evolution of this enzymatic complex. The molecular phylogenetic analyses and the organizations of the nqr operon indicate that Na+-NQR evolved within the Chlorobi/Bacteroidetes group, after the duplication and subsequent neofunctionalization of the operon that encodes the homolog RNF complex. Subsequently, the nqr operon dispersed through multiple horizontal transfer events to other bacterial lineages such as Chlamydiae, Planctomyces and α, β, γ and δ -proteobacteria. Considering the biochemical properties of the Na+-NQR complex and its physiological role in different bacteria, we propose a detailed scenario to explain the molecular mechanisms that gave rise to its novel redox- dependent sodium -pumping activity. Our model postulates that the evolution of the Na+-NQR complex involved a functional divergence from its RNF homolog, following the duplication of the rnf operon, the loss of the rnfB gene and the recruitment of the reductase subunit of an aromatic monooxygenase. PMID:24809444

MOJAVE: MONITORING OF JETS IN ACTIVE GALACTIC NUCLEI WITH VLBA EXPERIMENTS. VIII. FARADAY ROTATION IN PARSEC-SCALE AGN JETS

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

Hovatta, Talvikki; Lister, Matthew L.; Aller, Margo F.

2012-10-01

We report observations of Faraday rotation measures for a sample of 191 extragalactic radio jets observed within the MOJAVE program. Multifrequency Very Long Baseline Array observations were carried out over 12 epochs in 2006 at four frequencies between 8 and 15 GHz. We detect parsec-scale Faraday rotation measures in 149 sources and find the quasars to have larger rotation measures on average than BL Lac objects. The median core rotation measures are significantly higher than in the jet components. This is especially true for quasars where we detect a significant negative correlation between the magnitude of the rotation measure andmore » the de-projected distance from the core. We perform detailed simulations of the observational errors of total intensity, polarization, and Faraday rotation, and concentrate on the errors of transverse Faraday rotation measure gradients in unresolved jets. Our simulations show that the finite image restoring beam size has a significant effect on the observed rotation measure gradients, and spurious gradients can occur due to noise in the data if the jet is less than two beams wide in polarization. We detect significant transverse rotation measure gradients in four sources (0923+392, 1226+023, 2230+114, and 2251+158). In 1226+023 the rotation measure is for the first time seen to change sign from positive to negative over the transverse cuts, which supports the presence of a helical magnetic field in the jet. In this source we also detect variations in the jet rotation measure over a timescale of three months, which are difficult to explain with external Faraday screens and suggest internal Faraday rotation. By comparing fractional polarization changes in jet components between the four frequency bands to depolarization models, we find that an external purely random Faraday screen viewed through only a few lines of sight can explain most of our polarization observations, but in some sources, such as 1226+023 and 2251+158, internal Faraday rotation is needed.« less

Power Stroke Angular Velocity Profiles of Archaeal A-ATP Synthase Versus Thermophilic and Mesophilic F-ATP Synthase Molecular Motors.

PubMed

Sielaff, Hendrik; Martin, James; Singh, Dhirendra; Biuković, Goran; Grüber, Gerhard; Frasch, Wayne D

2016-12-02

The angular velocities of ATPase-dependent power strokes as a function of the rotational position for the A-type molecular motor A 3 B 3 DF, from the Methanosarcina mazei Gö1 A-ATP synthase, and the thermophilic motor α 3 β 3 γ, from Geobacillus stearothermophilus (formerly known as Bacillus PS3) F-ATP synthase, are resolved at 5 μs resolution for the first time. Unexpectedly, the angular velocity profile of the A-type was closely similar in the angular positions of accelerations and decelerations to the profiles of the evolutionarily distant F-type motors of thermophilic and mesophilic origins, and they differ only in the magnitude of their velocities. M. mazei A 3 B 3 DF power strokes occurred in 120° steps at saturating ATP concentrations like the F-type motors. However, because ATP-binding dwells did not interrupt the 120° steps at limiting ATP, ATP binding to A 3 B 3 DF must occur during the catalytic dwell. Elevated concentrations of ADP did not increase dwells occurring 40° after the catalytic dwell. In F-type motors, elevated ADP induces dwells 40° after the catalytic dwell and slows the overall velocity. The similarities in these power stroke profiles are consistent with a common rotational mechanism for A-type and F-type rotary motors, in which the angular velocity is limited by the rotary position at which ATP binding occurs and by the drag imposed on the axle as it rotates within the ring of stator subunits. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Specific Roles of NMDA Receptor Subunits in Mental Disorders.

PubMed

Yamamoto, H; Hagino, Y; Kasai, S; Ikeda, K

2015-01-01

N-methyl-D-aspartate (NMDA) receptor plays important roles in learning and memory. NMDA receptors are a tetramer that consists of two glycine-binding subunits GluN1, two glutamate-binding subunits (i.e., GluN2A, GluN2B, GluN2C, and GluN2D), a combination of a GluN2 subunit and glycine-binding GluN3 subunit (i.e., GluN3A or GluN3B), or two GluN3 subunits. Recent studies revealed that the specific expression and distribution of each subunit are deeply involved in neural excitability, plasticity, and synaptic deficits. The present article summarizes reports on the dysfunction of NMDA receptors and responsible subunits in various neurological and psychiatric disorders, including schizophrenia, autoimmune-induced glutamatergic receptor dysfunction, mood disorders, and autism. A key role for the GluN2D subunit in NMDA receptor antagonist-induced psychosis has been recently revealed.

Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR Is Essential for PROTEIN PHOSPHATASE 2A Holoenzyme Assembly and Plays Important Roles in Hormone Signaling, Salt Stress Response, and Plant Development1[W][OPEN

PubMed Central

Chen, Jian; Hu, Rongbin; Zhu, Yinfeng; Shen, Guoxin; Zhang, Hong

2014-01-01

PROTEIN PHOSPHATASE 2A (PP2A) is a major group of serine/threonine protein phosphatases in eukaryotes. It is composed of three subunits: scaffolding subunit A, regulatory subunit B, and catalytic subunit C. Assembly of the PP2A holoenzyme in Arabidopsis (Arabidopsis thaliana) depends on Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR (AtPTPA). Reduced expression of AtPTPA leads to severe defects in plant development, altered responses to abscisic acid, ethylene, and sodium chloride, and decreased PP2A activity. In particular, AtPTPA deficiency leads to decreased methylation in PP2A-C subunits (PP2Ac). Complete loss of PP2Ac methylation in the suppressor of brassinosteroid insensitive1 mutant leads to 30% reduction of PP2A activity, suggesting that PP2A with a methylated C subunit is more active than PP2A with an unmethylated C subunit. Like AtPTPA, PP2A-A subunits are also required for PP2Ac methylation. The interaction between AtPTPA and PP2Ac is A subunit dependent. In addition, AtPTPA deficiency leads to reduced interactions of B subunits with C subunits, resulting in reduced functional PP2A holoenzyme formation. Thus, AtPTPA is a critical factor for committing the subunit A/subunit C dimer toward PP2A heterotrimer formation. PMID:25281708

Assembly of the epithelial Na+ channel evaluated using sucrose gradient sedimentation analysis.

PubMed

Cheng, C; Prince, L S; Snyder, P M; Welsh, M J

1998-08-28

Three subunits, alpha, beta, and gamma, contribute to the formation of the epithelial Na+ channel. To investigate the oligomeric assembly of the channel complex, we used sucrose gradient sedimentation analysis to determine the sedimentation properties of individual subunits and heteromultimers comprised of multiple subunits. When the alpha subunit was expressed alone, it first formed an oligomeric complex with a sedimentation coefficient of 11 S, and then generated a higher order multimer of 25 S. In contrast, individual beta and gamma subunits predominately assembled into 11 S complexes. We obtained similar results with expression in cells and in vitro. When we co-expressed beta with alpha or with alpha plus gamma, the beta subunit assembled into a 25 S complex. Glycosylation of the alpha subunit was not required for assembly into a 25 S complex. We found that the alpha subunit formed intra-chain disulfide bonds. Although such bonds were not required to generate an oligomeric complex, under nonreducing conditions the alpha subunit formed a complex that migrated more homogeneously at 25 S. This suggests that intra-chain disulfide bonds may stabilize the complex. These data suggest that the epithelial Na+ channel subunits form high order oligomeric complexes and that the alpha subunit contains the information that facilitates such formation. Interestingly, the ability of the alpha, but not the beta or gamma, subunit to assemble into a 25 S homomeric complex correlates with the ability of these subunits to generate functional channels when expressed alone.

Assembly and mechanism of a group II ECF transporter.

PubMed

Karpowich, Nathan K; Wang, Da-Neng

2013-02-12

Energy-coupling factor (ECF) transporters are a recently discovered family of primary active transporters for micronutrients and vitamins, such as biotin, thiamine, and riboflavin. Found exclusively in archaea and bacteria, including the human pathogens Listeria, Streptococcus, and Staphylococcus, ECF transporters may be the only means of vitamin acquisition in these organisms. The subunit composition of ECF transporters is similar to that of ATP binding cassette (ABC) importers, whereby both systems share two homologous ATPase subunits (A and A'), a high affinity substrate-binding subunit (S), and a transmembrane coupling subunit (T). However, the S subunit of ECF transporters is an integral membrane protein, and the transmembrane coupling subunits do not share an obvious sequence homology between the two transporter families. Moreover, the subunit stoichiometry of ECF transporters is controversial, and the detailed molecular interactions between subunits and the conformational changes during substrate translocation are unknown. We have characterized the ECF transporters from Thermotoga maritima and Streptococcus thermophilus. Our data suggests a subunit stoichiometry of 2S:2T:1A:1A' and that S subunits for different substrates can be incorporated into the same transporter complex simultaneously. In the first crystal structure of the A-A' heterodimer, each subunit contains a novel motif called the Q-helix that plays a key role in subunit coupling with the T subunits. Taken together, these findings suggest a mechanism for coupling ATP binding and hydrolysis to transmembrane transport by ECF transporters.

Visual Vestibular Interaction in the Dynamic Visual Acuity Test during Voluntary Head Rotation

NASA Technical Reports Server (NTRS)

Lee, Moo Hoon; Durnford, Simon; Crowley, John; Rupert, Angus

1996-01-01

Although intact vestibular function is essential in maintaining spatial orientation, no good screening tests of vestibular function are available to the aviation community. High frequency voluntary head rotation was selected as a vestibular stimulus to isolate the vestibulo-ocular reflex (VOR) from visual influence. A dynamic visual acuity test that incorporates voluntary head rotation was evaluated as a potential vestibular function screening tool. Twenty-seven normal subjects performed voluntary sinusoidal head rotation at frequencies from 0.7-4.0 Hz under three different visual conditions: visually-enhanced VOR, normal VOR, and visually suppressed VOR. Standardized Baily-Lovie chart letters were presented on a computer monitor in front of the subject, who then was asked to read the letters while rotating his head horizontally. The electro-oculogram and dynamic visual acuity score were recorded and analyzed. There were no significant differences in gain or phase shift among three visual conditions in the frequency range of 2.8 to 4.0 Hz. The dynamic visual acuity score shifted less than 0.3 logMAR at frequencies under 2.0 Hz. The dynamic visual acuity test at frequencies a round 2.0 Hz can be recommended for evaluating vestibular function.

Production of rotational parts in small-series and computer-aided planning of its production engineering

NASA Astrophysics Data System (ADS)

Dudas, Illes; Berta, Miklos; Cser, Istvan

1998-12-01

Up-to-date manufacturing equipments of production of rotational parts in small series are lathe-centers and CNC grinding machines with high concentration of manufacturing operations. By the use of these machine tools it can be produced parts with requirements of increased accuracy and surface quality. In the lathe centers, which contain the manufacturing procedures of lathes using stationary tools and of drilling-milling machine tools using rotational tools, non-rotational surfaces of rotational parts can also be produced. The high concentration of manufacturing operations makes necessary the planning and programing of the measuring, monitoring and quality control into the technological process during manufacturing operation. In this way, taking into consideration the technological possibilities of lathe canters, the scope of computer aided technological planning duties significantly increases. It is trivial requirement to give only once the descriptions of the prefabricated parts and ready made parts. Starting taking into account these careful considerations we have been developing the planning system of technology of body of revolution on the base of GTIPROG/EC system which useful for programming of lathe centers. Out paper deals with the results of development and the occurring problems.

Use of Faraday-rotation data from beacon satellites to determine ionospheric corrections for interplanetary spacecraft navigation

NASA Technical Reports Server (NTRS)

Royden, H. N.; Green, D. W.; Walson, G. R.

1981-01-01

Faraday-rotation data from the linearly polarized 137-MHz beacons of the ATS-1, SIRIO, and Kiku-2 geosynchronous satellites are used to determine the ionospheric corrections to the range and Doppler data for interplanetary spacecraft navigation. The JPL operates the Deep Space Network of tracking stations for NASA; these stations monitor Faraday rotation with dual orthogonal, linearly polarized antennas, Teledyne polarization tracking receivers, analog-to-digital converter/scanners, and other support equipment. Computer software examines the Faraday data, resolves the pi ambiguities, constructs a continuous Faraday-rotation profile and converts the profile to columnar zenith total electron content at the ionospheric reference point; a second program computes the line-of-sight ionospheric correction for each pass of the spacecraft over each tracking complex. Line-of-sight ionospheric electron content using mapped Faraday-rotation data is compared with that using dispersive Doppler data from the Voyager spacecraft; a difference of about 0.4 meters, or 5 x 10 to the 16th electrons/sq m is obtained. The technique of determining the electron content of interplanetary plasma by subtraction of the ionospheric contribution is demonstrated on the plasma torus surrounding the orbit of Io.

Eye-Hand Coordination during Visuomotor Adaptation with Different Rotation Angles: Effects of Terminal Visual Feedback

PubMed Central

Rand, Miya K.; Rentsch, Sebastian

2016-01-01

This study examined adaptive changes of eye-hand coordination during a visuomotor rotation task under the use of terminal visual feedback. Young adults made reaching movements to targets on a digitizer while looking at targets on a monitor where the rotated feedback (a cursor) of hand movements appeared after each movement. Three rotation angles (30°, 75° and 150°) were examined in three groups in order to vary the task difficulty. The results showed that the 30° group gradually reduced direction errors of reaching with practice and adapted well to the visuomotor rotation. The 75° group made large direction errors of reaching, and the 150° group applied a 180° reversal shift from early practice. The 75°and 150° groups, however, overcompensated the respective rotations at the end of practice. Despite these group differences in adaptive changes of reaching, all groups gradually adapted gaze directions prior to reaching from the target area to the areas related to the final positions of reaching during the course of practice. The adaptive changes of both hand and eye movements in all groups mainly reflected adjustments of movement directions based on explicit knowledge of the applied rotation acquired through practice. Only the 30° group showed small implicit adaptation in both effectors. The results suggest that by adapting gaze directions from the target to the final position of reaching based on explicit knowledge of the visuomotor rotation, the oculomotor system supports the limb-motor system to make precise preplanned adjustments of reaching directions during learning of visuomotor rotation under terminal visual feedback. PMID:27812093

Method for measuring tri-axial lumbar motion angles using wearable sheet stretch sensors

PubMed Central

Nakamoto, Hiroyuki; Yamaji, Tokiya; Ootaka, Hideo; Bessho, Yusuke; Nakamura, Ryo; Ono, Rei

2017-01-01

Background Body movements, such as trunk flexion and rotation, are risk factors for low back pain in occupational settings, especially in healthcare workers. Wearable motion capture systems are potentially useful to monitor lower back movement in healthcare workers to help avoid the risk factors. In this study, we propose a novel system using sheet stretch sensors and investigate the system validity for estimating lower back movement. Methods Six volunteers (female:male = 1:1, mean age: 24.8 ± 4.0 years, height 166.7 ± 5.6 cm, weight 56.3 ± 7.6 kg) participated in test protocols that involved executing seven types of movements. The movements were three uniaxial trunk movements (i.e., trunk flexion-extension, trunk side-bending, and trunk rotation) and four multiaxial trunk movements (i.e., flexion + rotation, flexion + side-bending, side-bending + rotation, and moving around the cranial–caudal axis). Each trial lasted for approximately 30 s. Four stretch sensors were attached to each participant’s lower back. The lumbar motion angles were estimated using simple linear regression analysis based on the stretch sensor outputs and compared with those obtained by the optical motion capture system. Results The estimated lumbar motion angles showed a good correlation with the actual angles, with correlation values of r = 0.68 (SD = 0.35), r = 0.60 (SD = 0.19), and r = 0.72 (SD = 0.18) for the flexion-extension, side bending, and rotation movements, respectively (all P < 0.05). The estimation errors in all three directions were less than 3°. Conclusion The stretch sensors mounted on the back provided reasonable estimates of the lumbar motion angles. The novel motion capture system provided three directional angles without capture space limits. The wearable system possessed great potential to monitor the lower back movement in healthcare workers and helping prevent low back pain. PMID:29020053

The Role of RUB (related to ubiquitin) Family of Proteins in the Hormone Response. Final Report.

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

Callis, Judy

2013-03-22

The Rub pathway is a conserved protein modification pathway. RUB (called Rubp1 in budding yeast, Nedd8 in animals and RUB in plants) is a ubiquitin-like 76-amino acid protein. It covalently attaches to protein using an enzymatic machinery analogous to the enzymes that attach ubiquitin to its substrate proteins. However, the nature of the complement of Rub-modified proteins in organisms was not clear. From bioinformatics analyses, one can identify a Rub activating enzymes and Rub conjugating enzymes. However, in many cases, their biochemical properties were not described. In DOE-funded work, we made major advances in our understanding of the Rub pathwaymore » in yeast and plants, work that is applicable to other organisms as well. There is a multi-subunit enzyme called SCF in all eukaryotes. The SCF consists of several subunits that serve as a scaffold (the cullin, SKP and RBX subunits) and one subunit that interacts with the substrate. This cullin protein (called Cdc53p in yeast and CULLIN 1 in plants and animals) was a known Rub target. In this work, we identified additional Rub targets in yeast as the other cullin-like proteins Cul3p and Rtt101p. Additionally we described the conservation of the Rub pathway because plant RUB1 can conjugated to yeast Cdc53p- in yeast. In the model plant Arabidopsis thaliana, we characterized the Rub activating enzymes and showed that they are not biochemically equivalent. We also showed that the Rub pathway is essential in plants and characterized plants with reduced levels of rub proteins. These plants are affected in multiple developmental processes. We discovered that they over-produce ethylene as dark-grown seedlings. We characterized a mutant allele of CULLIN1 in Arabidopsis with impaired interaction with RBX and showed that it is unstable in vivo. We used our knowledge of monitoring protein degradation to map the degradation determinants in a plant transcription factor. Finally, we took a mass spectrometric approach to identify novel Rub targets in plants and identified DDB1a, a subunit of an different ubiquitin ligase as a potential Rub-modified protein. Altogether, these studies have advanced our knowledge of the Rub pathway in all organisms.« less

Development of a compact superconducting rotating-gantry for heavy-ion therapy

PubMed Central

Iwata, Yoshiyuki; Noda, K.; Murakami, T.; Shirai, T.; Furukawa, T.; Fujita, T.; Mori, S.; Sato, S.; Mizushima, K.; Shouda, K.; Fujimoto, T.; Arai, H.; Ogitsu, T.; Obana, T.; Amemiya, N.; Orikasa, T.; Takami, S.; Takayama, S.

2014-01-01

An isocentric superconducting rotating-gantry for heavy-ion therapy is being developed [ 1]. This rotating gantry can transport heavy ions having 430 MeV/u to an isocenter with irradiation angles of over ±180°, and is further capable of performing fast raster-scanning irradiation [ 2]. A layout of the beam-transport line for the compact rotating-gantry is presented in Fig. 1. The rotating gantry has 10 superconducting magnets (BM01-10), a pair of the scanning magnets (SCM-X and SCM-Y) and two pairs of beam profile- monitor and steering magnets (ST01-02 and PRN01-02). For BM01-BM06 and BM09-BM10, the combined-function superconducting magnets are employed. Further, these superconducting magnets are designed for fast slewing of the magnetic field to follow the multiple flattop operation of the synchrotron [ 3]. The use of the combined-function superconducting magnets with optimized beam optics allows a compact gantry design with a large scan size at the isocenter; the length and the radius of the gantry will be to be ∼13 and 5.5 m, respectively, which are comparable to those for the existing proton gantries. Furthermore, the maximum scan size at the isocenter is calculated to be as large as ∼200 mm square for heavy-ion beams at the maximum energy of 430 MeV/u. All of the superconducting magnets were designed, and their magnetic fields were calculated using the Opera-3d code [ 4]. With the calculated magnetic fields, beam-tracking simulations were made. The simulation results agreed well with those of the linear beam-optics calculation, proving validity of the final design for the superconducting magnets. The five out of 10 superconducting magnets, as well as the model magnet were currently manufactured. With these magnets, rotation tests, magnetic field measurements and fast slewing tests were conducted. However, we did not observe any significant temperature increase, which may cause a quench problem. Further, results of the magnetic field measurements roughly agreed with those calculated by the Opera-3d code. The design study as well as major tests of the superconducting magnets was completed, and the construction of the superconducting rotating-gantry is in progress. The construction of the superconducting rotating-gantry will be completed at the end of FY2014, and be commissioned within FY2015. Fig. 1.Layout of the superconducting rotating-gantry. The gantry consists of 10 superconducting magnets (BM01–BM10), a pair of the scanning magnets (SCM-X and SCMY), and two pairs of beam profile-monitor and steering magnets (STR01–STR02 and PRN01–PRN02).

Risk Assessment of Face Skin Exposure to UV Irradiance from Different Rotation Angle Ranges

PubMed Central

Wang, Fang; Gao, Qian; Deng, Yan; Chen, Rentong; Liu, Yang

2017-01-01

Ultraviolet (UV) is one of the environmental pathogenic factors causing skin damage. Aiming to assess the risk of face skin exposure to UV irradiance from different rotation angles, a rotating model was used to monitor the exposure of the skin on the face to UV irradiance, with skin damage action spectra used to determine the biologically effective UV irradiance (UVBEskin) and UVBEskin radiant exposure (HBEskin) causing skin damage. The results indicate that the UVBEskin is directly influenced by variations in rotation angles. A significant decrease of approximately 52.70% and 52.10% in UVBEskin was found when the cheek and nose measurement sites was rotated from 0° to 90°, while a decrease of approximately 62.70% was shown when the forehead measurement sites was rotated from an angle of 0° to 108°. When HBEskin was compared to the exposure limits (ELs; 30 J·m−2), the maximum relative risk ratios (RR) for cheek, nose, and forehead were found to be approximately 2.01, 2.40, and 2.90, respectively, which were all measured at a rotation angle of 0°. The maximal increase in the percentage of the average HBEskin for rotation angles of 60°, 120°, 180°, and 360° facing the sun to ELs were found to be approximately 62.10%, 52.72%, 43.43%, and 26.27% for the cheek; approximately 130.61%, 109.68%, 86.43%, and 50.06% for the nose; and approximately 178.61%, 159.19%, 134.38%, and 83.41% for the forehead, respectively. PMID:28587318

Risk Assessment of Face Skin Exposure to UV Irradiance from Different Rotation Angle Ranges.

PubMed

Wang, Fang; Gao, Qian; Deng, Yan; Chen, Rentong; Liu, Yang

2017-06-06

Ultraviolet (UV) is one of the environmental pathogenic factors causing skin damage. Aiming to assess the risk of face skin exposure to UV irradiance from different rotation angles, a rotating model was used to monitor the exposure of the skin on the face to UV irradiance, with skin damage action spectra used to determine the biologically effective UV irradiance (UVBE skin ) and UVBE skin radiant exposure (HBE skin ) causing skin damage. The results indicate that the UVBE skin is directly influenced by variations in rotation angles. A significant decrease of approximately 52.70% and 52.10% in UVBE skin was found when the cheek and nose measurement sites was rotated from 0° to 90°, while a decrease of approximately 62.70% was shown when the forehead measurement sites was rotated from an angle of 0° to 108°. When HBE skin was compared to the exposure limits (ELs; 30 J·m -2 ), the maximum relative risk ratios (RR) for cheek, nose, and forehead were found to be approximately 2.01, 2.40, and 2.90, respectively, which were all measured at a rotation angle of 0°. The maximal increase in the percentage of the average HBE skin for rotation angles of 60°, 120°, 180°, and 360° facing the sun to ELs were found to be approximately 62.10%, 52.72%, 43.43%, and 26.27% for the cheek; approximately 130.61%, 109.68%, 86.43%, and 50.06% for the nose; and approximately 178.61%, 159.19%, 134.38%, and 83.41% for the forehead, respectively.

Lumbar Corsets Can Decrease Lumbar Motion in Golf Swing

PubMed Central

Hashimoto, Koji; Miyamoto, Kei; Yanagawa, Takashi; Hattori, Ryo; Aoki, Takaaki; Matsuoka, Toshio; Ohno, Takatoshi; Shimizu, Katsuji

2013-01-01

Swinging a golf club includes the rotation and extension of the lumbar spine. Golf-related low back pain has been associated with degeneration of the lumbar facet and intervertebral discs, and with spondylolysis. Reflective markers were placed directly onto the skin of 11young male amateur golfers without a previous history of back pain. Using a VICON system (Oxford Metrics, U.K.), full golf swings were monitored without a corset (WOC), with a soft corset (SC), and with a hard corset (HC), with each subject taking 3 swings. Changes in the angle between the pelvis and the thorax (maximum range of motion and angular velocity) in 3 dimensions (lumbar rotation, flexion-extension, and lateral tilt) were analyzed, as was rotation of the hip joint. Peak changes in lumbar extension and rotation occurred just after impact with the ball. The extension angle of the lumbar spine at finish was significantly lower under SC (38°) or HC (28°) than under WOC (44°) conditions (p < 0.05). The maximum angular velocity after impact was significantly smaller under HC (94°/sec) than under SC (177°/sec) and WOC (191° /sec) conditions, as were the lumbar rotation angles at top and finish. In contrast, right hip rotation angles at top showed a compensatory increase under HC conditions. Wearing a lumbar corset while swinging a golf club can effectively decrease lumbar extension and rotation angles from impact until the end of the swing. These effects were significantly enhanced while wearing an HC. Key points Rotational and extension forces on the lumbar spine may cause golf-related low back pain Wearing lumbar corsets during a golf swing can effectively decrease lumbar extension and rotation angles and angular velocity. Wearing lumbar corsets increased the rotational motion of the hip joint while reducing the rotation of the lumbar spine. PMID:24149729

Lumbar corsets can decrease lumbar motion in golf swing.

PubMed

Hashimoto, Koji; Miyamoto, Kei; Yanagawa, Takashi; Hattori, Ryo; Aoki, Takaaki; Matsuoka, Toshio; Ohno, Takatoshi; Shimizu, Katsuji

2013-01-01

Swinging a golf club includes the rotation and extension of the lumbar spine. Golf-related low back pain has been associated with degeneration of the lumbar facet and intervertebral discs, and with spondylolysis. Reflective markers were placed directly onto the skin of 11young male amateur golfers without a previous history of back pain. Using a VICON system (Oxford Metrics, U.K.), full golf swings were monitored without a corset (WOC), with a soft corset (SC), and with a hard corset (HC), with each subject taking 3 swings. Changes in the angle between the pelvis and the thorax (maximum range of motion and angular velocity) in 3 dimensions (lumbar rotation, flexion-extension, and lateral tilt) were analyzed, as was rotation of the hip joint. Peak changes in lumbar extension and rotation occurred just after impact with the ball. The extension angle of the lumbar spine at finish was significantly lower under SC (38°) or HC (28°) than under WOC (44°) conditions (p < 0.05). The maximum angular velocity after impact was significantly smaller under HC (94°/sec) than under SC (177°/sec) and WOC (191° /sec) conditions, as were the lumbar rotation angles at top and finish. In contrast, right hip rotation angles at top showed a compensatory increase under HC conditions. Wearing a lumbar corset while swinging a golf club can effectively decrease lumbar extension and rotation angles from impact until the end of the swing. These effects were significantly enhanced while wearing an HC. Key pointsRotational and extension forces on the lumbar spine may cause golf-related low back painWearing lumbar corsets during a golf swing can effectively decrease lumbar extension and rotation angles and angular velocity.Wearing lumbar corsets increased the rotational motion of the hip joint while reducing the rotation of the lumbar spine.

Method and apparatus for monitoring the rotating frequency of de-energized induction motors

DOEpatents

Mikesell, H.E.; Lucy, E.

1998-02-03

The rotational speed of a coasting induction motor is measured by sensing e residual electrical voltages at the power terminals of the motor, thus eliminating the need for conventional tachometer equipment, additional mechanical components or modifications to the induction motor itself. The power terminal voltage signal is detected and transformed into a DC voltage proportional to the frequency of the signal. This DC voltage can be input to the control system of a variable frequency motor controller to regulate the output characteristics thereof relative to the speed of the coasting motor. 6 figs.

Method and apparatus for monitoring the rotating frequency of de-energized induction motors

DOEpatents

Mikesell, Harvey E.; Lucy, Eric

1998-01-01

The rotational speed of a coasting induction motor is measured by sensing e residual electrical voltages at the power terminals of the motor, thus eliminating the need for conventional tachometer equipment, additional mechanical components or modifications to the induction motor itself. The power terminal voltage signal is detected and transformed into a DC voltage proportional to the frequency of the signal. This DC voltage can be input to the control system of a variable frequency motor controller to regulate the output characteristics thereof relative to the speed of the coasting motor.

The equine LH/CGβ subunit combines divergent intracellular traits of the human LHβ and CGβ subunits

PubMed Central

Cohen, Limor; Bousfield, George R; Ben-Menahem, David

2017-01-01

The pituitary LHβ and placental CGβ subunits are products of different genes in primates. The major structural difference between the two subunits is in the carboxy-terminal region, where the short carboxyl sequence of hLHβ is replaced by a longer O-glycosylated carboxy-terminal peptide (CTP) in hCGβ. In association with this structural deviation, there are marked differences in the secretion kinetics and polarized routing of the two subunits. In equids, however, the CGβ and LHβ subunits are products of the same gene expressed in the placenta and pituitary (eLH/CGβ), and both contain a CTP. This unusual expression pattern intrigued us and led to our study of eLH/CGβ subunit secretion by transfected CHO and MDCK cells. In continuous labeling and pulse chase experiments, the secretion of the eLH/CGβ subunit from the transfected CHO cells was inefficient (medium recovery of 16–25%) and slow (t1/2 >6.5 hrs). This indicated that, the secretion of the eLH/CGβ subunit resembles that of hLHβ rather than hCGβ. In MDCK cells grown on Transwell filters, the eLH/CGβ subunit was preferentially secreted from the apical side, similar to the hCGβ subunit secretory route (~65% of the total protein secreted). Taken together, these data suggested that secretion of the eLH/CGβ subunit integrates features of both hLHβ and hCGβ subunits. We propose that the evolution of this intracellular behavior may fulfill the physiological demands for biosynthesis of the eLH/CGβ subunit in the pituitary as well as in the placenta. PMID:25796287

Dissociation energy and dynamics of water clusters

NASA Astrophysics Data System (ADS)

Ch'ng, Lee Chiat

The state-to-state vibrational predissociation (VP) dynamics of water clusters were studied following excitation of a vibrational mode of each cluster. Velocity-map imaging (VMI) and resonance-enhanced multiphoton ionization (REMPI) were used to determine pair-correlated center-of-mass translational energy distributions. Product energy distributions and dissociation energies were determined. Following vibrational excitation of the HCl stretch fundamental of the HCl-H2O dimer, HCl fragments were detected by 2 + 1 REMPI via the f 3□2(nu' = 0) ← X 1Sigma+(nu'' = 0) and V1Sigma + (nu' = 11 and 12) ← X1Sigma+ (nu'' = 0) transitions. REMPI spectra clearly show HCl from dissociation produced in the ground vibrational state with J'' up to 11. The fragments' center-of-mass translational energy distributions were determined from images of selected rotational states of HCl and were converted to rotational state distributions of the water cofragment. All the distributions could be fit well when using a dimer dissociation energy of bond dissociation energy D0 = 1334 +/- 10 cm--1. The rotational distributions in the water cofragment pair-correlated with specific rotational states of HCl appear nonstatistical when compared to predictions of the statistical phase space theory. A detailed analysis of pair-correlated state distributions was complicated by the large number of water rotational states available, but the data show that the water rotational populations increase with decreasing translational energy. H2O fragments of this dimer were detected by 2 + 1 REMPI via the C˜1B1(000) ← X˜1A1(000) transition. REMPI clearly shows that H2O from dissociation is produced in the ground vibrational state. The fragment's center-of-mass translational energy distributions were determined from images of selected rotational states of H2O and were converted to rotational state distributions of the HCl cofragment. The distributions gave D0 = 1334 +/- 10 cm --1 and show a clear preference for rotational levels in the HCl fragment that minimize translational energy release. The usefulness of 2 + 1 REMPI detection of water fragment is discussed. The hydrogen bonding in water is dominated by pair-wise dimer interactions, and the predissociation of the water dimer following vibrational excitation is reported. The measured D0 values of (H 2O)2 and (D2O)2, 1105 and 1244 +/- 10 cm--1, respectively, are in excellent agreement with the calculated values of 1103 and 1244 +/- 5 cm--1. Pair-correlated water fragment rovibrational state distributions following vibrational predissociation of (H2O)2 and (D2O) 2 were obtained upon excitation of the hydrogen bonded OH and OD stretch fundamentals, respectively. Quasiclassical trajectory calculations, using an accurate full-dimensional potential energy surface, are in accord with and help to elucidate experiment. Experiment and theory find predominant excitation of the fragment bending mode upon hydrogen bond breaking. A minor channel is also observed in which both fragments are in the ground vibrational state and are highly rotationally excited. The theoretical calculations reveal equal probability of bending excitation in the donor and acceptor subunits, which is a result of interchange of donor and acceptor roles. The rotational distributions associated with the major channel, in which one water fragment has one quantum of bend, and the minor channel with both water fragments in the ground vibrational state are calculated, and are in agreement with experiment. (Abstract shortened by UMI.)

Rotational Periods and Starspot Activity of Young Solar-Type Dwarfs in the Open Cluster IC 4665

NASA Technical Reports Server (NTRS)

Allain, S.; Bouvier, J.; Prosser, C.; Marschall, L. A.; Laaksonen, B. D.

1995-01-01

We present the results of a V-band photometric monitoring survey of 15 late-type dwarfs in the young open cluster IC 4665. Low-amplitude periodic light variations are found for 8 stars and ascribed to the modulation by starspots that cover typically a few percent of the stellar disk. Periods range from 0.6 to 3.7 d, translating to equatorial velocities between 13 and 93 km/s. That no period longer than 4 d was detected suggests a relative paucity of extremely slow rotators (V(sub eq) much less than 10 km/s) among late-type dwarfs in IC 4665. The fractional number of slow rotators in IC 4665 is similar to that of Alpha Per cluster, suggesting that IC 4665 is close in age to Alpha Per (approx. 50 Myr).

SPECTRAL VARIATIONS OF Of?p OBLIQUE MAGNETIC ROTATOR CANDIDATES IN THE MAGELLANIC CLOUDS

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

Walborn, Nolan R.; Morrell, Nidia I.; Nazé, Yaël

2015-10-15

Optical spectroscopic monitoring has been conducted of two O stars in the SMC and one in the LMC, the spectral characteristics of which place them in the Of?p category, which has been established in the Galaxy to consist of oblique magnetic rotators. All of these Magellanic stars show systematic spectral variations typical of the Of?p class, further strengthening their magnetic candidacy to the point of virtual certainty. The spectral variations are related to photometric variations derived from Optical Gravitational Lensing Experiment data by Nazé et al. in a parallel study, which yields rotational periods for two of them. Now circularmore » spectropolarimetry is required to measure their fields, and ultraviolet spectroscopy to further characterize their low-metallicity, magnetically confined winds, in support of hydrodynamical analyses.« less

Probing the gravitational Faraday rotation using quasar X-ray microlensing

PubMed Central

Chen, Bin

2015-01-01

The effect of gravitational Faraday rotation was predicted in the 1950s, but there is currently no practical method for measuring this effect. Measuring this effect is important because it will provide new evidence for correctness of general relativity, in particular, in the strong field limit. We predict that the observed degree and angle of the X-ray polarization of a cosmologically distant quasar microlensed by the random star field in a foreground galaxy or cluster lens vary rapidly and concurrently with flux during caustic-crossing events using the first simulation of quasar X-ray microlensing polarization light curves. Therefore, it is possible to detect gravitational Faraday rotation by monitoring the X-ray polarization of gravitationally microlensed quasars. Detecting this effect will also confirm the strong gravity nature of quasar X-ray emission. PMID:26574051

Semiconductor sensor for optically measuring polarization rotation of optical wavefronts using rare earth iron garnets

DOEpatents

Duncan, Paul G.

2002-01-01

Described are the design of a rare earth iron garnet sensor element, optical methods of interrogating the sensor element, methods of coupling the optical sensor element to a waveguide, and an optical and electrical processing system for monitoring the polarization rotation of a linearly polarized wavefront undergoing external modulation due to magnetic field or electrical current fluctuation. The sensor element uses the Faraday effect, an intrinsic property of certain rare-earth iron garnet materials, to rotate the polarization state of light in the presence of a magnetic field. The sensor element may be coated with a thin-film mirror to effectively double the optical path length, providing twice the sensitivity for a given field strength or temperature change. A semiconductor sensor system using a rare earth iron garnet sensor element is described.

Light-curve Modulation of Low-mass Stars in K2. I. Identification of 481 Fast Rotators in the Solar Neighborhood

NASA Astrophysics Data System (ADS)

Saylor, Dicy; Lepine, Sebastien; Crossfield, Ian; Petigura, Erik A.

2018-01-01

The K2 mission is targeting large numbers of nearby (d < 100 pc) GKM dwarfs selected from the SUPERBLINK proper motion survey (μ > 40 mas yr‑1, V < 20). Additionally, the mission is targeting low-mass, high proper motion stars associated with the local (d < 500 pc) Galactic halo population also selected from SUPERBLINK. K2 campaigns 0 through 8 monitored a total of 26,518 of these cool main-sequence stars. We used the auto-correlation function to search for fast rotators by identifying short-period photometric modulations in the K2 light curves. We identified 481 candidate fast rotators with rotation periods <4 days that show light-curve modulations consistent with starspots. Their kinematics show low average transverse velocities, suggesting that they are part of the young disk population. A subset (13) of the fast rotators is found among those targets with colors and kinematics consistent with the local Galactic halo population and may represent stars spun up by tidal interactions in close binary systems. We further demonstrate that the M dwarf fast rotators selected from the K2 light curves are significantly more likely to have UV excess and discuss the potential of the K2 mission to identify new nearby young GKM dwarfs on the basis of their fast rotation rates. Finally, we discuss the possible use of local halo stars as fiducial, non-variable sources in the Kepler fields.

Blood flow/pump rotation ratio as an artificial lung performance monitoring tool during extracorporeal respiratory support using centrifugal pumps.

PubMed

Park, Marcelo; Mendes, Pedro Vitale; Hirota, Adriana Sayuri; dos Santos, Edzangela Vasconcelos; Costa, Eduardo Leite Vieira; Azevedo, Luciano Cesar Pontes

2015-01-01

To analyze the correlations of the blood flow/pump rotation ratio and the transmembrane pressure, CO2 and O2 transfer during the extracorporeal respiratory support. Five animals were instrumented and submitted to extracorporeal membrane oxygenation in a five-step protocol, including abdominal sepsis and lung injury. This study showed that blood flow/pump rotations ratio variations are dependent on extracorporeal membrane oxygenation blood flow in a positive logarithmic fashion. Blood flow/pump rotation ratio variations are negatively associated with transmembrane pressure (R2 = 0.5 for blood flow = 1500mL/minute and R2 = 0.4 for blood flow = 3500mL/minute, both with p < 0.001) and positively associated with CO2 transfer variations (R2 = 0.2 for sweep gas flow ≤ 6L/minute, p < 0.001, and R2 = 0.1 for sweep gas flow > 6L/minute, p = 0.006), and the blood flow/pump rotation ratio is not associated with O2 transfer variations (R2 = 0.01 for blood flow = 1500mL/minute, p = 0.19, and R2 = - 0.01 for blood flow = 3500 mL/minute, p = 0.46). Blood flow/pump rotation ratio variation is negatively associated with transmembrane pressure and positively associated with CO2 transfer in this animal model. According to the clinical situation, a decrease in the blood flow/pump rotation ratio can indicate artificial lung dysfunction without the occurrence of hypoxemia.

Postural stability of biped robots and the foot-rotation indicator (FRI) point

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

Goswami, A.

1999-06-01

The focus of this paper is the problem of foot rotation in biped robots during the single-support phase. Foot rotation is an indication of postural instability, which should be carefully treated in a dynamically stable walk and avoided altogether in a statically stable walk. The author introduces the foot-rotation indicator (FRI) point, which is a point on the foot/ground-contact surface where the net ground-reaction force would have to act to keep the foot stationary. To ensure no foot rotation, the FRI point must remain within the convex hull of the foot-support area. In contrast with the ground projection of themore » center of mass (GCoM), which is a static criterion, the FRI point incorporates robot dynamics. As opposed to the center of pressure (CoP) -- better known as the zero-moment point (ZMP) in the robotics literature -- which may not leave the support area, the FRI point may leave the area. In fact, the position of the FRI point outside the footprint indicates the direction of the impending rotation and the magnitude of rotational moment acting on the foot. Owing to these important properties, the FRI point helps not only to monitor the state of postural stability of a biped robot during the entire gait cycle, but indicates the severity of instability of the gait as well. In response to a recent need, the paper also resolves the misconceptions surrounding the CoP/ZMP equivalence.« less

Determining the Rotation Periods of an Inactive LEO Satellite and the First Korean Space Debris on GEO, KOREASAT 1

NASA Astrophysics Data System (ADS)

Choi, Jin; Jo, Jung Hyun; Kim, Myung-Jin; Roh, Dong-Goo; Park, Sun-Youp; Lee, Hee-Jae; Park, Maru; Choi, Young-Jun; Yim, Hong-Suh; Bae, Young-Ho; Park, Young-Sik; Cho, Sungki; Moon, Hong-Kyu; Choi, Eun-Jung; Jang, Hyun-Jung; Park, Jang-Hyun

2016-06-01

Inactive space objects are usually rotating and tumbling as a result of internal or external forces. KOREASAT 1 has been inactive since 2005, and its drift trajectory has been monitored with the optical wide-field patrol network (OWL-Net). However, a quantitative analysis of KOREASAT 1 in regard to the attitude evolution has never been performed. Here, two optical tracking systems were used to acquire raw measurements to analyze the rotation period of two inactive satellites. During the optical campaign in 2013, KOREASAT 1 was observed by a 0.6 m class optical telescope operated by the Korea Astronomy and Space Science Institute (KASI). The rotation period of KOREASAT 1 was analyzed with the light curves from the photometry results. The rotation periods of the low Earth orbit (LEO) satellite ASTRO-H after break-up were detected by OWL-Net on April 7, 2016. We analyzed the magnitude variation of each satellite by differential photometry and made comparisons with the star catalog. The illumination effect caused by the phase angle between the Sun and the target satellite was corrected with the system tool kit (STK) and two line element (TLE) technique. Finally, we determined the rotation period of two inactive satellites on LEO and geostationary Earth orbit (GEO) with light curves from the photometry. The main rotation periods were determined to be 5.2 sec for ASTRO-H and 74 sec for KOREASAT 1.

Subunit association of gamma-glutamyltranspeptidase of Escherichia coli K-12.

PubMed

Hashimoto, W; Suzuki, H; Nohara, S; Tachi, H; Yamamoto, K; Kumagai, H

1995-12-01

gamma-Glutamyltranspeptidase [EC 2.3.2.2] of Escherichia coli K-12 consists of one large subunit and one small subunit, which can be separated from each other by high-performance liquid chromatography. Using ion spray mass spectrometry, the masses of the large and the small subunit were determined to be 39,207 and 20,015, respectively. The large subunit exhibited no gamma-glutamyltranspeptidase activity and the small subunit had little enzymatic activity, but a mixture of the two subunits showed partial recovery of the enzymatic activity. The results of native-polyacrylamide gel electrophoresis suggested that they could partially recombine, and that the recombined dimer exhibited enzymatic activity. The gene of gamma-glutamyltranspeptidase encoded a signal peptide, and the large and small subunits in a single open reading frame in that order. Two kinds of plasmid were constructed encoding the signal peptide and either the large or the small subunit. A gamma-glutamyltranspeptidase-less mutant of E. coli K-12 was transformed with each plasmid or with both of them. The strain harboring the plasmid encoding each subunit produced a small amount of the corresponding subunit protein in the periplasmic space but exhibited no enzymatic activity. The strain transformed with both plasmids together exhibited the enzymatic activity, but its specific activity was approximately 3% of that of a strain harboring a plasmid encoding the intact structural gene. These results indicate that a portion of the separated large and small subunits can be reconstituted in vitro and exhibit the enzymatic activity, and that the expressed large and small subunits independently are able to associate in vivo and be folded into an active structure, though the specific activity of the associated subunits was much lower than that of native enzyme. This suggests that the synthesis of gamma-glutamyltranspeptidase in a single precursor polypeptide and subsequent processing are more effective to construct the intact structure of gamma-glutamyltranspeptidase than the association of the separated large and small subunits.

A non-contacting approach for full field dynamic strain monitoring of rotating structures using the photogrammetry, finite element, and modal expansion techniques

NASA Astrophysics Data System (ADS)

Baqersad, Javad

Health monitoring of rotating structures such as wind turbines and helicopter rotors is generally performed using conventional sensors that provide a limited set of data at discrete locations near or on the hub. These sensors usually provide no data on the blades or interior locations where failures may occur. Within this work, an unique expansion algorithm was extended and combined with finite element (FE) modeling and an optical measurement technique to identify the dynamic strain in rotating structures. The merit of the approach is shown by using the approach to predict the dynamic strain on a small non-rotating and rotating wind turbine. A three-bladed wind turbine having 2.3-meter long blades was placed in a semi-built-in boundary condition using a hub, a machining chuck, and a steel block. A finite element model of the three wind turbine blades assembled to the hub was created and used to extract resonant frequencies and mode shapes. The FE model was validated and updated using experimental modal tests. For the non-rotating optical test, the turbine was excited using a sinusoidal excitation, a pluck test, arbitrary impacts on three blades, and random force excitations with a mechanical shaker. The response of the structure to the excitations was measured using three-dimensional point tracking. A pair of high-speed cameras was used to measure the displacement of optical targets on the structure when the blades were vibrating. The measured displacements at discrete locations were expanded and applied to the finite element model of the structure to extract the full-field dynamic strain. The results of the work show an excellent correlation between the strain predicted using the proposed approach and the strain measured with strain-gages for all of the three loading conditions. Similar to the non-rotating case, optical measurements were also preformed on a rotating wind turbine. The point tracking technique measured both rigid body displacement and flexible deformation of the blades at target locations. The measured displacements were expanded and applied to the finite element model of the turbine to extract full-field dynamic strain on the structure. In order to validate the results for the rotating turbine, the predicted strain was compared to strain measured at four locations on the spinning blades using a wireless strain-gage system. The approach used in this work to predict the strain showed higher accuracy than measurements obtainable by using the digital image correlation technique. The new expansion approach is able to extract dynamic strain all over the entire structure, even inside the structure beyond the line of sight of the measurement system. Because the method is based on a non-contacting measurement approach, it can be readily applied to a variety of structures having different boundary and operating conditions, including rotating blades.

The NMDA receptor NR2A subunit regulates proliferation of MKN45 human gastric cancer cells

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

Watanabe, Kanako; Department of Anesthesiology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya 663-8501; Kanno, Takeshi

2008-03-07

The present study investigated proliferation of MKN28 and MKN45 human gastric cancer cells regulated by the N-methyl-D-aspartate (NMDA) receptor subunit. The NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (AP5) inhibited proliferation of MKN45 cells, but not MKN28 cells. Of the NMDA subunits such as NR1, NR2 (2A, 2B, 2C, and 2D), and NR3 (3A and 3B), all the NMDA subunit mRNAs except for the NR2B subunit mRNA were expressed in both MKN28 and MKN45 cells. MKN45 cells were characterized by higher expression of the NR2A subunit mRNA and lower expression of the NR1 subunit mRNA, but MKN28 otherwise by higher expression ofmore » the NR1 subunit mRNA and lower expression of the NR2A subunit mRNA. MKN45 cell proliferation was also inhibited by silencing the NR2A subunit-targeted gene. For MKN45 cells, AP5 or knocking-down the NR2A subunit increased the proportion of cells in the G{sub 1} phase of cell cycling and decreased the proportion in the S/G{sub 2} phase. The results of the present study, thus, suggest that blockage of NMDA receptors including the NR2A subunit suppresses MKN45 cell proliferation due to cell cycle arrest at the G{sub 1} phase; in other words, the NR2A subunit promotes MKN45 cell proliferation by accelerating cell cycling.« less

An X-ray monitor for measurement of a titanium tritide target thickness

NASA Technical Reports Server (NTRS)

Alger, D. L.; Steinberg, R.

1972-01-01

An X-ray device capable of measuring titanium tritide film thickness from 0.1 to 30 micrometers has been built and tested. The monitor was designed for use in a rotating target system which used thick targets and incorporated a sputtering electrode to remove depleted layers from the target surface. The thickness measurement can be done in the presence of an intense background of bremsstrahlung and characteristic titanium X-radiation. A measurement can be accomplished in situ in two hours with reasonable accuracy.

Sacramento Municipal Utility District (SMUD): Rotating Shadowband Radiometer (RSR); Anatolia - Rancho Cordova, California (Data)

DOE Data Explorer

Maxey, C.; Andreas, A.

2009-02-03

A partnership with industry and U.S. Department of Energy's National Renewable Energy Laboratory (NREL) to collect solar data to support future solar power generation in the United States. The measurement station monitors global horizontal, direct normal, and diffuse horizontal irradiance to define the amount of solar energy that hits this particular location. The solar measurement instrumentation is also accompanied by meteorological monitoring equipment to provide scientists with a complete picture of the solar power possibilities.

Overexpression of PP2A-C5 that encodes the catalytic subunit 5 of protein phosphatase 2A in Arabidopsis confers better root and shoot development under salt conditions

USDA-ARS?s Scientific Manuscript database

Protein phosphatase 2A (PP2A) is an enzyme consisting of three subunits: a scaffolding A subunit, a regulatory B subunit and a catalytic C subunit. PP2As were shown to play diverse roles in eukaryotes. In this study, the function of the Arabidopsis PP2A-C5 gene that encodes the catalytic subunit 5 o...

Estimating carbon fluxes on small rotationally grazed pastures

USDA-ARS?s Scientific Manuscript database

Satellite-based Normalized Difference Vegetation Index (NDVI) data have been extensively used for estimating gross primary productivity (GPP) and yield of grazing lands throughout the world. Large-scale estimates of GPP are a necessary component of efforts to monitor the soil carbon balance of grazi...

Weather and Rotation on Young Brown Dwarfs

NASA Astrophysics Data System (ADS)

Vos, Johanna; Biller, Beth; Allers, Katelyn; Manjavacas, Elena; Liu, Michael; Best, William; Metchev, Stanimir; Buenzli, Esther; Bonavita, Mariangela; Eriksson, Simon; Dupuy, Trent; Kopytova, Taisiya; Brandner, Wolfgang; Henning, Thomas; Bonnefoy, Mickael; Crossfield, Ian; Schlieder, Joshua; Homeier, Derek; Janson, Markus; Radigan, Jacqueline

2018-05-01

As part of a large, ground-based survey for weather patterns on exoplanet analogues, we have detected J-band variability in 5 young exoplanet analogues. We have already carried out followup Spitzer monitoring of two objects and here we propose Spitzer 3.6um and 4.5um monitoring of three early-mid-L detections in our survey. The proposed observations will enable us to assess the role of gravity in the variability properties of these young objects by providing a full measure of mid-IR amplitude across the full L spectral sequence for low-gravity objects. The proposed observations will also allow us to measure the rotational periods of our three targets. This will provide vital information on the angular momentum of young brown dwarfs, while enabling us to correct for geometric effects when considering the variability properties of our targets. This study will act as a necessary pathfinder for future variability studies of free-floating and companion exoplanets with JWST.

Modelling of wicking and moisture interactions of flax and viscose fibres.

PubMed

Stuart, T; McCall, R D; Sharma, H S S; Lyons, G

2015-06-05

Methods for assessing the wicking properties of individual fibre bundles have been developed from models based on the original Washburn equation (WE) and the modified Washburn equation (MWE), which also accounts for swelling. Both models gave indication of differences in wicking properties of flax and the viscose fibres, though MWE gave additional information that could be interpreted in terms of the physical model. Wicking of the viscose fibres is mainly via inter-fibre capillaries while that of flax is a combination of inter-fibre capillaries and lumen present in some elementary fibres. The degree of swelling and associated rotation of flax fibre in a vapour pressure range of 1-6torr were monitored using an environmental scanning electron microscope (ESEM). Viscose fibre exhibited swelling under the same conditions but did not rotate. The two techniques highlighted different mechanisms of wicking which can be used for monitoring moisture uptake/swelling of treated fibres for fabrication of composites. Copyright © 2015 Elsevier Ltd. All rights reserved.

Arrangement of RecA protein in its active filament determined by polarized-light spectroscopy.

PubMed

Morimatsu, Katsumi; Takahashi, Masayuki; Nordén, Bengt

2002-09-03

Linear dichroism (LD) polarized-light spectroscopy is used to determine the arrangement of RecA in its large filamentous complex with DNA, active in homologous recombination. Angular orientation data for two tryptophan and seven tyrosine residues, deduced from differential LD of wild-type RecA vs. mutants that were engineered to attenuate the UV absorption of selected residues, revealed a rotation by some 40 degrees of the RecA subunits relative to the arrangement in crystal without DNA. In addition, conformational changes are observed for tyrosine residues assigned to be involved in DNA binding and in RecA-RecA contacts, thus potentially related to the global structure of the filament and its biological function. The presented spectroscopic approach, called "Site-Specific Linear Dichroism" (SSLD), may find forceful applications also to other biologically important fibrous complexes not amenable to x-ray crystallographic or NMR structural analysis.

Constructing biomolecular motor-powered hybrid NEMS devices

NASA Astrophysics Data System (ADS)

Bachand, George D.; Montemagno, Carlo D.

1999-10-01

The recognition of many enzymes as nanoscale molecular motors has allowed for the potential creation of hybrid organic/inorganic nano-electro-mechanical (NEMS) devices. The long-range goal of this research is the integration of F1-ATPase with NEMS to produce useful nanoscale devices. A thermostable F1-ATPase coding sequence has been isolated, cloned, and engineered for high-level protein expression. Precise positioning, spacing, and orientation of single F1-ATPase molecules were achieved using patterned nickel arrays. An efficient, accurate, and adaptable assay was developed to assess the performance of single F1- ATPase motors, and confirmed a three-step mechanism of (gamma) subunit rotation during ATP hydrolysis. Further evaluation of the bioengineering and biophysical properties of F1-ATPase currently are being conducted, as well as the construction of an F1-ATPase-powered, hybrid NEMS device. The evolution of this technology will permit the creation of novel classes of nanoscale, hybrid devices.

Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity.

PubMed

Higgins, N P; Peebles, C L; Sugino, A; Cozzarelli, N R

1978-04-01

Extensively purified DNA gyrase from Escherichia coli is inhibited by nalidixic acid and by novobiocin. The enzyme is composed of two subunits, A and B, which were purified as separate components. Subunit A is the product of the gene controlling sensitivity to nalidixic acid (nalA) because: (i) the electrophoretic mobility of subunit A in the presence of sodium dodecyl sulfate is identical to that of the 105,000-dalton nalA gene product; (ii) mutants that are resistant to nalidixic acid (nalA(r)) produce a drug-resistant subunit A; and (iii) wild-type subunit A confers drug sensitivity to in vitro synthesis of varphiX174 DNA directed by nalA(r) mutants. Subunit B contains a 95,000-dalton polypeptide and is controlled by the gene specifying sensitivity to novobiocin (cou) because cou(r) mutants produce a novobiocin-resistant subunit B and novobiocin-resitant gyrase is made drug sensitive by wild-type subunit B. Subunits A and B associate, so that gyrase was also purified as a complex containing 105,000- and 95,000-dalton polypeptides. This enzyme and gyrase reconstructed from subunits have the same drug sensitivity, K(m) for ATP, and catalytic properties. The same ratio of subunits gives efficient reconstitution of the reactions intrinsic to DNA gyrase, including catalysis of supercoiling of closed duplex DNA, relaxation of supercoiled DNA in the absence of ATP, and site-specific cleavage of DNA induced by sodium dodecyl sulfate.

Soybean glycinin subunits: Characterization of physicochemical and adhesion properties.

PubMed

Mo, Xiaoqun; Zhong, Zhikai; Wang, Donghai; Sun, Xiuzhi

2006-10-04

Soybean proteins have shown great potential for applications as renewable and environmentally friendly adhesives. The objective of this work was to study physicochemical and adhesion properties of soy glycinin subunits. Soybean glycinin was extracted from soybean flour and then fractionated into acidic and basic subunits with an estimated purity of 90 and 85%, respectively. Amino acid composition of glycinin subunits was determined. The high hydrophobic amino acid content is a major contributor to the solubility behavior and water resistance of the basic subunits. Acidic subunits and glycinin had similar solubility profiles, showing more than 80% solubility at pH 2.0-4.0 or 6.5-12.0, whereas basic subunits had considerably lower solubility with the minimum at pH 4.5-8.0. Thermal analysis using a differential scanning calorimeter suggested that basic subunits form new oligomeric structures with higher thermal stability than glycinin but no highly ordered structures present in isolated acidic subunits. The wet strength of basic subunits was 160% more than that of acidic subunits prepared at their respective isoelectric points (pI) and cured at 130 degrees C. Both pH and the curing temperature significantly affected adhesive performance. High-adhesion water resistance was usually observed for adhesives from protein prepared at their pI values and cured at elevated temperatures. Basic subunits are responsible for the water resistance of glycinin and are a good starting material for the development of water-resistant adhesives.

The role of upper torso and pelvis rotation in driving performance during the golf swing.

PubMed

Myers, Joseph; Lephart, Scott; Tsai, Yung-Shen; Sell, Timothy; Smoliga, James; Jolly, John

2008-01-15

While the role of the upper torso and pelvis in driving performance is anecdotally appreciated by golf instructors, their actual biomechanical role is unclear. The aims of this study were to describe upper torso and pelvis rotation and velocity during the golf swing and determine their role in ball velocity. One hundred recreational golfers underwent a biomechanical golf swing analysis using their own driver. Upper torso and pelvic rotation and velocity, and torso-pelvic separation and velocity, were measured for each swing. Ball velocity was assessed with a golf launch monitor. Group differences (groups based on ball velocity) and moderate relationships (r > or = 0.50; P < 0.001) were observed between an increase in ball velocity and the following variables: increased torso-pelvic separation at the top of the swing, maximum torso-pelvic separation, maximum upper torso rotation velocity, upper torso rotational velocity at lead arm parallel and last 40 ms before impact, maximum torso-pelvic separation velocity and torso-pelvic separation velocity at both lead arm parallel and at the last 40 ms before impact. Torso-pelvic separation contributes to greater upper torso rotation velocity and torso-pelvic separation velocity during the downswing, ultimately contributing to greater ball velocity. Golf instructors can consider increasing ball velocity by maximizing separation between the upper torso and pelvis at the top of and initiation of the downswing.

Initial treatment of complete rotator cuff tear and transition to surgical treatment: systematic review of the evidence

PubMed Central

Abdul-Wahab, Taiceer A.; Betancourt, Jean P.; Hassan, Fadi; Thani, Saeed Al.; Choueiri, Hened; Jain, Nitin B.; Malanga, Gerard A.; Murrell, William D.; Prasad, Anil; Verborgt, Olivier

2016-01-01

Summary Background rotator cuff tear affects many people. Natural history, and evidence for non-operative treatment remains limited. Our objective is to assess evidence available for the efficacy and morbidity of commonly used systemic medications, physiotherapy, and injections alongside evaluating any negative long-term effects. Methods a systematic search was performed of PubMed, Cochrane, EMBASE and CINAHL dates (1 January 1960 – 1 December 2014), search terms: ‘rotator cuff tear’, ‘natural history’, ‘atraumatic’, ‘injection’, ‘physiotherapy’ or ‘physical therapy’, ‘injection’, ‘corticosteroid’, ‘PRP‘, ‘MSC’, risk of conservative treatment’, and ‘surgical indication’. Results eleven studies were included. The mean Coleman Methodology Score modified for conservative therapy is 69.21 (range 88–44) (SD 12.31). This included 2 RCTs, 7 prospective, and 2 retrospective studies. Evidence suggests it is safe to monitor symptomatic rotator cuff tears, as tear size and symptoms are not correlated with pain, function, and/or ultimate outcome. Conclusions complete rotator cuff tears may be effectively treated with injections, exercise in the short and intermediate terms respectively. Negative effect of corticosteroids on rotator cuff tissue has not been demonstrated. Timing to end conservative treatment is unknown, but likely indicated when a patient demonstrates increased weakness and loss of function not recoverable by physiotherapy. PMID:27331030

Comprehensive Characterization of AMP-activated Protein Kinase Catalytic Domain by Top-down Mass Spectrometry

PubMed Central

Yu, Deyang; Peng, Ying; Ayaz-Guner, Serife; Gregorich, Zachery R.; Ge, Ying

2015-01-01

AMP-activated protein kinase (AMPK) is a serine/threonine protein kinase that is essential in regulating energy metabolism in all eukaryotic cells. It is a heterotrimeric protein complex composed of a catalytic subunit (α) and two regulatory subunits (β and γ. C-terminal truncation of AMPKα at residue 312 yielded a protein that is active upon phosphorylation of Thr172 in the absence of β and γ subunits, which is refered to as the AMPK catalytic domain and commonly used to substitute for the AMPK heterotrimeric complex in in vitro kinase assays. However, a comprehensive characterization of the AMPK catalytic domain is lacking. Herein, we expressed a His-tagged human AMPK catalytic domin (denoted as AMPKΔ) in E. coli, comprehensively characterized AMPKΔ in its basal state and after in vitro phosphorylation using top-down mass spectrometry (MS), and assessed how phosphorylation of AMPKΔ affects its activity. Unexpectedly, we found that bacterially-expressed AMPKΔ was basally phosphorylated and localized the phosphorylation site to the His-tag. We found that AMPKΔ has noticeable basal activity and was capable of phosphorylating itself and its substrates without activating phosphorylation at Thr172. Moreover, our data suggested that Thr172 is the only site phosphorylated by its upstream kinase, liver kinase B1, and that this phosphorylation dramatically increases the kinase activity of AMPKΔ. Importantly, we demonstrated that top-down MS in conjunction with in vitro phosphorylation assay is a powerful approach for monitoring phosphorylation reaction and determining sequential order of phosphorylation events in kinase-substrate systems. PMID:26489410

CD94-NKG2A recognition of human leukocyte antigen (HLA)-E bound to an HLA class I leader sequence.

PubMed

Petrie, Emma J; Clements, Craig S; Lin, Jie; Sullivan, Lucy C; Johnson, Darryl; Huyton, Trevor; Heroux, Annie; Hoare, Hilary L; Beddoe, Travis; Reid, Hugh H; Wilce, Matthew C J; Brooks, Andrew G; Rossjohn, Jamie

2008-03-17

The recognition of human leukocyte antigen (HLA)-E by the heterodimeric CD94-NKG2 natural killer (NK) receptor family is a central innate mechanism by which NK cells monitor the expression of other HLA molecules, yet the structural basis of this highly specific interaction is unclear. Here, we describe the crystal structure of CD94-NKG2A in complex with HLA-E bound to a peptide derived from the leader sequence of HLA-G. The CD94 subunit dominated the interaction with HLA-E, whereas the NKG2A subunit was more peripheral to the interface. Moreover, the invariant CD94 subunit dominated the peptide-mediated contacts, albeit with poor surface and chemical complementarity. This unusual binding mode was consistent with mutagenesis data at the CD94-NKG2A-HLA-E interface. There were few conformational changes in either CD94-NKG2A or HLA-E upon ligation, and such a "lock and key" interaction is typical of innate receptor-ligand interactions. Nevertheless, the structure also provided insight into how this interaction can be modulated by subtle changes in the peptide ligand or by the pairing of CD94 with other members of the NKG2 family. Differences in the docking strategies used by the NKG2D and CD94-NKG2A receptors provided a basis for understanding the promiscuous nature of ligand recognition by NKG2D compared with the fidelity of the CD94-NKG2 receptors.

CD94-NKG2A recognition of human leukocyte antigen (HLA)-E bound to an HLA class I leader sequence

PubMed Central

Petrie, Emma J.; Clements, Craig S.; Lin, Jie; Sullivan, Lucy C.; Johnson, Darryl; Huyton, Trevor; Heroux, Annie; Hoare, Hilary L.; Beddoe, Travis; Reid, Hugh H.; Wilce, Matthew C.J.; Brooks, Andrew G.; Rossjohn, Jamie

2008-01-01

The recognition of human leukocyte antigen (HLA)-E by the heterodimeric CD94-NKG2 natural killer (NK) receptor family is a central innate mechanism by which NK cells monitor the expression of other HLA molecules, yet the structural basis of this highly specific interaction is unclear. Here, we describe the crystal structure of CD94-NKG2A in complex with HLA-E bound to a peptide derived from the leader sequence of HLA-G. The CD94 subunit dominated the interaction with HLA-E, whereas the NKG2A subunit was more peripheral to the interface. Moreover, the invariant CD94 subunit dominated the peptide-mediated contacts, albeit with poor surface and chemical complementarity. This unusual binding mode was consistent with mutagenesis data at the CD94-NKG2A–HLA-E interface. There were few conformational changes in either CD94-NKG2A or HLA-E upon ligation, and such a “lock and key” interaction is typical of innate receptor–ligand interactions. Nevertheless, the structure also provided insight into how this interaction can be modulated by subtle changes in the peptide ligand or by the pairing of CD94 with other members of the NKG2 family. Differences in the docking strategies used by the NKG2D and CD94-NKG2A receptors provided a basis for understanding the promiscuous nature of ligand recognition by NKG2D compared with the fidelity of the CD94-NKG2 receptors. PMID:18332182

AMPA Receptors Mediate Acetylcholine Release from Starburst Amacrine Cells in the Rabbit Retina

PubMed Central

FIRTH, SALLY I.; LI, WEI; MASSEY, STEPHEN C.; MARSHAK, DAVID W.

2012-01-01

The light response of starburst amacrine cells is initiated by glutamate released from bipolar cells. To identify the receptors that mediate this response, we used a combination of anatomical and physiological techniques. An in vivo, rabbit eyecup was preloaded with [3H]-choline, and the [3H]-acetylcholine (ACh) released into the superfusate was monitored. A photopic, 3 Hz flashing light increased ACh release, and the selective AMPA receptor antagonist, GYKI 53655, blocked this light-evoked response. Nonselective AMPA/kainate agonists increased the release of ACh, but the specific kainate receptor agonist, SYM 2081, did not increase ACh release. Selective AMPA receptor antagonists, GYKI 53655 or GYKI 52466, also blocked the responses to agonists. We conclude that the predominant excitatory input to starburst amacrine cells is mediated by AMPA receptors. We also labeled lightly fixed rabbit retinas with antisera to choline acetyltransferase (ChAT), AMPA receptor subunits GluR1, GluR2/3, or GluR4, and kainate receptor subunits GluR6/7 and KA2. Labeled puncta were observed in the inner plexiform layer with each of these antisera to glutamate receptors, but only GluR2/3-IR puncta and GluR4-IR puncta were found on the ChAT-IR processes. The same was true of starburst cells injected intracellularly with Neurobiotin, and these AMPA receptor subunits were localized to two populations of puncta. The AMPA receptors are expected to desensitize rapidly, enhancing the sensitivity of starburst amacrine cells to moving or other rapidly changing stimuli. PMID:14515241

The Pleckstrin Homology Domain of Phospholipase Cβ Transmits Enzymatic Activation through Modulation of Membrane - Domain Orientation§

PubMed Central

Drin, Guillaume; Douguet, Dominique; Scarlata, Suzanne

2008-01-01

Phospholipase C-beta (PLCβ) enzymes are activated by Gαq and Gβγ subunits and catalyze the hydrolysis of the minor membrane lipid phosphatidylinositol 4,5 bisphosphate (PI(4,5)P2). Activation of PLCβ2 by Gβγ subunits has been shown to be conferred through its N-terminal pleckstrin homology (PH) domain although the underlying mechanism is unclear. Also unclear are observations that the extent of Gβγ activation differs on different membrane surfaces. In this study, we have identified a unique region of the PH domain of PLCβ2 domain (residues 71-88) which, when added to the enzyme as a peptide, causes enzyme activation similar to Gβγ subunits. This PH domain segment interacts strongly with membranes composed of lipid mixtures but not those containing lipids with electrically neutral zwitterionic head groups. Moreso, addition of this segment perturbs interaction of the catalytic domain, but not the PH domain, with membrane surfaces. We monitored the orientation of the PH and catalytic domains of PLC by intermolecular fluorescence resonance energy transfer (FRET) using the Gβγ activatable mutant, PLCβ2/δ1(C193S). We find an increase in FRET with binding to membranes with mixed lipids but not to those containing only lipids with electrically neutral head groups. These results suggest that enzymatic activation can be conferred through optimal association of the PHβ71-88 region to specific membrane surfaces. These studies allow us to understand the basis of variations of Gβγ activation on different membrane surfaces. PMID:16669615

Specific phosphopeptide binding regulates a conformational change in the PI 3-kinase SH2 domain associated with enzyme activation.

PubMed Central

Shoelson, S E; Sivaraja, M; Williams, K P; Hu, P; Schlessinger, J; Weiss, M A

1993-01-01

SH2 (src-homology 2) domains define a newly recognized binding motif that mediates the physical association of target phosphotyrosyl proteins with downstream effector enzymes. An example of such phosphoprotein-effector coupling is provided by the association of phosphatidylinositol 3-kinase (PI 3-kinase) with specific phosphorylation sites within the PDGF receptor, the c-Src/polyoma virus middle T antigen complex and the insulin receptor substrate IRS-1. Notably, phosphoprotein association with the SH2 domains of p85 also stimulates an increase in catalytic activity of the PI 3-kinase p110 subunit, which can be mimicked by phosphopeptides corresponding to targeted phosphoprotein phosphorylation sites. To investigate how phosphoprotein binding to the p85 SH2 domain stimulates p110 catalytic activation, we have examined the differential effects of phosphotyrosine and PDGF receptor-, IRS-1- and c-Src-derived phosphopeptides on the conformation of an isolated SH2 domain of PI 3-kinase. Although phosphotyrosine and both activating and non-activating phosphopeptides bind to the SH2 domain, activating phosphopeptides bind with higher affinity and induce a qualitatively distinct conformational change as monitored by CD and NMR spectroscopy. Amide proton exchange and protease protection assays further show that high affinity, specific phosphopeptide binding induces non-local dynamic SH2 domain stabilization. Based on these findings we propose that specific phosphoprotein binding to the p85 subunit induces a change in SH2 domain structure which is transmitted to the p110 subunit and regulates enzymatic activity by an allosteric mechanism. Images PMID:8382612

Localization of yeast RNA polymerase I core subunits by immunoelectron microscopy.

PubMed Central

Klinger, C; Huet, J; Song, D; Petersen, G; Riva, M; Bautz, E K; Sentenac, A; Oudet, P; Schultz, P

1996-01-01

Immunoelectron microscopy was used to determine the spatial organization of the yeast RNA polymerase I core subunits on a three-dimensional model of the enzyme. Images of antibody-labeled enzymes were compared with the native enzyme to determine the localization of the antibody binding site on the surface of the model. Monoclonal antibodies were used as probes to identify the two largest subunits homologous to the bacterial beta and beta' subunits. The epitopes for the two monoclonal antibodies were mapped using subunit-specific phage display libraries, thus allowing a direct correlation of the structural data with functional information on conserved sequence elements. An epitope close to conserved region C of the beta-like subunit is located at the base of the finger-like domain, whereas a sequence between conserved regions C and D of the beta'-like subunit is located in the apical region of the enzyme. Polyclonal antibodies outlined the alpha-like subunit AC40 and subunit AC19 which were found co-localized also in the apical region of the enzyme. The spatial location of the subunits is correlated with their biological activity and the inhibitory effect of the antibodies. Images PMID:8887555

Investigation of pH and temperature on optical rotatory dispersion for noninvasive glucose monitoring

NASA Astrophysics Data System (ADS)

Baba, Justin S.; Meledeo, Adam; Cameron, B. D.; Cote, Gerard L.

2001-06-01

The widespread occurrence of diabetes mellitus and the severity of its associated complications necessitate the development of non-invasive blood glucose measurement devices in an attempt to improve treatment regimens and curb the complications associated with this disease. One method showing promise in this endeavor utilizes optical polarimetry to monitor blood glucose levels indirectly by measuring glucose rotation of polarized light, which is a direct indication of glucose concentration, in the aqueous humor of the eye. The presence of other optically active (chiral) components in the aqueous humor of the eye have the potential to confound the glucose measurement of optical rotation when using a single wavelength polarimeter. Thus, this has led to the recent investigation of multispectral polarimetric systems which have the potential to enable the removal of confounder contributions to the net observed optical rotation, therefore, increasing glucose specificity and reducing glucose prediction errors. Such polarimetric systems take advantage of the uniqueness in the rotation of polarized light, as a function of wavelength, by the chiral molecule of interest. This is commonly referred to as the optical rotatory dispersion (ORD) spectra of the chiral molecule. ORD characterization of the chiral molecules within the aqueous humor is necessary for determining the optimum number of wavelengths needed to reduce glucose prediction errors; however, this information is often only given at the sodium-D line (589 nm) in the literature. This report describes the system we designed and built to measure ORD spectra for glucose and for albumin, the main optical confounder within the aqueous humor, as well as our investigation of the effects of temperature and pH on these ORD spectra.

Chromospheric Variability: Analysis of 36 years of Time Series from the National Solar Observatory/Sacramento Peak Ca II K-line Monitoring Program

NASA Technical Reports Server (NTRS)

Scargle, Jeffrey D.; Keil, Stephen L.; Worden, Simon P.

2014-01-01

Analysis of more than 36 years of time series of seven parameters measured in the NSO/AFRL/Sac Peak K-line monitoring program elucidates five elucidates five components of the variation: (1) the solar cycle (period approx. 11 years), (2) quasi-periodic variations (periods approx 100 days), (3) a broad band stochastic process (wide range of periods), (4) rotational modulation, and (5) random observational errors. Correlation and power spectrum analyses elucidate periodic and aperiodic variation of the chromospheric parameters. Time-frequency analysis illuminates periodic and quasi periodic signals, details of frequency modulation due to differential rotation, and in particular elucidates the rather complex harmonic structure (1) and (2) at time scales in the range approx 0.1 - 10 years. These results using only full-disk data further suggest that similar analyses will be useful at detecting and characterizing differential rotation in stars from stellar light-curves such as those being produced by NASA's Kepler observatory. Component (3) consists of variations over a range of timescales, in the manner of a 1/f random noise process. A timedependent Wilson-Bappu effect appears to be present in the solar cycle variations (1), but not in the stochastic process (3). Component (4) characterizes differential rotation of the active regions, and (5) is of course not characteristic of solar variability, but the fact that the observational errors are quite small greatly facilitates the analysis of the other components. The recent data suggest that the current cycle is starting late and may be relatively weak. The data analyzed in this paper can be found at the National Solar Observatory web site http://nsosp.nso.edu/cak_mon/, or by file transfer protocol at ftp://ftp.nso.edu/idl/cak.parameters.

The Role of Auxiliary Subunits for the Functional Diversity of Voltage-Gated Calcium Channels

PubMed Central

Campiglio, Marta; Flucher, Bernhard E

2015-01-01

Voltage-gated calcium channels (VGCCs) represent the sole mechanism to convert membrane depolarization into cellular functions like secretion, contraction, or gene regulation. VGCCs consist of a pore-forming α1 subunit and several auxiliary channel subunits. These subunits come in multiple isoforms and splice-variants giving rise to a stunning molecular diversity of possible subunit combinations. It is generally believed that specific auxiliary subunits differentially regulate the channels and thereby contribute to the great functional diversity of VGCCs. If auxiliary subunits can associate and dissociate from pre-existing channel complexes, this would allow dynamic regulation of channel properties. However, most auxiliary subunits modulate current properties very similarly, and proof that any cellular calcium channel function is indeed modulated by the physiological exchange of auxiliary subunits is still lacking. In this review we summarize available information supporting a differential modulation of calcium channel functions by exchange of auxiliary subunits, as well as experimental evidence in support of alternative functions of the auxiliary subunits. At the heart of the discussion is the concept that, in their native environment, VGCCs function in the context of macromolecular signaling complexes and that the auxiliary subunits help to orchestrate the diverse protein–protein interactions found in these calcium channel signalosomes. Thus, in addition to a putative differential modulation of current properties, differential subcellular targeting properties and differential protein–protein interactions of the auxiliary subunits may explain the need for their vast molecular diversity. J. Cell. Physiol. 999: 00–00, 2015. © 2015 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. J. Cell. Physiol. 230: 2019–2031, 2015. © 2015 Wiley Periodicals, Inc. PMID:25820299

Subunits of the Snf1 kinase heterotrimer show interdependence for association and activity.

PubMed

Elbing, Karin; Rubenstein, Eric M; McCartney, Rhonda R; Schmidt, Martin C

2006-09-08

The Snf1 kinase and its mammalian orthologue, the AMP-activated protein kinase (AMPK), function as heterotrimers composed of a catalytic alpha-subunit and two non-catalytic subunits, beta and gamma. The beta-subunit is thought to hold the complex together and control subcellular localization whereas the gamma-subunit plays a regulatory role by binding to and blocking the function of an auto-inhibitory domain (AID) present in the alpha-subunit. In addition, catalytic activity requires phosphorylation by a distinct upstream kinase. In yeast, any one of three Snf1-activating kinases, Sak1, Tos3, or Elm1, can fulfill this role. We have previously shown that Sak1 is the only Snf1-activating kinase that forms a stable complex with Snf1. Here we show that the formation of the Sak1.Snf1 complex requires the beta- and gamma-subunits in vivo. However, formation of the Sak1.Snf1 complex is not necessary for glucose-regulated phosphorylation of the Snf1 activation loop. Snf1 kinase purified from cells lacking the beta-subunits do not contain any gamma-subunit, indicating that the Snf1 kinase does not form a stable alphagamma dimer in vivo. In vitro kinase assays using purified full-length and truncated Snf1 proteins demonstrate that the kinase domain, which lacks the AID, is significantly more active than the full-length Snf1 protein. Addition of purified beta- and gamma-subunits could stimulate the kinase activity of the full-length alpha-subunit but only when all three subunits were present, suggesting an interdependence of all three subunits for assembly of a functional complex.

Tripeptidyl peptidase I, the late infantile neuronal ceroid lipofuscinosis gene product, initiates the lysosomal degradation of subunit c of ATP synthase.

PubMed

Ezaki, J; Takeda-Ezaki, M; Kominami, E

2000-09-01

The specific accumulation of a hydrophobic protein, subunit c of ATP synthase, in lysosomes from the cells of patients with the late infantile form of NCL (LINCL) is caused by a defect in the CLN2 gene product, tripeptidyl peptidase I (TPP-I). The data here show that TPP-I is involved in the initial degradation of subunit c in lysosomes and suggest that its absence leads directly to the lysosomal accumulation of subunit c. The inclusion of a specific inhibitor of TPP-I, Ala-Ala-Phe-chloromethylketone (AAF-CMK), in the culture medium of normal fibroblasts induced the lysosomal accumulation of subunit c. In an in vitro incubation experiment the addition of AAF-CMK to mitochondrial-lysosomal fractions from normal cells inhibited the proteolysis of subunit c, but not the b-subunit of ATP synthase. The use of two antibodies that recognize the aminoterminal and the middle portion of subunit c revealed that the subunit underwent aminoterminal proteolysis, when TPP-I, purified from rat spleen, was added to the mitochondrial fractions. The addition of both purified TPP-I and the soluble lysosomal fractions, which contain various proteinases, to the mitochondrial fractions resulted in rapid degradation of the entire molecule of subunit c, whereas the degradation of subunit c was markedly delayed through the specific inhibition of TPP-I in lysosomal extracts by AAF-CMK. The stable subunit c in the mitochondrial-lysosomal fractions from cells of a patient with LINCL was degraded on incubation with purified TPP-I. The presence of TPP-I led to the sequential cleavage of tripeptides from the N-terminus of the peptide corresponding to the amino terminal sequence of subunit c.

GABAA receptors: Various stoichiometrics of subunit arrangement in α1β3 and α1β3ε receptors.

PubMed

Has, Ahmad Tarmizi Che; Chebib, Mary

2018-05-15

GABAA receptors (GABAARs) are members of the Cys-loop ligand-gated ion channel (LGIC) superfamily, which includes nicotinic acetylcholine, glycine, and serotonin (5HT3) receptors [1,2,3,4]. LGICs typically mediate fast synaptic transmission via the movement of ions through channels gated by neurotransmitters, such as acetylcholine for nicotinic receptors and GABA for GABAARs [5]. The term Cys-loop receptors originates from the presence of a conserved disulphide bond (or bridge) which holds together two cysteine amino acids of the loop that forms from the structure of polypeptides in the extracellular domain of the receptor's subunit [6]. GABAARs are pentameric transmembrane protein complexes consisting of five subunits from a variety of polypeptide subunits [7,8]. All of these subunits are pseudo-symmetrically organized in the plane of the membrane, with a Cl--selective channel in the middle of the complex. To date, nineteen GABAAR subunits have been identified and categorized into eight classes, α1-6, β1-3, γ1-3, δ, ε, θ, π and ρ1-3, but their variety is further broadened by the existence of several splice forms for certain subunits (e.g., α6, β2 and γ2) [9,10,11,12]. The subunits within each class have an amino acid sequence homology of 70% or more, whereas those with a sequence homology of 30% or less are grouped into different classes [13,14]. A subunit consists of four transmembrane domains (TM1-TM4), each forming an α-helix; a large extracellular N-terminal domain that incorporates part of the orthosteric agonist/antagonist binding site; a large intracellular loop between the TM3 and TM4; a small intracellular loop between TM1 and TM2; a small extracellular loop between TM2 and TM3; and a C-terminal extracellular domain [15,16]. Each subunit is arranged in such a way as to create principal and complementary interfaces with the other subunits, and in a position such that the TM2 of each subunit forms the wall of the channel pore [17,18,19]. The major subunit combination found in the brain comprises α1, β2 and γ2 subunits with the stoichiometry 2α1:2β2:1γ2 [18,20]. For the GABAA α1β2γ2 receptors, the subunits form a specific arrangement in which α1 and β2 subunits alternate with each other and are connected by a γ2 subunit (Figure A) [16,20,21]. For minor subtypes, different α and β subunits have been detected to co-exist as proven by the existence of GABAARs containing α1α2, α1α3, α1α5, α2α3, α3α5, α4α1, α4α2 and α4α3 in the central nervous system [22,23]. Meanwhile, the same pattern has been detected with β and γ subunits, at least the co-occurrence of β in the same GABAAR as well as γ2 with γ3, indicating that these subunits have the capacity to co-exist with each other [24,25,26]. Since different subunits can actually occur in one receptor, GABAARs are considered to exist in a multi-subunit arrangement, leading to ambiguity in the determination of a receptor's stoichiometry. In this review, we first briefly discuss the different subunit arrangements of α1 and β3 subunits in the binary α1β3 receptors. Then we review the GABAA ε-containing receptors predominantly in terms of the ability of ε subunit to present in different position in the ternary α1β3ε receptors, which introduce distinct populations of receptor. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Viral and cellular SOS-regulated motor proteins: dsDNA translocation mechanisms with divergent functions.

PubMed

Wolfe, Annie; Phipps, Kara; Weitao, Tao

2014-01-01

DNA damage attacks on bacterial cells have been known to activate the SOS response, a transcriptional response affecting chromosome replication, DNA recombination and repair, cell division and prophage induction. All these functions require double-stranded (ds) DNA translocation by ASCE hexameric motors. This review seeks to delineate the structural and functional characteristics of the SOS response and the SOS-regulated DNA translocases FtsK and RuvB with the phi29 bacteriophage packaging motor gp16 ATPase as a prototype to study bacterial motors. While gp16 ATPase, cellular FtsK and RuvB are similarly comprised of hexameric rings encircling dsDNA and functioning as ATP-driven DNA translocases, they utilize different mechanisms to accomplish separate functions, suggesting a convergent evolution of these motors. The gp16 ATPase and FtsK use a novel revolution mechanism, generating a power stroke between subunits through an entropy-DNA affinity switch and pushing dsDNA inward without rotation of DNA and the motor, whereas RuvB seems to employ a rotation mechanism that remains to be further characterized. While FtsK and RuvB perform essential tasks during the SOS response, their roles may be far more significant as SOS response is involved in antibiotic-inducible bacterial vesiculation and biofilm formation as well as the perspective of the bacteria-cancer evolutionary interaction.

Na+, K+-ATPase β1 subunit associates with α1 subunit modulating a "higher-NKA-in-hyposmotic media" response in gills of euryhaline milkfish, Chanos chanos.

PubMed

Hu, Yau-Chung; Chu, Keng-Fu; Yang, Wen-Kai; Lee, Tsung-Han

2017-10-01

The euryhaline milkfish (Chanos chanos) is a popular aquaculture species that can be cultured in fresh water, brackish water, or seawater in Southeast Asia. In gills of the milkfish, Na + , K + -ATPase (i.e., NKA; sodium pump) responds to salinity challenges including changes in mRNA abundance, protein amount, and activity. The functional pump is composed of a heterodimeric protein complex composed of α- and β-subunits. Among the NKA genes, α1-β1 isozyme comprises the major form of NKA subunits in mammalian osmoregulatory organs; however, most studies on fish gills have focused on the α1 subunit and did not verify the α1-β1 isozyme. Based on the sequenced milkfish transcriptome, an NKA β1 subunit gene was identified that had the highest amino acid homology to β233, a NKA β1 subunit paralog originally identified in the eel. Despite this high level of homology to β233, phylogenetic analysis and the fact that only a single NKA β1 subunit gene exists in the milkfish suggest that the milkfish gene should be referred to as the NKA β1 subunit gene. The results of accurate domain prediction of the β1 subunit, co-localization of α1 and β1 subunits in epithelial ionocytes, and co-immunoprecipitation of α1 and β1 subunits, indicated the formation of a α1-β1 complex in milkfish gills. Moreover, when transferred to hyposmotic media (fresh water) from seawater, parallel increases in branchial mRNA and protein expression of NKA α1 and β1 subunits suggested their roles in hypo-osmoregulation of euryhaline milkfish. This study molecularly characterized the NKA β1 subunit and provided the first evidence for an NKA α1-β1 association in gill ionocytes of euryhaline teleosts.

Instantaneous speed jitter detection via encoder signal and its application for the diagnosis of planetary gearbox

NASA Astrophysics Data System (ADS)

Zhao, Ming; Jia, Xiaodong; Lin, Jing; Lei, Yaguo; Lee, Jay

2018-01-01

In modern rotating machinery, rotary encoders have been widely used for the purpose of positioning and dynamic control. The study in this paper indicates that, the encoder signal, after proper processing, can be also effectively used for the health monitoring of rotating machines. In this work, a Kurtosis-guided local polynomial differentiator (KLPD) is proposed to estimate the instantaneous angular speed (IAS) of rotating machines based on the encoder signal. Compared with the central difference method, the KLPD is more robust to noise and it is able to precisely capture the weak speed jitters introduced by mechanical defects. The fault diagnosis of planetary gearbox has proven to be a challenging issue in both industry and academia. Based on the proposed KLPD, a systematic method for the fault diagnosis of planetary gearbox is proposed. In this method, residual time synchronous time averaging (RTSA) is first employed to remove the operation-related IAS components that come from normal gear meshing and non-stationary load variations, KLPD is then utilized to detect and enhance the speed jitter from the IAS residual in a data-driven manner. The effectiveness of proposed method has been validated by both simulated data and experimental data. The results demonstrate that the proposed KLPD-RTSA could not only detect fault signatures but also identify defective components, thus providing a promising tool for the health monitoring of planetary gearbox.

Use of a purified and functional recombinant calcium-channel beta4 subunit in surface-plasmon resonance studies.

PubMed Central

Geib, Sandrine; Sandoz, Guillaume; Mabrouk, Kamel; Matavel, Alessandra; Marchot, Pascale; Hoshi, Toshinori; Villaz, Michel; Ronjat, Michel; Miquelis, Raymond; Lévêque, Christian; de Waard, Michel

2002-01-01

Native high-voltage-gated calcium channels are multi-subunit complexes comprising a pore-forming subunit Ca(v) and at least two auxiliary subunits alpha(2)delta and beta. The beta subunit facilitates cell-surface expression of the channel and contributes significantly to its biophysical properties. In spite of its importance, detailed structural and functional studies are hampered by the limited availability of native beta subunit. Here, we report the purification of a recombinant calcium-channel beta(4) subunit from bacterial extracts by using a polyhistidine tag. The purified protein is fully functional since it binds on the alpha1 interaction domain, its main Ca(v)-binding site, and regulates the activity of P/Q calcium channel expressed in Xenopus oocytes in a similar way to the beta(4) subunit produced by cRNA injection. We took advantage of the functionality of the purified material to (i) develop an efficient surface-plasmon resonance assay of the interaction between two calcium channel subunits and (ii) measure, for the first time, the affinity of the recombinant His-beta(4) subunit for the full-length Ca(v)2.1 channel. The availability of this purified material and the development of a surface-plasmon resonance assay opens two immediate research perspectives: (i) drug screening programmes applied to the Ca(v)/beta interaction and (ii) crystallographic studies of the calcium-channel beta(4) subunit. PMID:11988102

Electrophysiological and biochemical evidence that DEG/ENaC cation channels are composed of nine subunits.

PubMed

Snyder, P M; Cheng, C; Prince, L S; Rogers, J C; Welsh, M J

1998-01-09

Members of the DEG/ENaC protein family form ion channels with diverse functions. DEG/ENaC subunits associate as hetero- and homomultimers to generate channels; however the stoichiometry of these complexes is unknown. To determine the subunit stoichiometry of the human epithelial Na+ channel (hENaC), we expressed the three wild-type hENaC subunits (alpha, beta, and gamma) with subunits containing mutations that alter channel inhibition by methanethiosulfonates. The data indicate that hENaC contains three alpha, three beta, and three gamma subunits. Sucrose gradient sedimentation of alphahENaC translated in vitro, as well as alpha-, beta-, and gammahENaC coexpressed in cells, was consistent with complexes containing nine subunits. FaNaCh and BNC1, two related DEG/ENaC channels, produced complexes of similar mass. Our results suggest a novel nine-subunit stoichiometry for the DEG/ENaC family of ion channels.

Rat lung glutathione S-transferases. Evidence for two distinct types of 22000-Mr subunits.

PubMed Central

Singh, S V; Partridge, C A; Awasthi, Y C

1984-01-01

Two immunologically distinct types of 22000-Mr subunits are present in rat lung glutathione S-transferases. One of these subunits is probably similar to Ya subunits of rat liver glutathione S-transferases, whereas the other subunit Ya' is immunologically distinct. Glutathione S-transferase II (pI7.2) of rat lung is a heterodimer (YaYa') of these subunits, and glutathione S-transferase VI (pI4.8) of rat lung is a homodimer of Ya' subunits. On hybridization in vitro of the subunits of glutathione S-transferase II of rat lung three active dimers having pI values 9.4, 7.2 and 4.8 are obtained. Immunological properties and substrate specificities indicate that the hybridized enzymes having pI7.2 and 4.8 correspond to glutathione S-transferases II and VI of rat lung respectively. Images Fig. 1. Fig. 5. PMID:6433888

SUGGESTIONS FOR OPTIMIZED PLANNING OF MULTIVARIATE MONITORING OF ATMOSPHERIC POLLUTION

EPA Science Inventory

Recent work in factor analysis of multivariate data sets has shown that variables with little signal should not be included in the factor analysis. Work also shows that rotational ambiguity is reduced if sources impacting a receptor have both large and small contributions. Thes...

Small subunits of RNA polymerase: localization, levels and implications for core enzyme composition.

PubMed

Doherty, Geoff P; Fogg, Mark J; Wilkinson, Anthony J; Lewis, Peter J

2010-12-01

Bacterial RNA polymerases (RNAPs) contain several small auxiliary subunits known to co-purify with the core α, β and β' subunits. The ω subunit is conserved between Gram-positive and Gram-negative bacteria, while the δ subunit is conserved within, but restricted to, Gram-positive bacteria. Although various functions have been assigned to these subunits via in vitro assays, very little is known about their in vivo roles. In this work we constructed a pair of vectors to investigate the subcellular localization of the δ and ω subunits in Bacillus subtilis with respect to the core RNAP. We found these subunits to be closely associated with RNAP involved in transcribing both mRNA and rRNA operons. Quantification of these subunits revealed δ to be present at equimolar levels with RNAP and ω to be present at around half the level of core RNAP. For comparison, the localization and quantification of RNAP β' and ω subunits in Escherichia coli was also investigated. Similar to B. subtilis, β' and ω closely associated with the nucleoid and formed subnucleoid regions of high green fluorescent protein intensity, but, unlike ω in B. subtilis, ω levels in E. coli were close to parity with those of β'. These results indicate that δ is likely to be an integral RNAP subunit in Gram-positives, whereas ω levels differ substantially between Gram-positives and -negatives. The ω subunit may be required for RNAP assembly and subsequently be turned over at different rates or it may play roles in Gram-negative bacteria that are performed by other factors in Gram-positives.

The carboxyl-terminal region is a determinant for the intracellular behavior of the chorionic gonadotropin beta subunit: effects on the processing of the Asn-linked oligosaccharides.

PubMed

Muyan, M; Boime, I

1998-05-01

The placental hormone human CG (hCG) consists of two noncovalently linked alpha- and beta-subunits similar to the other glycoprotein hormones LH, FSH, and TSH. These heterodimers share a common alpha subunit but differ in their structurally distinct beta subunits. The CGbeta subunit is distinguished among the beta subunits by the presence of a C-terminal extension with four serine-linked oligosaccharides (carboxyl terminal peptide or CTP). In previous studies we observed that deleting this sequence decreased assembly of the truncated CGbeta subunit (CGbeta114) with the alpha-subunit and increased the heterogeneity of the secreted forms of the uncombined subunit synthesized in transfected Chinese hamster ovary (CHO) cells. The latter result was attributed to alterations in the processing of the two N-linked oligosaccharides. To examine at what step this heterogeneity occurs, the CGbeta and CGbeta114 genes were transfected into wild-type and mutant CHO cell lines that are defective in the late steps of the N-linked carbohydrate-processing pathway. We show here that removal of the CTP alters the processing of the core mannosyl unit of the subunit to complex forms at both glycosylation sites and that the oligosaccharides contain polylactosamine. Although it has been presumed that there is little intramolecular interaction between the CTP and the proximal domains of the subunit, our data suggest that the CTP sequence participates in the folding of the newly synthesized subunit, which is manifest by the posttranslational changes observed here.

Structural Determinants for Functional Coupling Between the β and α Subunits in the Ca2+-activated K+ (BK) Channel

PubMed Central

Orio, Patricio; Torres, Yolima; Rojas, Patricio; Carvacho, Ingrid; Garcia, Maria L.; Toro, Ligia; Valverde, Miguel A.; Latorre, Ramon

2006-01-01

High conductance, calcium- and voltage-activated potassium (BK, MaxiK) channels are widely expressed in mammals. In some tissues, the biophysical properties of BK channels are highly affected by coexpression of regulatory (β) subunits. The most remarkable effects of β1 and β2 subunits are an increase of the calcium sensitivity and the slow down of channel kinetics. However, the detailed characteristics of channels formed by α and β1 or β2 are dissimilar, the most remarkable difference being a reduction of the voltage sensitivity in the presence of β1 but not β2. Here we reveal the molecular regions in these β subunits that determine their differential functional coupling with the pore-forming α-subunit. We made chimeric constructs between β1 and β2 subunits, and BK channels formed by α and chimeric β subunits were expressed in Xenopus laevis oocytes. The electrophysiological characteristics of the resulting channels were determined using the patch clamp technique. Chimeric exchange of the different regions of the β1 and β2 subunits demonstrates that the NH3 and COOH termini are the most relevant regions in defining the behavior of either subunit. This strongly suggests that the intracellular domains are crucial for the fine tuning of the effects of these β subunits. Moreover, the intracellular domains of β1 are responsible for the reduction of the BK channel voltage dependence. This agrees with previous studies that suggested the intracellular regions of the α-subunit to be the target of the modulation by the β1-subunit. PMID:16446507

Differential plasma membrane targeting of voltage-dependent calcium channel subunits expressed in a polarized epithelial cell line

PubMed Central

Brice, Nicola L; Dolphin, Annette C

1999-01-01

Voltage-dependent calcium channels (VDCCs) show a highly non-uniform distribution in many cell types, including neurons and other polarized secretory cells. We have examined whether this can be mimicked in a polarized epithelial cell line (Madin-Darby canine kidney), which has been used extensively to study the targeting of proteins. We expressed the VDCC α1A, α1B or α1C subunits either alone or in combination with accessory subunits α2-δ and the different β subunits, and examined their localization immunocytochemically. An α1 subunit was only targeted to the plasma membrane if co-expressed with the accessory subunits. The combination α1C/α2-δ and all β subunits was always localized predominantly to the basolateral membrane. It has been suggested that this is equivalent to somatodendritic targeting in neurons. In contrast, the α1B subunit was expressed at the apical membrane with all the accessory subunit combinations, by 24 h after microinjection. This membrane destination shows some parallels with axonal targeting in neurons. The α1A subunit was consistently observed at the apical membrane in the combinations α1A/α2-δ/β1b or β4. In contrast, when co-expressed with α2-δ/β2a, α1A was clearly targeted to the basolateral membrane. In conclusion, the VDCC α1 subunit appears to be the primary determinant for targeting the VDCC complex, but the β subunit can modify this destination, particularly for α1A. PMID:10066897

In situ synchrotron study of electromigration induced grain rotations in Sn solder joints

DOE PAGES

Shen, Hao; Zhu, Wenxin; Li, Yao; ...

2016-04-18

In this paper we report an in situ study of the early stage of microstructure evolution induced by electromigration in a Pb-free β-Sn based solder joint by synchrotron polychromatic X-ray microdiffraction. With this technique, crystal orientation evolution is monitored at intragranular levels with high spatial and angular resolution. During the entire experiment, no crystal growth is detected, and rigid grain rotation is observed only in the two grains within the current crowding region, where high density and divergence of electric current occur. Theoretical calculation indicates that the trend of electrical resistance drop still holds under the present conditions in themore » grain with high electrical resistivity, while the other grain with low resistivity reorients to align its a-axis more parallel with the ones of its neighboring grains. A detailed study of dislocation densities and subgrain boundaries suggests that grain rotation in β-Sn, unlike grain rotation in high melting temperature metals which undergo displacive deformation, is accomplished via diffusional process mainly, due to the high homologous temperature.« less

Diurnal and Semidiurnal Variations in Earth Rotation

NASA Astrophysics Data System (ADS)

Weijing, Q.; Xu, X.; Dong, D.; Zhou, Y.

2016-12-01

In recent decades, earth orientation has been monitored with increasing accuracy by advanced space-geodetic techniques, including Satellite Laser ranging (SLR), Very Long Baseline Interferometry (VLBI) and the Global Positioning System (GPS). We are able to obtain the Earth Rotation Parameters (ERP, polar motion and rotation rate changes) by even 1 to 2 hours observation data, form which obvious diurnal and semidiurnal signals can be detected, and compare them with the predicted results by the ocean model. Both the amplitude and phase are in good agreement in the main diurnal and semidiurnal wave frequency, especially for the UT1 with Consistency of 90% , and 60% for polar motion, there are 30% motivating factor of the diurnal and semidiurnal polar motion have not been identified. This work add the motivating term libration to the empirical tidal models, which can reduce the difference between the high frequency earth rotation model and observations. Then the numerical simulated ocean tidal model is obtained with the newest ERP datas from GPS, and the Scaled Sensitivity Matrix (SSM) approach is used to separate the sidebands in major ocean tides.

Monitoring ibuprofen-nicotinamide cocrystal formation during solvent free continuous cocrystallization (SFCC) using near infrared spectroscopy as a PAT tool.

PubMed

Kelly, A L; Gough, T; Dhumal, R S; Halsey, S A; Paradkar, A

2012-04-15

The purpose of this work was to explore NIR spectroscopy as a PAT tool to monitor the formation of ibuprofen and nicotinamide cocrystals during extrusion based solvent free continuous cocrystallization (SFCC). Drug and co-former were gravimetrically fed into a heated co-rotating twin screw extruder to form cocrystals. Real-time process monitoring was performed using a high temperature NIR probe in the extruder die to assess cocrystal content and subsequently compared to off-line powder X-ray diffraction measurements. The effect of processing variables, such as temperature and mixing intensity, on the extent of cocrystal formation was investigated. NIR spectroscopy was sensitive to cocrystal formation with the appearance of new peaks and peak shifts, particularly in the 4800-5200 cm(-1) wave-number region. PXRD confirmed an increased conversion of the mixture into cocrystal with increase in barrel temperature and screw mixing intensity. A decrease in screw rotation speed also provided improved cocrystal yield due to the material experiencing longer residence times within the process. A partial least squares analysis in this region of NIR spectrum correlated well with PXRD data, providing a best fit with cocrystal conversion when a limited range of process conditions were considered, for example a single set temperature. The study suggests that NIR spectroscopy could be used to monitor cocrystal purity on an industrial scale using this continuous, solvent-free process. Copyright © 2011 Elsevier B.V. All rights reserved.

A polar-region-adaptable systematic bias collaborative measurement method for shipboard redundant rotational inertial navigation systems

NASA Astrophysics Data System (ADS)

Wang, Lin; Wu, Wenqi; Wei, Guo; Lian, Junxiang; Yu, Ruihang

2018-05-01

The shipboard redundant rotational inertial navigation system (RINS) configuration, including a dual-axis RINS and a single-axis RINS, can satisfy the demand of marine INSs of especially high reliability as well as achieving trade-off between position accuracy and cost. Generally, the dual-axis RINS is the master INS, and the single-axis RINS is the hot backup INS for high reliability purposes. An integrity monitoring system performs a fault detection function to ensure sailing safety. However, improving the accuracy of the backup INS in case of master INS failure has not been given enough attention. Without the aid of any external information, a systematic bias collaborative measurement method based on an augmented Kalman filter is proposed for the redundant RINSs. Estimates of inertial sensor biases can be used by the built-in integrity monitoring system to monitor the RINS running condition. On the other hand, a position error prediction model is designed for the single-axis RINS to estimate the systematic error caused by its azimuth gyro bias. After position error compensation, the position information provided by the single-axis RINS still remains highly accurate, even if the integrity monitoring system detects a dual-axis RINS fault. Moreover, use of a grid frame as a navigation frame makes the proposed method applicable in any area, including the polar regions. Semi-physical simulation and experiments including sea trials verify the validity of the method.

Applicability of a Crack-Detection System for Use in Rotor Disk Spin Test Experiments Being Evaluated

NASA Technical Reports Server (NTRS)

Abdul-Aziz, Ali; Baaklini, George Y.; Roth, Don J.

2004-01-01

Engine makers and aviation safety government institutions continue to have a strong interest in monitoring the health of rotating components in aircraft engines to improve safety and to lower maintenance costs. To prevent catastrophic failure (burst) of the engine, they use nondestructive evaluation (NDE) and major overhauls for periodic inspections to discover any cracks that might have formed. The lowest cost fluorescent penetrant inspection NDE technique can fail to disclose cracks that are tightly closed during rest or that are below the surface. The NDE eddy current system is more effective at detecting both crack types, but it requires careful setup and operation and only a small portion of the disk can be practically inspected. So that sensor systems can sustain normal function in a severe environment, health-monitoring systems require the sensor system to transmit a signal if a crack detected in the component is above a predetermined length (but below the length that would lead to failure) and lastly to act neutrally upon the overall performance of the engine system and not interfere with engine maintenance operations. Therefore, more reliable diagnostic tools and high-level techniques for detecting damage and monitoring the health of rotating components are very essential in maintaining engine safety and reliability and in assessing life.

Ergonomic assessment of neck posture in the minimally invasive surgery suite during laparoscopic cholecystectomy.

PubMed

van Det, M J; Meijerink, W J H J; Hoff, C; van Veelen, M A; Pierie, J P E N

2008-11-01

With the expanding implementation of minimally invasive surgery, the operating team is confronted with challenges in the field of ergonomics. Visual feedback is derived from a monitor placed outside the operating field. This crossover trial was conducted to evaluate and compare neck posture in relation to monitor position in a dedicated minimally invasive surgery (MIS) suite and a conventional operating room. Assessment of the neck was conducted for 16 surgeons, assisting surgeons, and scrub nurses performing a laparoscopic cholecystectomy in both types of operating room. Flexion and rotation of the cervical spine were measured intraoperatively using a video analysis system. A two-question visual analog scale (VAS) questionnaire was used to evaluate posture in relation to the monitor position. Neck rotation was significantly reduced in the MIS suite for the surgeon (p = 0.018) and the assisting surgeon (p < 0.001). Neck flexion was significantly improved in the MIS suite for the surgeon (p < 0.001) and the scrub nurse (p = 0.018). On the questionnaire, the operating room team scored their posture significantly higher in the MIS suite and also indicated fewer musculoskeletal complaints. The ergonomic quality of the neck posture is significantly improved in the MIS suite for the entire operating room team.

A Novel Laser and Video-Based Displacement Transducer to Monitor Bridge Deflections

PubMed Central

Vicente, Miguel A.; Gonzalez, Dorys C.; Minguez, Jesus; Schumacher, Thomas

2018-01-01

The measurement of static vertical deflections on bridges continues to be a first-level technological challenge. These data are of great interest, especially for the case of long-term bridge monitoring; in fact, they are perhaps more valuable than any other measurable parameter. This is because material degradation processes and changes of the mechanical properties of the structure due to aging (for example creep and shrinkage in concrete bridges) have a direct impact on the exhibited static vertical deflections. This paper introduces and evaluates an approach to monitor displacements and rotations of structures using a novel laser and video-based displacement transducer (LVBDT). The proposed system combines the use of laser beams, LED lights, and a digital video camera, and was especially designed to capture static and slow-varying displacements. Contrary to other video-based approaches, the camera is located on the bridge, hence allowing to capture displacements at one location. Subsequently, the sensing approach and the procedure to estimate displacements and the rotations are described. Additionally, laboratory and in-service field testing carried out to validate the system are presented and discussed. The results demonstrate that the proposed sensing approach is robust, accurate, and reliable, and also inexpensive, which are essential for field implementation. PMID:29587380

A Novel Laser and Video-Based Displacement Transducer to Monitor Bridge Deflections.

PubMed

Vicente, Miguel A; Gonzalez, Dorys C; Minguez, Jesus; Schumacher, Thomas

2018-03-25

The measurement of static vertical deflections on bridges continues to be a first-level technological challenge. These data are of great interest, especially for the case of long-term bridge monitoring; in fact, they are perhaps more valuable than any other measurable parameter. This is because material degradation processes and changes of the mechanical properties of the structure due to aging (for example creep and shrinkage in concrete bridges) have a direct impact on the exhibited static vertical deflections. This paper introduces and evaluates an approach to monitor displacements and rotations of structures using a novel laser and video-based displacement transducer (LVBDT). The proposed system combines the use of laser beams, LED lights, and a digital video camera, and was especially designed to capture static and slow-varying displacements. Contrary to other video-based approaches, the camera is located on the bridge, hence allowing to capture displacements at one location. Subsequently, the sensing approach and the procedure to estimate displacements and the rotations are described. Additionally, laboratory and in-service field testing carried out to validate the system are presented and discussed. The results demonstrate that the proposed sensing approach is robust, accurate, and reliable, and also inexpensive, which are essential for field implementation.

Morphodynamic rotation of an embayed sandy beach in a mud-dominated setting

NASA Astrophysics Data System (ADS)

Brunier, Guillaume; Anthony, Edward; Gardel, Antoine; Millet, Bertrand; Fleury, Jules; Dussouillez, Philippe

2016-04-01

The morphodynamics of beaches between bedrock headlands along the muddy French Guiana coast in South America are controlled by rotation induced by the alongshore migration of mud banks from the mouths of the Amazon River. As they migrate alongshore, these mud banks generate changes in shore-incident wave angles, resulting in reversals in longshore drift. A poor appreciation of the problems caused by this process has resulted in the past in damages to the highly urbanized sea-fronts on these beaches, including erosion and flooding. This work enhances our understanding of this rather unusual type of mud-induced rotation based on surveys of the 4 km-long Montjoly beach near Cayenne, in French Guiana, in the course of an approaching mud bank between October 2013 and October 2014. Our method was based on innovative high-resolution topographic surveys from airborne Structure-from-Motion (SfM) photogrammetry over the beach in October 2013, March 2014 and October 2014. We produced digital surface models (DSM) with a resolution of 10 cm/pixel and an accuracy less than 10 cm from RTK-GPS measurements. We further measured incident wave heights from pressure sensors and conducted a bathymetric survey of the nearshore zone in October 2014. We also modelled high-tide wave propagation over the bathymetry using the REF/DIF v2.5 model. The results show the transfer of sand from the northern part of beach to the south between October 2013 and March 2014. The October 2013 DSM shows a reflective beach in the north indicative of erosion, with a narrow 50 m-wide upper beach. The southern sector was smoother and up to 90 m-wide. Between October 2013 and March 2014, the beach rotated under the influence of a mud bank, with a 30-m retreat of the berm in the north and an advance of 40 m in the south. We quantified a loss of ≈66,000 m³ of sand in the north and a gain of ≈22,000 m³ in the south over this six-month period. The October 2014 DSM shows minor morphological changes, thus indicating the end of the rotation around March 2014 and the beginning of a phase of nearshore mud-bounding of the beach as the mud bank migrated westward. The bathymetric survey showed a 2 m-high and 100 m-wide mud bar in the nearshore zone that dissipated wave energy. Hydrodynamic monitoring between March and October 2014 confirmed this low wave-energy regime (Hs < 0.5 m) compared to a more energetic regime in the neighbouring Montabo Bay beach (Hs up to 1.5 m) located downdrift of the mud bank. The REF/DIF model confirmed the strong wave energy dissipation over the mud bar. Our study highlights the rapidity and intensity of the morphodynamic changes these beaches can undergo in a muddy context, and the need for continuous and accurate monitoring in order to assess the erosion hazard caused by rotation. Keywords: Beach rotation; Mud banks; aerial SFM photogrammetry; bathymetric monitoring; hydrodynamic modelling; REF/DIF model.

Microwave Sensor for Blade Tip Clearance and Structural Health Measurements

NASA Technical Reports Server (NTRS)

Woike, Mark R.; Bencic, Timothy J.

2008-01-01

The use of microwave based sensors for the health monitoring of rotating machinery is being explored at the NASA Glenn Research Center. The microwave sensor works on the principle of sending a continuous signal towards a rotating component and measuring the reflected signal. The phase shift of the reflected signal is proportional to the distance between the sensor and the component that is being measured. This type of sensor is beneficial in that it has the ability to operate at extremely high temperatures and is unaffected by contaminants that may be present in the rotating machinery. It is intended to use these probes in the hot sections of turbine engines for closed loop turbine clearance control and structural health measurements. Background on the sensors, an overview of their calibration and preliminary results from using them to make blade tip clearance and health measurements on a large axial vane fan will be presented.

Continuous wave dye-laser technique for simultaneous, spatially resolved measurements of temperature, pressure, and velocity of NO in an underexpanded free jet

NASA Technical Reports Server (NTRS)

Di Rosa, Michael D.; Chang, Albert Y.; Hanson, Ronald K.

1993-01-01

Gas dynamic quantities within an underexpanded nitrogen free jet, seeded with 0.5 percent NO, were measured nonintrusively by using an intracavity-doubled, rapid-tuning, CW ring dye laser. The UV beam passed obliquely through the jet axis, and its frequency repetitively scanned across adjacent rotational lines in the NO gamma band near 225 nm at a rate of 4 kHz. Spatially resolved excitation scans were obtained by monitoring the induced broadband fluoresence. Modeling the Doppler-shifted excitation scans with Voigt profiles permitted simultaneous determinations of NO velocity, rotational temperature, and pressure. Zero Doppler shift was referenced to an absorption trace obtained across a static cell and recorded concurrently with the excitation scan. Typically, the measured and predicted axial distributions agreed within 10 percent. At high Mach numbers there was evidence of rotational freezing of NO.

Short-rotation management of Eucalyptus: Guidelines for plantations in Hawaii. Forest Service general technical report (Final)

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

Whitesell, C.D.; DeBell, D.S.; Schubert, T.H.

1992-11-01

A 10-year research and development program was conducted on the island of Hawaii, where nearly 230,000 acres are suitable for growing biomass in short-rotation Eucalyptus plantations. Successful techniques are described for seedling production, plantation establishment (site preparation, weed control, planting), maintenance (weed control, fertilization), biomass yield estimation, and harvest. Basic biological relationships are described to aid decisions on site selection, initial spacing, fertilizer schedules, and rotation length. Environmental issues likely to be faced by growers of Eucalyptus plantations are discussed, including soil erosion, nutrient depletion, and monocultures. Continuing programs for tree improvement, monitoring, and silviculture research are recommeded. Production costsmore » for biomass yields are estimated for three promising management regimes, representing pure Eucalyptus plantings at dense and wide spacings and a mixed species plantation where Albizia is used as a nurse crop to provide nitrogen needed for optimum Eucalyptus growth.« less

Currents in the DIII-D Tokamak

NASA Astrophysics Data System (ADS)

Azari, A.; Eidietis, N. W.

2012-10-01

Loss of vertical control of an elongated tokamak plasma results in a vertical displacement event (VDE) which can induce large currents on open field lines and exert high JxB forces on in-vessel components. An array of first-wall tile current monitors on DIII-D provides direct measurement of the poloidal halo currents. These measurements are analyzed to create a database of halo current magnitude and asymmetry, which are found to lie within the ranges seen by numerous other tokamaks in the ITPA Disruption Database. In addition, an analysis of halo asymmetry rotation is presented, as rotation at the resonance frequencies of in-vessel components could lead to significant amplification of the halo forces. Halo current rotation is found to be far more prevalent in old (1997-2002) DIII-D halo current data than recent data (2009), perhaps due to a change in divertor geometry over that time.

Rotational homogeneity in graphene grown on Au(111)

NASA Astrophysics Data System (ADS)

Wofford, Joseph; Starodub, Elena; Walter, Andrew; Nie, Shu; Bostwick, Aaron; Bartelt, Norman; Thürmer, Konrad; Rotenberg, Eli; McCarty, Kevin; Dubon, Oscar

2012-02-01

The set of properties offered by the (111) surface of gold makes it intriguing as a platform on which to study the fundamental processes that underpin graphene growth on metals. Among these are the low carbon solubility and an interaction strength with graphene that is predicted to be smaller than most transition metals. We have investigated this synthesis process using low-energy electron microscopy and diffraction to monitor the sample surface in real time, and found that the resulting graphene film possesses a remarkable degree of rotational homogeneity. The dominant orientation of the graphene is aligned with the Au lattice, with a small minority rotated by 30 degrees. The origins of this in-plane structuring are puzzling because angularly resolved photo-emission spectroscopy and scanning tunneling microscopy experiments both suggest only a relatively small interaction between the two materials. Finally, the implications of these findings for the growth of high structural-quality graphene films are discussed.

The Influence of Antenna Pattern on Faraday Rotation in Remote Sensing at L-band

NASA Technical Reports Server (NTRS)

LeVine, David M.; Jacob, S. Daniel

2007-01-01

Faraday rotation is a change in the polarization vector of electromagnetic radiation that occurs as the waves propagate from the Earth surface through the ionosphere to a spaceborne sensor. This change can cause errors in monitoring parameters at the surface such as soil moisture and sea surface salinity and it is an important consideration for radiometers on future missions in space such as NASA's Aquarius mission and ESA's SMOS mission. Two prominent strategies for compensating for Faraday rotation are using a sum of the signal at two polarizations and using the correlation between the signals at the two polarizations. These strategies work for an idealized antenna. This paper evaluates the strategies in the context of realistic antennas such as will be built for the Aquarius radiometer. Realistic antennas will make small differences that need to be included in planning for retrieval algorithms in future missions.

Distinctive interactions of the Arabidopsis homolog of the 30 kD subunit of the cleavage and polyadenylation specificity factor (AtCPSF30) with other polyadenylation factor subunits

USDA-ARS?s Scientific Manuscript database

Background: The Arabidopsis ortholog of the 30 kD subunit of the mammalian Cleavage and Polyadenylation Specificity Factor (AtCPSF30) is an RNA-binding endonuclease that is associated with other Arabidopsis CPSF subunits (orthologs of the 160, 100, and 73 kD subunits of CPSF). In order to better u...

Inactivation properties of voltage-gated K+ channels altered by presence of beta-subunit.

PubMed

Rettig, J; Heinemann, S H; Wunder, F; Lorra, C; Parcej, D N; Dolly, J O; Pongs, O

1994-05-26

Structural and functional diversity of voltage-gated Kv1-type potassium channels in rat brain is enhanced by the association of two different types of subunits, the membrane-bound, poreforming alpha-subunits and a peripheral beta-subunit. We have cloned a beta-subunit (Kv beta 1) that is specifically expressed in the rat nervous system. Association of Kv beta 1 with alpha-subunits confers rapid A-type inactivation on non-inactivating Kv1 channels (delayed rectifiers) in expression systems in vitro. This effect is mediated by an inactivating ball domain in the Kv beta 1 amino terminus.

Analysis of the interaction mode between hyperthermophilic archaeal group II chaperonin and prefoldin using a platform of chaperonin oligomers of various subunit arrangements.

PubMed

Sahlan, Muhamad; Kanzaki, Taro; Zako, Tamotsu; Maeda, Mizuo; Yohda, Masafumi

2010-09-01

Prefoldin is a co-chaperone that captures an unfolded protein substrate and transfers it to the group II chaperonin for completion of protein folding. Group II chaperonin of a hyperthermophilic archaeon, Thermococcus strain KS-1, interacts and cooperates with archaeal prefoldins. Although the interaction sites within chaperonin and prefoldin have been analyzed, the binding mode between jellyfish-like hexameric prefoldin and the double octameric ring group II chaperonin remains unclear. As prefoldin binds the chaperonin beta subunit more strongly than the alpha subunit, we analyzed the binding mode between prefoldin and chaperonin in the context of Thermococcus group II chaperonin complexes of various subunit compositions and arrangements. The oligomers exhibited various affinities for prefoldins according to the number and order of subunits. Binding affinity increased with the number of Cpnbeta subunits. Interestingly, chaperonin complexes containing two beta subunits adjacently exhibited stronger affinities than other chaperonin complexes containing the same number of beta subunits. The result suggests that all four beta tentacles of prefoldin interact with the helical protrusions of CPN in the PFD-CPN complex as the previously proposed model that two adjacent PFD beta subunits seem to interact with two CPN adjacent subunits. Copyright © 2010 Elsevier B.V. All rights reserved.

Genetic analysis of neuronal ionotropic glutamate receptor subunits

PubMed Central

Granger, Adam J; Gray, John A; Lu, Wei; Nicoll, Roger A

2011-01-01

Abstract In the brain, fast, excitatory synaptic transmission occurs primarily through AMPA- and NMDA-type ionotropic glutamate receptors. These receptors are composed of subunit proteins that determine their biophysical properties and trafficking behaviour. Therefore, determining the function of these subunits and receptor subunit composition is essential for understanding the physiological properties of synaptic transmission. Here, we discuss and evaluate various genetic approaches that have been used to study AMPA and NMDA receptor subunits. These approaches have demonstrated that the GluA1 AMPA receptor subunit is required for activity-dependent trafficking and contributes to basal synaptic transmission, while the GluA2 subunit regulates Ca2+ permeability, homeostasis and trafficking to the synapse under basal conditions. In contrast, the GluN2A and GluN2B NMDA receptor subunits regulate synaptic AMPA receptor content, both during synaptic development and plasticity. Ongoing research in this field is focusing on the molecular interactions and mechanisms that control these functions. To accomplish this, molecular replacement techniques are being used, where native subunits are replaced with receptors containing targeted mutations. In this review, we discuss a single-cell molecular replacement approach which should arguably advance our physiological understanding of ionotropic glutamate receptor subunits, but is generally applicable to study of any neuronal protein. PMID:21768264