Force feedback effects on single molecule hopping and pulling experiments.

PubMed

Rico-Pasto, M; Pastor, I; Ritort, F

2018-03-28

Single-molecule experiments with optical tweezers have become an important tool to study the properties and mechanisms of biological systems, such as cells and nucleic acids. In particular, force unzipping experiments have been used to extract the thermodynamics and kinetics of folding and unfolding reactions. In hopping experiments, a molecule executes transitions between the unfolded and folded states at a preset value of the force [constant force mode (CFM) under force feedback] or trap position [passive mode (PM) without feedback] and the force-dependent kinetic rates extracted from the lifetime of each state (CFM) and the rupture force distributions (PM) using the Bell-Evans model. However, hopping experiments in the CFM are known to overestimate molecular distances and folding free energies for fast transitions compared to the response time of the feedback. In contrast, kinetic rate measurements from pulling experiments have been mostly done in the PM while the CFM is seldom implemented in pulling protocols. Here, we carry out hopping and pulling experiments in a short DNA hairpin in the PM and CFM at three different temperatures (6 °C, 25 °C, and 45 °C) exhibiting largely varying kinetic rates. As expected, we find that equilibrium hopping experiments in the CFM and PM perform well at 6 °C (where kinetics are slow), whereas the CFM overestimates molecular parameters at 45 °C (where kinetics are fast). In contrast, nonequilibrium pulling experiments perform well in both modes at all temperatures. This demonstrates that the same kind of feedback algorithm in the CFM leads to more reliable determination of the folding reaction parameters in irreversible pulling experiments.

Do isometric pull-down exercises increase the acromio-humeral distance?

PubMed

Sealey, P; Critchley, D

2017-06-01

To evaluate the effect of isometric shoulder extension in 90° shoulder flexion on the acromio-humeral distance, to establish the force required to achieve a clinically important increase in the acromio-humeral distance, and to investigate the practicality and reliability of real-time ultrasound measurement of the acromio-humeral distance in 90° shoulder forward flexion. Prospective single-group intervention. King's College London, Guy's Campus. Twenty healthy volunteers [five males and 15 females (40 shoulders)] with a mean age of 32 (standard deviation 10, range 19 to 55) years were recruited from the faculty and staff at King's College London. The acromio-humeral distance in asymptomatic participants was measured using real-time ultrasound in the neutral position at rest, at 90° shoulder flexion at rest, and while performing an isometric pull-down exercise at 100%, 50%, 30% and 10% maximal voluntary isometric contraction. Real-time ultrasound measures of the acromio-humeral distance. Of the 20 participants, 38 shoulders were imaged. In 90° shoulder flexion, pull-down exercises at all levels of force increased the acromio-humeral distance compared with no pull-down (P<0.05), but this was only clinically significant in males. Measures had excellent short-term intra-operator reliability. Isometric pull-down exercises lead to an increase in the acromio-humeral distance in asymptomatic males that may be clinically important, and therefore may be an appropriate exercise for patients with shoulder pathology. Ultrasound measurement of the acromio-humeral distance in 90° shoulder flexion is practical and reliable. Copyright © 2016 Chartered Society of Physiotherapy. Published by Elsevier Ltd. All rights reserved.

Linker histone H1.0 interacts with an extensive network of proteins found in the nucleolus

PubMed Central

Kalashnikova, Anna A.; Winkler, Duane D.; McBryant, Steven J.; Henderson, Ryan K.; Herman, Jacob A.; DeLuca, Jennifer G.; Luger, Karolin; Prenni, Jessica E.; Hansen, Jeffrey C.

2013-01-01

The H1 linker histones are abundant chromatin-associated DNA-binding proteins. Recent evidence suggests that linker histones also may function through protein–protein interactions. To gain a better understanding of the scope of linker histone involvement in protein–protein interactions, we used a proteomics approach to identify H1-binding proteins in human nuclear extracts. Full-length H1.0 and H1.0 lacking its C-terminal domain (CTD) were used for protein pull-downs. A total of 107 candidate H1.0 binding proteins were identified by LC-MS/MS. About one-third of the H1.0-dependent interactions were mediated by the CTD, and two-thirds by the N-terminal domain-globular domain fragment. Many of the proteins pulled down by H1.0 were core splicing factors. Another group of H1-binding proteins functions in rRNA biogenesis. H1.0 also pulled down numerous ribosomal proteins and proteins involved in cellular transport. Strikingly, nearly all of the H1.0-binding proteins are found in the nucleolus. Quantitative biophysical studies with recombinant proteins confirmed that H1.0 directly binds to FACT and the splicing factors SF2/ASF and U2AF65. Our results demonstrate that H1.0 interacts with an extensive network of proteins that function in RNA metabolism in the nucleolus, and suggest that a new paradigm for linker histone action is in order. PMID:23435226

Kinetics of molecular transitions with dynamic disorder in single-molecule pulling experiments

NASA Astrophysics Data System (ADS)

Zheng, Yue; Li, Ping; Zhao, Nanrong; Hou, Zhonghuai

2013-05-01

Macromolecular transitions are subject to large fluctuations of rate constant, termed as dynamic disorder. The individual or intrinsic transition rates and activation free energies can be extracted from single-molecule pulling experiments. Here we present a theoretical framework based on a generalized Langevin equation with fractional Gaussian noise and power-law memory kernel to study the kinetics of macromolecular transitions to address the effects of dynamic disorder on barrier-crossing kinetics under external pulling force. By using the Kramers' rate theory, we have calculated the fluctuating rate constant of molecular transition, as well as the experimentally accessible quantities such as the force-dependent mean lifetime, the rupture force distribution, and the speed-dependent mean rupture force. Particular attention is paid to the discrepancies between the kinetics with and without dynamic disorder. We demonstrate that these discrepancies show strong and nontrivial dependence on the external force or the pulling speed, as well as the barrier height of the potential of mean force. Our results suggest that dynamic disorder is an important factor that should be taken into account properly in accurate interpretations of single-molecule pulling experiments.

Pulling Down the Clouds: The O'odham Intellectual Tradition during the "Time of Famine"

ERIC Educational Resources Information Center

Martinez, David

2010-01-01

Members of the Pima, or Akimel O'odham, community, despite their experiment with a pre-1934 constitutional government, not to mention their conversion to Christianity and sending their children to school, have not generated writers and activists as did their tribal peers in other parts of the United States such as Oklahoma, the Upper Plains, and…

The association of metabotropic glutamate receptor type 5 with the neuronal Ca2+-binding protein 2 modulates receptor function.

PubMed

Canela, Laia; Fernández-Dueñas, Víctor; Albergaria, Catarina; Watanabe, Masahiko; Lluís, Carme; Mallol, Josefa; Canela, Enric I; Franco, Rafael; Luján, Rafael; Ciruela, Francisco

2009-10-01

Metabotropic glutamate (mGlu) receptors mediate in part the CNS effects of glutamate. These receptors interact with a large array of intracellular proteins in which the final role is to regulate receptor function. Here, using co-immunoprecipitation and pull-down experiments we showed a close and specific interaction between mGlu(5) receptor and NECAB2 in both transfected human embryonic kidney cells and rat hippocampus. Interestingly, in pull-down experiments increasing concentrations of calcium drastically reduced the ability of these two proteins to interact, suggesting that NECAB2 binds to mGlu(5) receptor in a calcium-regulated manner. Immunoelectron microscopy detection of NECAB2 and mGlu(5) receptor in the rat hippocampal formation indicated that both proteins are codistributed in the same subcellular compartment of pyramidal cells. In addition, the NECAB2/mGlu(5) receptor interaction regulated mGlu(5b)-mediated activation of both inositol phosphate accumulation and the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway. Overall, these findings indicate that NECAB2 by its physical interaction with mGlu(5b) receptor modulates receptor function.

A three-dimensional computerized isometric strength measurement system.

PubMed

Black, Nancy L; Das, Biman

2007-05-01

The three-dimensional Computerized Isometric Strength Measurement System (CISMS) reliably and accurately measures isometric pull and push strengths in work spaces of paraplegic populations while anticipating comparative studies with other populations. The main elements of the system were: an extendable arm, a vertical supporting track, a rotating platform, a force transducer, stability sensors and a computerized data collection interface. The CISMS with minor modification was successfully used to measure isometric push-up and pull-down strengths of paraplegics and isometric push, pull, push-up and pull-down strength in work spaces for seated and standing able-bodied populations. The instrument has satisfied criteria of versatility, safety and comfort, ease of operation, and durability. Results are accurate within 2N for aligned forces. Costing approximately $1,500 (US) including computer, the system is affordable and accurate for aligned isometric strength measurements.

Nuclear pool of phosphatidylinositol 4 phosphate 5 kinase 1α is modified by polySUMO-2 during apoptosis.

PubMed

Chakrabarti, Rajarshi; Bhowmick, Debajit; Bhargava, Varsha; Bhar, Kaushik; Siddhanta, Anirban

2013-09-20

Phosphatidylinositol 4 phosphate 5 kinase 1α (PIP5K) is mainly localized in the cytosol and plasma membrane. Studies have also indicated its prominent association with nuclear speckles. The exact nature of this nuclear pool of PIP5K is not clear. Using biochemical and microscopic techniques, we have demonstrated that the nuclear pool of PIP5K is modified by SUMO-1 in HEK-293 cells stably expressing PIP5K. Moreover, this SUMOylated pool of PIP5K increased during apoptosis. PolySUMO-2 chain conjugated PIP5K was detected by pull-down experiment using affinity-tagged RNF4, a polySUMO-2 binding protein, during late apoptosis. Copyright © 2013 Elsevier Inc. All rights reserved.

Pulling the River: The Interactions of Local and Global Influences in Chinese Early Childhood Music Education

ERIC Educational Resources Information Center

Chen-Hafteck, Lily; Zhuoya, X. U.

2008-01-01

Educators in China are facing challenges as a tug-of-war between local culture and global influences in Chinese early childhood music educations plays out. By exploring the situations of Hong Kong and Nanjing, the authors demonstrate a wide gap between policy and practice. The top-down policy from government officials is based on global views of…

Einstein's Elevator in Class: A Self-Construction by Students for the Study of the Equivalence Principle

NASA Astrophysics Data System (ADS)

Kapotis, Efstratios; Kalkanis, George

2016-10-01

According to the principle of equivalence, it is impossible to distinguish between gravity and inertial forces that a noninertial observer experiences in his own frame of reference. For example, let's consider an elevator in space that is being accelerated in one direction. An observer inside it would feel as if there was gravity force pulling him toward the opposite direction. The same holds for a person in a stationary elevator located in Earth's gravitational field. No experiment enables us to distinguish between the accelerating elevator in space and the motionless elevator near Earth's surface. Strictly speaking, when the gravitational field is non-uniform (like Earth's), the equivalence principle holds only for experiments in elevators that are small enough and that take place over a short enough period of time (Fig. 1). However, performing an experiment in an elevator in space is impractical. On the other hand, it is easy to combine both forces on the same observer, i.e., gravity and a fictitious inertial force due to acceleration. Imagine an observer in an elevator that falls freely within Earth's gravitational field. The observer experiences gravity pulling him down while it might be said that the inertial force due to gravity acceleration g pulls him up. Gravity and inertial force cancel each other, (mis)leading the observer to believe there is no gravitational field. This study outlines our implementation of a self-construction idea that we have found useful in teaching introductory physics students (undergraduate, non-majors).

Centrosome Defects, Genetic Instability and Breast Cancer Progression

DTIC Science & Technology

2006-08-01

midbody-ring localization of GFP-GAPCenA onfirmed the ring structure seen by immunofluores- ence microscopy and demonstrated that there were o antibody ...sec8 (upper panels). Graph shows sec8 levels, iodixanol density, and total protein. (C) Immunoprecipitation (IP) of Nud1-DBD (DBD antibody ) pulls down...immunoprecipi- tated endogenous centriolin from HeLa cell lysates with affinity-purified centriolin antibodies and showed that sec8 and sec5 coprecipitated

Protein Aggregation Inhibitors for ALS Therapy

DTIC Science & Technology

2013-07-01

mechanisms of neuronal degeneration remain unknown in ALS, it has been postulated that protein misfolding and aggregation may be an early event that...our best compounds from last year at different doses in the ALS mouse model, and investigating possible mechanisms of action of the compounds...attachment of groups for pull-down mechanism of action experiments. Table 1. SAR studies of substituted pyrazolones. Entry R1 R2 EC50 (M) a

DNA-PKcs, a novel functional target of acriflavine, mediates acriflavine's p53-dependent synergistic anti-tumor efficiency with melphalan.

PubMed

Cao, Ji; Lin, Guanyu; Gong, Yanling; Pan, Peichen; Ma, Yaxi; Huang, Ping; Ying, Meidan; Hou, Tingjun; He, Qiaojun; Yang, Bo

2016-12-01

Acriflavine (ACF), a known antibacterial drug, has recently been recognized as a suitable candidate for cancer chemotherapy. However, the molecular target of ACF is not fully understood, which limits its application in cancer therapy. In this study, we established a structure-specific probe-based pull-down approach to comprehensively profile the potential target of ACF, and we identified DNA dependent protein kinase catalytic subunit (DNA-PKcs) as the direct target of ACF. Since DNA-PKcs facilitates the repair process following DNA double-strand breaks, we further developed a drug combination strategy that combined ACF with the bifunctional alkylating agent melphalan, which exerted a p53-dependent synergistic efficacy against human cancer cells both in vitro and in vivo. With these findings, our study demonstrated that structure-specific probe-based pull-down approaches can be used to identify new functional target of drug, and provided novel opportunities for the development of ACF-based antitumor chemotherapies. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Measurement of O-GlcNAcylated endothelial nitric oxide synthase by using 2',5'-ADP-Sepharose pull-down assay.

PubMed

Long, Yang; Yan, Jianghong; Luo, Suxin; Liu, Zhenguo; Xia, Yong

2017-11-15

Endothelial nitric oxide synthase (eNOS) plays central roles in cardiovascular regulation and disease. eNOS function is critically affected by O-linked N-acetylglucosamine (O-GlcNAc) modification. The present method for measuring O-GlcNAcylated eNOS relies on immunoprecipitation. Such method exhibits low detection efficiency and is also costly. We here report a simplified assay by employing the high binding affinity of eNOS with the 2',5'-ADP-Sepharose resins. Together with the O-GlcNAc antibody, this assay readily allows the detection of O-GlcNAcylated eNOS in both cultured endothelial cells and rat vascular tissues. By using this assay, we demonstrate that eNOS O-GlcNAcylation is markedly elevated in the vessels of diabetic rats. Thus, a 2',5'-ADP-Sepharose-based pull-down assay is developed to measure O-GlcNAcylated eNOS. This assay is simple and efficient in detecting O-GlcNAcylated eNOS in cultured cells and animal tissues under both normal and disease conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

High Speed Modulation, Beam Steering and Control of High Power Diode Lasers

DTIC Science & Technology

2000-03-03

designing the interactive simulator in such a way that the user will be able to probe the system and change parameters on the fly. Pull -down menus will...AR and HR coatings, beam splitters, external cavities, etc) can be built and accessed via pull -down menus within an optical systems graphical GUI...supported in part by the National Science Foundation’s GOALI program under grant number DM59811466 and AFOSR under contract number F49620-97-1-0002 and

Identifying chromatin readers using a SILAC-based histone peptide pull-down approach.

PubMed

Vermeulen, Michiel

2012-01-01

Posttranslational modifications (PTMs) on core histones regulate essential processes inside the nucleus such as transcription, replication, and DNA repair. An important function of histone PTMs is the recruitment or stabilization of chromatin-modifying proteins, which are also called chromatin "readers." We have developed a generic SILAC-based peptide pull-down approach to identify such readers for histone PTMs in an unbiased manner. In this chapter, the workflow behind this method will be presented in detail. Copyright © 2012 Elsevier Inc. All rights reserved.

Binding Assays Using Recombinant SH2 Domains: Far-Western, Pull-Down, and Fluorescence Polarization.

PubMed

Machida, Kazuya; Liu, Bernard

2017-01-01

Recognition of phosphotyrosine-containing sequences by SH2 domains confers specificity in tyrosine kinase pathways. By assessing interactions between isolated SH2 domains and their binding proteins, it is possible to gain insight into otherwise inaccessible complex cellular systems. Far-Western, pull-down, and fluorescence polarization (FP) have been frequently used for characterization of phosphotyrosine signaling. Here, we outline standard protocols for these established assays using recombinant SH2 domain, emphasizing the importance of appropriate sample preparation and assay controls.

Combat Fitness a Concept Vital to National Defense

DTIC Science & Technology

2010-06-18

Physical fitness testing has traditionally been focused on a 1.5- to 3-mile run, push-ups, sit-ups, and, in some Services pull -ups, flexibility, and...Performance 6 Shoot Physical Requirements Employ hand grenades Run under load, jump, bound, high/low crawl, climb, push, pull , squat, lunge, roll...jump, bound, high/low crawl, climb, push, pull , squat, lunge, roll, stop, start, change direction and get up/down. Navigate from one point to

2005 5th Annual CMMI Technology Conference and User Group. Volume 3 - Wednesday

DTIC Science & Technology

2005-11-17

Product-Related Mistakes 28. Requirements gold-plating 29. Feature creep 30. Developer gold-plating 31. Push me, pull me negotiation 32. Research...STATE UNIVERSITY 14 IV&V Layer – Select Criticality Levels for IV&V Techniques using pull -down menus PORTLAND STATE UNIVERSITY 15...of time • Develop a proposal describing how to accomplish the goal and identifying what resources would be required Look for better solutions! • Pull

Burning in Outer Space: Microgravity

NASA Technical Reports Server (NTRS)

Matkowsky, Bernard; Aldushin, Anatoly

2000-01-01

A better understanding of combustion can lead to significant technological advances, such as less polluting, more fuel-efficient vehicles. Unfortunately, gravity can interfere with the study of combustion. Gravity drags down gases that are cooler- and, therefore, denser-than heated gases. This movement mixes the fuel and the oxidizer substance that promotes burning. Because of this mixing, an observer cannot necessarily distinguish what is happening as a result of the natural combustion process and what is caused, by the pull of gravity. To remove this uncertainty, scientists can conduct experiments that simulate the negation of gravity through freefall. This condition is known as a microgravity environment. A micro-gravity experiment may take place in a chamber that is dropped down a hole or from a high-speed drop tower. The experiment also be conducted in an airplane or a rocket during freefall in a parabolic flight path. This method provides less than a minute of microgravity at most. An experiment that requires the prolonged absence of gravity may necessitate the use of an orbiting spacecraft as a venue. However, access to an orbital laboratory is difficult to acquire. High-end computing centers such as the NCCS can provide a practical alternative to operating in microgravity. Scientists can model phenomena such as combustion without gravitys observational interference. The study of microgravity combustion produces important benefits beyond increased observational accuracy. Certain valuable materials that are produced through combustion can be formed with a more uniform crystal structure-and, therefore, improved structural quality-when the pull of gravity is removed. Furthermore, understanding how fires propagate in the absence of gravity can improve fire safety aboard spacecraft.

Measurement of Vehicle Air Conditioning Pull-Down Period

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

Thomas, John F.; Huff, Shean P.; Moore, Larry G.

2016-08-01

Air conditioner usage was characterized for high heat-load summer conditions during short driving trips using a 2009 Ford Explorer and a 2009 Toyota Corolla. Vehicles were parked in the sun with windows closed to allow the cabin to become hot. Experiments were conducted by entering the instrumented vehicles in this heated condition and driving on-road with the windows up and the air conditioning set to maximum cooling, maximum fan speed and the air flow setting to recirculate cabin air rather than pull in outside humid air. The main purpose was to determine the length of time the air conditioner systemmore » would remain at or very near maximum cooling power under these severe-duty conditions. Because of the variable and somewhat uncontrolled nature of the experiments, they serve only to show that for short vehicle trips, air conditioning can remain near or at full cooling capacity for 10-minutes or significantly longer and the cabin may be uncomfortably warm during much of this time.« less

Aldolase directly interacts with ARNO and modulates cell morphology and acidic vesicle distribution

PubMed Central

Merkulova, Maria; Hurtado-Lorenzo, Andrés; Hosokawa, Hiroyuki; Zhuang, Zhenjie; Brown, Dennis; Ausiello, Dennis A.

2011-01-01

Previously, we demonstrated that the vacuolar-type H+-ATPase (V-ATPase) a2-subunit functions as an endosomal pH sensor that interacts with the ADP-ribosylation factor (Arf) guanine nucleotide exchange factor, ARNO. In the present study, we showed that ARNO directly interacts not only with the a2-subunit but with all a-isoforms (a1–a4) of the V-ATPase, indicating a widespread regulatory interaction between V-ATPase and Arf GTPases. We then extended our search for other ARNO effectors that may modulate V-ATPase-dependent vesicular trafficking events and actin cytoskeleton remodeling. Pull-down experiments using cytosol of mouse proximal tubule cells (MTCs) showed that ARNO interacts with aldolase, but not with other enzymes of the glycolytic pathway. Direct interaction of aldolase with the pleckstrin homology domain of ARNO was revealed by pull-down assays using recombinant proteins, and surface plasmon resonance revealed their high avidity interaction with a dissociation constant: KD = 2.84 × 10−10 M. MTC cell fractionation revealed that aldolase is also associated with membranes of early endosomes. Functionally, aldolase knockdown in HeLa cells produced striking morphological changes accompanied by long filamentous cell protrusions and acidic vesicle redistribution. However, the 50% knockdown we achieved did not modulate the acidification capacity of endosomal/lysosomal compartments. Finally, a combination of small interfering RNA knockdown and overexpression revealed that the expression of aldolase is inversely correlated with gelsolin levels in HeLa cells. In summary, we have shown that aldolase forms a complex with ARNO/Arf6 and the V-ATPase and that it may contribute to remodeling of the actin cytoskeleton and/or the trafficking and redistribution of V-ATPase-dependent acidic compartments via a combination of protein-protein interaction and gene expression mechanisms. PMID:21307348

How does a soap film burst during generation?

NASA Astrophysics Data System (ADS)

Rio, Emmanuelle; Saulnier, Laurie; Restagno, Frederic; Langevin, Dominique

2011-11-01

Foams are dispersions of bubbles in a liquid matrix in the presence of stabilizing surfactants. Even if foams are ubiquitous, the ability of a solution to create a certain foam quantity is still not fully understood. As a first step, we choose to work on a simplified system and studied the stability of a soap film during its generation. We have built an experiment, in which we determine simultaneously the velocity of a frame pulled out of a soapy solution and the entire shape of the liquid film. We found that the film is made of two parts: the bottom part is of uniform and stationary thickness, well described by the classical Frankel's law; in the top part, the film drains until a black film appears near the frame upper boundary frame, and then bursts. In this study, we characterize both part of the films and show that the Frankel law breaks down at high capillary number due to surfactants confinement. We also explain why films pulled at high velocity have a shorter lifetime than those pulled at low velocity. L. Saulnier is funded by CNES.

Electromyographic analysis of three different types of lat pull-down.

PubMed

Sperandei, Sandro; Barros, Marcos A P; Silveira-Júnior, Paulo C S; Oliveira, Carlos G

2009-10-01

The purpose of this work was to evaluate the activity of the primary motor muscles during the performance of 3 lat pull-down techniques through surface electromyography (EMG). Twenty-four trained adult men performed 5 repetitions of behind-the-neck (BNL), front-of-the-neck (FNL), and V-bar exercises at 80% of 1 repetition maximum. For each technique, the root mean square from the EMG signal was registered from the pectoralis major (PM), latissimus dorsi (LD), posterior deltoid (PD), and biceps brachii (BB) and further normalized in respect to that which presented the highest value of all the techniques. A series of two-way repeated measures analysis of variance was used to compare the results, with Tukey-Kramer as the post hoc test and alpha = 0.05. During the concentric phase, PM value showed the FNL to be significantly higher than V-bar/BNL and V-bar higher than BNL. During the eccentric phase, FNL/V-bar was higher than BNL. For LD, there was no difference between techniques. PD presented BNL higher than FNL/V-bar and FNL higher than V-bar in the concentric phase and BNL higher than V-bar in the eccentric phase. BB exhibited BNL higher than V-bar/FNL and V-bar higher than FNL in both concentric and eccentric phases. Considering the main objectives of lat pull-down, we concluded that FNL is the better choice, whereas BNL is not a good lat pull-down technique and should be avoided. V-bar could be used as an alternative.

From pull-down data to protein interaction networks and complexes with biological relevance.

PubMed

Zhang, Bing; Park, Byung-Hoon; Karpinets, Tatiana; Samatova, Nagiza F

2008-04-01

Recent improvements in high-throughput Mass Spectrometry (MS) technology have expedited genome-wide discovery of protein-protein interactions by providing a capability of detecting protein complexes in a physiological setting. Computational inference of protein interaction networks and protein complexes from MS data are challenging. Advances are required in developing robust and seamlessly integrated procedures for assessment of protein-protein interaction affinities, mathematical representation of protein interaction networks, discovery of protein complexes and evaluation of their biological relevance. A multi-step but easy-to-follow framework for identifying protein complexes from MS pull-down data is introduced. It assesses interaction affinity between two proteins based on similarity of their co-purification patterns derived from MS data. It constructs a protein interaction network by adopting a knowledge-guided threshold selection method. Based on the network, it identifies protein complexes and infers their core components using a graph-theoretical approach. It deploys a statistical evaluation procedure to assess biological relevance of each found complex. On Saccharomyces cerevisiae pull-down data, the framework outperformed other more complicated schemes by at least 10% in F(1)-measure and identified 610 protein complexes with high-functional homogeneity based on the enrichment in Gene Ontology (GO) annotation. Manual examination of the complexes brought forward the hypotheses on cause of false identifications. Namely, co-purification of different protein complexes as mediated by a common non-protein molecule, such as DNA, might be a source of false positives. Protein identification bias in pull-down technology, such as the hydrophilic bias could result in false negatives.

Transanal pull-through procedure for Hirschsprung's disease: a 5-year experience.

PubMed

Jester, I; Holland-Cunz, S; Loff, S; Hosie, S; Reinshagen, K; Wirth, H; Ali, M; Waag, K-L

2009-04-01

Transanal endorectal pull-through (TEPT) has become a widely used approach for the treatment of Hirschsprung's Disease. The technique is safe and, according to previous reports, it has a good clinical outcome. In this study our experience with TEPT in the early postoperative period is evaluated. The clinical course of 34 children (28 boys and 6 girls) who underwent one-stage pull-through operation according to De la Torre for Hirschsprung's disease from January 2003 to December 2007 was reviewed. Their ages ranged from 2 months to 4 years. Complications occurring within the first four weeks after operation were analyzed. Eight of 34 children (24 %) had early complications in the form of dehiscences of the anastomosis. Two children (6 %) had symptomatic anastomotic dehiscences. One child had an almost full retraction of the colon that had to be pulled down and resutured. One child developed a retrorectal abscess three weeks postoperatively due to anastomotic leakage. The dehiscences of 6 children (18 %) were asymptomatic. These dehiscences were detected only with standardized routine examination. The dehiscences healed uneventfully after resuturing. Two other patients (6 %) developed an anastomotic stricture that could be treated with rectal dilatations. Four children (12 %) showed a single episode of postoperative enterocolitis. The rate of early clinical and particularly subclinical complications such as anastomotic dehiscences after TEPT is higher than previously estimated. Patients should be monitored carefully during the early postoperative period. Severe complications can only be avoided with a thorough examination. Early resuturing of dehiscences might be helpful to prevent hazardous sequelae.

Noxa/Bcl-2 Protein Interactions Contribute to Bortezomib Resistance in Human Lymphoid Cells*

PubMed Central

Smith, Alyson J.; Dai, Haiming; Correia, Cristina; Takahashi, Rie; Lee, Sun-Hee; Schmitz, Ingo; Kaufmann, Scott H.

2011-01-01

Previous studies have suggested that the BH3 domain of the proapoptotic Bcl-2 family member Noxa only interacts with the anti-apoptotic proteins Mcl-1 and A1 but not Bcl-2. In view of the similarity of the BH3 binding domains of these anti-apoptotic proteins as well as recent evidence that studies of isolated BH3 domains can potentially underestimate the binding between full-length Bcl-2 family members, we examined the interaction of full-length human Noxa with anti-apoptotic human Bcl-2 family members. Surface plasmon resonance using bacterially expressed proteins demonstrated that Noxa binds with mean dissociation constants (KD) of 3.4 nm for Mcl-1, 70 nm for Bcl-xL, and 250 nm for wild type human Bcl-2, demonstrating selectivity but not absolute specificity of Noxa for Mcl-1. Further analysis showed that the Noxa/Bcl-2 interaction reflected binding between the Noxa BH3 domain and the Bcl-2 BH3 binding groove. Analysis of proteins expressed in vivo demonstrated that Noxa and Bcl-2 can be pulled down together from a variety of cells. Moreover, when compared with wild type Bcl-2, certain lymphoma-derived Bcl-2 mutants bound Noxa up to 20-fold more tightly in vitro, pulled down more Noxa from cells, and protected cells against killing by transfected Noxa to a greater extent. When killing by bortezomib (an agent whose cytotoxicity in Jurkat T-cell leukemia cells is dependent on Noxa) was examined, apoptosis was enhanced by the Bcl-2/Bcl-xL antagonist ABT-737 or by Bcl-2 down-regulation and diminished by Bcl-2 overexpression. Collectively, these observations not only establish the ability of Noxa and Bcl-2 to interact but also identify Bcl-2 overexpression as a potential mechanism of bortezomib resistance. PMID:21454712

Identification of GPCR-Interacting Cytosolic Proteins Using HDL Particles and Mass Spectrometry-Based Proteomic Approach

PubMed Central

Chung, Ka Young; Day, Peter W.; Vélez-Ruiz, Gisselle; Sunahara, Roger K.; Kobilka, Brian K.

2013-01-01

G protein-coupled receptors (GPCRs) have critical roles in various physiological and pathophysiological processes, and more than 40% of marketed drugs target GPCRs. Although the canonical downstream target of an agonist-activated GPCR is a G protein heterotrimer; there is a growing body of evidence suggesting that other signaling molecules interact, directly or indirectly, with GPCRs. However, due to the low abundance in the intact cell system and poor solubility of GPCRs, identification of these GPCR-interacting molecules remains challenging. Here, we establish a strategy to overcome these difficulties by using high-density lipoprotein (HDL) particles. We used the β2-adrenergic receptor (β2AR), a GPCR involved in regulating cardiovascular physiology, as a model system. We reconstituted purified β2AR in HDL particles, to mimic the plasma membrane environment, and used the reconstituted receptor as bait to pull-down binding partners from rat heart cytosol. A total of 293 proteins were identified in the full agonist-activated β2AR pull-down, 242 proteins in the inverse agonist-activated β2AR pull-down, and 210 proteins were commonly identified in both pull-downs. A small subset of the β2AR-interacting proteins isolated was confirmed by Western blot; three known β2AR-interacting proteins (Gsα, NHERF-2, and Grb2) and 3 newly identified known β2AR-interacting proteins (AMPKα, acetyl-CoA carboxylase, and UBC-13). Profiling of the identified proteins showed a clear bias toward intracellular signal transduction pathways, which is consistent with the role of β2AR as a cell signaling molecule. This study suggests that HDL particle-reconstituted GPCRs can provide an effective platform method for the identification of GPCR binding partners coupled with a mass spectrometry-based proteomic analysis. PMID:23372797

Capture of microRNA-bound mRNAs identifies the tumor suppressor miR-34a as a regulator of growth factor signaling.

PubMed

Lal, Ashish; Thomas, Marshall P; Altschuler, Gabriel; Navarro, Francisco; O'Day, Elizabeth; Li, Xiao Ling; Concepcion, Carla; Han, Yoon-Chi; Thiery, Jerome; Rajani, Danielle K; Deutsch, Aaron; Hofmann, Oliver; Ventura, Andrea; Hide, Winston; Lieberman, Judy

2011-11-01

A simple biochemical method to isolate mRNAs pulled down with a transfected, biotinylated microRNA was used to identify direct target genes of miR-34a, a tumor suppressor gene. The method reidentified most of the known miR-34a regulated genes expressed in K562 and HCT116 cancer cell lines. Transcripts for 982 genes were enriched in the pull-down with miR-34a in both cell lines. Despite this large number, validation experiments suggested that ~90% of the genes identified in both cell lines can be directly regulated by miR-34a. Thus miR-34a is capable of regulating hundreds of genes. The transcripts pulled down with miR-34a were highly enriched for their roles in growth factor signaling and cell cycle progression. These genes form a dense network of interacting gene products that regulate multiple signal transduction pathways that orchestrate the proliferative response to external growth stimuli. Multiple candidate miR-34a-regulated genes participate in RAS-RAF-MAPK signaling. Ectopic miR-34a expression reduced basal ERK and AKT phosphorylation and enhanced sensitivity to serum growth factor withdrawal, while cells genetically deficient in miR-34a were less sensitive. Fourteen new direct targets of miR-34a were experimentally validated, including genes that participate in growth factor signaling (ARAF and PIK3R2) as well as genes that regulate cell cycle progression at various phases of the cell cycle (cyclins D3 and G2, MCM2 and MCM5, PLK1 and SMAD4). Thus miR-34a tempers the proliferative and pro-survival effect of growth factor stimulation by interfering with growth factor signal transduction and downstream pathways required for cell division.

Tectonic Evolution of the Izmir Ankara Suture Zone in Northwest Turkey Using Zircon U-Pb Geochronology and Zircon Lu-Hf Isotopic Tracers

NASA Astrophysics Data System (ADS)

Campbell, C.; Taylor, M. H.; Licht, A.; Mueller, M.; Ocakglu, F.; Moeller, A.; Metais, G.; Beard, K. C.

2017-12-01

Detrital zircons from a Cretaceous forearc basin and Tertiary foreland basin located along the Sakarya Zone of the Western Pontides were analyzed to better understand the closure history of the Tethyan oceans. The Variscan Orogeny is characterized by abundant 350-300 Ma U-Pb ages and vertical ɛHf arrays, consistent with a mature magmatic arc that emplaced plutons through a southward growing accretionary margin. An ɛHf pull-up is observed from 300-250 Ma interpreted as rifting of the Intra-Pontide Ocean. The Cimmerian Orogeny is characterized by a 250-230 Ma ɛHf pull-down, followed by a 230-200 Ma magmatic gap consistent with underthrusting of the Karakaya Complex. From 200-120 Ma another magmatic lull is observed. The Alpine Orogeny is characterized by an ɛHf pull-down from 120-85 Ma within Cretaceous forearc sediments and a 100 Ma deviant ɛHf vertical array within Tertiary foreland basin sediments. Minor zircon U-Pb age peaks and contrasting inter-basinal ɛHf evolution are interpreted to represent onset of Andean-style subduction along the southern margin of the Sakarya Zone at 120 Ma followed by crustal thickening until 85 Ma. The deviant 100 Ma ɛHf vertical array within foreland basin detritus is interpreted as initiation of intra-oceanic subduction within the Izmir-Ankara Ocean. An 85-75 Ma ɛHf pull-up from forearc basin sediments is interpreted as slab roll-back along the southern margin of the Sakarya Zone, responsible for final rifting of the Western Black Sea. At 80 Ma, a vertical ɛHf array from Tertiary foreland basin deposits is interpreted to represent synchronous melting of the Tavsanli Zone and intra-oceanic slab break-off. A single 66 Myr pre-collisional grain defines a sharp ɛHf pull-down immediately prior to total arc shut-off, interpreted to represent incipient collision between the Sakarya and Tavsanli zones. A 52 Ma syn-collisional tuff yields minimally intermediate ɛHf values followed by a slight 48 Ma ɛHf pull-down, interpreted as a second episode of slab break-off followed by crustal thickening, a result of renewed underthrusting.

Mapping mechanical force propagation through biomolecular complexes

DOE PAGES

Schoeler, Constantin; Bernardi, Rafael C.; Malinowska, Klara H.; ...

2015-08-11

In this paper, we employ single-molecule force spectroscopy with an atomic force microscope (AFM) and steered molecular dynamics (SMD) simulations to reveal force propagation pathways through a mechanically ultrastable multidomain cellulosome protein complex. We demonstrate a new combination of network-based correlation analysis supported by AFM directional pulling experiments, which allowed us to visualize stiff paths through the protein complex along which force is transmitted. Finally, the results implicate specific force-propagation routes nonparallel to the pulling axis that are advantageous for achieving high dissociation forces.

Interaction of cationic surfactants with DNA: a single-molecule study

PubMed Central

Husale, Sudhir; Grange, Wilfried; Karle, Marc; Bürgi, Stephan; Hegner, Martin

2008-01-01

The interaction of cationic surfactants with single dsDNA molecules has been studied using force-measuring optical tweezers. For hydrophobic chains of length 12 and greater, pulling experiments show characteristic features (e.g. hysteresis between the pulling and relaxation curves, force-plateau along the force curves), typical of a condensed phase (compaction of a long DNA into a micron-sized particle). Depending on the length of the hydrophobic chain of the surfactant, we observe different mechanical behaviours of the complex (DNA-surfactants), which provide evidence for different binding modes. Taken together, our measurements suggest that short-chain surfactants, which do not induce any condensation, could lie down on the DNA surface and directly interact with the DNA grooves through hydrophobic–hydrophobic interactions. In contrast, long-chain surfactants could have their aliphatic tails pointing away from the DNA surface, which could promote inter-molecular interactions between hydrophobic chains and subsequently favour DNA condensation. PMID:18203749

High-power laser with Nd:YAG single-crystal fiber grown by the micro-pulling-down technique

NASA Astrophysics Data System (ADS)

Didierjean, Julien; Castaing, Marc; Balembois, François; Georges, Patrick; Perrodin, Didier; Fourmigué, Jean Marie; Lebbou, Kherreddine; Brenier, Alain; Tillement, Olivier

2006-12-01

We present optical characterization and laser results achieved with single-crystal fibers directly grown by the micro-pulling-down technique. We investigate the spectroscopic and optical quality of the fiber, and we present the first laser results. We achieved a cw laser power of 10 W at 1064 nm for an incident pump power of 60 W at 808 nm and 360 kW peak power for 12 ns pulses at 1 kHz in the Q-switched regime. It is, to the best of our knowledge, the highest laser power ever achieved with directly grown single-crystal fibers.

How far will a behaviourally flexible invasive bird go to innovate?

PubMed Central

Logan, Corina J.

2016-01-01

Behavioural flexibility is considered a key factor in the ability to adapt to changing environments. A traditional way of characterizing behavioural flexibility is to determine whether individuals invent solutions to novel problems, termed innovativeness. Great-tailed grackles are behaviourally flexible in that they can change their preferences when a task changes using existing behaviours; however, it is unknown how far they will go to invent solutions to novel problems. To begin to answer this question, I gave grackles two novel tests that a variety of other species can perform: stick tool use and string pulling. No grackle used a stick to access out-of-reach food, even after seeing a human demonstrate the solution. No grackle spontaneously pulled a vertically oriented string, but one did pull a horizontally oriented string twice. Additionally, a third novel test was previously conducted on these individuals and it was found that no grackle spontaneously dropped stones down a platform apparatus to release food, but six out of eight did become proficient after training. These results support the idea that behavioural flexibility is a multi-faceted trait because grackles are flexible, but not particularly innovative. This contradicts the idea that behavioural flexibility and innovativeness are interchangeable terms. PMID:27429781

Cryogenic insulation strength and bond tester

NASA Technical Reports Server (NTRS)

Schuerer, P. H.; Ehl, J. H.; Prasthofer, W. P. (Inventor)

1985-01-01

A method and apparatus for testing the tensile strength and bonding strength of sprayed-on foam insulation attached to metal cryogenic fuel tanks is described. A circular cutter is used to cut the insulation down to the surface of the metal tank to form plugs of the insulation for testing in situ on the tank. The apparatus comprises an electromechanical pulling device powered by a belt battery pack. The pulling device comprises a motor driving a mechanical pulling structure comprising a horizontal shaft connected to two bell cracks which are connected to a central member. When the lower end of member is attached to a fitting, which in turn is bonded to a plug, a pulling force is exerted on the plug sufficient to rupture it. The force necessary to rupture the plug or pull it loose is displayed as a digital read-out.

Leveling up and down: the experiences of benign and malicious envy.

PubMed

van de Ven, Niels; Zeelenberg, Marcel; Pieters, Rik

2009-06-01

Envy is the painful emotion caused by the good fortune of others. This research empirically supports the distinction between two qualitatively different types of envy, namely benign and malicious envy. It reveals that the experience of benign envy leads to a moving-up motivation aimed at improving one's own position, whereas the experience of malicious envy leads to a pulling-down motivation aimed at damaging the position of the superior other. Study 1 used guided recall of the two envy types in a culture (the Netherlands) that has separate words for benign and malicious envy. Analyses of the experiential content of these emotions found the predicted differences. Study 2 and 3 used one sample from the United States and one from Spain, respectively, where a single word exists for both envy types. A latent class analysis based on the experiential content of envy confirmed the existence of separate experiences of benign and malicious envy in both these cultures as well. The authors discuss the implications of distinguishing the two envy types for theories of cooperation, group performance, and Schadenfreude.

Slab-pull and slab-push earthquakes in the Mexican, Chilean and Peruvian subduction zones

NASA Astrophysics Data System (ADS)

Lemoine, A.; Madariaga, R.; Campos, J.

2002-09-01

We studied intermediate depth earthquakes in the Chile, Peru and Mexican subduction zones, paying special attention to slab-push (down-dip compression) and slab-pull (down-dip extension) mechanisms. Although, slab-push events are relatively rare in comparison with slab-pull earthquakes, quite a few have occurred recently. In Peru, a couple slab-push events occurred in 1991 and one slab-pull together with several slab-push events occurred in 1970 near Chimbote. In Mexico, several slab-push and slab-pull events occurred near Zihuatanejo below the fault zone of the 1985 Michoacan event. In central Chile, a large M=7.1 slab-push event occurred in October 1997 that followed a series of four shallow Mw>6 thrust earthquakes on the plate interface. We used teleseismic body waveform inversion of a number of Mw>5.9 slab-push and slab-pull earthquakes in order to obtain accurate mechanisms, depths and source time functions. We used a master event method in order to get relative locations. We discussed the occurrence of the relatively rare slab-push events in the three subduction zones. Were they due to the geometry of the subduction that produces flexure inside the downgoing slab, or were they produced by stress transfer during the earthquake cycle? Stress transfer can not explain the occurence of several compressional and extensional intraplate intermediate depth earthquakes in central Chile, central Mexico and central Peru. It seemed that the heterogeneity of the stress field produced by complex slab geometry has an important influence on intraplate intermediate depth earthquakes.

Analogue modelling for localization of deformation in the extensional pull-apart basins: comparison with the west part of NAF, Turkey

NASA Astrophysics Data System (ADS)

Bulkan, Sibel; Storti, Fabrizio; Cavozzi, Cristian; Vannucchi, Paola

2017-04-01

Analogue modelling remains one of the best methods for investigating progressive deformation of pull apart systems in strike slip faults that are poorly known. Analogue model experiments for the North Anatolian Fault (NAF) system around the Sea of Marmara are extremely rare in the geological literature. Our purpose in this work is to monitor the relation between the horizontal propagation and branching of the strike slip fault, and the structural and topographic expression resulting from this process. These experiments may provide insights into the geometric evolution and kinematic of west part of the NAF system. For this purpose, we run several 3D sand box experiments, appropriately scaled. Plexiglass sheets were purposely cut to simulate the geometry of the NAF. Silicone was placed on the top of these to simulate the viscous lower crust, while the brittle upper crust was simulated with pure dry sand. Dextral relative fault motion was imposed as well using different velocities to reproduce different strain rates and pull apart formation at the releasing bend. Our experiments demonstrate the variation of the shear zone shapes and how the master-fault propagates during the deformation, helping to cover the gaps between geodetic and geologic slip information. Lower crustal flow may explain how the deformation is transferred to the upper crust, and stress partitioned among the strike slip faults and pull-apart basin systems. Stress field evolution seems to play an interesting role to help strain localization. We compare the results of these experiments with natural examples around the western part of NAF and with seismic observations.

Pulling monatomic gold wires with single molecules: an Ab initio simulation.

PubMed

Krüger, Daniel; Fuchs, Harald; Rousseau, Roger; Marx, Dominik; Parrinello, Michele

2002-10-28

Car-Parrinello molecular dynamics simulations demonstrate that pulling a single thiolate molecule anchored on a stepped gold surface does not preferentially break the sulfur-gold chemical bond. Instead, it is found that this process leads to the formation of a monoatomic gold nanowire, followed by breaking a gold-gold bond with a rupture force of about 1.2 nN. The simulations also indicate that previous single-molecule thiolate-gold and gold-gold rupture experiments both probe the same phenomenon, namely, the breaking of a gold-gold bond within a gold nanowire.

Development of advanced methods for continuous Czochralski growth. Silicon sheet growth development for the large area silicon sheet task of the low cost silicon solar array project

NASA Technical Reports Server (NTRS)

Wolfson, R. G.; Sibley, C. B.

1978-01-01

The three components required to modify the furnace for batch and continuous recharging with granular silicon were designed. The feasibility of extended growth cycles up to 40 hours long was demonstrated by a recharge simulation experiment; a 6 inch diameter crystal was pulled from a 20 kg charge, remelted, and pulled again for a total of four growth cycles, 59-1/8 inch of body length, and approximately 65 kg of calculated mass.

Development and melt growth of novel scintillating halide crystals

NASA Astrophysics Data System (ADS)

Yoshikawa, Akira; Yokota, Yuui; Shoji, Yasuhiro; Kral, Robert; Kamada, Kei; Kurosawa, Shunsuke; Ohashi, Yuji; Arakawa, Mototaka; Chani, Valery I.; Kochurikhin, Vladimir V.; Yamaji, Akihiro; Andrey, Medvedev; Nikl, Martin

2017-12-01

Melt growth of scintillating halide crystals is reviewed. The vertical Bridgman growth technique is still considered as very popular method that enables production of relatively large and commercially attractive crystals. On the other hand, the micro-pulling-down method is preferable when fabrication of small samples, sufficient for preliminary characterization of their optical and/or scintillation performance, is required. Moreover, bulk crystal growth is also available using the micro-pulling-down furnace. The examples of growths of various halide crystals by industrially friendly melt growth techniques including Czochralski and edge-defined film-fed growth methods are also discussed. Finally, traveling molten zone growth that in some degree corresponds to horizontal zone melting is briefly overviewed.

The C-type Arabidopsis thioredoxin reductase ANTR-C acts as an electron donor to 2-Cys peroxiredoxins in chloroplasts

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

Moon, Jeong Chan; Jang, Ho Hee; Chae, Ho Byoung

2006-09-22

2-Cys peroxiredoxins (Prxs) play important roles in the antioxidative defense systems of plant chloroplasts. In order to determine the interaction partner for these proteins in Arabidopsis, we used a yeast two-hybrid screening procedure with a C175S-mutant of Arabidopsis 2-Cys Prx-A as bait. A cDNA encoding an NADPH-dependent thioredoxin reductase (NTR) isotype C was identified and designated ANTR-C. We demonstrated that this protein effected efficient transfer of electrons from NADPH to the 2-Cys Prxs of chloroplasts. Interaction between 2-Cys Prx-A and ANTR-C was confirmed by a pull-down experiment. ANTR-C contained N-terminal TR and C-terminal Trx domains. It exhibited both TR andmore » Trx activities and co-localized with 2-Cys Prx-A in chloroplasts. These results suggest that ANTR-C functions as an electron donor for plastidial 2-Cys Prxs and represents the NADPH-dependent TR/Trx system in chloroplasts.« less

USP17 is upregulated in osteosarcoma and promotes cell proliferation, metastasis, and epithelial-mesenchymal transition through stabilizing SMAD4.

PubMed

Song, Chenyang; Liu, Wenge; Li, Jiandong

2017-07-01

USP17 is upregulated in several cancers, indicating that USP17 might play essential functions in tumor development. However, the function of USP17 in osteosarcoma is still unknown. Our work aimed to investigate the function of USP17 in osteosarcoma. We found that the expression of USP17 was upregulated in osteosarcoma tissues and cell lines, including MG-63 and U2OS. Several functional experiments, such as colony formation analysis, Cell Counting Kit-8 assay, wound healing analysis, and transwell assay, showed that USP17 promoted cell proliferation, migration, and invasion. Moreover, we found that USP17 facilitated migration and invasion through promoting epithelial-mesenchymal transition. SMAD4 has been found to regulate epithelial-mesenchymal transition, co-immunopurification, and glutathione S-transferase pull-down analysis demonstrated that USP17 interacted with SMAD4. Furthermore, USP17 stabilized SMAD4 through its deubiquitinase activity. In conclusion, this study shows that USP17 enhances osteosarcoma cell proliferation and invasion through stabilizing SMAD4.

Nuclear pool of phosphatidylinositol 4 phosphate 5 kinase 1α is modified by polySUMO-2 during apoptosis

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

Chakrabarti, Rajarshi; Bhowmick, Debajit; Bhargava, Varsha

2013-09-20

Highlights: •Nuclear pool of PIP5K is SUMOylated. •Enhancement of SUMOylated nuclear PIP5K during apoptosis. •Nuclear PIP5K is modified by polySUMO-1 during apoptosis. •Nuclear PIP5K is modified by polySUMO-2 chain during apoptosis. -- Abstract: Phosphatidylinositol 4 phosphate 5 kinase 1α (PIP5K) is mainly localized in the cytosol and plasma membrane. Studies have also indicated its prominent association with nuclear speckles. The exact nature of this nuclear pool of PIP5K is not clear. Using biochemical and microscopic techniques, we have demonstrated that the nuclear pool of PIP5K is modified by SUMO-1 in HEK-293 cells stably expressing PIP5K. Moreover, this SUMOylated pool ofmore » PIP5K increased during apoptosis. PolySUMO-2 chain conjugated PIP5K was detected by pull-down experiment using affinity-tagged RNF4, a polySUMO-2 binding protein, during late apoptosis.« less

Identification of RNAIII-binding proteins in Staphylococcus aureus using tethered RNAs and streptavidin aptamers based pull-down assay.

PubMed

Zhang, Xu; Zhu, Qing; Tian, Tian; Zhao, Changlong; Zang, Jianye; Xue, Ting; Sun, Baolin

2015-05-15

It has been widely recognized that small RNAs (sRNAs) play important roles in physiology and virulence control in bacteria. In Staphylococcus aureus, many sRNAs have been identified and some of them have been functionally studied. Since it is difficult to identify RNA-binding proteins (RBPs), very little has been known about the RBPs in S. aureus, especially those associated with sRNAs. Here we adopted a tRNA scaffold streptavidin aptamer based pull-down assay to identify RBPs in S. aureus. The tethered RNA was successfully captured by the streptavidin magnetic beads, and proteins binding to RNAIII were isolated and analyzed by mass spectrometry. We have identified 81 proteins, and expressed heterologously 9 of them in Escherichia coli. The binding ability of the recombinant proteins with RNAIII was further analyzed by electrophoresis mobility shift assay, and the result indicates that proteins CshA, RNase J2, Era, Hu, WalR, Pyk, and FtsZ can bind to RNAIII. This study suggests that some proteins can bind to RNA III in S. aureus, and may be involved in RNA III function. And tRSA based pull-down assay is an effective method to search for RBPs in bacteria, which should facilitate the identification and functional study of RBPs in diverse bacterial species.

FACTOR FINDER CD-ROM | Science Inventory | US EPA

EPA Pesticide Factsheets

The Factor Finder CD-ROM is a user-friendly, searchable tool used to locate exposure factors and sociodemographic data for user-defined populations. Factor Finder improves the exposure assessors and risk assessors (etc.) ability to efficiently locate exposure-related information for a population of concern. Users can either enter keywords into a user-defined search box or use pull-down menus to help pinpoint specific information. The pull-down menu features general categories such as chemicals of concern, contaminated media, geographic region, exposure pathways and routes, age, food categories, and activities to name just a few. Numerous subcategories are available for selection from the pull down menu as well. Factor Finder searches both documents to retrieve the specified data and displays the information on the user's personal computer (PC) screen. Factor Finder is used by exposure assessors, risk assessors, and other concerned communities to locate exposure-related data contained within the Exposure Factors Handbook (EFH) and Sociodemographic Data Used in Identifying Potentially Highly Exposed Populations (HEP). The EFH and the HEP are companion guidance documents produced by the National Center for Environmental Assessment (NCEA) within EPA's Office of Research and Development. The Exposure Factors Handbook (EFH) summarizes data on exposure factors (values that describe human behaviors and characteristics that affect exposure to environmental cont

Identification and Characterization of Novel FMRP-Associated miRNAs

DTIC Science & Technology

2014-12-01

further - primarily because of advantages of the approaches described below (Figures 6 and 7). We next tried FMRP antibody covalently crosslinked to...hands, crosslinked FMRP antibody did not always efficiently pull down FMRP from fly brain extract (as shown in Figure 3A) and inconsistently pulled...these goals has shed light on the molecular pathogenesis of FXS. Progress towards completion of aims: First, after significant setbacks developing novel

Manipulating motions of targeted single cells in solution by an integrated double-ring magnetic tweezers imaging microscope.

PubMed

Wu, Meiling; Yadav, Rajeev; Pal, Nibedita; Lu, H Peter

2017-07-01

Controlling and manipulating living cell motions in solution hold a high promise in developing new biotechnology and biological science. Here, we developed a magnetic tweezers device that employs a combination of two permanent magnets in up-down double-ring configuration axially fitting with a microscopic objective, allowing a picoNewton (pN) bidirectional force and motion control on the sample beyond a single upward pulling direction. The experimental force calibration and magnetic field simulation using finite element method magnetics demonstrate that the designed magnetic tweezers covers a linear-combined pN force with positive-negative polarization changes in a tenability of sub-pN scale, which can be utilized to further achieve motion manipulation by shifting the force balance. We demonstrate an application of the up-down double-ring magnetic tweezers for single cell manipulation, showing that the cells with internalized paramagnetic beads can be selectively picked up and guided in a controlled fine motion.

Manipulating motions of targeted single cells in solution by an integrated double-ring magnetic tweezers imaging microscope

NASA Astrophysics Data System (ADS)

Wu, Meiling; Yadav, Rajeev; Pal, Nibedita; Lu, H. Peter

2017-07-01

Controlling and manipulating living cell motions in solution hold a high promise in developing new biotechnology and biological science. Here, we developed a magnetic tweezers device that employs a combination of two permanent magnets in up-down double-ring configuration axially fitting with a microscopic objective, allowing a picoNewton (pN) bidirectional force and motion control on the sample beyond a single upward pulling direction. The experimental force calibration and magnetic field simulation using finite element method magnetics demonstrate that the designed magnetic tweezers covers a linear-combined pN force with positive-negative polarization changes in a tenability of sub-pN scale, which can be utilized to further achieve motion manipulation by shifting the force balance. We demonstrate an application of the up-down double-ring magnetic tweezers for single cell manipulation, showing that the cells with internalized paramagnetic beads can be selectively picked up and guided in a controlled fine motion.

High performance printed oxide field-effect transistors processed using photonic curing.

PubMed

Garlapati, Suresh Kumar; Marques, Gabriel Cadilha; Gebauer, Julia Susanne; Dehm, Simone; Bruns, Michael; Winterer, Markus; Tahoori, Mehdi Baradaran; Aghassi-Hagmann, Jasmin; Hahn, Horst; Dasgupta, Subho

2018-06-08

Oxide semiconductors are highly promising candidates for the most awaited, next-generation electronics, namely, printed electronics. As a fabrication route for the solution-processed/printed oxide semiconductors, photonic curing is becoming increasingly popular, as compared to the conventional thermal curing method; the former offers numerous advantages over the latter, such as low process temperatures and short exposure time and thereby, high throughput compatibility. Here, using dissimilar photonic curing concepts (UV-visible light and UV-laser), we demonstrate facile fabrication of high performance In 2 O 3 field-effect transistors (FETs). Beside the processing related issues (temperature, time etc.), the other known limitation of oxide electronics is the lack of high performance p-type semiconductors, which can be bypassed using unipolar logics from high mobility n-type semiconductors alone. Interestingly, here we have found that our chosen distinct photonic curing methods can offer a large variation in threshold voltage, when they are fabricated from the same precursor ink. Consequently, both depletion and enhancement-mode devices have been achieved which can be used as the pull-up and pull-down transistors in unipolar inverters. The present device fabrication recipe demonstrates fast processing of low operation voltage, high performance FETs with large threshold voltage tunability.

High performance printed oxide field-effect transistors processed using photonic curing

NASA Astrophysics Data System (ADS)

Garlapati, Suresh Kumar; Cadilha Marques, Gabriel; Gebauer, Julia Susanne; Dehm, Simone; Bruns, Michael; Winterer, Markus; Baradaran Tahoori, Mehdi; Aghassi-Hagmann, Jasmin; Hahn, Horst; Dasgupta, Subho

2018-06-01

Oxide semiconductors are highly promising candidates for the most awaited, next-generation electronics, namely, printed electronics. As a fabrication route for the solution-processed/printed oxide semiconductors, photonic curing is becoming increasingly popular, as compared to the conventional thermal curing method; the former offers numerous advantages over the latter, such as low process temperatures and short exposure time and thereby, high throughput compatibility. Here, using dissimilar photonic curing concepts (UV–visible light and UV-laser), we demonstrate facile fabrication of high performance In2O3 field-effect transistors (FETs). Beside the processing related issues (temperature, time etc.), the other known limitation of oxide electronics is the lack of high performance p-type semiconductors, which can be bypassed using unipolar logics from high mobility n-type semiconductors alone. Interestingly, here we have found that our chosen distinct photonic curing methods can offer a large variation in threshold voltage, when they are fabricated from the same precursor ink. Consequently, both depletion and enhancement-mode devices have been achieved which can be used as the pull-up and pull-down transistors in unipolar inverters. The present device fabrication recipe demonstrates fast processing of low operation voltage, high performance FETs with large threshold voltage tunability.

Quantitative identification of proteins that influence miRNA biogenesis by RNA pull-down-SILAC mass spectrometry (RP-SMS).

PubMed

Choudhury, Nila Roy; Michlewski, Gracjan

2018-06-08

RNA-binding proteins mediate and control gene expression. As some examples, they regulate pre-mRNA synthesis and processing; mRNA localisation, translation and decay; and microRNA (miRNA) biogenesis and function. Here, we present a detailed protocol for RNA pull-down coupled to stable isotope labelling by amino acids in cell culture (SILAC) mass spectrometry (RP-SMS) that enables quantitative, fast and specific detection of RNA-binding proteins that regulate miRNA biogenesis. In general, this method allows for the identification of RNA-protein complexes formed using in vitro or chemically synthesized RNAs and protein extracts derived from cultured cells. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Rod pumping and proppant flowback at the Lost Hills Field

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

Roberts, I.G.

1995-12-31

Proppant flowback from hydraulically fractured wells can lead to sand wear on the pump barrel and plunger and increased pulling costs on rod pumped wells. Two approaches for lengthening run times of the pumps were tried. One approach was to install pumps that will allow production of a sand laden fluid. Pressure actuated plunger (PAP) pumps were field tested and showed an average increase of 81.6% in run time. These split ring wiper pumps clean the barrel of sand prior to the passing of the plunger. The other approach was to keep the sand and from entering the pumps. Whenmore » down hole filters were utilized, run life of the pumps with the filters increases 135%. Well pulling cost savings of $11.91 per well-day and $9.24 per well-day are documented for the PAP pumps and filters, respectively. Application guidelines based on the sand loading rate and gross liquid production of the wells are presented, as well as some operational experiences.« less

Confirmation of translatability and functionality certifies the dual endothelin1/VEGFsp receptor (DEspR) protein.

PubMed

Herrera, Victoria L M; Steffen, Martin; Moran, Ann Marie; Tan, Glaiza A; Pasion, Khristine A; Rivera, Keith; Pappin, Darryl J; Ruiz-Opazo, Nelson

2016-06-14

In contrast to rat and mouse databases, the NCBI gene database lists the human dual-endothelin1/VEGFsp receptor (DEspR, formerly Dear) as a unitary transcribed pseudogene due to a stop [TGA]-codon at codon#14 in automated DNA and RNA sequences. However, re-analysis is needed given prior single gene studies detected a tryptophan [TGG]-codon#14 by manual Sanger sequencing, demonstrated DEspR translatability and functionality, and since the demonstration of actual non-translatability through expression studies, the standard-of-excellence for pseudogene designation, has not been performed. Re-analysis must meet UNIPROT criteria for demonstration of a protein's existence at the highest (protein) level, which a priori, would override DNA- or RNA-based deductions. To dissect the nucleotide sequence discrepancy, we performed Maxam-Gilbert sequencing and reviewed 727 RNA-seq entries. To comply with the highest level multiple UNIPROT criteria for determining DEspR's existence, we performed various experiments using multiple anti-DEspR monoclonal antibodies (mAbs) targeting distinct DEspR epitopes with one spanning the contested tryptophan [TGG]-codon#14, assessing: (a) DEspR protein expression, (b) predicted full-length protein size, (c) sequence-predicted protein-specific properties beyond codon#14: receptor glycosylation and internalization, (d) protein-partner interactions, and (e) DEspR functionality via DEspR-inhibition effects. Maxam-Gilbert sequencing and some RNA-seq entries demonstrate two guanines, hence a tryptophan [TGG]-codon#14 within a compression site spanning an error-prone compression sequence motif. Western blot analysis using anti-DEspR mAbs targeting distinct DEspR epitopes detect the identical glycosylated 17.5 kDa pull-down protein. Decrease in DEspR-protein size after PNGase-F digest demonstrates post-translational glycosylation, concordant with the consensus-glycosylation site beyond codon#14. Like other small single-transmembrane proteins, mass spectrometry analysis of anti-DEspR mAb pull-down proteins do not detect DEspR, but detect DEspR-protein interactions with proteins implicated in intracellular trafficking and cancer. FACS analyses also detect DEspR-protein in different human cancer stem-like cells (CSCs). DEspR-inhibition studies identify DEspR-roles in CSC survival and growth. Live cell imaging detects fluorescently-labeled anti-DEspR mAb targeted-receptor internalization, concordant with the single internalization-recognition sequence also located beyond codon#14. Data confirm translatability of DEspR, the full-length DEspR protein beyond codon#14, and elucidate DEspR-specific functionality. Along with detection of the tryptophan [TGG]-codon#14 within an error-prone compression site, cumulative data demonstrating DEspR protein existence fulfill multiple UNIPROT criteria, thus refuting its pseudogene designation.

Akt1 binds focal adhesion kinase via the Akt1 kinase domain independently of the pleckstrin homology domain.

PubMed

Basson, M D; Zeng, B; Wang, S

2015-10-01

Akt1 and focal adhesion kinase (FAK) are protein kinases that play key roles in normal cell signaling. Individually, aberrant expression of these kinases has been linked to a variety of cancers. Together, Akt1/FAK interactions facilitate cancer metastasis by increasing cell adhesion under conditions of increased extracellular pressure. Pathological and iatrogenic sources of pressure arise from tumor growth against constraining stroma or direct perioperative manipulation. We previously reported that 15 mmHg increased extracellular pressure causes Akt1 to both directly interact with FAK and to phosphorylate and activate it. We investigated the nature of the Akt1/FAK binding by creating truncations of recombinant FAK, conjugated to glutathione S-transferase (GST), to pull down full-length Akt1. Western blots probing for Akt1 showed that FAK/Akt1 binding persisted in FAK truncations consisting of only amino acids 1-126, FAK(NT1), which contains the F1 subdomain of its band 4.1, ezrin, radixin, and moesin (FERM) domain. Using FAK(NT1) as bait, we then pulled down truncated versions of recombinant Akt1 conjugated to HA (human influenza hemagglutinin). Probes for GST-FAK(NT1) showed Akt1-FAK binding to occur in the absence of the both the Akt1 (N)-terminal pleckstrin homology (PH) domain and its adjacent hinge region. The Akt1 (C)-terminal regulatory domain was equally unnecessary for Akt1/FAK co-immunoprecipitation. Truncations involving the Akt1 catalytic domain showed that the domain by itself was enough to pull down FAK. Additionally, a fragment spanning from the PH domain to half way through the catalytic domain demonstrated increased FAK binding compared to full length Akt1. These results begin to delineate the Akt1/FAK interaction and can be used to manipulate their force-activated signal interactions. Furthermore, the finding that the N-terminal half of the Akt1 catalytic domain binds so strongly to FAK when cleaved from the rest of the protein may suggest a means for developing novel inhibitors that target this specific Akt1/FAK interaction.

8-Nitro-cGMP Attenuates the Interaction between SNARE Complex and Complexin through S-Guanylation of SNAP-25.

PubMed

Kishimoto, Yusuke; Kunieda, Kohei; Kitamura, Atsushi; Kakihana, Yuki; Akaike, Takaaki; Ihara, Hideshi

2018-02-21

8-Nitroguanosine 3',5'-cyclic monophosphate (8-nitro-cGMP) is the second messenger in nitric oxide/reactive oxygen species redox signaling. This molecule covalently binds to protein thiol groups, called S-guanylation, and exerts various biological functions. Recently, we have identified synaptosomal-associated protein 25 (SNAP-25) as a target of S-guanylation, and demonstrated that S-guanylation of SNAP25 enhanced SNARE complex formation. In this study, we have examined the effects of S-guanylation of SNAP-25 on the interaction between the SNARE complex and complexin (cplx), which binds to the SNARE complex with a high affinity. Pull-down assays and coimmunoprecipitation experiments have revealed that S-guanylation of Cys90 in SNAP-25 attenuates the interaction between the SNARE complex and cplx. In addition, blue native-PAGE followed by Western blot analysis revealed that the amount of cplx detected at a high molecular weight decreased upon 8-nitro-cGMP treatment in SH-SY5Y cells. These results demonstrated for the first time that S-guanylation of SNAP-25 attenuates the interaction between the SNARE complex and cplx.

Data in support of the identification of neuronal and astrocyte proteins interacting with extracellularly applied oligomeric and fibrillar α-synuclein assemblies by mass spectrometry

PubMed Central

Shrivastava, Amulya Nidhi; Redeker, Virginie; Fritz, Nicolas; Pieri, Laura; Almeida, Leandro G.; Spolidoro, Maria; Liebmann, Thomas; Bousset, Luc; Renner, Marianne; Léna, Clément; Aperia, Anita; Melki, Ronald; Triller, Antoine

2016-01-01

α-Synuclein (α-syn) is the principal component of Lewy bodies, the pathophysiological hallmark of individuals affected by Parkinson disease (PD). This neuropathologic form of α-syn contributes to PD progression and propagation of α-syn assemblies between neurons. The data we present here support the proteomic analysis used to identify neuronal proteins that specifically interact with extracellularly applied oligomeric or fibrillar α-syn assemblies (conditions 1 and 2, respectively) (doi: 10.15252/embj.201591397[1]). α-syn assemblies and their cellular partner proteins were pulled down from neuronal cell lysed shortly after exposure to exogenous α-syn assemblies and the associated proteins were identified by mass spectrometry using a shotgun proteomic-based approach. We also performed experiments on pure cultures of astrocytes to identify astrocyte-specific proteins interacting with oligomeric or fibrillar α-syn (conditions 3 and 4, respectively). For each condition, proteins interacting selectively with α-syn assemblies were identified by comparison to proteins pulled-down from untreated cells used as controls. The mass spectrometry data, the database search and the peak lists have been deposited to the ProteomeXchange Consortium database via the PRIDE partner repository with the dataset identifiers PRIDE: PXD002256 to PRIDE: PXD002263 and doi: 10.6019/PXD002256 to 10.6019/PXD002263. PMID:26958642

Data in support of the identification of neuronal and astrocyte proteins interacting with extracellularly applied oligomeric and fibrillar α-synuclein assemblies by mass spectrometry.

PubMed

Shrivastava, Amulya Nidhi; Redeker, Virginie; Fritz, Nicolas; Pieri, Laura; Almeida, Leandro G; Spolidoro, Maria; Liebmann, Thomas; Bousset, Luc; Renner, Marianne; Léna, Clément; Aperia, Anita; Melki, Ronald; Triller, Antoine

2016-06-01

α-Synuclein (α-syn) is the principal component of Lewy bodies, the pathophysiological hallmark of individuals affected by Parkinson disease (PD). This neuropathologic form of α-syn contributes to PD progression and propagation of α-syn assemblies between neurons. The data we present here support the proteomic analysis used to identify neuronal proteins that specifically interact with extracellularly applied oligomeric or fibrillar α-syn assemblies (conditions 1 and 2, respectively) (doi: 10.15252/embj.201591397[1]). α-syn assemblies and their cellular partner proteins were pulled down from neuronal cell lysed shortly after exposure to exogenous α-syn assemblies and the associated proteins were identified by mass spectrometry using a shotgun proteomic-based approach. We also performed experiments on pure cultures of astrocytes to identify astrocyte-specific proteins interacting with oligomeric or fibrillar α-syn (conditions 3 and 4, respectively). For each condition, proteins interacting selectively with α-syn assemblies were identified by comparison to proteins pulled-down from untreated cells used as controls. The mass spectrometry data, the database search and the peak lists have been deposited to the ProteomeXchange Consortium database via the PRIDE partner repository with the dataset identifiers PRIDE: PXD002256 to PRIDE: PXD002263 and doi: 10.6019/PXD002256 to 10.6019/PXD002263.

Msx1 and Msx2 are functional interacting partners of T-box factors in the regulation of Connexin43.

PubMed

Boogerd, Kees-Jan; Wong, L Y Elaine; Christoffels, Vincent M; Klarenbeek, Meinke; Ruijter, Jan M; Moorman, Antoon F M; Barnett, Phil

2008-06-01

T-box factors Tbx2 and Tbx3 play key roles in the development of the cardiac conduction system, atrioventricular canal, and outflow tract of the heart. They regulate the gap-junction-encoding gene Connexin43 (Cx43) and other genes critical for heart development and function. Discovering protein partners of Tbx2 and Tbx3 will shed light on the mechanisms by which these factors regulate these gene programs. Employing an yeast 2-hybrid screen and subsequent in vitro pull-down experiments we demonstrate that muscle segment homeobox genes Msx1 and Msx2 are able to bind the cardiac T-box proteins Tbx2, Tbx3, and Tbx5. This interaction, as that of the related Nkx2.5 protein, is supported by the T-box and homeodomain alone. Overlapping spatiotemporal expression patterns of Msx1 and Msx2 together with the T-box genes during cardiac development in mouse and chicken underscore the biological significance of this interaction. We demonstrate that Msx proteins together with Tbx2 and Tbx3 suppress Cx43 promoter activity and down regulate Cx43 gene activity in a rat heart-derived cell line. Using chromatin immunoprecipitation analysis we demonstrate that Msx1 can bind the Cx43 promoter at a conserved binding site located in close proximity to a previously defined T-box binding site, and that the activity of Msx proteins on this promoter appears dependent in the presence of Tbx3. Msx1 and Msx2 can function in concert with the T-box proteins to suppress Cx43 and other working myocardial genes.

Reviews

NASA Astrophysics Data System (ADS)

2008-05-01

WE RECOMMEND Why the Sky is Blue This book gives an excellent answer to the age-old question Science Magic Book of experiments finds the fun in physics Function Generator Kit Build your own simple wave generator Dent pullers Instead of using them to pull out dents, get your pupils to pull them apart Rocket Tracker Launch and track rockets with this kit Stephen Hawking, A biograpy This book looks at both the science and the personal life of the famous physicist WORTH A LOOK The Universe and the Atom All-encompassing but uninspiring physics book Sizzling Magnets Another cheap toy proves its usefulness in the physics lab Efergy Energy-saving meter is easy to use but may not save you energy Experiments and Demonstrations in Physics This book is full of interesting experiments but skewed to a particular hardware system WEB WATCH Gary Williams recounts the valuable lessons he learned at the Software 4 Skint Schools workshop

Improving estimation of kinetic parameters in dynamic force spectroscopy using cluster analysis

NASA Astrophysics Data System (ADS)

Yen, Chi-Fu; Sivasankar, Sanjeevi

2018-03-01

Dynamic Force Spectroscopy (DFS) is a widely used technique to characterize the dissociation kinetics and interaction energy landscape of receptor-ligand complexes with single-molecule resolution. In an Atomic Force Microscope (AFM)-based DFS experiment, receptor-ligand complexes, sandwiched between an AFM tip and substrate, are ruptured at different stress rates by varying the speed at which the AFM-tip and substrate are pulled away from each other. The rupture events are grouped according to their pulling speeds, and the mean force and loading rate of each group are calculated. These data are subsequently fit to established models, and energy landscape parameters such as the intrinsic off-rate (koff) and the width of the potential energy barrier (xβ) are extracted. However, due to large uncertainties in determining mean forces and loading rates of the groups, errors in the estimated koff and xβ can be substantial. Here, we demonstrate that the accuracy of fitted parameters in a DFS experiment can be dramatically improved by sorting rupture events into groups using cluster analysis instead of sorting them according to their pulling speeds. We test different clustering algorithms including Gaussian mixture, logistic regression, and K-means clustering, under conditions that closely mimic DFS experiments. Using Monte Carlo simulations, we benchmark the performance of these clustering algorithms over a wide range of koff and xβ, under different levels of thermal noise, and as a function of both the number of unbinding events and the number of pulling speeds. Our results demonstrate that cluster analysis, particularly K-means clustering, is very effective in improving the accuracy of parameter estimation, particularly when the number of unbinding events are limited and not well separated into distinct groups. Cluster analysis is easy to implement, and our performance benchmarks serve as a guide in choosing an appropriate method for DFS data analysis.

XRayView: a teaching aid for X-ray crystallography.

PubMed

Phillips, G N

1995-10-01

A software package, XRayView, has been developed that uses interactive computer graphics to introduce basic concepts of x-ray diffraction by crystals, including the reciprocal lattice, the Ewald sphere construction, Laue cones, the wavelength dependence of the reciprocal lattice, primitive and centered lattices and systematic extinctions, rotation photography. Laue photography, space group determination and Laue group symmetry, and the alignment of crystals by examination of reciprocal space. XRayView is designed with "user-friendliness" in mind, using pull-down menus to control the program. Many of the experiences of using real x-ray diffraction equipment to examine crystalline diffraction can be simulated. Exercises are available on-line to guide the users through many typical x-ray diffraction experiments.

A global view of Escherichia coli Rsd protein and its interactions.

PubMed

Piper, Sarah E; Mitchell, Jennie E; Lee, David J; Busby, Stephen J W

2009-12-01

The Escherichia coli Rsd protein forms 1 : 1 complexes with sigma(70) protein, which is the major factor in determining promoter recognition by RNA polymerase. Here we describe measurements of the levels of Rsd, RNA polymerase, sigma(70) and the alternative sigma(38) factor. Rsd levels are sufficient to sequester a significant proportion of sigma(70) and immunoaffinity pull-down experiments show that this occurs in stationary phase but not in exponentially growing cells. Rsd expression is controlled by two promoters, P1 and P2. Experiments with lac fusions show that the P2 promoter is stronger than P1, that P2 is active in all phases of growth, and that this accounts for the high levels of Rsd.

Micro-pulling-down furnace modification and single crystal fibers growth

NASA Astrophysics Data System (ADS)

Yuan, Dongsheng; Jia, Zhitai; Li, Yang; Wu, Baiyi; Tao, Xutang

2016-03-01

Single crystal fiber (SCF) combines the excellent instinct properties of conventional bulk laser crystals, and the special geometry advantage of active optical fibers. YAG and LuAG are proper host candidates for single crystal fiber laser with high thermal conductivity. Despite a lower thermal conductivity for pure crystal than YAG, LuAG crystal is easier to obtain homogeneous optical quality, and has a thermal conductivity nearly independent from the doping level. Micropulling- down (μ-PD) has relatively small thermal gradient, and here we use μ-PD to carry out high quality SCFs. Through the μ-PD furnace manufactured by ourselves, crystal fibers with different diameters have been grown successfully. We designed and fabricated a method to adjust the thermal distribution, and with the favor of pulling-down rate, the specific diameter can be controlled perfectly. The crystalline quality and homogeneity along the whole fiber were investigated, and LuAG SCF was confirmed to have a fine crystal quality for laser.

Yeast two-hybrid and pull-down assays propose an interaction between P50 of apple chlorotic leaf spot virus and PR-10 of Malus sylvestris cv. R12740-7A.

PubMed

Wang, Y; Li, N; Zhao, X; Hu, J; He, Y; Hu, T; Wang, S; Wang, Y; Cao, K

Apple chlorotic leaf spot virus (ACLSV) movement protein (P50) is involved in cell-to-cell transport and influences the long-distance spread of silencing activity. Previously, we obtained 69 P50-interacting proteins from Malus sylvestris cv. R12740-7A and using bioinformatics analyzed their biological functions. In this study, we used the GAL4-based two-hybrid yeast system and His pull-down assays to confirm an interaction between PR-10 of M. sylvestris cv. R12740-7A and ACLSV P50. Our results provide a theoretical basis for further research on the biological function of PR-10 in ACLSV infection and the interacting mechanism between host and virus.

Triptolide inhibits COX-2 expression by regulating mRNA stability in TNF-{alpha}-treated A549 cells

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

Sun, Lixin; Zhang, Shuang; Jiang, Zhenzhou

2011-12-09

Highlights: Black-Right-Pointing-Pointer Triptolide inhibited COX-2 expression and the half-life of COX-2 mRNA is decreased. Black-Right-Pointing-Pointer The HuR protein shuttling from nucleus to cytoplasm is inhibited by triptolide. Black-Right-Pointing-Pointer Triptolide inhibited 3 Prime -UTR fluorescence reporter gene activity. Black-Right-Pointing-Pointer COX-2 mRNA binding to HuR is decreased by triptolide in pull-down experiments. -- Abstract: Cyclooxygenase-2 (COX-2) over-expression is frequently associated with human non-small-cell lung cancer (NSCLC) and involved in tumor proliferation, invasion, angiogenesis and resistance to apoptosis. In the present study, the effects of triptolide on COX-2 expression in A549 cells were investigated and triptolide was found to inhibit TNF-{alpha}-induced COX-2 expression.more » In our further studies, it was found that triptolide decreased the half-life of COX-2 mRNA dramatically and that it inhibited 3 Prime -untranslated region (3 Prime -UTR) fluorescence reporter gene activity. Meanwhile, triptolide inhibited the HuR shuttling from nucleus to cytoplasm. After triptolide treatment, decreased COX-2 mRNA in pull-down experiments with anti-HuR antibodies was observed, indicating that the decreased cytoplasmic HuR is responsible for the decreased COX-2 mRNA. Taken together, our results provided evidence for the first time that triptolide inhibited COX-2 expression by COX-2 mRNA stability modulation and post-transcriptional regulation. These results provide a novel mechanism of action for triptolide which may be important in the treatment of lung cancer.« less

Effects of mastic gum Pistacia lentiscus var. Chia on innate cellular immune effectors.

PubMed

Kottakis, Filippos; Kouzi-Koliakou, Kokona; Pendas, Stefanos; Kountouras, Jannis; Choli-Papadopoulou, Theodora

2009-02-01

The essential oil and Chios mastic gum (CMG) are natural antimicrobial agents currently broadly used in medicine owing to their antimicrobial, antioxidant, and hepatoprotective properties. The aim of this study was to investigate the effect of CMG-extracted arabinogalactan proteins (AGPs/CMG) both in vitro and in vivo, under the presence of Helicobacter pylori neutrophil-activating protein (HP-NAP), on the innate cellular immune effectors (neutrophils activations) comparing H. pylori-infected patients and healthy controls. The in-vivo effect of AGPs/CMG under the presence of HP-NAP in neutrophil activation was investigated in five H. pylori-infected patients and three healthy volunteers who received 1 g daily consumption of CMG for 2 months. All participants did not receive any immunosuppressive medication before or during the trial; patients with infectious diseases that could modify their immunologic status were excluded. In-vitro studies with pull-down experiments to assess the effect of AGPs/CMG under the presence of HP-NAP on the neutrophil activation were also carried out. Neutrophil activation was estimated by nicotinamide adenine dinucleotide phosphate-oxidase assays and optical microscopy methods by measurement of cytochrome C reduction. Neutrophil activation was reduced when incubated in vitro with HP-NAP (P=0.0027) and AGP plus HP-NAP (P=0.0004) in H. pylori-positive patients who consumed AGP for 2 months. Similar results were also obtained when neutrophils were incubated with AGP plus HP-NAP (P=0.0038) in controls. Pull-down experiments showed a specific binding of AGPs to two membrane proteins of neutrophils, possibly suggesting inhibition of neutrophil activation. AGPs/CMG inhibit neutrophil activation in the presence of HP-NAP, playing a crucial role in H. pylori-associated pathologies in gastric mucosa.

Photoaffinity Labeling Studies on a Promoter of Dendritic Spine Formation

NASA Astrophysics Data System (ADS)

Sibucao, Kevin Carlo Abril

The small molecule BTA-EG4 has been shown to be a promoter of dendritic spine formation. The mechanism behind this phenomenon, however, is not well understood. The work in this dissertation is motivated by this gap in knowledge. The first part of this dissertation focuses on photoaffinity labeling studies to identify the cellular targets of BTA-EG4. Chapter 1 provides a summary of Alzheimer's disease, the rational design of BTA-EG 4, and methods to determine targets of small molecules. In Chapter 2, the synthesis of a BTA-EG4-based photoaffinity labeling probe and photodegradation studies are presented. Kinetic studies demonstrate that the probe photolyzes rapidly under UV light. In Chapter 3, photoaffinity labeling studies and subsequent protein identification experiments are reported. Competition experiments with the photoaffinity labeling probe and BTA-EG4 demonstrate that the probe labels a 55-kDa protein specifically. Tandem mass spectrometry revealed that the 55-kDa protein is the actin binding protein fascin 1. The second part of this dissertation focuses on the major protein identified from photoaffinity labeling studies, fascin 1. Chapter 4 provides a brief survey of the structure and function of fascin 1. In Chapter 5, characterizations of the interaction between BTA-EG4 and fascin 1 are reported. Isothermal titration calorimetry confirms the physical binding between fascin 1 and BTA-EG6, a BTA-EG4 analog. Slow speed sedimentation assays reveal that BTA-EG4 does not affect the actin-bundling activity of fascin 1. However, GST pull-down experiments show that BTA-EG4 inhibits the binding of fascin 1 with the GTPase Rab35. In addition, this work demonstrates that BTA-EG4 may be mechanistically distinct from the known fascin inhibitor G2.

The PDZ protein tax-interacting protein-1 inhibits beta-catenin transcriptional activity and growth of colorectal cancer cells.

PubMed

Kanamori, Mutsumi; Sandy, Peter; Marzinotto, Stefania; Benetti, Roberta; Kai, Chikatoshi; Hayashizaki, Yoshihide; Schneider, Claudio; Suzuki, Harukazu

2003-10-03

Wnt signaling is essential during development while deregulation of this pathway frequently leads to the formation of various tumors including colorectal carcinomas. A key component of the pathway is beta-catenin that, in association with TCF-4, directly regulates the expression of Wnt-responsive genes. To identify novel binding partners of beta-catenin that may control its transcriptional activity, we performed a mammalian two-hybrid screen and isolated the Tax-interacting protein (TIP-1). The in vivo complex formation between beta-catenin and TIP-1 was verified by coimmunoprecipitation, and a direct physical association was revealed by glutathione S-transferase pull-down experiments in vitro. By using a panel of deletion mutants of both proteins, we demonstrate that the interaction is mediated by the PDZ (PSD-95/DLG/ZO-1 homology) domain of TIP-1 and requires primarily the last four amino acids of beta-catenin. TIP-1 overexpression resulted in a dose-dependent decrease in the transcriptional activity of beta-catenin when tested on the TOP/FOPFLASH reporter system. Conversely, siRNA-mediated knock-down of endogenous TIP-1 slightly increased endogenous beta-catenin transactivation function. Moreover, we show that overexpression of TIP-1 reduced the proliferation and anchorage-independent growth of colorectal cancer cells. These data suggest that TIP-1 may represent a novel regulatory element in the Wnt/beta-catenin signaling pathway.

Proceedings of the 20th International Conference on Defects in Semiconductors Held in Berkeley, CA, USA, 26-30 July 1999

DTIC Science & Technology

1999-07-30

National Science Foundation through the GOALI Program, under grant number ECS-9705134. References [1] T. Ogino, M. Aoki, Jap. J. Appl. Phys. 19 (1980... pulled from molten silicon through a graphite slot for solar cell production in economical way [8]. It was observed that EFG silicon contains high...samples the closest resem- blance to our observations is found in the Au-Hj config- uration where the --/- gold acceptor level is pulled down in the

Instructor Manual for Social Awareness and Influence Workshop

DTIC Science & Technology

2008-07-01

28 15 Module 2: Planning Phase Influence GoalI fl c l Evaluation of Situation v l ti f it ti Influence Strategy I fl c tr t y Phase 1...Inner corners of eyebrows raised, eyebrows drawn together Corner of lips pulled down Corner of lips tightened and pressed Eyebrows...being naïve, but I suppose I could pull a few strings and send in a requisition for more funding,” said James. “Can you guarantee me that I won’t be

Dopamine D1A directly interacts with otoferlin synaptic pathway proteins: Ca2+ and phosphorylation underlie an NSF-to-AP2mu1 molecular switch.

PubMed

Selvakumar, Dakshnamurthy; Drescher, Marian J; Deckard, Nathan A; Ramakrishnan, Neeliyath A; Morley, Barbara J; Drescher, Dennis G

2017-01-01

Dopamine receptors regulate exocytosis via protein-protein interactions (PPIs) as well as via adenylyl cyclase transduction pathways. Evidence has been obtained for PPIs in inner ear hair cells coupling D1A to soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE)-related proteins snapin, otoferlin, N-ethylmaleimide-sensitive factor (NSF), and adaptor-related protein complex 2, mu 1 (AP2mu1), dependent on [Ca 2+ ] and phosphorylation. Specifically, the carboxy terminus of dopamine D1A was found to directly bind t-SNARE-associated protein snapin in teleost and mammalian hair cell models by yeast two-hybrid (Y2H) and pull-down assays, and snapin directly interacts with hair cell calcium-sensor otoferlin. Surface plasmon resonance (SPR) analysis, competitive pull-downs, and co-immunoprecipitation indicated that these interactions were promoted by Ca 2+ and occur together. D1A was also found to separately interact with NSF, but with an inverse dependence on Ca 2+ Evidence was obtained, for the first time, that otoferlin domains C2A, C2B, C2D, and C2F interact with NSF and AP2mu1, whereas C2C or C2E do not bind to either protein, representing binding characteristics consistent with respective inclusion or omission in individual C2 domains of the tyrosine motif YXXΦ. In competitive pull-down assays, as predicted by K D values from SPR (+Ca 2+ ), C2F pulled down primarily NSF as opposed to AP2mu1. Phosphorylation of AP2mu1 gave rise to a reversal: an increase in binding by C2F to phosphorylated AP2mu1 was accompanied by a decrease in binding to NSF, consistent with a molecular switch for otoferlin from membrane fusion (NSF) to endocytosis (AP2mu1). An increase in phosphorylated AP2mu1 at the base of the cochlear inner hair cell was the observed response elicited by a dopamine D1A agonist, as predicted. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

Multilevel samplers as microcosms to assess microbial response to biostimulation

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

Baldwin, Brett R.; Peacock, Aaron D.; Park, Melora M.

Passive multilevel samplers (MLS) containing a solid matrix for microbial colonization were used in conjunction with a push-pull biostimulation experiment designed to promote biological U(VI) and Tc(VII) reduction. MLS were deployed at 24 elevations in the injection well and two down gradient wells to investigate the spatial variability in microbial community composition and growth prior to and following biostimulation. The microbial community was characterized by real-time PCR (Q-PCR) quantification of Bacteria, NO3- reducing bacteria (nirS and nirK), δ-proteobacteria, Geobacter sp., and methanogens (mcrA). Pretest cell densities were low overall but varied substantially with significantly greater Bacterial populations detected at circumneutralmore » pH (T-test, α=0.05) suggesting carbon substrate and low pH limitations of microbial activity. Although pretest cell densities were low, denitrifying bacteria were dominant members of the microbial community. Biostimulation with an ethanol amended groundwater resulted in concurrent NO3- and Tc(VII) reduction followed by U(VI) reduction. Q-PCR analysis of MLS revealed significant (1-2 orders of magnitude, T-test, α=0.05) increases in cell densities of Bacteria, denitrifiers, δ-proteobacteria, Geobacter sp., and methanogens in response to biostimulation. Traditionally characterization of sediment samples has been used to investigate the microbial community response to biostimulation, however, collection of sediment samples is expensive and not conducive to deep aquifers or temporal studies. The results presented demonstrate that push-pull tests with passive MLS provide an inexpensive approach to determine the effect of biostimulation on contaminant concentrations, geochemical conditions, and the microbial community composition and function.« less

Retroperitoneal laparoscopic dismembered pyeloplasty with a novel technique of JJ stenting in children.

PubMed

Yu, Jianhua; Wu, Zhonghua; Xu, Youming; Li, Zhuo; Wang, Jiansong; Qi, Fan; Chen, Xiang

2011-09-01

• To report our experience with retroperitoneal laparoscopic dismembered pyeloplasty for pelvi-ureteric junction (PUJ) obstruction in children. • Between March 2007 and December 2009, 38 children with PUJ obstruction (mean age 8.3 years, range 3-14) underwent retroperitoneal laparoscopic dismembered pyeloplasty. • A ureteric catheter was inserted into the mid-ureter cystoscopically. During pyeloplasty, the proximal end of the ureteric catheter was extracorporeally sutured to the distal end of the JJ stent with silk. • The ureteric catheter was then pulled down and the stent was pulled antegrade into the ureter and bladder. • The approach was retroperitoneal in all patients except one who required open conversion. The overall mean operative time was 162 min (range 145-210 min) and this appeared to decrease with experience. Mean hospital stay was 4 days (range 3-7 days). • Mean follow-up was 20.2 months (range 6-32 months). Satisfactory drainage with decreased hydronephrosis was documented in all patients on ultrasonography and intravenous urography. • Our study shows that retroperitoneal laparoscopic dismembered pyeloplasty is a feasible and effective alternative to open pyeloplasty with a relatively minimal complication rate in children 3 years of age and older, but it should be undertaken by experienced laparoscopic surgeons. © 2011 THE AUTHORS. BJU INTERNATIONAL © 2011 BJU INTERNATIONAL.

Cardiovirus Leader proteins bind exportins: Implications for virus replication and nucleocytoplasmic trafficking inhibition

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

Ciomperlik, Jessica J.; Basta, Holly A.; Palmenberg, Ann C., E-mail: acpalmen@wisc.edu

2016-01-15

Cardiovirus Leader proteins (L{sub X}) inhibit cellular nucleocytoplasmic trafficking by directing host kinases to phosphorylate Phe/Gly-containing nuclear pore proteins (Nups). Resolution of the Mengovirus L{sub M} structure bound to Ran GTPase, suggested this complex would further recruit specific exportins (karyopherins), which in turn mediate kinase selection. Pull-down experiments and recombinant complex reconstitution now confirm that Crm1 and CAS exportins form stable dimeric complexes with encephalomyocarditis virus L{sub E}, and also larger complexes with L{sub E}:Ran. shRNA knockdown studies support this idea. Similar activities could be demonstrated for recombinant L{sub S} and L{sub T} from Theiloviruses. When mutations were introduced tomore » alter the L{sub E} zinc finger domain, acidic domain, or dual phosphorylation sites, there was reduced exportin selection. These regions are not involved in Ran interactions, so the Ran and Crm1 binding sites on L{sub E} must be non-overlapping. The involvement of exportins in this mechanism is important to viral replication and the observation of trafficking inhibition by L{sub E}.« less

Apple RING E3 ligase MdMIEL1 inhibits anthocyanin accumulation by ubiquitinating and degrading MdMYB1 protein.

PubMed

An, Jian-Ping; Liu, Xin; Li, Hao-Hao; You, Chun-Xiang; Wang, Xiao-Fei; Hao, Yu-Jin

2017-11-01

MdMYB1 is an important regulator for anthocyanin accumulation in apple (Malus × domestica). Here, an apple RING E3 ligase, MdMIEL1, was screened out as a partner of MdMYB1 with a yeast two-hybrid approach. Pull-down, bimolecular fluorescence complementation and coimmunoprecipitation assays further verified the interaction between MdMIEL1 and MdMYB1 proteins. Subsequently, in vitro and in vivo experiments indicated that MdMIEL1 functioned as a ubiquitin E3 ligase to ubiquitinate MdMYB1 protein, followed by degradation through a 26S proteasome pathway. Furthermore, transgenic studies in apple calli and Arabidopsis demonstrated that MdMIEL1 negatively regulated anthocyanin accumulation by modulating the degradation of MdMYB1 protein. Taken together, our findings provide a new insight into the molecular mechanism by which MdMIEL1 negatively regulates anthocyanin biosynthesis by ubiquitinating and degrading MdMYB1 protein. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Determination of Rab5 activity in the cell by effector pull-down assay.

PubMed

Qi, Yaoyao; Liang, Zhimin; Wang, Zonghua; Lu, Guodong; Li, Guangpu

2015-01-01

Rab5 targets to early endosomes and is a master regulator of early endosome fusion and endocytosis in all eukaryotic cells. Like other GTPases, Rab5 functions as a molecular switch by alternating between GTP-bound and GDP-bound forms, with the former being biologically active via interactions with multiple effector proteins. Thus the Rab5-GTP level in the cell reflects Rab5 activity in promoting endosome fusion and endocytosis and is indicative of cellular endocytic activity. In this chapter, we describe a Rab5 activity assay by using GST fusion proteins with the Rab5 effectors such as Rabaptin-5, Rabenosyn-5, and EEA1 that specifically bind to GTP-bound Rab5. We compare the efficiencies of the three GST fusion proteins in the pull-down of mammalian and fungal Rab5 proteins.

BiFC Assay to Detect Calmodulin Binding to Plant Receptor Kinases.

PubMed

Fischer, Cornelia; Sauter, Margret; Dietrich, Petra

2017-01-01

Plant receptor-like kinases (RLKs) are regulated at various levels including posttranscriptional modification and interaction with regulatory proteins. Calmodulin (CaM) is a calcium-sensing protein that was shown to bind to some RLKs such as the PHYTOSULFOKINE RECEPTOR1 (PSKR1). The CaM-binding site is embedded in subdomain VIa of the kinase domain. It is possible that many more of RLKs interact with CaM than previously described. To unequivocally confirm CaM binding, several methods exist. Bimolecular fluorescence complementation (BiFC) and pull-down assays have been successfully used to study CaM binding to PSKR1 and are described in this chapter (BiFC) and in Chapter 15 (pull down). The two methods are complementary. BiFC is useful to show localization and interaction of soluble as well as of membrane-bound proteins in planta.

Selective activation of the latissimus dorsi and the inferior fibers of trapezius at various shoulder angles during isometric pull-down exertion.

PubMed

Park, Se-yeon; Yoo, Won-gyu

2013-12-01

The aim of this study was to determine the effect of isometric pull down exercise on muscle activity with shoulder elevation angles of 60°, 90°, and 120° and sagittal, scapular, and frontal movement planes, by electromyography (EMG) of the latissimus dorsi, inferior fibers of trapezius, and latissimus dorsi/inferior fibers of trapezius activity ratio. Fourteen men performed nine conditions of isometric pull down exercise (three conditions of shoulder elevation × three conditions of movement planes). Surface EMG was used to collect data from the latissimus dorsi and inferior fibers of trapezius during exercise. Two-way repeated analysis of variance with two within-subject factors (shoulder elevation angles and planes of movement) was used to determine the significance of the latissimus dorsi and inferior fibers of trapezius activity and latissimus dorsi/inferior fibers of trapezius activity ratio. The latissimus dorsi activity and ratio between the latissimus dorsi and the inferior fibers of trapezius were significantly decreased as shoulder elevation angle increased from 60° to 120°. The inferior fibers of trapezius activity was significantly increased with shoulder elevation angle. The EMG activity and the ratios were not affected by changes in movement planes. This study suggests that selective activation of the latissimus dorsi is accomplished with a low shoulder elevation angle, while the inferior fibers of the trapezius are activated with high shoulder elevation angles. Copyright © 2013 Elsevier Ltd. All rights reserved.

Drug-induced keratin 9 interaction with Hsp70 in bladder cancer cells.

PubMed

Andolino, C; Hess, C; Prince, T; Williams, H; Chernin, M

2018-05-25

A pull-down experiment (co-immunoprecipitation) was performed on a T24 human bladder cancer cell lysate treated with the Hsp inhibitor VER155008 using an Hsp70 antibody attached to Dynabeads. Keratin 9, a cytoskeleton intermediate filament protein, was identified by LC MS/MS analysis. This novel finding was confirmed by Western blotting, RT-PCR, and immunocytochemistry. Other members of the keratin family of proteins have been shown to be involved in cancer progression, most recently identified to be associated with cell invasion and metastasis. The specific role of keratin 9 expression in these cells is yet to be determined.

Bone sialoprotein does not interact with pro-gelatinase A (MMP-2) or mediate MMP-2 activation.

PubMed

Hwang, Queena; Cheifetz, Sela; Overall, Christopher M; McCulloch, Christopher A; Sodek, Jaro

2009-04-22

A recent model for activation of the zymogen form of matrix metalloproteinase 2 (MMP-2, also known as gelatinase A) has suggested that interactions between the SIBLING protein bone sialoprotein (BSP) and MMP-2 leads to conformational change in MMP-2 that initiates the conversion of the pro-enzyme into a catalytically active form. This model is particularly relevant to cancer cell metastasis to bone since BSP, bound to the alphavbeta3 integrin through its arginine-glycine-aspartic acid motif, could recruit MMP-2 to the cell surface. We critically assessed the relationship between BSP and proMMP-2 and its activation using various forms of recombinant and purified BSP and MMP-2. Gelatinase and collagenase assays, fluorescence binding assays, real-time PCR, cell culture and pull-down assays were employed to test the model. Studies with a fluorogenic substrate for MMP-2 showed no activation of proMMP-2 by BSP. Binding and pull-down assays demonstrated no interaction between MMP-2 and BSP. While BSP-mediated invasiveness has been shown to depend on its integrin-binding RGD sequence, analysis of proMMP-2 activation and the level of membrane type 1 (MT1)-MMP in cells grown on a BSP substratum showed that the BSP-alphavbeta3 integrin interaction does not induce the expression of MT1-MMP. These studies do not support a role for BSP in promoting metastasis through interactions with pro-MMP-2.

Proteomic identification of Profilin1 as a corepressor of estrogen receptor alpha in MCF7 breast cancer cells.

PubMed

Kanaujiya, Jitendra Kumar; Lochab, Savita; Kapoor, Isha; Pal, Pooja; Datta, Dipak; Bhatt, Madan L B; Sanyal, Sabyasachi; Behre, Gerhard; Trivedi, Arun Kumar

2013-07-01

Nuclear receptor coregulators play an important role in the transcriptional regulation of nuclear receptors. In the present study, we aimed to identify estrogen receptor α (ERα) interacting proteins in Tamoxifen treated MCF7 cells. Using in vitro GST-pull down assay with ERα ligand-binding domain (ERα-LBD) and MS-based proteomics approach we identified Profilin1 as a novel ERα interacting protein. Profilin1 contains I/LXX/L/H/I amino acid signature motif required for corepressor interaction with ERα. We show that these two proteins physically interact with each other both in vitro as well as in vivo by GST-pull down and coimmunoprecipitation, respectively. We further show that these two proteins also colocalize together in the nucleus. Previous studies have reported reduced expression of Profilin1 in breast cancer; and here we found that Tamoxifen increases Profilin1 expression in MCF7 cells. Our data demonstrate that over expression of Profilin1 inhibits ERα-mediated transcriptional activation as well as its downstream target genes in ERα positive breast cancer cells MCF7. In addition, Profilin1 overexpression in MCF7 cells leads to inhibition of cell proliferation that apparently is due to enhanced apoptosis. In nutshell, these data indicate that MS-based proteomics approach identifies a novel ERα interacting protein Profilin1 that serves as a putative corepressor of ERα functions. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Causal assessment of occupational pushing or pulling and low back pain: results of a systematic review.

PubMed

Roffey, Darren M; Wai, Eugene K; Bishop, Paul; Kwon, Brian K; Dagenais, Simon

2010-06-01

Low back pain (LBP) is a prevalent and expensive musculoskeletal condition that predominantly occurs in working-age individuals of industrialized nations. Although numerous occupational physical activities have been implicated in its etiology, determining the causation of occupational LBP still remains a challenge. To conduct a systematic review evaluating the causal relationship between occupational pushing or pulling and LBP. Systematic review of the literature. Studies reporting an association between occupational pushing or pulling and LBP. Numerical association between exposure to pushing or pulling and the presence of LBP. A systematic review was performed to identify, evaluate, and summarize the literature related to establishing a causal relationship, according to Bradford-Hill criteria for causation for occupational pushing or pulling and LBP. A search was conducted using Medline, EMBASE, CINAHL, Cochrane Library, and OSH-ROM, gray literature, hand-searching occupational health journals, reference lists of included studies, and expert knowledge. Methodological quality was assessed using a modified Newcastle-Ottawa Scale. This search yielded 2,766 citations. Thirteen studies met the inclusion criteria. Eight were high-quality studies and five were low-quality studies. There was conflicting evidence with one high-quality study demonstrating a positive association between occupational pushing or pulling and LBP and five studies showing no relationship. One study reported a nonstatistically significant dose-response trend, four studies discussed temporality of which one indicated a positive finding, two studies discussed the biological plausibility of a causal link between occupational pushing or pulling and LBP, and no evidence was uncovered to assess the experiment criterion. A qualitative summary of existing studies was not able to find any high-quality studies that fully satisfied any of the Bradford-Hill causation criteria for occupational pushing or pulling and LBP. Based on the evidence reviewed, it is unlikely that occupational pushing or pulling is independently causative of LBP in the populations of workers studied. Copyright 2010 Elsevier Inc. All rights reserved.

Methylation Integration (Mint) | Informatics Technology for Cancer Research (ITCR)

Cancer.gov

A comprehensive software pipeline and set of Galaxy tools/workflows for integrative analysis of genome-wide DNA methylation and hydroxymethylation data. Data types can be either bisulfite sequencing and/or pull-down methods.

Characterization of MRP RNA–protein interactions within the perinucleolar compartment

PubMed Central

Pollock, Callie; Daily, Kelly; Nguyen, Van Trung; Wang, Chen; Lewandowska, Marzena Anna; Bensaude, Olivier; Huang, Sui

2011-01-01

The perinucleolar compartment (PNC) forms in cancer cells and is highly enriched with a subset of polymerase III RNAs and RNA-binding proteins. Here we report that PNC components mitochondrial RNA–processing (MRP) RNA, pyrimidine tract–binding protein (PTB), and CUG-binding protein (CUGBP) interact in vivo, as demonstrated by coimmunoprecipitation and RNA pull-down experiments. Glycerol gradient analyses show that this complex is large and sediments at a different fraction from known MRP RNA–containing complexes, the MRP ribonucleoprotein ribozyme and human telomerase reverse transcriptase. Tethering PNC components to a LacO locus recruits other PNC components, further confirming the in vivo interactions. These interactions are present both in PNC-containing and -lacking cells. High-resolution localization analyses demonstrate that MRP RNA, CUGBP, and PTB colocalize at the PNC as a reticulated network, intertwining with newly synthesized RNA. Furthermore, green fluorescent protein (GFP)–PTB and GFP-CUGBP show a slower rate of fluorescence recovery after photobleaching at the PNC than in the nucleoplasm, illustrating the different molecular interaction of the complexes associated with the PNC. These findings support a working model in which the MRP RNA–protein complex becomes nucleated at the PNC in cancer cells and may play a role in gene expression regulation at the DNA locus that associates with the PNC. PMID:21233287

Characterization of MRP RNA-protein interactions within the perinucleolar compartment.

PubMed

Pollock, Callie; Daily, Kelly; Nguyen, Van Trung; Wang, Chen; Lewandowska, Marzena Anna; Bensaude, Olivier; Huang, Sui

2011-03-15

The perinucleolar compartment (PNC) forms in cancer cells and is highly enriched with a subset of polymerase III RNAs and RNA-binding proteins. Here we report that PNC components mitochondrial RNA-processing (MRP) RNA, pyrimidine tract-binding protein (PTB), and CUG-binding protein (CUGBP) interact in vivo, as demonstrated by coimmunoprecipitation and RNA pull-down experiments. Glycerol gradient analyses show that this complex is large and sediments at a different fraction from known MRP RNA-containing complexes, the MRP ribonucleoprotein ribozyme and human telomerase reverse transcriptase. Tethering PNC components to a LacO locus recruits other PNC components, further confirming the in vivo interactions. These interactions are present both in PNC-containing and -lacking cells. High-resolution localization analyses demonstrate that MRP RNA, CUGBP, and PTB colocalize at the PNC as a reticulated network, intertwining with newly synthesized RNA. Furthermore, green fluorescent protein (GFP)-PTB and GFP-CUGBP show a slower rate of fluorescence recovery after photobleaching at the PNC than in the nucleoplasm, illustrating the different molecular interaction of the complexes associated with the PNC. These findings support a working model in which the MRP RNA-protein complex becomes nucleated at the PNC in cancer cells and may play a role in gene expression regulation at the DNA locus that associates with the PNC.

Selective recruitment of the lower fibers of the trapezius muscle.

PubMed

Arlotta, Melissa; Lovasco, Gina; McLean, Linda

2011-06-01

We aimed to determine the effectiveness of five isometric exercises at maximally activating the lower trapezius muscle in healthy subjects. Surface electromyography data were recorded from the upper, middle, and lower fibers of the trapezius muscle bilaterally while 18 healthy subjects performed five different exercises: Latissimus Pull-down, Prone Row, Prone V-Raise, Posterior Fly and Modified Prone Cobra. The peak activation was determined from the rectified and smoothed data to determine which exercise generated the highest amount of lower trapezius activity, and to determine which exercise best resulted in activation of the lower fibers of trapezius while minimizing activation of the upper and middle fibers of trapezius. Males and females demonstrated different patterns of lower trapezius recruitment and therefore the data were analyzed separately for each sex. For the males, the Prone Row exercise (2.84 ± 1.67 mV), the Posterior Fly (2.23 ± 1.00 mV) and the Modified Prone Cobra (2.26 ± 1.19 mV) exercises generated the highest EMG activity in the lower trapezius muscle. For the females, the Modified Prone Cobra (2.40 ± 1.32 mV) and the Prone Row (2.37 ± 1.14 mV) exercises generated higher activation than the Latissimus Pull Down (1.04 ± 0.56 mV), the Posterior Fly (1.62 ± 1.044 mV) and the Prone V-Raise (1.32 ± 1.07 mV). In both sexes, the Modified Prone Cobra, the Prone Row and the Latissimus Pull Down outperformed the other exercises in terms of maximizing lower trapezius activation while minimizing activation of the upper and middle fibers of trapezius. The Modified Prone Cobra showed lower relative activation of the upper trapezius muscle than did the Prone Row exercise. The Modified Prone Cobra and Prone Row exercises are the most effective exercises for targeted strengthening of the lower trapezius muscle in both sexes. The Modified Prone Cobra is somewhat better than the Prone Row due to the low activation of the upper trapezius muscle during this exercise. The Modified Prone Cobra exercise should therefore be considered as a manual muscle test position and as a strengthening exercise for the lower trapezius muscle fibers. Copyright © 2010 Elsevier Ltd. All rights reserved.

What is the mechanism of soap film entrainment?

PubMed

Saulnier, Laurie; Restagno, Frédéric; Delacotte, Jérôme; Langevin, Dominique; Rio, Emmanuelle

2011-11-15

Classical Frankel's law describes the formation of soap films and their evolution upon pulling, a model situation of film dynamics in foams (formation, rheology, and destabilization). With the purpose of relating film pulling to foam dynamics, we have built a new setup able to give an instantaneous measurement of film thickness, thus allowing us to determine film thickness profile during pulling. We found that only the lower part of the film is of uniform thickness and follows Frankel's law, provided the entrainment velocity is small. We show that this is due to confinement effects: there is not enough surfactant in the bulk to fully cover the newly created surfaces which results in immobile film surfaces. At large velocities, surfaces become mobile and then Frankel's law breaks down, leading to a faster drainage and thus to a nonstationary thickness at the bottom of the film. These findings should help in understanding the large dispersion of previous experimental data reported during the last 40 years and clarifying the pulling phenomenon of thin liquid films.

Neutrally buoyant tracers in hydrogeophysics: Field demonstration in fractured rock

NASA Astrophysics Data System (ADS)

Shakas, Alexis; Linde, Niklas; Baron, Ludovic; Selker, John; Gerard, Marie-Françoise; Lavenant, Nicolas; Bour, Olivier; Le Borgne, Tanguy

2017-04-01

Electrical and electromagnetic methods are extensively used to map electrically conductive tracers within hydrogeologic systems. Often, the tracers used consist of dissolved salt in water, leading to a denser mixture than the ambient formation water. Density effects are often ignored and rarely modeled but can dramatically affect transport behavior and introduce dynamics that are unrepresentative of the response obtained with classical tracers (e.g., uranine). We introduce a neutrally buoyant tracer consisting of a mixture of salt, water, and ethanol and monitor its movement during push-pull experiments in a fractured rock aquifer using ground-penetrating radar. Our results indicate a largely reversible transport process and agree with uranine-based push-pull experiments at the site, which is in contrast to results obtained using dense saline tracers. We argue that a shift toward neutrally buoyant tracers in both porous and fractured media would advance hydrogeophysical research and enhance its utility in hydrogeology.

Growth of Nd doped (Lu, Gd)3(Ga, Al)5O12 single crystal by the micro pulling down method and their scintillation properties

NASA Astrophysics Data System (ADS)

Kamada, Kei; Kurosawa, Shunsuke; Yamaji, Akihiro; Shoji, Yasuhiro; Pejchal, Jan; Ohashi, Yuji; Yokota, Yuui; Yoshikawa, Akira

2015-03-01

Nd 1 mol% doped (Lu, Gd)3(Ga, Al)5O12 (LGGAG) single crystals were grown by the micro-pulling down (μ-PD) method. Luminescence and scintillation properties such as absorption, excitation and emission spectra, light yield and decay time were evaluated. Nd1%:Lu3Al5O12 showed the highest light output of around 8200 photons/MeV among the grown crystals. Scintillation decay time of Nd:Y3Al5O12 was 1.32 μs (36%) 2.02 μs (64%). Nd:Lu3Ga3Al2O12 was relatively high dense scintillator of 7.38 g/cm3 with good light yield of 6800 photons/MeV and scintillation decay time of 0.20 μs (5%) 2.60 μs (95%).

Mg,Ce co-doped Lu2Gd1(Ga,Al)5O12 by micro-pulling down method and their luminescence properties

NASA Astrophysics Data System (ADS)

Kamada, Kei; Yamaguchi, Hiroaki; Yoshino, Masao; Kurosawa, Shunsuke; Shoji, Yasuhiro; Yokota, Yuui; Ohashi, Yuji; Pejchal, Jan; Nikl, Martin; Yoshikawa, Akira

2018-04-01

The effects of Mg co-doping on the scintillation properties of Ce:Lu2Gd1(Ga,Al)5O12 (LGGAG) single crystals with different Ga/Al ratios were investigated. Mg co-doped and non co-doped Ce:LGGAG single crystals were grown by the micro-pulling down (µ-PD) method and then cut, polished and annealed for each measurement. Absorption spectra, radioluminescence (RL) spectra, pulse height spectra, and scintillation decay were measured to reveal the effect of Mg co-doping. Ce4+ charge transfer (CT) absorption band peaking at ∼260 nm was observed in Mg co-doped samples, which is in good agreement with previous reports for the Ce4+ CT absorption band in other garnet-based crystals. The scintillation decay time tended to be accelerated and the light yield tended to be decreased by Mg co-doping at higher Ga concentrations.

8. VIEW SHOWING DEMOSSING WITH A DRAGLINE ON THE WESTERN ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. VIEW SHOWING DEMOSSING WITH A DRAGLINE ON THE WESTERN CANAL. THE WEEDS WERE LOOSENED UPSTREAM BY PULLING A CHAIN OR CABLE CROSSWAYS DOWN THE CANAL BED. THE LOOSENED WEEDS THEN FLOATED DOWN TO A GRATE, WHERE THEY ACCUMULATED INTO THE SOLID MASS WHICH WAS REMOVED BY DRAGLINE BUCKET. Photographer: Unknown. July 30, 1947 - Western Canal, South side of Salt River between Tempe, Phoenix & Mesa, Mesa, Maricopa County, AZ

12. ENCLOSED SLEEPING PORCH INTERIOR DETAIL SHOWING PULLDOWN STAIRCASE TO ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

12. ENCLOSED SLEEPING PORCH INTERIOR DETAIL SHOWING PULL-DOWN STAIRCASE TO ATTIC. VIEW TO SOUTHEAST. - Big Creek Hydroelectric System, Big Creek Town, Operator House, Orchard Avenue south of Huntington Lake Road, Big Creek, Fresno County, CA

Approaches to Validate and Manipulate RNA Targets with Small Molecules in Cells.

PubMed

Childs-Disney, Jessica L; Disney, Matthew D

2016-01-01

RNA has become an increasingly important target for therapeutic interventions and for chemical probes that dissect and manipulate its cellular function. Emerging targets include human RNAs that have been shown to directly cause cancer, metabolic disorders, and genetic disease. In this review, we describe various routes to obtain bioactive compounds that target RNA, with a particular emphasis on the development of small molecules. We use these cases to describe approaches that are being developed for target validation, which include target-directed cleavage, classic pull-down experiments, and covalent cross-linking. Thus, tools are available to design small molecules to target RNA and to identify the cellular RNAs that are their targets.

Akt recruits Dab2 to albumin endocytosis in the proximal tubule.

PubMed

Koral, Kelly; Li, Hui; Ganesh, Nandita; Birnbaum, Morris J; Hallows, Kenneth R; Erkan, Elif

2014-12-15

Proximal tubule epithelial cells have a highly sophisticated endocytic machinery to retrieve the albumin in the glomerular filtrate. The megalin-cubilin complex and the endocytic adaptor disabled-2 (Dab2) play a pivotal role in albumin endocytosis. We previously demonstrated that protein kinase B (Akt) regulates albumin endocytosis in the proximal tubule through an interaction with Dab2. Here, we examined the nature of Akt-Dab2 interaction. The pleckstrin homology (PH) and catalytic domains (CD) of Akt interacted with the proline-rich domain (PRD) of Dab2 based on yeast-two hybrid (Y2H) experiments. Pull-down experiments utilizing the truncated constructs of Dab2 demonstrated that the initial 11 amino acids of Dab2-PRD were sufficient to mediate the interaction between Akt and Dab2. Endocytosis experiments utilizing Akt1- and Akt2-silencing RNA revealed that both Akt1 and Akt2 mediate albumin endocytosis in proximal tubule epithelial cells; therefore, Akt1 and Akt2 may play a compensatory role in albumin endocytosis. Furthermore, both Akt isoforms phosphorylated Dab2 at Ser residues 448 and 449. Ser-to-Ala mutations of these Dab2 residues inhibited albumin endocytosis and resulted in a shift in location of Dab2 from the peripheral to the perinuclear area, suggesting the physiological relevance of these phosphorylation sites in albumin endocytosis. We conclude that both Akt1 and Akt2 are involved in albumin endocytosis, and phosphorylation of Dab2 by Akt induces albumin endocytosis in proximal tubule epithelial cells. Further delineation of how Akt affects expression/phosphorylation of endocytic adaptors and receptors will enhance our understanding of the molecular network triggered by albumin overload in the proximal tubule. Copyright © 2014 the American Physiological Society.

A new one-man submarine is tested as vehicle for solid rocket booster retrieval

NASA Technical Reports Server (NTRS)

2000-01-01

A Diver Operator Plug (DOP) is being pulled down into the ocean by a newly designed one-man submarine known as DeepWorker 2000. The activity is part of an operation to attach the plug to a mockup of a solid rocket booster nozzle. DeepWorker 2000 is being tested on its ability to duplicate the sometimes hazardous job United Space Alliance (USA) divers perform to recover the expended boosters in the ocean after a launch. The boosters splash down in an impact area about 140 miles east of Jacksonville and after recovery are towed back to KSC for refurbishment by the specially rigged recovery ships. DeepWorker 2000 will be used in a demonstration during retrieval operations after the upcoming STS-101 launch. The submarine pilot will demonstrate capabilities to cut tangled parachute riser lines using a manipulator arm and attach the DOP to extract water and provide flotation for the booster. DeepWorker 2000 was built by Nuytco Research Ltd., North Vancouver, British Columbia. It is 8.25 feet long, 5.75 feet high, and weighs 3,800 pounds. USA is a prime contractor to NASA for the Space Shuttle program.

KSC-00padig012

NASA Image and Video Library

2000-04-22

KENNEDY SPACE CENTER, FLA. -- A Diver Operator Plug (DOP) is being pulled down into the ocean by a newly designed one-man submarine known as DeepWorker 2000. The activity is part of an operation to attach the plug to a mockup of a solid rocket booster nozzle. DeepWorker 2000 is being tested on its ability to duplicate the sometimes hazardous job United Space Alliance (USA) divers perform to recover the expended boosters in the ocean after a launch. The boosters splash down in an impact area about 140 miles east of Jacksonville and after recovery are towed back to KSC for refurbishment by the specially rigged recovery ships. DeepWorker 2000 will be used in a demonstration during retrieval operations after the upcoming STS-101 launch. The submarine pilot will demonstrate capabilities to cut tangled parachute riser lines using a manipulator arm and attach the DOP to extract water and provide flotation for the booster. DeepWorker 2000 was built by Nuytco Research Ltd., North Vancouver, British Columbia. It is 8.25 feet long, 5.75 feet high, and weighs 3,800 pounds. USA is a prime contractor to NASA for the Space Shuttle program

KSC00padig012

NASA Image and Video Library

2000-04-22

KENNEDY SPACE CENTER, FLA. -- A Diver Operator Plug (DOP) is being pulled down into the ocean by a newly designed one-man submarine known as DeepWorker 2000. The activity is part of an operation to attach the plug to a mockup of a solid rocket booster nozzle. DeepWorker 2000 is being tested on its ability to duplicate the sometimes hazardous job United Space Alliance (USA) divers perform to recover the expended boosters in the ocean after a launch. The boosters splash down in an impact area about 140 miles east of Jacksonville and after recovery are towed back to KSC for refurbishment by the specially rigged recovery ships. DeepWorker 2000 will be used in a demonstration during retrieval operations after the upcoming STS-101 launch. The submarine pilot will demonstrate capabilities to cut tangled parachute riser lines using a manipulator arm and attach the DOP to extract water and provide flotation for the booster. DeepWorker 2000 was built by Nuytco Research Ltd., North Vancouver, British Columbia. It is 8.25 feet long, 5.75 feet high, and weighs 3,800 pounds. USA is a prime contractor to NASA for the Space Shuttle program

Bone sialoprotein does not interact with pro-gelatinase A (MMP-2) or mediate MMP-2 activation

PubMed Central

2009-01-01

Background A recent model for activation of the zymogen form of matrix metalloproteinase 2 (MMP-2, also known as gelatinase A) has suggested that interactions between the SIBLING protein bone sialoprotein (BSP) and MMP-2 leads to conformational change in MMP-2 that initiates the conversion of the pro-enzyme into a catalytically active form. This model is particularly relevant to cancer cell metastasis to bone since BSP, bound to the αvβ3 integrin through its arginine-glycine-aspartic acid motif, could recruit MMP-2 to the cell surface. Methods We critically assessed the relationship between BSP and proMMP-2 and its activation using various forms of recombinant and purified BSP and MMP-2. Gelatinase and collagenase assays, fluorescence binding assays, real-time PCR, cell culture and pull-down assays were employed to test the model. Results Studies with a fluorogenic substrate for MMP-2 showed no activation of proMMP-2 by BSP. Binding and pull-down assays demonstrated no interaction between MMP-2 and BSP. While BSP-mediated invasiveness has been shown to depend on its integrin-binding RGD sequence, analysis of proMMP-2 activation and the level of membrane type 1 (MT1)-MMP in cells grown on a BSP substratum showed that the BSP-αvβ3 integrin interaction does not induce the expression of MT1-MMP. Conclusion These studies do not support a role for BSP in promoting metastasis through interactions with pro-MMP-2. PMID:19386107

In Vitro Identification of Histatin 5 Salivary Complexes

PubMed Central

Moffa, Eduardo B.; Machado, Maria A. A. M.; Mussi, Maria C. M.; Xiao, Yizhi; Garrido, Saulo S.; Giampaolo, Eunice T.; Siqueira, Walter L.

2015-01-01

With recent progress in the analysis of the salivary proteome, the number of salivary proteins identified has increased dramatically. However, the physiological functions of many of the newly discovered proteins remain unclear. Closely related to the study of a protein’s function is the identification of its interaction partners. Although in saliva some proteins may act primarily as single monomeric units, a significant percentage of all salivary proteins, if not the majority, appear to act in complexes with partners to execute their diverse functions. Coimmunoprecipitation (Co-IP) and pull-down assays were used to identify the heterotypic complexes between histatin 5, a potent natural antifungal protein, and other salivary proteins in saliva. Classical protein–protein interaction methods in combination with high-throughput mass spectrometric techniques were carried out. Co-IP using protein G magnetic Sepharose TM beads suspension was able to capture salivary complexes formed between histatin 5 and its salivary protein partners. Pull-down assay was used to confirm histatin 5 protein partners. A total of 52 different proteins were identified to interact with histatin 5. The present study used proteomic approaches in conjunction with classical biochemical methods to investigate protein–protein interaction in human saliva. Our study demonstrated that when histatin 5 is complexed with salivary amylase, one of the 52 proteins identified as a histatin 5 partner, the antifungal activity of histatin 5 is reduced. We expected that our proteomic approach could serve as a basis for future studies on the mechanism and structural-characterization of those salivary protein interactions to understand their clinical significance. PMID:26544073

Interplay between intergrin-linked kinase and ribonuclease inhibitor affects growth and metastasis of bladder cancer through signaling ILK pathways.

PubMed

Zhuang, Xiang; Lv, Mengxin; Zhong, Zhenyu; Zhang, Luyu; Jiang, Rong; Chen, Junxia

2016-08-30

Integrin-linked kinase (ILK) is a multifunctional adaptor protein which is involved with protein signalling within cells to modulate malignant (cancer) cell movement, cell cycle, metastasis and epithelial-mesenchymal transition (EMT). Our previous experiment demonstrated that ILK siRNA inhibited the growth and induced apoptosis of bladder cancer cells as well as increased the expression of Ribonuclease inhibitor (RI), an important cytoplasmic protein with many functions. We also reported that RI overexpression inhibited ILK and phosphorylation of AKT and GSK3β. ILK and RI gene both locate on chromosome 11p15 and the two genes are always at the adjacent position of same chromosome during evolution, which suggest that ILK and RI could have some relationship. However, underlying interacting mechanisms remain unclear between them. Here, we postulate that RI might regulate ILK signaling pathway via interacting with ILK. Co-immunoprecipitation, GST pull-down and co-localization under laser confocal microscope assay were used to determine the interaction between ILK and RI exogenously and endogenously. Furthermore, we further verified that there is a direct binding between the two proteins by fluorescence resonance energy transfer (FRET) in cells. Next, The effects of interplay between ILK and RI on the key target protein expressions of PI3K/AKT/mTOR signaling pathway were determined by western blot, immunohistochemistry and immunofluorescence assay in vivo and in vitro. Finally, the interaction was assessed using nude mice xenograft model. We first found that ILK could combine with RI both in vivo and in vitro by GST pull-down, co-immunoprecipitation (Co-IP) and FRET. The protein levels of ILK and RI revealed a significant inverse correlation in vivo and in vitro. Subsequently, The results showed that up-regulating ILK could increase cell proliferation, change cell morphology and regulate cell cycle. We also demonstrated that the overexpression of ILK remarkably promoted EMT and expressions of target molecules of ILK signaling pathways in vitro and in vivo. Finally, we found that ILK overexpression significantly enhanced growth, metastasis and angiogenesis of xenograft tumor; Whereas, RI has a contrary role compared to ILK in vivo and in vitro. Our findings, for the first time, directly proved that the interplay between ILK and RI regulated EMT via ILK/PI3K/AKT signaling pathways for bladder cancer, which highlights the possibilities that ILK/RI could be valuable markers together for the therapy and diagnosis of human carcinoma of urinary bladder.

The Ca{sup 2+} channel TRPML3 specifically interacts with the mammalian ATG8 homologue GATE16 to regulate autophagy

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

Choi, Suzy; Kim, Hyun Jin, E-mail: kimhyunjin@skku.edu

2014-01-03

Highlights: •Split-ubiquitin MY2H screen identified GATE16 as an interacting protein of TRPML3. •TRPML3 specifically binds to a mammalian ATG8 homologue GATE16, not to LC3B. •The interaction of TRPML3 with GATE16 facilitates autophagosome formation. •GATE16 is expressed in both autophagosome and extra-autophagosomal compartments. -- Abstract: TRPML3 is a Ca{sup 2+} permeable cation channel expressed in multiple intracellular compartments. Although TRPML3 is implicated in autophagy, how TRPML3 can regulate autophagy is not understood. To search interacting proteins with TRPML3 in autophagy, we performed split-ubiquitin membrane yeast two-hybrid (MY2H) screening with TRPML3-loop as a bait and identified GATE16, a mammalian ATG8 homologue. GSTmore » pull-down assay revealed that TRPML3 and TRPML3-loop specifically bind to GATE16, not to LC3B. Co-immunoprecipitation (co-IP) experiments showed that TRPML3 and TRPML3-loop pull down only the lipidated form of GATE16, indicating that the interaction occurs exclusively at the organellar membrane. The interaction of TRPML3 with GATE16 and GATE16-positive vesicle formation were increased in starvation induced autophagy, suggesting that the interaction facilitates the function of GATE16 in autophagosome formation. However, GATE16 was not required for TRPML3 trafficking to autophagosomes. Experiments using dominant-negative (DN) TRPML3(D458K) showed that GATE16 is localized not only in autophagosomes but also in extra-autophagosomal compartments, by contrast with LC3B. Since GATE16 acts at a later stage of the autophagosome biogenesis, our results suggest that TRPML3 plays a role in autophagosome maturation through the interaction with GATE16, by providing Ca{sup 2+} in the fusion process.« less

The EICP22 Protein of Equine Herpesvirus 1 Physically Interacts with the Immediate-Early Protein and with Itself To Form Dimers and Higher-Order Complexes

PubMed Central

Derbigny, Wilbert A.; Kim, Seong K.; Caughman, Gretchen B.; O'Callaghan, Dennis J.

2000-01-01

The EICP22 protein (EICP22P) of Equine herpesvirus 1 (EHV-1) is an early protein that functions synergistically with other EHV-1 regulatory proteins to transactivate the expression of early and late viral genes. We have previously identified EICP22P as an accessory regulatory protein that has the ability to enhance the transactivating properties and the sequence-specific DNA-binding activity of the EHV-1 immediate-early protein (IEP). In the present study, we identify EICP22P as a self-associating protein able to form dimers and higher-order complexes during infection. Studies with the yeast two-hybrid system also indicate that physical interactions occur between EICP22P and IEP and that EICP22P self-aggregates. Results from in vitro and in vivo coimmunoprecipitation experiments and glutathione S-transferase (GST) pull-down studies confirmed a direct protein-protein interaction between EICP22P and IEP as well as self-interactions of EICP22P. Analyses of infected cells by laser-scanning confocal microscopy with antibodies specific for IEP and EICP22P revealed that these viral regulatory proteins colocalize in the nucleus at early times postinfection and form aggregates of dense nuclear structures within the nucleoplasm. Mutational analyses with a battery of EICP22P deletion mutants in both yeast two-hybrid and GST pull-down experiments implicated amino acids between positions 124 and 143 as the critical domain mediating the EICP22P self-interactions. Additional in vitro protein-binding assays with a library of GST-EICP22P deletion mutants identified amino acids mapping within region 2 (amino acids [aa] 65 to 196) and region 3 (aa 197 to 268) of EICP22P as residues that mediate its interaction with IEP. PMID:10627553

Can Energy Cost During Low-Intensity Resistance Exercise be Predicted by the OMNI-RES Scale?

PubMed Central

Vianna, Jefferson M.; Reis, Victor M.; Saavedra, Francisco; Damasceno, Vinicius; Silva, Sérgio G.; Goss, Fredric

2011-01-01

The aim of the present study was to assess the precision of the OMNI-RES scale to predict energy cost (EC) at low intensity in four resistance exercises (RE). 17 male recreational body builders (age = 26.6 ± 4.9 years; height = 177.7 ± 0.1 cm; body weight = 79.0 ± 11.1 kg and percent body fat = 10.5 ± 4.6%) served as subjects. Initially tests to determine 1RM for four resistance exercises (bench press, half squat, lat pull down and triceps extension) were administered. Subjects also performed resistance exercise at 12, 16, 20, and 24% of 1RM at a rate of 40 bpm until volitional exhaustion. Oxygen uptake (VO2) and rate of perceived exertion (RPE) using the OMNI-RES were obtained during and after all RE. EC was calculated using VO2 and the caloric values of VO2 for non-protein RER. Regression analyses were performed for every RE, using EC as the dependent and RPE as the predictor variable. The triceps extension, lat pull down and bench press, RPE correlated strongly with EC (R > 0.97) and predicted EC with a error of less than 0.2 kcal.min−1. In conclusion, RPE using the OMNI-RES scale can be considered as an accurate indicator of EC in the bench press, lat pull down and triceps extension performed by recreational bodybuilders, provided lower intensities are used (up to 24% of 1-RM) and provided each set of exercise is performed for the maximal sustainable duration. It would be interesting in future studies to consider having the subjects exercise at low intensities for longer durations than those in the present study. PMID:23486188

Impact of dehydration on a full body resistance exercise protocol.

PubMed

Kraft, Justin A; Green, James M; Bishop, Phillip A; Richardson, Mark T; Neggers, Yasmin H; Leeper, James D

2010-05-01

This study examined effects of dehydration on a full body resistance exercise workout. Ten males completed two trials: heat exposed (with 100% fluid replacement) (HE) and dehydration (approximately 3% body mass loss with no fluid replacement) (DEHY) achieved via hot water bath (approximately 39 degrees C). Following HE and DEHY, participants performed three sets to failure (using predetermined 12 repetition maximum) of bench press, lat pull down, overhead press, barbell curl, triceps press, and leg press with a 2-min recovery between each set and 2 min between exercises. A paired t test showed total repetitions (all sets combined) were significantly lower for DEHY: (144.1 +/- 26.6 repetitions) versus HE: (169.4 +/- 29.1 repetitions). ANOVAs showed significantly lower repetitions (approximately 1-2 repetitions on average) per exercise for DEHY versus HE (all exercises). Pre-set rate of perceived exertion (RPE) and pre-set heart rate (HR) were significantly higher [approximately 0.6-1.1 units on average in triceps press, leg press, and approached significance in lat pull down (P = 0.14) and approximately 6-13 b min(-1) on average in bench press, lat pull down, triceps press, and approached significance for overhead press (P = 0.10)] in DEHY versus HE. Session RPE difference approached significance (DEHY: 8.6 +/- 1.9, HE: 7.4 +/- 2.3) (P = 0.12). Recovery HR was significantly higher for DEHY (116 +/- 15 b min(-1)) versus HE (105 +/- 13 b min(-1)). Dehydration (approximately 3%) impaired resistance exercise performance, decreased repetitions, increased perceived exertion, and hindered HR recovery. Results highlight the importance of adequate hydration during full body resistance exercise sessions.

Fibroblast growth factor homologous factor 1 interacts with NEMO to regulate NF-κB signaling in neurons.

PubMed

König, Hans-Georg; Fenner, Beau J; Byrne, Jennifer C; Schwamborn, Robert F; Bernas, Tytus; Jefferies, Caroline A; Prehn, Jochen H M

2012-12-15

Neuronal survival and plasticity critically depend on constitutive activity of the transcription factor nuclear factor-κB (NF-κB). We here describe a role for a small intracellular fibroblast growth factor homologue, the fibroblast growth factor homologous factor 1 (FHF1/FGF12), in the regulation of NF-κB activity in mature neurons. FHFs have previously been described to control neuronal excitability, and mutations in FHF isoforms give rise to a form of progressive spinocerebellar ataxia. Using a protein-array approach, we identified FHF1b as a novel interactor of the canonical NF-κB modulator IKKγ/NEMO. Co-immunoprecipitation, pull-down and GAL4-reporter experiments, as well as proximity ligation assays, confirmed the interaction of FHF1 and NEMO and demonstrated that a major site of interaction occurred within the axon initial segment. Fhf1 gene silencing strongly activated neuronal NF-κB activity and increased neurite lengths, branching patterns and spine counts in mature cortical neurons. The effects of FHF1 on neuronal NF-κB activity and morphology required the presence of NEMO. Our results imply that FHF1 negatively regulates the constitutive NF-κB activity in neurons.

Drosophila RISC component VIG and its homolog Vig2 impact heterochromatin formation.

PubMed

Gracheva, Elena; Dus, Monica; Elgin, Sarah C R

2009-07-08

Heterochromatin formation plays an important role in gene regulation and the maintenance of genome integrity. Here we present results from a study of the D. melanogaster gene vig, encoding an RNAi complex component and its homolog vig2 (CG11844) that support their involvement in heterochromatin formation and/or maintenance. Protein null mutations vig(EP812) and vig2(PL470) act as modifiers of Position Effect Variegation (PEV). VIG and Vig2 are present in polytene chromosomes and partially overlap with HP1. Quantitative immunoblots show depletion of HP1 and HP2 (large isoform) in isolated nuclei from the vig(EP812) mutant. The vig2(PL470) mutant strain demonstrates a decreased level of H3K9me2. Pull-down experiments using antibodies specific to HP1 recovered both VIG and Vig2. The association between HP1 and both VIG and Vig2 proteins depends on an RNA component. The above data and the developmental profiles of the two genes suggest that Vig2 may be involved in heterochromatin targeting and establishment early in development, while VIG may have a role in stabilizing HP1/HP2 chromatin binding during later stages.

Beyond genome-wide scan: Association of a cis-regulatory NCR3 variant with mild malaria in a population living in the Republic of Congo.

PubMed

Baaklini, Sabrina; Afridi, Sarwat; Nguyen, Thy Ngoc; Koukouikila-Koussounda, Felix; Ndounga, Mathieu; Imbert, Jean; Torres, Magali; Pradel, Lydie; Ntoumi, Francine; Rihet, Pascal

2017-01-01

Linkage studies have revealed a linkage of mild malaria to chromosome 6p21 that contains the NCR3 gene encoding a natural killer cell receptor, whereas NCR3-412G>C (rs2736191) located in its promoter region was found to be associated with malaria in Burkina Faso. Here we confirmed the association of rs2736191 with mild malaria in a Congolese cohort and investigated its potential cis-regulatory effect. Luciferase assay results indicated that rs2736191-G allele had a significantly increased promoter activity compared to rs2736191-C allele. Furthermore, EMSAs demonstrated an altered binding of two nuclear protein complexes to the rs2736191-C allele in comparison to rs2736191-G allele. Finally, after in silico identification of transcription factor candidates, pull-down western blot experiments confirmed that both STAT4 and RUNX3 bind the region encompassing rs2736191 with a higher affinity for the G allele. To our knowledge, this is the first report that explored the functional role of rs2736191. These results support the hypothesis that genetic variation within natural killer cell receptors alters malaria resistance in humans.

Sticking properties of ice grains

NASA Astrophysics Data System (ADS)

Jongmanns, M.; Kumm, M.; Wurm, G.; Wolf, D. E.; Teiser, J.

2017-06-01

We study the size dependence of pull-off forces of water ice in laboratory experiments and numerical simulations. To determine the pull-off force in our laboratory experiments, we use a liquid nitrogen cooled centrifuge. Depending on its rotation frequency, spherical ice grains detach due to the centrifugal force which is related to the adhesive properties. Numerical simulations are conducted by means of molecular dynamics simulations of hexagonal ice using a standard coarse-grained water potential. The pull-off force of a single contact between two spherical ice grains is measured due to strain controlled simulations. Both, the experimental study and the simulations reveal a dependence between the pull-off force and the (reduced) particle radii, which differ significantly from the linear dependence of common contact theories.

Identification of the C-terminal domain of Daxx acts as a potential regulator of intracellular cholesterol synthesis in HepG2 cells

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

Sun, Shaowei; Medical School, Hunan University of Chinese Medicine, Changsha 410208, Hunan; Wen, Juan

Daxx is a highly conserved nuclear transcriptional factor, which has been implicated in many nuclear processes including transcription and cell cycle regulation. Our previous study demonstrated Daxx also plays a role in regulation of intracellular cholesterol content. Daxx contains several domains that are essential for interaction with a growing number of proteins. To delineate the underlying mechanism of hypocholesterolemic activity of Daxx, we constructed a set of plasmids which can be used to overexpress different fragments of Daxx and transfected to HepG2 cells. We found that the C- terminal region Daxx626–740 clearly reduced intracellular cholesterol levels and inhibited the expressionmore » of SREBPs and SCAP. In GST pull-down experiments and Double immunofluorescence assays, Daxx626–740 was demonstrated to bind directly to androgen receptor (AR). Our findings suggest that the interaction of Daxx626-740 and AR abolishes the AR-mediated activation of SCAP/SREBPs pathway, which suppresses the de novo cholesterol synthesis. Thus, C-terminal domain of Daxx acts as a potential regulator of intracellular cholesterol content in HepG2 cells. - Highlights: • Daxx C-terminal domain reduces cholesterol levels. • Daxx C-terminal domain binds directly to AR. • The interaction of Daxx C-terminal domain and AR suppresses cholesterol synthesis.« less

Receptacle for Optical-Fiber Scraps

NASA Technical Reports Server (NTRS)

Nevin, R.

1986-01-01

Small pieces of glass trapped by moving air. Device traps fibers in section of black air-conditioner filter material. Filter section rests on metal screen above axial fan, which pulls air down through filter. Fan is small, quiet unit of type ordinarily used to cool electronic equipment.

Stalled Momentum

ERIC Educational Resources Information Center

Agron, Joe

2006-01-01

The old adage, "what goes up must come down," can be used to describe education construction spending in 2005. After impressive gains in expenditures during the first half of the decade, total construction spending by schools and universities pulled back from historic highs. A combination of factors, including skyrocketing fuel and…

A Ureteral Inguinoscrotal Hernia from a Pelvic Kidney

PubMed Central

Dikmen, Ayse V.; Guneri, Cagri; Yalcin, Serdar; Acikgoz, Onur; Ak, Esat; Cetiner, Sadettin

2017-01-01

A 74-year-old male patient with prostate cancer under remission was admitted with left inguinoscrotal swelling. He underwent scrotal ultrasound demonstrating a giant in-guinoscrotal hernia. Contrast-enhanced computerized tomography of the abdomen and pelvis demonstrated a left pelvic kidney associated with severe hydroureteronephrosis secondary to a ureteral inguinoscrotal hernia. Upon exploration with left inguinal incision, a paraperitoneal ureteral in-guinoscrotal hernia and a hypertrophic left spermatic cord were observed. The elongated and tortuous left ureter, being pulled down to the scrotum by the hernia, was released from the herniating tissues fullfilling left hemiscrotum. The ureter was tapered followed by ureteroureterostomy. The accompanying left spermatic cord was excessively elongated and curled, necessitating cordectomy. The hernia was repaired with prolene mesh after removal of herniating peritoneal tissue. This is a rare case of a paraperitoneal ureteral inguinoscrotal hernia of the left pelvic kidney. PMID:29463977

Mechanical load on the low back and shoulders during pushing and pulling of two-wheeled waste containers compared with lifting and carrying of bags and bins.

PubMed

Schibye, B; Søgaard, K; Martinsen, D; Klausen, K

2001-08-01

Compare the mechanical load on the low back and shoulders during pushing and pulling a two-wheeled container with the load during lifting and carrying the same amount of waste. Only little is known about risk factors and mechanical loads during push/pull operations. A complete 2(3) factor push/pull experiment. A two-wheeled container with 25 or 50 kg was pushed in front of and pulled behind the body by seven waste collectors. Further, the same subjects lifted and carried a paper bag and a dustbin both loaded with 7 and 25 kg. All operations were video recorded and the push/pull force was measured by means of a three-dimensional force transducer. Peak Motus and Watbak software were used for digitising and calculation of torque at L4/L5 and the shoulder joints and compression and shear forces at L4/L5. During pushing and pulling the compression at L4/L5 is from 605 to 1445 N. The extension torque at L4/L5 produced by the push/pull force is counteracted by the forward leaning of the upper body. The shear force is below 202 N in all situations. The torque at the shoulders is between 1 and 38 Nm. In the present experiments the torques at the low back and the shoulders are low during pushing and pulling. No relation exists between the size of the external force and the torque at the low back and the shoulder. Pushing and pulling are common in many workplaces and have often replaced lifting and carrying situations. This has emphasised the need for more knowledge of the internal mechanical load on the body during these activities.

Membrane-based actuation for high-speed single molecule force spectroscopy studies using AFM.

PubMed

Sarangapani, Krishna; Torun, Hamdi; Finkler, Ofer; Zhu, Cheng; Degertekin, Levent

2010-07-01

Atomic force microscopy (AFM)-based dynamic force spectroscopy of single molecular interactions involves characterizing unbinding/unfolding force distributions over a range of pulling speeds. Owing to their size and stiffness, AFM cantilevers are adversely affected by hydrodynamic forces, especially at pulling speeds >10 microm/s, when the viscous drag becomes comparable to the unbinding/unfolding forces. To circumvent these adverse effects, we have fabricated polymer-based membranes capable of actuating commercial AFM cantilevers at speeds >or=100 microm/s with minimal viscous drag effects. We have used FLUENT, a computational fluid dynamics (CFD) software, to simulate high-speed pulling and fast actuation of AFM cantilevers and membranes in different experimental configurations. The simulation results support the experimental findings on a variety of commercial AFM cantilevers and predict significant reduction in drag forces when membrane actuators are used. Unbinding force experiments involving human antibodies using these membranes demonstrate that it is possible to achieve bond loading rates >or=10(6) pN/s, an order of magnitude greater than that reported with commercial AFM cantilevers and systems.

Associative Mechanisms Allow for Social Learning and Cultural Transmission of String Pulling in an Insect

PubMed Central

Zhu, Xingfu; Ingraham, Thomas; Søvik, Eirik

2016-01-01

Social insects make elaborate use of simple mechanisms to achieve seemingly complex behavior and may thus provide a unique resource to discover the basic cognitive elements required for culture, i.e., group-specific behaviors that spread from “innovators” to others in the group via social learning. We first explored whether bumblebees can learn a nonnatural object manipulation task by using string pulling to access a reward that was presented out of reach. Only a small minority “innovated” and solved the task spontaneously, but most bees were able to learn to pull a string when trained in a stepwise manner. In addition, naïve bees learnt the task by observing a trained demonstrator from a distance. Learning the behavior relied on a combination of simple associative mechanisms and trial-and-error learning and did not require “insight”: naïve bees failed a “coiled-string experiment,” in which they did not receive instant visual feedback of the target moving closer when tugging on the string. In cultural diffusion experiments, the skill spread rapidly from a single knowledgeable individual to the majority of a colony’s foragers. We observed that there were several sequential sets (“generations”) of learners, so that previously naïve observers could first acquire the technique by interacting with skilled individuals and, subsequently, themselves become demonstrators for the next “generation” of learners, so that the longevity of the skill in the population could outlast the lives of informed foragers. This suggests that, so long as animals have a basic toolkit of associative and motor learning processes, the key ingredients for the cultural spread of unusual skills are already in place and do not require sophisticated cognition. PMID:27701411

78 FR 40074 - Airworthiness Directives; Airbus Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-03

... certain batch of passenger emergency oxygen container assemblies might become detached by extreme pulling of the mask tube at the end of oxygen supply causing a high temperature oxygen generator and mask to fall down. This proposed AD would require modifying the passenger emergency oxygen container assembly...

10 CFR 431.62 - Definitions concerning commercial refrigerators, freezers and refrigerator-freezers.

Code of Federal Regulations, 2014 CFR

2014-01-01

... occupancy sensor means a device which uses passive infrared, ultrasonic, or other motion-sensing technology..., frozen, combination chilled and frozen, or variable temperature; (4) Displays or stores merchandise and... doors; (6) Is designed for pull-down temperature applications or holding temperature applications; and...

My Project. In: SMARTe20ll, EPA/600/C-10/007

EPA Science Inventory

SMARTe's "My Project" is intended to allow stakeholders to work together in a project "team room" and evaluate different reuse options for their specific situation. "My Project" is a password protected version of SMARTe. This personal SMARTe site has pull down menus for access ...

Optical pulling of airborne absorbing particles and smut spores over a meter-scale distance with negative photophoretic force

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

Lin, Jinda; Hart, Adam G.; Li, Yong-qing, E-mail: liy@ecu.edu

2015-04-27

We demonstrate optical pulling of single light-absorbing particles and smut spores in air over a meter-scale distance using a single collimated laser beam based on negative photophoretic force. The micron-sized particles are pulled towards the light source at a constant speed of 1–10 cm/s in the optical pulling pipeline while undergoing transverse rotation at 0.2–10 kHz. The pulled particles can be manipulated and precisely positioned on the entrance window with an accuracy of ∼20 μm, and their chemical compositions can be characterized with micro-Raman spectroscopy.

Push-pull aminobithiophenes--highly fluorescent stable fluorophores.

PubMed

Dong, Yanmei; Bolduc, Andréanne; McGregor, Nicholas; Skene, W G

2011-04-01

Stable 2-aminobithiophenes were prepared using the Gewald reaction. The resulting push-pull bithiophenes exhibited both unprecedented high fluorescence yields and stability in addition to demonstrating fluorescence on-off properties.

The forming simulation of flexible glass with silt down draw method

NASA Astrophysics Data System (ADS)

Yansheng, Hou; Jinshu, Cheng; Junfeng, Kang; Jing, Cui

2018-03-01

The slit down draw method is the main manufacturing process of flexible glass. In this study, Flow3DTM software was used to simulate the process of drawing and thinning glass slits during the slit down draw process. The influence of glass viscosity, initial plate thickness and initial plate speed on the glass spreading process was studied. The maximum pull-down force that the root can bear is linearly proportional to the viscosity, the initial thickness of 1.3837 power and the initial plate speed, respectively. The best way to improve the tensile strength of flexible glass is to increase the viscosity. Flexible glass was more easily to obtain with low viscosity, low thickness and low drawing speed.

Near Surface Geophysical Investigations of Potential Direct Recharge Zones in the Biscayne Aquifer within Everglades National Park, Florida.

NASA Astrophysics Data System (ADS)

Mount, G.; Comas, X.

2017-12-01

The karstic Miami Limestone of the Biscayne aquifer is characterized as having water flow that is controlled by the presence of dissolution enhanced porosity and mega-porous features. The dissolution features and other high porosity areas create horizontal preferential flow paths and high rates of ground water velocity, which may not be accurately conceptualized in groundwater flow models. In addition, recent research suggests the presence of numerous vertical dissolution features across Everglades National Park at Long Pine Key Trail, that may act as areas of direct recharge to the aquifer. These vertical features have been identified through ground penetrating radar (GPR) surveys as areas of velocity pull-down which have been modeled to have porosity values higher than the surrounding Miami Limestone. As climate change may induce larger and longer temporal variability between wet and dry times in the Everglades, a more comprehensive understanding of preferential flow pathways from the surface to the aquifer would be a great benefit to modelers and planners. This research utilizes near surface geophysical techniques, such as GPR, to identify these vertical dissolution features and then estimate the spatial variability of porosity using petrophysical models. GPR transects that were collected for several kilometers along the Long Pine Key Trail, show numerous pull down areas that correspond to dissolution enhanced porosity zones within the Miami Limestone. Additional 3D GPR surveys have attempted to delineate the boundaries of these features to elucidate their geometry for future modelling studies. We demonstrate the ability of near surface geophysics and petrophysical models to identify dissolution enhanced porosity in shallow karstic limestones to better understand areas that may act as zones of direct recharge into the Biscayne Aquifer.

Directional Flow of Summer Aeration to Manage Insect Pests in Stored Wheat

USDA-ARS?s Scientific Manuscript database

Field trials were conducted in metal wheat storage bins to determine whether pressure aeration, pushing ambient air from the bottom, or suction aeration, pulling air down from the top, would be more efficient at cooling the wheat mass and thereby limiting insect population growth. Aeration was accom...

10 CFR 431.62 - Definitions concerning commercial refrigerators, freezers and refrigerator-freezers.

Code of Federal Regulations, 2013 CFR

2013-01-01

... measurements taken during the test. Lighting occupancy sensor means a device which uses passive infrared...) Operates at a chilled, frozen, combination chilled and frozen, or variable temperature; (4) Displays or... doors, or no doors; (6) Is designed for pull-down temperature applications or holding temperature...

Material test machine for tension-compression tests at high temperature

DOEpatents

Cioletti, Olisse C.

1988-01-01

Apparatus providing a device for testing the properties of material specimens at high temperatures and pressures in controlled water chemistries includes, inter alia, an autoclave housing the specimen which is being tested. The specimen is connected to a pull rod which couples out of the autoclave to an external assembly which includes one or more transducers, a force balance chamber and a piston type actuator. The pull rod feeds through the force balance chamber and is compensated thereby for the pressure conditions existing within the autoclave and tending to eject the pull rod therefrom. The upper end of the push rod is connected to the actuator through elements containing a transducer comprising a linear variable differential transformer (LVDT). The housing and coil assembly of the LVDT is coupled to a tube which runs through a central bore of the pull rod into the autoclave where it is connected to one side of the specimen. The movable core of the LVDT is coupled to a stem which runs through the tube where it is then connected to the other side of the specimen through a coupling member. A transducer in the form of a load cell including one or more strain gages is located on a necked-down portion of the upper part of the pull rod intermediate the LVDT and force balance chamber.

Large-Scale CTRW Analysis of Push-Pull Tracer Tests and Other Transport in Heterogeneous Porous Media

NASA Astrophysics Data System (ADS)

Hansen, S. K.; Berkowitz, B.

2014-12-01

Recently, we developed an alternative CTRW formulation which uses a "latching" upscaling scheme to rigorously map continuous or fine-scale stochastic solute motion onto discrete transitions on an arbitrarily coarse lattice (with spacing potentially on the meter scale or more). This approach enables model simplification, among many other things. Under advection, for example, we see that many relevant anomalous transport problems may be mapped into 1D, with latching to a sequence of successive, uniformly spaced planes. On this formulation (which we term RP-CTRW), the spatial transition vector may generally be made deterministic, with CTRW waiting time distributions encapsulating all the stochastic behavior. We demonstrate the excellent performance of this technique alongside Pareto-distributed waiting times in explaining experiments across a variety of scales using only two degrees of freedom. An interesting new application of the RP-CTRW technique is the analysis of radial (push-pull) tracer tests. Given modern computational power, random walk simulations are a natural fit for the inverse problem of inferring subsurface parameters from push-pull test data, and we propose them as an alternative to the classical type curve approach. In particular, we explore the visibility of heterogeneity through non-Fickian behavior in push-pull tests, and illustrate the ability of a radial RP-CTRW technique to encapsulate this behavior using a sparse parameterization which has predictive value.

Push versus pull gastrostomy in cancer patients: A single center retrospective analysis of complications and technical success rates.

PubMed

Currie, B M; Getrajdman, G I; Covey, A M; Alago, W; Erinjeri, J P; Maybody, M; Boas, F E

2018-04-28

To compare the technical success and complication rates of push versus pull gastrostomy tubes in cancer patients, and to examine their dependence on operator experience. A retrospective review was performed of 304 cancer patients (170 men, 134 women; mean age 60.3±12.6 [SD], range: 19-102 years) referred for primary gastrostomy tube placement, 88 (29%) of whom had a previously unsuccessful attempt at percutaneous endoscopic gastrostomy (PEG) placement. Analyzed variables included method of insertion (push versus pull), indication for gastrostomy, technical success, operator experience, and procedure-related complications within 30 days of placement. Gastrostomy tubes were placed for feeding in 189 patients and palliative decompression in 115 patients. Technical success was 91%: 78% after endoscopy had previously been unsuccessful and 97% when excluding failures associated with prior endoscopy. In the first 30 days, there were 29 minor complications (17.2%) associated with push gastrostomies, and only 8 minor complications (7.5%) with pull gastrostomies (P<0.05). There was no significant difference in major complications (push gastrostomy 5.3%, pull gastrostomy 5.6%). For decompressive gastrostomy tubes, the pull technique resulted in lower rates of both minor and major complications. There was no difference in complications or technical success rates for more versus less experienced operators. Pull gastrostomy tube placement had a lower rate of complications than push gastrostomy tube placement, especially when the indication was decompression. The technical success rate was high, even after a failed attempt at endoscopic placement. Both the rates of success and complications were independent of operator experience. Copyright © 2018 Société française de radiologie. Published by Elsevier Masson SAS. All rights reserved.

Gravity Driven Universe: Energy from a Unified Field

NASA Astrophysics Data System (ADS)

Masters, Roy

2012-10-01

One way or another, whether push or pull, we know for sure that gravity is omnidirectional with identical mathematics. With PULL, gravity can be seen as as a property of matter. If so something is wrong. The Moon, lifting the tides twice-daily, should have fallen into orbital decay, with Earth having pulled it down eons ago. It is puzzling that physicists are not troubled by the fact that the Moon not only insists on forever lifting the tides, but, adding insult to injury, keeps moving it about 4 cm further away from Earth each year. Now if instead, we consider gravity as driven by an omnidirectional pressure--a PUSH force, another possibility arises. We can consider that it is mysteriously infusing energy into the Earth-Moon system, sustaining the Moon's orbit with the appearance of raising the tides and actually pushing it away from Earth. Here we can show push and pull, while being identical in their mathematics, have different outcomes. With push, gravity is a property of the universe. If this is true, then gravitation is flowing from an everlasting source, and the Earth/Moon system is one example of many other vacuum energy machines in the universe.

Applications of micro-SAXS/WAXS to study polymer fibers

NASA Astrophysics Data System (ADS)

Riekel, C.

2003-01-01

Instrumentation and selected applications for X-ray microdiffraction experiments on polymer and biopolymer fibers at the European Synchrotron Radiation Facility (ESRF) microfocus beamline are reviewed. Combined SAXS/WAXS experiments can be performed on single fibers with a beam size down to about 5 μm. WAXS experiments can be performed down to about 2 μm and in exceptional cases down to 0.1 μm beam size. The instrumental possibilities are demonstrated for the production line of spider silk.

Pulling of 3 mm diameter AlSb rods by micro-pulling down method

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

Bourret-Courchesne Ph.D., Edith; Perrodin, Didier

2009-05-14

We designed and supplied special crucibles for AlSb material. Thermal insulation and limitation of Sb losses were our first work. The protection of the growth environment was also one of our priority to avoid any pollution of the Fibercryst {mu}PD facility. When this work was achieved, the next step was the calibration of the heating power for these new crucibles. Then, it was the definition of single crystal growth conditions that oriented our research. Following our proposal, many growths attempts were performed. We started from Al & Sb pure powder or from LBNL AlSb crystal as expected. We used differentmore » crucibles and different seeds.« less

On artifacts in single-molecule force spectroscopy

PubMed Central

Cossio, Pilar; Hummer, Gerhard; Szabo, Attila

2015-01-01

In typical force spectroscopy experiments, a small biomolecule is attached to a soft polymer linker that is pulled with a relatively large bead or cantilever. At constant force, the total extension stochastically changes between two (or more) values, indicating that the biomolecule undergoes transitions between two (or several) conformational states. In this paper, we consider the influence of the dynamics of the linker and mesoscopic pulling device on the force-dependent rate of the conformational transition extracted from the time dependence of the total extension, and the distribution of rupture forces in force-clamp and force-ramp experiments, respectively. For these different experiments, we derive analytic expressions for the observables that account for the mechanical response and dynamics of the pulling device and linker. Possible artifacts arise when the characteristic times of the pulling device and linker become comparable to, or slower than, the lifetimes of the metastable conformational states, and when the highly anharmonic regime of stretched linkers is probed at high forces. We also revisit the problem of relating force-clamp and force-ramp experiments, and identify a linker and loading rate-dependent correction to the rates extracted from the latter. The theory provides a framework for both the design and the quantitative analysis of force spectroscopy experiments by highlighting, and correcting for, factors that complicate their interpretation. PMID:26540730

Comparative study of optical and scintillation properties of Tm3+:YAG, and Tm3+:LuAG single crystals

NASA Astrophysics Data System (ADS)

Fujimoto, Yutaka; Sugiyama, Makoto; Yanagida, Takayuki; Wakahara, Shingo; Suzuki, Shotaro; Kurosawa, Shunsuke; Chani, Valery; Yoshikawa, Akira

2013-09-01

The optical and scintillation properties of Tm3+-doped yttrium aluminum garnet Y3Al5O12 (YAG) and Tm3+-doped lutetium aluminum garnet Lu3Al5O12 (LuAG) are compared. The Tm3+-doped single crystals were grown by the micro-pulling down (μ-PD) technique. Both crystals demonstrated some emission peaks originated from 4f-4f forbidden transition of Tm3+ under 241Am alpha-ray excitation. The scintillation decay time of Tm3+-doped YAG was similar to that of LuAG. When irradiated by the gamma-rays from a 137Cs source, the relative scintillation light yields of Tm:YAG was 90% greater than that of Tm:LuAG.

Hyperpolarizabilities of Push-Pull Polyenes: Experimental Results and a New Two-State Model

NASA Technical Reports Server (NTRS)

Perry, J.; Bourhill, G.; Marder, S.; Lu, D.; Chen, G.; Goddard, W.

1994-01-01

Recent work has demonstrated a correlation between the geometry (i.e., the bond length alteration, BLA) and the first and second hyperpolarizabilities (betta and gamma) of donor-acceptor substituted (push-pull)polyenes.

Envy and admiration: emotion and motivation following upward social comparison.

PubMed

van de Ven, Niels

2017-01-01

Two key emotions people can experience when someone else is better than them are envy and admiration. There are conflicting findings in the scientific literature on which behaviour is elicited by these emotions. In one study (with two samples, total N = 345), we test which motivations are triggered by envy and admiration. The main finding is that (benign) envy and admiration both lead to a motivation to improve oneself. This confirms earlier findings that admiration leads to a motivation to affiliate with the admired other and a motivation to improve one's own position. Furthermore, it supports the idea that envy can lead to both a motivation to improve oneself and a motivation to pull down the envied other, finding support for a subtypes theory of envy.

The effect of a slack-pulling device in reducing operator physiological workload during log winching operations.

PubMed

Spinelli, Raffaele; Aalmo, Giovanna Ottaviani; Magagnotti, Natascia

2015-01-01

The authors conducted a comparative test to determine whether the introduction of a hydraulic slack puller allowed reducing the physiological workload of operators assigned to log winching tasks. The tests were conducted in northern Italy, on the mountains near Como. The study involved five volunteer subjects, considered representatives of the regional logging workforce. Physiological workload was determined by measuring the operators' heart rate upon completion of specific tasks. The slack puller improved the efficiency of downhill winching, since it allowed a single operator to pull out the cable on his own, without requiring the assistance of a colleague. However, introduction of the slack puller did not result in any reductions of operator physiological workload. The main stressor when working on a steep slope is moving up and down the slope: pulling a cable is only a secondary stressor. Any measures targeting secondary stressors are unlikely to produce dramatic reductions of operator workload.

Inclusion at Risk? Push- and Pull-Out Phenomena in Inclusive School Systems: The Italian and Norwegian Experiences

ERIC Educational Resources Information Center

Nes, Kari; Demo, Heidrun; Ianes, Dario

2018-01-01

The main objective of this article is to explore and compare research data on pull-out and push-out phenomena within inclusive school systems, discussing if and how they represent a risk for inclusion. The terms pull-out and push-out refer to situations in which some groups of students in regular schools learn in settings apart from their peers.…

Advanced Team Decision Making: A Developmental Model

DTIC Science & Technology

1992-08-03

role - right wing, center, goalie - but as they set up plays and bring the STRONG puck down the ice, those individuals begin to function TEAM... pulled the team member’s attention away from assigned work - Unwise use of a member’s expertise in designating roles or functions In these cases, advanced

Special Education Is Funding Early Help

ERIC Educational Resources Information Center

Samuels, Christina A.

2008-01-01

Bit by bit, the U.S. Department of Education is trying to pull down the walls that have traditionally separated general and special education. One facet of the plan is the department's support of "response to intervention," or RTI, an educational technique that bolsters the skills of academically struggling students before they fall so…

77 FR 10291 - Energy Conservation Program: Test Procedure for Commercial Refrigeration Equipment

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-21

... designed for holding temperature applications, as well as self-contained refrigerators with transparent doors designed for pull-down applications. (42 U.S.C. 6313(c)(2)-(3)) EPCA also requires DOE to develop... designed for ice-cream application. 71 FR 71357 (Dec. 8, 2006). Therefore, in the December 2006 en masse...

Influential Observations in Time Series.

DTIC Science & Technology

1984-07-01

to appear). 20. Treadway, A. B. (1978). Ifectos sabre la economia espanola de usa devaluacion da Ia "eseta, undacion Ramon-Areces, Madrid. . 21...Also, w (X) z and ;(Xw’X) + 0. in practice this result means that lA . when w is large, all the estimated coefficients w0 are pulled down towards

The geometry of pull-apart basins in the southern part of Sumatran strike-slip fault zone

NASA Astrophysics Data System (ADS)

Aribowo, Sonny

2018-02-01

Models of pull-apart basin geometry have been described by many previous studies in a variety tectonic setting. 2D geometry of Ranau Lake represents a pull-apart basin in the Sumatran Fault Zone. However, there are unclear geomorphic traces of two sub-parallel overlapping strike-slip faults in the boundary of the lake. Nonetheless, clear geomorphic traces that parallel to Kumering Segment of the Sumatran Fault are considered as inactive faults in the southern side of the lake. I demonstrate the angular characteristics of the Ranau Lake and Suoh complex pull-apart basins and compare with pull-apart basin examples from published studies. I use digital elevation model (DEM) image to sketch the shape of the depression of Ranau Lake and Suoh Valley and measure 2D geometry of pull-apart basins. This study shows that Ranau Lake is not a pull-apart basin, and the pull-apart basin is actually located in the eastern side of the lake. Since there is a clear connection between pull-apart basin and volcanic activity in Sumatra, I also predict that the unclear trace of the pull-apart basin near Ranau Lake may be covered by Ranau Caldera and Seminung volcanic products.

Initial atomic coherences and Ramsey frequency pulling in fountain clocks

NASA Astrophysics Data System (ADS)

Gerginov, Vladislav; Nemitz, Nils; Weyers, Stefan

2014-09-01

In the uncertainty budget of primary atomic cesium fountain clocks, evaluations of frequency-pulling shifts of the hyperfine clock transition caused by unintentional excitation of its nearby transitions (Rabi and Ramsey pulling) have been based so far on an approach developed for cesium beam clocks. We re-evaluate this type of frequency pulling in fountain clocks and pay particular attention to the effect of initial coherent atomic states. We find significantly enhanced frequency shifts caused by Ramsey pulling due to sublevel population imbalance and corresponding coherences within the state-selected hyperfine component of the initial atom ground state. Such shifts are experimentally investigated in an atomic fountain clock and quantitative agreement with the predictions of the model is demonstrated.

Two-Photon Entanglement and EPR Experiments Using Type-2 Spontaneous Parametric Down Conversion

NASA Technical Reports Server (NTRS)

Sergienko, A. V.; Shih, Y. H.; Pittman, T. B.; Rubin, M. H.

1996-01-01

Simultaneous entanglement in spin and space-time of a two-photon quantum state generated in type-2 spontaneous parametric down-conversion is demonstrated by the observation of quantum interference with 98% visibility in a simple beam-splitter (Hanburry Brown-Twiss) anticorrelation experiment. The nonlocal cancellation of two-photon probability amplitudes as a result of this double entanglement allows us to demonstrate two different types of Bell's inequality violations in one experimental setup.

Glucose-6-phosphate dehydrogenase and Trypanothione reductase interaction protects Leishmania donovani from metalloid mediated oxidative stress.

PubMed

Ghosh, Ayan Kumar; Saini, Savita; Das, Sushmita; Mandal, Abhishek; Sardar, Abul Hasan; Ansari, Md Yousuf; Abhishek, Kumar; Kumar, Ajay; Singh, Ruby; Verma, Sudha; Equbal, Asif; Ali, Vahab; Das, Pradeep

2017-05-01

Exploration of metabolons as viable drug target is rare in kinetoplastid biology. Here we present a novel protein-protein interaction among Glucose-6-phosphate dehydrogenase (LdG6PDH) and Trypanothione reductase (LdTryR) of Leishmania donovani displaying interconnection between central glucose metabolism and thiol metabolism of this parasite. Digitonin fractionation patterns observed through immunoblotting indicated localisation of both LdG6PDH and LdTryR in cytosol. In-silico and in-vitro interaction observed by size exclusion chromatography, co-purification, pull-down assay and spectrofluorimetric analysis revealed LdG6PDH and LdTryR physically interact with each other in a NADPH dependent manner. Coupled enzymatic assay displayed that NADPH generation was severely impaired by addition of Sb III , As III and Te IV extraneously, which hint towards metalloid driven structural changes of the interacting proteins. Co-purification patterns and pull-down assays also depicted that metalloids (Sb III , As III and Te IV ) hinder the in-vitro interaction of these two enzymes. Surprisingly, metalloids at sub-lethal concentrations induced the in-vivo interaction of LdG6PDH and LdTryR, as analyzed by pull-down assays and fluorescence microscopy signifying protection against metalloid mediated ROS. Inhibition of LdTryR by thioridazine in LdG6PDH -/- parasites resulted in metalloid induced apoptotic death of the parasites due to abrupt fall in reduced thiol content, disrupted NADPH/NADP + homeostasis and lethal oxidative stress. Interestingly, clinical isolates of L.donovani resistant to SAG exhibited enhanced interaction between LdG6PDH and LdTryR and showed cross resistivity towards As III and Te IV . Thus, our findings propose the metabolon of LdG6PDH and LdTryR as an alternate therapeutic target and provide mechanistic insight about metalloid resistance in Visceral Leishmaniasis. Copyright © 2017. Published by Elsevier Inc.

The effect of load reductions on repetition performance for commonly performed multijoint resistance exercises.

PubMed

Willardson, Jeffrey M; Simão, Roberto; Fontana, Fabio E

2012-11-01

The purpose of this study was to compare 4 different loading schemes for the free weight bench press, wide grip front lat pull-down, and free weight back squat to determine the extent of progressive load reductions necessary to maintain repetition performance. Thirty-two recreationally trained women (age = 29.34 ± 4.58 years, body mass = 59.61 ± 4.72 kg, height = 162.06 ± 4.04 cm) performed 4 resistance exercise sessions that involved 3 sets of the free weight bench press, wide grip front lat pull-down, and free weight back squat, performed in this exercise order during all 4 sessions. Each of the 4 sessions was conducted under different randomly ordered loading schemes, including (a) a constant 10 repetition maximum (RM) load for all 3 sets and for all 3 exercises, (b) a 5% reduction after the first and second sets for all the 3 exercises, (c) a 10% reduction after the first and second sets for all the 3 exercises, and (d) a 15% reduction after the first and second sets for all the 3 exercises. The results indicated that for the wide grip front lat pull-down and free weight back squat, a 10% load reduction was necessary after the first and second sets to accomplish 10 repetitions on all the 3 sets. For the free weight bench press, a load reduction between 10 and 15% was necessary; specifically, a 10% reduction was insufficient and a 15% reduction was excessive, as evidenced by significantly >10 repetitions on the second and third sets for this exercise (p ≤ 0.05). In conclusion, the results of this study indicate that a resistance training prescription that involves 1-minute rest intervals between multiple 10RM sets does require load reductions to maintain repetition performance. Practitioners might apply these results by considering an approximate 10% load reduction after the first and second sets for the exercises examined, when training women of similar characteristics as in this study.

Participation of the extracellular domain in (pro)renin receptor dimerization

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

Suzuki-Nakagawa, Chiharu; Nishimura, Misa; Tsukamoto, Tomoko

Highlights: • The (pro)renin receptor [(P)RR] is a regulator of the renin–angiotensin system. • The region responsible for (P)RR dimerization was investigated. • (P)RR extracellular domain constructs were retained intracellularly. • The extracellular domain of (P)RR is responsible for its dimerization. • Novel insight into the regulatory mechanism of soluble (P)RR secretion is provided. - Abstract: The (pro)renin receptor [(P)RR] induces the catalytic activation of prorenin, as well as the activation of the mitogen-activated protein kinase (MAPK) signaling pathway; as such, it plays an important regulatory role in the renin–angiotensin system. (P)RR is known to form a homodimer, but themore » region participating in its dimerization is unknown. Using glutathione S-transferase (GST) as a carrier protein and a GST pull-down assay, we investigated the interaction of several (P)RR constructs with full-length (FL) (P)RR in mammalian cells. GST fusion proteins with FL (P)RR (GST-FL), the C-terminal M8-9 fragment (GST-M8-9), the extracellular domain (ECD) of (P)RR (GST-ECD), and the (P)RR ECD with a deletion of 32 amino acids encoded by exon 4 (GST-ECDd4) were retained intracellularly, whereas GST alone was efficiently secreted into the culture medium when transiently expressed in COS-7 cells. Immunofluorescence microscopy showed prominent localization of GST-ECD to the endoplasmic reticulum. The GST pull-down analysis revealed that GST-FL, GST-ECD, and GST-ECDd4 bound FLAG-tagged FL (P)RR, whereas GST-M8-9 showed little or no binding when transiently co-expressed in HEK293T cells. Furthermore, pull-down analysis using His-tag affinity resin showed co-precipitation of soluble (P)RR with FL (P)RR from a stable CHO cell line expressing FL h(P)RR with a C-terminal decahistidine tag. These results indicate that the (P)RR ECD participates in dimerization.« less

Pushing, pulling and manoeuvring an industrial cart: a psychophysiological study.

PubMed

Giagloglou, Evanthia; Radenkovic, Milan; Brankovic, Sasa; Antoniou, Panagiotis; Zivanovic-Macuzic, Ivana

2017-09-18

One of the most frequent manual occupational tasks involves the pushing and pulling of a cart. Although several studies have associated health risks with pushing and pulling, the effects are not clear since occupational tasks have social, cognitive and physical components. The present work investigates a real case of a pushing and pulling occupational task from a manufacturing company. The study initially characterizes the case in accordance with Standard No. ISO 11228-2:2007 as low risk. An experiment with 14 individuals during three modalities of pushing and pulling was performed in order to further investigate the task with the application of electrophysiology. At the end, a simple questionnaire was given. The results show electrophysiological differences among the three modalities of pushing and pulling, with a major difference between action with no load and fully loaded with a full range of motions on the cart to handle.

Characterization and Effects of Fiber Pull-Outs in Hole Quality of Carbon Fiber Reinforced Plastics Composite.

PubMed

Alizadeh Ashrafi, Sina; Miller, Peter W; Wandro, Kevin M; Kim, Dave

2016-10-13

Hole quality plays a crucial role in the production of close-tolerance holes utilized in aircraft assembly. Through drilling experiments of carbon fiber-reinforced plastic composites (CFRP), this study investigates the impact of varying drilling feed and speed conditions on fiber pull-out geometries and resulting hole quality parameters. For this study, hole quality parameters include hole size variance, hole roundness, and surface roughness. Fiber pull-out geometries are quantified by using scanning electron microscope (SEM) images of the mechanically-sectioned CFRP-machined holes, to measure pull-out length and depth. Fiber pull-out geometries and the hole quality parameter results are dependent on the drilling feed and spindle speed condition, which determines the forces and undeformed chip thickness during the process. Fiber pull-out geometries influence surface roughness parameters from a surface profilometer, while their effect on other hole quality parameters obtained from a coordinate measuring machine is minimal.

Requirement of PDZ domains for the stimulation of the epithelial Ca2+ channel TRPV5 by the NHE regulating factor NHERF2 and the serum and glucocorticoid inducible kinase SGK1.

PubMed

Palmada, Monica; Poppendieck, Susanne; Embark, Hamdy M; van de Graaf, Stan F J; Boehmer, Christoph; Bindels, René J M; Lang, Florian

2005-01-01

Renal calcium reabsorption involves the epithelial calcium channel ECaC1 (TRPV5) which is tightly regulated by 1,25(OH)2D3. As shown recently, TRPV5 is activated by the serum and glucocorticoid inducible kinase SGK1, a kinase transcriptionally upregulated by 1,25(OH)2D3. This stimulatory effect is due to enhanced TRPV5 abundance in the plasma membrane and requires the presence of the scaffold protein NHERF2 (sodium hydrogen exchanger regulating factor 2). The present study aims to define the molecular requirements for the interaction of TRPV5 with SGK1 and NHERF2. Pull-down experiments and overlay assays revealed that the TRPV5 C-tail interacts in a Ca2+-independent manner with NHERF2. Deletion of the second but not of the first PDZ domain in NHERF2 abrogates the stimulating effect of SGK1/NHERF2 on TRPV5 protein abundance in the plasma membrane as quantified by chemiluminescence and electrophysiology. Thus, the second PDZ domain in NHERF2 is required for stabilization at or TRPV5 targeting to the plasma membrane. The experiments demonstrate the significance of SGK1 and NHERF2 as TRPV5 modulators which are likely to participate in the regulation of calcium homeostasis by 1,25(OH)2D3. Copyright 2005 S. Karger AG, Basel.

Mutations in the Nucleolar Phosphoprotein, Nucleophosmin, Promote the Expression of the Oncogenic Transcription Factor MEF/ELF4 in Leukemia Cells and Potentiates Transformation*

PubMed Central

Ando, Koji; Tsushima, Hideki; Matsuo, Emi; Horio, Kensuke; Tominaga-Sato, Shinya; Imanishi, Daisuke; Imaizumi, Yoshitaka; Iwanaga, Masako; Itonaga, Hidehiro; Yoshida, Shinichiro; Hata, Tomoko; Moriuchi, Ryozo; Kiyoi, Hitoshi; Nimer, Stephen; Mano, Hiroyuki; Naoe, Tomoki; Tomonaga, Masao; Miyazaki, Yasushi

2013-01-01

Myeloid ELF1-like factor (MEF/ELF4), a member of the ETS transcription factors, can function as an oncogene in murine cancer models and is overexpressed in various human cancers. Here, we report a mechanism by which MEF/ELF4 may be activated by a common leukemia-associated mutation in the nucleophosmin gene. By using a tandem affinity purification assay, we found that MEF/ELF4 interacts with multifactorial protein nucleophosmin (NPM1). Coimmunoprecipitation and GST pull-down experiments demonstrated that MEF/ELF4 directly forms a complex with NPM1 and also identified the region of NPM1 that is responsible for this interaction. Functional analyses showed that wild-type NPM1 inhibited the DNA binding and transcriptional activity of MEF/ELF4 on the HDM2 promoter, whereas NPM1 mutant protein (Mt-NPM1) enhanced these activities of MEF/ELF4. Induction of Mt-NPM1 into MEF/ELF4-overexpressing NIH3T3 cells facilitated malignant transformation. In addition, clinical leukemia samples with NPM1 mutations had higher human MDM2 (HDM2) mRNA expression. Our data suggest that enhanced HDM2 expression induced by mutant NPM1 may have a role in MEF/ELF4-dependent leukemogenesis. PMID:23393136

Mutations in the nucleolar phosphoprotein, nucleophosmin, promote the expression of the oncogenic transcription factor MEF/ELF4 in leukemia cells and potentiates transformation.

PubMed

Ando, Koji; Tsushima, Hideki; Matsuo, Emi; Horio, Kensuke; Tominaga-Sato, Shinya; Imanishi, Daisuke; Imaizumi, Yoshitaka; Iwanaga, Masako; Itonaga, Hidehiro; Yoshida, Shinichiro; Hata, Tomoko; Moriuchi, Ryozo; Kiyoi, Hitoshi; Nimer, Stephen; Mano, Hiroyuki; Naoe, Tomoki; Tomonaga, Masao; Miyazaki, Yasushi

2013-03-29

Myeloid ELF1-like factor (MEF/ELF4), a member of the ETS transcription factors, can function as an oncogene in murine cancer models and is overexpressed in various human cancers. Here, we report a mechanism by which MEF/ELF4 may be activated by a common leukemia-associated mutation in the nucleophosmin gene. By using a tandem affinity purification assay, we found that MEF/ELF4 interacts with multifactorial protein nucleophosmin (NPM1). Coimmunoprecipitation and GST pull-down experiments demonstrated that MEF/ELF4 directly forms a complex with NPM1 and also identified the region of NPM1 that is responsible for this interaction. Functional analyses showed that wild-type NPM1 inhibited the DNA binding and transcriptional activity of MEF/ELF4 on the HDM2 promoter, whereas NPM1 mutant protein (Mt-NPM1) enhanced these activities of MEF/ELF4. Induction of Mt-NPM1 into MEF/ELF4-overexpressing NIH3T3 cells facilitated malignant transformation. In addition, clinical leukemia samples with NPM1 mutations had higher human MDM2 (HDM2) mRNA expression. Our data suggest that enhanced HDM2 expression induced by mutant NPM1 may have a role in MEF/ELF4-dependent leukemogenesis.

Evaluation of Selected Binding Domains for the Analysis of Ubiquitinated Proteomes

NASA Astrophysics Data System (ADS)

Nakayasu, Ernesto S.; Ansong, Charles; Brown, Joseph N.; Yang, Feng; Lopez-Ferrer, Daniel; Qian, Wei-Jun; Smith, Richard D.; Adkins, Joshua N.

2013-08-01

Ubiquitination is an abundant post-translational modification that consists of covalent attachment of ubiquitin to lysine residues or the N-terminus of proteins. Mono- and polyubiquitination have been shown to be involved in many critical eukaryotic cellular functions and are often disrupted by intracellular bacterial pathogens. Affinity enrichment of ubiquitinated proteins enables global analysis of this key modification. In this context, the use of ubiquitin-binding domains is a promising but relatively unexplored alternative to more broadly used immunoaffinity or tagged affinity enrichment methods. In this study, we evaluated the application of eight ubiquitin-binding domains that have differing affinities for ubiquitination states. Small-scale proteomics analysis identified ~200 ubiquitinated protein candidates per ubiquitin-binding domain pull-down experiment. Results from subsequent Western blot analyses that employed anti-ubiquitin or monoclonal antibodies against polyubiquitination at lysine 48 and 63 suggest that ubiquitin-binding domains from Dsk2 and ubiquilin-1 have the broadest specificity in that they captured most types of ubiquitination, whereas the binding domain from NBR1 was more selective to polyubiquitination. These data demonstrate that with optimized purification conditions, ubiquitin-binding domains can be an alternative tool for proteomic applications. This approach is especially promising for the analysis of tissues or cells resistant to transfection, of which the overexpression of tagged ubiquitin is a major hurdle.

The Tandem CARDs of NOD2: Intramolecular Interactions and Recognition of RIP2

PubMed Central

Fridh, Veronica; Rittinger, Katrin

2012-01-01

Caspase recruitment domains (CARDs) are homotypic protein interaction modules that link the stimulus-dependent assembly of large signaling platforms such as inflammasomes to the activation of downstream effectors that often include caspases and kinases and thereby play an important role in the regulation of inflammatory and apoptotic signaling pathways. NOD2 belongs to the NOD-like (NLR) family of intracellular pattern recognition receptors (PRR) and induces activation of the NF-κB pathway in response to the recognition of bacterial components. This process requires the specific recognition of the CARD of the protein kinase RIP2 by the tandem CARDs of NOD2. Here we demonstrate that the tandem CARDs of NOD2 are engaged in an intramolecular interaction that is important for the structural stability of this region. Using a combination of ITC and pull-down experiments we identify distinct surface areas that are involved in the intramolecular tandem CARD interaction and the interaction with the downstream effector RIP2. Our findings indicate that while CARDa of NOD2 might be the primary binding partner of RIP2 the two CARDs of NOD2 do not act independently of one another but may cooperate to from a binding surface that is distinct from that of single CARDs. PMID:22470564

Identification of four type II toxin-antitoxin systems in Actinobacillus pleuropneumoniae.

PubMed

Zheng, Chengkun; Zhao, Xigong; Zeng, Ting; Cao, Manman; Xu, Jiali; Shi, Guolin; Li, Jinquan; Chen, Huanchun; Bei, Weicheng

2017-07-03

Toxin-antitoxin (TA) systems are small genetic elements that are widely prevalent in the genomes of bacteria and archaea. These modules have been identified in various bacteria and proposed to play an important role in bacterial physiology and virulence. However, their presence in the genomes of Actinobacillus species has received no attention. In this study, we describe the identification of four type II TA systems in Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Reverse transcription PCR analysis revealed that the genes encoding the toxin and antitoxin are co-transcribed. Overexpression of each toxin inhibited the growth of Escherichia coli, and the toxic effect could be counteracted by its cognate antitoxin. The pull-down experiments demonstrated that each toxin interacts with its cognate antitoxin in vivo. The promoter activity assays showed that each antitoxin could autoregulate either positively or negatively the TA operon transcription. In addition, the APJL_0660/0659 TA system is present in half of the detected serovars of A. pleuropneumoniae, while the others are present in all. Collectively, we identified four type II TA systems in A. pleuropneumoniae, and this study has laid the foundation for further functional study of these TA systems. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Long noncoding RNA OR3A4 promotes metastasis and tumorigenicity in gastric cancer

PubMed Central

Guo, Xiaobo; Yang, Ziguo; Zhi, Qiaoming; Wang, Dan; Guo, Lei; Li, Guimei; Miao, Ruizhen; Shi, Yulong; Kuang, Yuting

2016-01-01

The contribution of long noncoding RNAs (lncRNAs) to metastasis of gastric cancer remains largely unknown. We used microarray analysis to identify lncRNAs differentially expressed between normal gastric tissues and gastric cancer tissues and validated these differences in quantitative real-time (qRT)-PCR experiments. The expression levels of lncRNA olfactory receptor, family 3, subfamily A, member 4 (OR3A4) were significantly associated with lymphatic metastasis, the depth of cancer invasion, and distal metastasis in 130 paired gastric cancer tissues. The effects of OR3A4 were assessed by overexpressing and silencing OR3A4 in gastric cancer cells. OR3A4 promoted cancer cell growth, angiogenesis, metastasis, and tumorigenesis in vitro and in vivo. Global microarray analysis combined with RT-PCR, RNA immunoprecipitation, and RNA pull-down analyses after OR3A4 transfection demonstrated that OR3A4 influenced biologic functions in gastric cancer cells via regulating the activation of PDLIM2, MACC1, NTN4, and GNB2L1. Our results reveal OR3A4 as an oncogenic lncRNA that promotes tumor progression, Therefore, lncRNAs might function as key regulatory hubs in gastric cancer progression. PMID:26863570

Containerless glass fiber processing

NASA Technical Reports Server (NTRS)

Ethridge, E. C.; Naumann, R. J.

1986-01-01

An acoustic levitation furnace system is described that was developed for testing the feasibility of containerless fiber pulling experiments. It is possible to levitate very dense materials such as platinum at room temperature. Levitation at elevated temperatures is much more difficult. Samples of dense heavy metal fluoride glass were levitated at 300 C. It is therefore possible that containerless fiber pulling experiments could be performed. Fiber pulling from the melt at 650 C is not possible at unit gravity but could be possible at reduced gravities. The Acoustic Levitation Furnace is described, including engineering parameters and processing information. It is illustrated that a shaped reflector greatly increases the levitation force aiding the levitation of more dense materials.

Micro pulling down growth of very thin shape memory alloys single crystals

NASA Astrophysics Data System (ADS)

López-Ferreño, I.; Juan, J. San; Breczewski, T.; López, G. A.; Nó, M. L.

Shape memory alloys (SMAs) have attracted much attention in the last decades due to their thermo-mechanical properties such as superelasticity and shape memory effect. Among the different families of SMAs, Cu-Al-Ni alloys exhibit these properties in a wide range of temperatures including the temperature range of 100-200∘C, where there is a technological demand of these functional materials, and exhibit excellent behavior at small scale making them more competitive for applications in Micro Electro-Mechanical Systems (MEMS). However, polycrystalline alloys of Cu-based SMAs are very brittle so that they show their best thermo-mechanical properties in single-crystal state. Nowadays, conventional Bridgman and Czochralski methods are being applied to elaborate single-crystal rods up to a minimum diameter of 1mm, but no works have been reported for smaller diameters. With the aim of synthesizing very thin single-crystals, the Micro-Pulling Down (μ-PD) technique has been applied, for which the capillarity and surface tension between crucible and the melt play a critical role. The μ-PD method has been successfully applied to elaborate several cylindrical shape thin single-crystals down to 200μm in diameter. Finally, the martensitic transformation, which is responsible for the shape memory properties of these alloys, has been characterized for different single-crystals. The experimental results evidence the good quality of the grown single-crystals.

An Automated Optical Fiber Puller for Use in Low-Earth Orbit

NASA Technical Reports Server (NTRS)

Tucker, Dennis S.; Smith, W. Scott (Technical Monitor)

2002-01-01

With the slowdown in space station construction, limiting astronaut time for scientific experiments, an effort is being made to automate certain experiments. One such experiment is production of heavy metal fluoride fibers in the microgravity environment. Previous work by this author and others have shown that microgravity inhibits crystallization of ZBLAN glass. Thus an automated experiment has been designed. This experiment will consist of several elements, one which includes the use of an autonomous robot to initiate fiber pulling. The first element will be to melt the preform to eliminate crystals. The preform tip will then be heated to the viscosity necessary for fiber drawing. The robot will initiate the draw and attach the fiber end to the take-up reel. Once fiber pulling has commenced, sensors will be used to detect a fiber break, whereupon the robot can re-initiate the pulling process. The fiber will be coated with a polymer and the polymer cured with ultraviolet light. A laser micrometer will be used to monitor fiber diameter. The experiment is designed so that up to 10 preforms can be pulled into fiber during one flight. The apparatus will be mounted on a free-flying carrier which will be placed into low-earth orbit from the cargo bay of the space shuttle by the shuttle robot arm. The experiment can be started by a signal from the shuttle or from the ground via telescience. The experiment will proceed automatically using specially designed algorithms and will be monitored from the ground. The carrier will be picked up by the shuttle before return to earth.

Detection of Lipomannan in Cattle Infected with Bovine Tuberculosis

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

Vu, Dung M.; Sakamuri, Rama M.; Waters, W. Ray

Early and rapid detection of bovine tuberculosis (bTB) is critical to controlling the spread of this disease in cattle and other animals. Here in this study, we demonstrate the development of an immunoassay for the direct detection of the bovine bTB biomarker, lipomannan (LM) in serum using a waveguide-based optical biosensor. We apply an ultra-sensitive detection strategy developed by our team, termed lipoprotein capture, that exploits the pull-down of high-density lipoprotein (HDL) nanodiscs from cattle blood that allows for the recovery and detection of associated LM. We also profile the change in the expression of these TB biomarkers as amore » function of time from a small set of samples collected from studies of bovine TB-infected cattle. Lastly, we demonstrate for the first time the direct detection of bovine LM in serum, and clearly show that the biomarker is expressed in detectable concentrations during the entire course of the infection.« less

Stimulus learning and response learning by observation in the European starling, in a two-object/two-action test.

PubMed

Campbell; Heyes; Goldsmith

1999-07-01

Juvenile European starlings, Sturnus vulgaris, were allowed to observe a conspecific demonstrator using its beak to remove one of two distinctively coloured objects (i.e. a red or a black plug) from a hole in the lid of a plastic box. Both plugs could be removed by either pulling up on a loop of string inserted through the centre of the plug, or pushing down on the plug. When subsequently allowed access to the plugs, and rewarded with food for all removal responses, regardless of the object to which they were made and their direction, observer birds removed the same plug in the same direction as their demonstrator. These results suggest that the two-object/two-action paradigm is a valuable procedure for testing for the simultaneous effects of learning about a stimulus and a response, an object and an action, through conspecific observation. Copyright 1999 The Association for the Study of Animal Behaviour.

Detection of Lipomannan in Cattle Infected with Bovine Tuberculosis

DOE PAGES

Vu, Dung M.; Sakamuri, Rama M.; Waters, W. Ray; ...

2017-04-10

Early and rapid detection of bovine tuberculosis (bTB) is critical to controlling the spread of this disease in cattle and other animals. Here in this study, we demonstrate the development of an immunoassay for the direct detection of the bovine bTB biomarker, lipomannan (LM) in serum using a waveguide-based optical biosensor. We apply an ultra-sensitive detection strategy developed by our team, termed lipoprotein capture, that exploits the pull-down of high-density lipoprotein (HDL) nanodiscs from cattle blood that allows for the recovery and detection of associated LM. We also profile the change in the expression of these TB biomarkers as amore » function of time from a small set of samples collected from studies of bovine TB-infected cattle. Lastly, we demonstrate for the first time the direct detection of bovine LM in serum, and clearly show that the biomarker is expressed in detectable concentrations during the entire course of the infection.« less

Combining affinity enrichment, cross-linking with photo-amino acids, and mass spectrometry for probing protein kinase D2 interactions.

PubMed

Häupl, Björn; Ihling, Christian H; Sinz, Andrea

2017-04-07

We present a novel approach that relies on the affinity capture of protein interaction partners from a complex mixture, followed by covalent fixation via UV-induced activation of incorporated diazirine photo-reactive amino acids (photo-methionine and photo-leucine). The captured protein complexes are enzymatically digested and interacting proteins are identified and quantified by label-free LC/MS analysis. Using HeLa cell lysates with photo-methionine and photo-leucine-labeled proteins, we were able to capture and preserve protein interactions that are otherwise elusive in conventional pull-down experiments. Our approach is exemplified for mapping the protein interaction network of protein kinase D2, but has the potential be applied to any protein system. Data are available via ProteomeXchange with identifiers PXD005346 (photo-amino acid incorporation) and PXD005349 (enrichment experiments). This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Alternate retrieval technology demonstrations program - test report (ARD Environmental, Inc.)

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

Berglin, E.J.

A prototype vehicle, control system, and waste and water scavenging system were designed and fabricated with essentially the full capabilities of the vehicle system proposed by ARD Environmental. A test tank mockup, including riser and decontamination chamber were designed and fabricated, and approximately 830 cubic feet of six varieties of waste simulants poured. The tests were performed by ARD Environmental personnel at its site in Laurel, Maryland, from 4/22/97 through 5/2/97. The capabilities tested were deployment and retrieval, extended mobility and productivity, the ability to operate the system using video viewing only, retrieval after simulated failure, and retrieval and decontamination.more » Testing commenced with deployment of the vehicle into the tank. Deployment was accomplished using a crane and auxiliary winch to position the vehicle and lower it through the decontamination chamber, into the 36`` diameter x 6` high riser, and touch down on the waste field in the tank. The initial mobility tests were conducted immediately after deployment, prior to sluicing, as the waste field exhibited the greatest amount of variation at this time. This test demonstrated the ability of the vehicle to maneuver over the simulated waste field, and the ability of the operator to work with only video viewing available. In addition, the ability of the vehicle to right itself after being turned on its side was demonstrated. The production rate was evaluated daily through the testing period by measuring the surface and estimating the amount of material removed. The test demonstrated the ability of the vehicle to reduce the waste surface using 400 psi (nominal) water jets, scavenge water and material from the work area, and move to any location, even in the relatively confined space of the 20` diameter test tank. In addition, the ability to sluice to a remote scavenging module was demonstrated. The failure mode test demonstrated the ability to retrieve a stuck vehicle by pulling on the tether, even if the vehicle wheels were locked or the vehicle was on its side. Line pull required to retrieve the vehicle was measured, and side load on the riser calculated from the line pull and line angles. Finally, the decontamination test demonstrated the ability to effectively clean the umbilical and vehicle. The issues addressed and resolved during the testing were: Feasibility of deploying a vehicle- based system, mobility, production rate and limitation of water in the tank during sluicing, mining strategy, operator efficiency, vehicle recovery, and decontamination. Water usage and waste removal rates were used to estimate the time and water usage requirements for cleaning a Hanford SST.« less

JPRS Report, East Europe.

DTIC Science & Technology

1988-05-31

anecdote. My son is 18 and a goalie for a B-League soccer team. In a recent game his team was losing 3 to nothing. In the stands they started yelling...recover. This is so not only because "the low level traps of a lack of balance" would constantly pull us down. It will not be easy to find a clear

75 FR 25853 - Agency Information Collection Activities; Submission to OMB for Review and Approval; Comment...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-10

....gov , to submit or view public comments, access the index listing of the contents of the docket, and...:8080/glas/login.htm . The Web-site will contain a user-friendly data entry interface for recipients to... series of screens containing pull-down menus and text boxes, where users can enter project specific...

An Empirical Comparison of Navigation Effect of Pull-Down Menu Style on The World Wide Web.

ERIC Educational Resources Information Center

Yu, Byeong-Min; Han, Sungwook

Effective navigation is becoming more and more critical to the success of electronic commerce (E-commerce). It remains a challenge for educational technologists and Web designers to develop Web systems that can help customers find products or services without experiencing disorientation problems and cognitive overload. Many E-commerce Web sites…

From Sea to Shining Sea

ERIC Educational Resources Information Center

Scott, Beverly

2005-01-01

Deep down in the depths of the sea, beautiful fish, mysterious ocean life, and unusual plants glimmer and glow in the eerie atmosphere of an ever-changing ocean. This article describes how, with this vision and a purpose in mind, three teachers pulled open classroom walls and joined forces so their second graders could create a mammoth 30 x 75"…

NAPI Stories. Blackfeet Heritage Program: Browning.

ERIC Educational Resources Information Center

Rides At The Door, Darnell Davis, Comp.

Napi stories have been passed down from generation to generation in the Blackfeet Nation. All Blackfeet people knew of Napi, from the serious side of his creation to the foolish and spiteful deeds he performed. At one time it is said that Napi could talk with all living things--the animals, plants, rocks, everything. He teased, pulled pranks, many…

A Theory of Inference Derivation for Qualitative Data: Development and Test with Application to Criminal and Terrorist Detection

DTIC Science & Technology

1991-07-01

me ’Don’t struggle or scream or cIzc you’]’ ’e a d-ad girl .’ He made me kneel with my back to him. He kept saying ’How old are you ?’ and he also kept...your boyfriend or the police. If I hear from anybody that you have told anybody you will be a dead girl ’. He then said ’don’t move until I’ve gone’. He...down. He pulled my tights and panties off completely. He undid his trousers and got back on top of me and had sex with me. He stood up and pulled me up

mRNA-Selective Translation Induced by FSH in Primary Sertoli Cells

PubMed Central

Musnier, Astrid; León, Kelly; Morales, Julia; Reiter, Eric; Boulo, Thomas; Costache, Vlad; Vourc'h, Patrick; Heitzler, Domitille; Oulhen, Nathalie; Poupon, Anne; Boulben, Sandrine; Cormier, Patrick

2012-01-01

FSH is a key hormonal regulator of Sertoli cell secretory activity, required to optimize sperm production. To fulfil its biological function, FSH binds a G protein-coupled receptor, the FSH-R. The FSH-R-transduced signaling network ultimately leads to the transcription or down-regulation of numerous genes. In addition, recent evidence has suggested that FSH might also regulate protein translation. However, this point has never been demonstrated conclusively yet. Here we have addressed this issue in primary rat Sertoli cells endogenously expressing physiological levels of FSH-R. We observed that, within 90 min of stimulation, FSH not only enhanced overall protein synthesis in a mammalian target of rapamycin-dependent manner but also increased the recruitment of mRNA to polysomes. m7GTP pull-down experiments revealed the functional recruitment of mammalian target of rapamycin and p70 S6 kinase to the 5′cap, further supported by the enhanced phosphorylation of one of p70 S6 kinase targets, the eukaryotic initiation factor 4B. Importantly, the scaffolding eukaryotic initiation factor 4G was also recruited, whereas eukaryotic initiation factor 4E-binding protein, the eukaryotic initiation factor 4E generic inhibitor, appeared to play a minor role in translational regulations induced by FSH, in contrast to what is generally observed in response to anabolic factors. This particular regulation of the translational machinery by FSH stimulation might support mRNA-selective translation, as shown here by quantitative RT-PCR amplification of the c-fos and vascular endothelial growth factor mRNA but not of all FSH target mRNA, in polysomal fractions. These findings add a new level of complexity to FSH biological roles in its natural target cells, which has been underappreciated so far. PMID:22383463

Intraoperative Physical Examination for Diagnosis of Interosseous Ligament Rupture-Cadaveric Study.

PubMed

Kachooei, Amir Reza; Rivlin, Michael; Wu, Fei; Faghfouri, Aram; Eberlin, Kyle R; Ring, David

2015-09-01

To study the intraobserver and interobserver reliability of the diagnosis of interosseous ligament (IOL) rupture in a cadaver model. On 12 fresh frozen cadavers, radial heads were cut using an identical incision and osteotomy. After randomization, the soft tissues of the limbs were divided into 4 groups: both IOL and triangular fibrocartilage (TFCC) intact; IOL disruption but TFCC intact; both IOL and TFCC divided; and IOL intact but TFCC divided. All incisions had identical suturing. After standard instruction and demonstration of radius pull-push and radius lateral pull tests, 10 physician evaluators with different levels of experience examined the cadaver limbs in a standardized way (elbow at 90° with the forearm held in both supination and pronation) and were asked to classify them into one of the 4 groups. Next, the same examiners were asked to re-examine the limbs after randomly changing the order of examination. The interobserver reliability of agreement for the diagnosis of IOL injury (groups 2 and 3) was fair in both rounds of examination and the intraobserver reliability was moderate. The intra- and interobserver reliabilities of agreement for the 4 groups of injuries among the examiners were fair in both rounds of examination. The sensitivity, specificity, accuracy, positive, and negative predictive values were all around 70%. The likelihood of a positive test corresponding with the presence of IOL rupture (positive likelihood ratio) was 2.2. The likelihood of a negative test correctly diagnosing an intact IOL was 0.40. In cadavers, intraoperative tests had fair reliability and 70% accuracy for the diagnosis of IOL rupture using the push-pull and lateral pull maneuvers. The level of experience did not have any effect on the correct diagnosis of intact versus disrupted IOL. Although not common, some failure of surgeries for traumatic elbow fracture-dislocations is because of failure in timely diagnosis of IOL disruption. Copyright © 2015 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

RNA-binding proteins regulate the expression of the immune activating ligand MICB

PubMed Central

Nachmani, Daphna; Gutschner, Tony; Reches, Adi

2014-01-01

The recognition of stress-induced ligands by the activating receptor NKG2D expressed on cytotoxic lymphocytes is crucial for the prevention and containment of various diseases and is also one of the best-studied examples of how danger is sensed by the immune system. Still, however, the mechanisms leading to the expression of the NKG2D ligands are far from being completely understood. Here, we use an unbiased and systematic RNA pull-down approach combined with mass spectrometry to identify six RNA-binding proteins (RBPs) that bind and regulate the expression of MICB, one of the major stress-induced ligands of NKG2D. We further demonstrate that at least two of the identified RBPs function during genotoxic stress. Our data provide insights into stress recognition and hopefully open new therapeutic venues. PMID:24924487

ZBLAN Fiber Phase B Study

NASA Technical Reports Server (NTRS)

Workman, Gary L.; Smith, Guy A.

1997-01-01

A Phase B feasibility study will be performed for the study of the effects of microgravity on the preform processing and fiber pulling of ZBLAN optical glass. Continuing from the positive results achieved in the fiber annealing experiments in 20 second intervals at 0.001 g on the KC-135 and the 5 minute experiments on the SPAR rocket, experiments will continue to work towards design of a fiber sting to initiate fiber pulling operations in space. Anticipated results include less homogeneous nucleation than ground-based annealed fibers. Infrared Fiber Systems and Galileo are the participating industrial investigators.

Mitigating Oscillator Pulling Due To Magnetic Coupling in Monolithic Mixed-Signal Radio-Frequency Integrated Circuits

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

Sobering, Ian David

2014-01-01

An analysis of frequency pulling in a varactor-tuned LC VCO under coupling from an on-chip PA is presented. The large-signal behavior of the VCO's inversion-mode MOS varactors is outlined, and the susceptibility of the VCO to frequency pulling from PA aggressor signals with various modulation schemes is discussed. We show that if the aggressor signal is aperiodic, band-limited, or amplitude-modulated, the varactor-tuned LC VCO will experience frequency pulling due to time-modulation of the varactor capacitance. However, if the aggressor signal has constant-envelope phase modulation, VCO pulling can be eliminated, even in the presence of coupling, through careful choice of VCOmore » frequency and divider ratio. Additional mitigation strategies, including new inductor topologies and system-level architectural choices, are also examined.« less

Characterization and Effects of Fiber Pull-Outs in Hole Quality of Carbon Fiber Reinforced Plastics Composite

PubMed Central

Alizadeh Ashrafi, Sina; Miller, Peter W.; Wandro, Kevin M.; Kim, Dave

2016-01-01

Hole quality plays a crucial role in the production of close-tolerance holes utilized in aircraft assembly. Through drilling experiments of carbon fiber-reinforced plastic composites (CFRP), this study investigates the impact of varying drilling feed and speed conditions on fiber pull-out geometries and resulting hole quality parameters. For this study, hole quality parameters include hole size variance, hole roundness, and surface roughness. Fiber pull-out geometries are quantified by using scanning electron microscope (SEM) images of the mechanically-sectioned CFRP-machined holes, to measure pull-out length and depth. Fiber pull-out geometries and the hole quality parameter results are dependent on the drilling feed and spindle speed condition, which determines the forces and undeformed chip thickness during the process. Fiber pull-out geometries influence surface roughness parameters from a surface profilometer, while their effect on other hole quality parameters obtained from a coordinate measuring machine is minimal. PMID:28773950

Enhanced isoprene biosynthesis in Saccharomyces cerevisiae by engineering of the native acetyl-CoA and mevalonic acid pathways with a push-pull-restrain strategy.

PubMed

Lv, Xiaomei; Xie, Wenping; Lu, Wenqiang; Guo, Fei; Gu, Jiali; Yu, Hongwei; Ye, Lidan

2014-09-30

To explore the capacity of isoprene production in Saccharomyces cerevisiae, a rational push-pull-restrain strategy was proposed to engineer the mevalonic acid (MVA) and acetyl-CoA pathways. The strategy can be decomposed into the up-regulation of precursor supply in the acetyl-CoA module and the MVA pathway (push-strategy), increase of the isoprene branch flux (pull-strategy), and down-regulation of the competing pathway (restrain-strategy). Furthermore, to reduce the production cost arising from galactose addition and meanwhile maintain the high expression of Gal promoters, the galactose regulatory network was modulated by Gal80p deletion. Finally, the engineered strain YXM10-ispS-ispS could accumulate up to 37 mg/L isoprene (about 782-fold increase compared to the parental strain) under aerobic conditions with glycerol-sucrose as carbon source. In this way, a new potential platform for isoprene production was established via metabolic engineering of the yeast native pathways. Copyright © 2014 Elsevier B.V. All rights reserved.

SpeedyTime-5_Water_In_Space

NASA Image and Video Library

2017-08-10

The International Space Station is a one-of-a-kind spot for scientists who want to do experiments where there’s no gravity, to find out how other natural forces function without gravity’s influence. In this “SpeedyTime” segment, Expedition 52 flight engineer Jack Fischer uses a few simple tools to demonstrate what happens to water in space when there’s no pull of gravity. _______________________________________ FOLLOW THE SPACE STATION! Twitter: https://twitter.com/Space_Station Facebook: https://www.facebook.com/ISS Instagram: https://instagram.com/iss/

Use an Interactive Whiteboard: Get a Handle on How This Technology Can Spice up the Classroom

ERIC Educational Resources Information Center

Branzburg, Jeffrey

2006-01-01

Interactive whiteboards are desirable peripherals these days. When hooked up to a computer, the whiteboard's screen becomes a "live" computer desktop, which can be tapped to pull down menus, highlight, and move or open files. Users can also circle relevant sections on the projected image, draw geometric figures, and underline. Then they can save…

70. DETAIL OF TRACTION CABLE ENGAGEMENT DEVICE. SMALL, KNOBBED LEVER ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

70. DETAIL OF TRACTION CABLE ENGAGEMENT DEVICE. SMALL, KNOBBED LEVER ON BUCKET HANGER WAS PULLED DOWN BY A CAMEL (FIXED CAM RAIL AT CENTER) AS BUCKET ROLLED PAST IT, CAUSING A CLAMP TO CLOSE AGAINST TRACTION CABLE. A SIMILAR CAMEL (NO LONGER EXTANT) DISENGAGED CLAMP ON RECEIVING SIDE. - Shenandoah-Dives Mill, 135 County Road 2, Silverton, San Juan County, CO

Argonaute pull-down and RISC analysis using 2'-O-methylated oligonucleotides affinity matrices.

PubMed

Jannot, Guillaume; Vasquez-Rifo, Alejandro; Simard, Martin J

2011-01-01

During the last decade, several novel small non-coding RNA pathways have been unveiled, which reach out to many biological processes. Common to all these pathways is the binding of a small RNA molecule to a protein member of the Argonaute family, which forms a minimal core complex called the RNA-induced silencing complex or RISC. The RISC targets mRNAs in a sequence-specific manner, either to induce mRNA cleavage through the intrinsic activity of the Argonaute protein or to abrogate protein synthesis by a mechanism that is still under investigation. We describe here, in details, a method for the affinity chromatography of the let-7 RISC starting from extracts of the nematode Caenorhabditis elegans. Our method exploits the sequence specificity of the RISC and makes use of biotinylated and 2'-O-methylated oligonucleotides to trap and pull-down small RNAs and their associated proteins. Importantly, this technique may easily be adapted to target other small RNAs expressed in different cell types or model organisms. This method provides a useful strategy to identify the proteins associated with the RISC, and hence gain insight in the functions of small RNAs.

Vacuum pull down method for an enhanced bonding process

DOEpatents

Davidson, James C.; Balch, Joseph W.

1999-01-01

A process for effectively bonding arbitrary size or shape substrates. The process incorporates vacuum pull down techniques to ensure uniform surface contact during the bonding process. The essence of the process for bonding substrates, such as glass, plastic, or alloys, etc., which have a moderate melting point with a gradual softening point curve, involves the application of an active vacuum source to evacuate interstices between the substrates while at the same time providing a positive force to hold the parts to be bonded in contact. This enables increasing the temperature of the bonding process to ensure that the softening point has been reached and small void areas are filled and come in contact with the opposing substrate. The process is most effective where at least one of the two plates or substrates contain channels or grooves that can be used to apply vacuum between the plates or substrates during the thermal bonding cycle. Also, it is beneficial to provide a vacuum groove or channel near the perimeter of the plates or substrates to ensure bonding of the perimeter of the plates or substrates and reduce the unbonded regions inside the interior region of the plates or substrates.

Low temperature-pressure batch experiments and field push-pull tests: Assessing potential effects of an unintended CO2 release from CCUS projects on groundwater chemistry

NASA Astrophysics Data System (ADS)

Mickler, P. J.; Yang, C.; Lu, J.; Reedy, R. C.; Scanlon, B. R.

2012-12-01

Carbon Capture Utilization and Storage projects (CCUS), where CO2 is captured at point sources such as power stations and compressed into a supercritical liquid for underground storage, has been proposed to reduce atmospheric CO2 and mitigate global climate change. Problems may arise from CO2 releases along discreet pathways such as abandoned wells and faults, upwards and into near surface groundwater. Migrating CO2 may inversely impact fresh water resources by increasing mineral solubility and dissolution rates and mobilizing harmful trace elements including As and Pb. This study addresses the impacts on fresh water resources through a combination of laboratory batch experiments, where aquifer sediment are reacted in their corresponding groundwater in 100% CO2 environments, and field push-pull tests where groundwater is equilibrated with 100% CO2, reacted in-situ in the groundwater system, and pulled out for analyses. Batch experiments were performed on aquifer material from carbonate dominated, mixed carbonate/silicalstic, and siliclastic dominated systems. A mixed silicalstic/carbonate system was chosen for the field based push-pull test. Batch experiment results suggest carbonate dissolution increased the concentration of Ca, Mg, Sr, Ba, Mn, U and HCO3- in groundwater. In systems with significant carbonate content, dissolution continued until carbonate saturation was achieved at approximately 1000 hr. Silicate dissolution increased the conc. of Si, K Ni and Co, but at much lower rates than carbonate dissolution. The elements As, Mo, V, Zn, Se and Cd generally show similar behavior where concentrations initially increase but soon drop to levels at or below the background concentrations (~48 hours). A Push-Pull test on one aquifer system produced similar geochemical behavior but observed reaction rates are higher in batch experiments relative to push-pull tests. Release of CO2 from CCUS sites into overlying aquifer systems may adversely impact groundwater quality primarily through carbonate dissolution which releases Ca and elements that substitute for Ca in crystal lattices. Silicate weathering releases primarily Si and K at lower rates. Chemical changes with the addition of CO2 may initially mobilize As, Mo, V, Zn, Se and Cd but these elements become immobile in the lowered pH water and sorb onto aquifer minerals. A combined laboratory batch experiment and field push-pull test in fresh water aquifers overlying CCUS projects will best characterize the response of the aquifer to increased pCO2. The long experimental duration of the batch experiments may allow reactions to reach equilibrium however; reaction rates may be artificially high due to increased mineral surface areas. Field based push-pull tests offer a more realistic water rock ratio and test a much larger volume of aquifer material but the test must be shorter in duration because the high pCO2 water is subject to mixing with low pCO2 background water and migration away from the test well with groundwater flow. A comparison of the two methods best characterizes the potential effects on groundwater chemistry

Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis

PubMed Central

Vara, Dina; Watt, Joanna M.; Fortunato, Tiago M.; Mellor, Harry; Burgess, Matthew; Wicks, Kate; Mace, Kimberly; Reeksting, Shaun; Lubben, Anneke; Wheeler-Jones, Caroline P.D.

2018-01-01

Abstract Aims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, electron paramagnetic resonance, pull-down of dRP-interacting proteins, followed by immunoblotting, gene silencing of different NADPH oxidases (NOXs), and their regulatory cosubunits by small interfering RNA (siRNA) transfection, and experiments with inhibitors of the sugar transporter glucose transporter 1 (GLUT1) were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NOX2 complex, but not NOX4. Increased reactive oxygen species generation in response to NOX2 activity leads to redox-dependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces vascular endothelial growth factor receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA, and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and NOX2-dependent stimulation of angiogenesis by dRP were confirmed in excisional wound and Matrigel plug vascularization assays in vivo using NOX2−/− mice. Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex. Conclusions: This study describes a novel molecular mechanism underlying the proangiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and upregulation of VEGFR2. Antioxid. Redox Signal. 28, 110–130. PMID:28793782

Choices of Destination for Transnational Higher Education: "Pull" Factors in an Asia Pacific Market

ERIC Educational Resources Information Center

Ahmad, Syed Zamberi; Buchanan, Frederick Robert

2016-01-01

Traditional assumptions favouring native English language countries in transnational higher education (TNHE) overlook experiences of international students in new emerging Asian education hubs. Specifically, there has been limited research relating to international students' choice for studying in Malaysia. Drawing from the "push-pull"…

The synthetic peptide P111-136 derived from the C-terminal domain of heparin affin regulatory peptide inhibits tumour growth of prostate cancer PC-3 cells

PubMed Central

2011-01-01

Background Heparin affin regulatory peptide (HARP), also called pleiotrophin, is a heparin-binding, secreted factor that is overexpressed in several tumours and associated to tumour growth, angiogenesis and metastasis. The C-terminus part of HARP composed of amino acids 111 to 136 is particularly involved in its biological activities and we previously established that a synthetic peptide composed of the same amino acids (P111-136) was capable of inhibiting the biological activities of HARP. Here we evaluate the ability of P111-136 to inhibit in vitro and in vivo the growth of a human tumour cell line PC-3 which possess an HARP autocrine loop. Methods A total lysate of PC-3 cells was incubated with biotinylated P111-136 and pulled down for the presence of the HARP receptors in Western blot. In vitro, the P111-136 effect on HARP autocrine loop in PC-3 cells was determined by colony formation in soft agar. In vivo, PC-3 cells were inoculated in the flank of athymic nude mice. Animals were treated with P111-136 (5 mg/kg/day) for 25 days. Tumour volume was evaluated during the treatment. After the animal sacrifice, the tumour apoptosis and associated angiogenesis were evaluated by immunohistochemistry. In vivo anti-angiogenic effect was confirmed using a mouse Matrigel™ plug assay. Results Using pull down experiments, we identified the HARP receptors RPTPβ/ζ, ALK and nucleolin as P111-136 binding proteins. In vitro, P111-136 inhibits dose-dependently PC-3 cell colony formation. Treatment with P111-136 inhibits significantly the PC-3 tumour growth in the xenograft model as well as tumour angiogenesis. The angiostatic effect of P111-136 on HARP was also confirmed using an in vivo Matrigel™ plug assay in mice Conclusions Our results demonstrate that P111-136 strongly inhibits the mitogenic effect of HARP on in vitro and in vivo growth of PC-3 cells. This inhibition could be linked to a direct or indirect binding of this peptide to the HARP receptors (ALK, RPTPβ/ζ, nucleolin). In vivo, the P111-136 treatment significantly inhibits both the PC-3 tumour growth and the associated angiogenesis. Thus, P111-136 may be considered as an interesting pharmacological tool to interfere with tumour growth that has now to be evaluated in other cancer types. PMID:21624116

Casingless down-hole for sealing an ablation volume and obtaining a sample for analysis

DOEpatents

Noble, Donald T.; Braymen, Steven D.; Anderson, Marvin S.

1996-10-01

A casing-less down hole sampling system for acquiring a subsurface sample for analysis using an inductively coupled plasma system is disclosed. The system includes a probe which is pushed into the formation to be analyzed using a hydraulic ram system. The probe includes a detachable tip member which has a soil point mad a barb, with the soil point aiding the penetration of the earth, and the barb causing the tip member to disengage from the probe and remain in the formation when the probe is pulled up. The probe is forced into the formation to be tested, and then pulled up slightly, to disengage the tip member and expose a column of the subsurface formation to be tested. An instrumentation tube mounted in the probe is then extended outward from the probe to longitudinally extend into the exposed column. A balloon seal mounted on the end of the instrumentation tube allows the bottom of the column to be sealed. A source of laser radiation is emitted from the instrumentation tube to ablate a sample from the exposed column. The instrumentation tube can be rotated in the probe to sweep the laser source across the surface of the exposed column. An aerosol transport system carries the ablated sample from the probe to the surface for testing in an inductively coupled plasma system. By testing at various levels in the down-hole as the probe is extracted from the soil, a profile of the subsurface formation may be obtained.

Pull-pull position control of dual motor wire rope transmission.

PubMed

Guo, Quan; Jiao, Zongxia; Yan, Liang; Yu, Qian; Shang, Yaoxing

2016-08-01

Wire rope transmission is very efficient because of the small total moving object mass. The wire rope could only transmit pulling force. Therefore it has to be kept in a tightened state during transmission; in high speed applications the dynamic performance depends on the rope's stiffness, which can be adjusted by the wire rope tension. To improve the system dynamic performance output, this paper proposes a novel pull-pull method based on dual motors connected by wire ropes, for precise, high speed position control applications. The method can regulate target position and wire rope tension simultaneously. Wire ropes remain in a pre-tightening state at all times, which prevents the influence of elasticity and reduces the position tracking error in the changing direction process. Simulations and experiments were conducted; the results indicate that both position precision and superior dynamic performance can be synchronously achieved. The research is relevant to space craft precision pointing instruments.

A pull out test to compare two riparian species, Phyllanthus sellowianus and Sebastiania schottiana in terms of root anchorage ability

NASA Astrophysics Data System (ADS)

Hörbinger, Stephan; Sutili, Fabricio J.; Rauch, Hans Peter

2013-04-01

Soil bioengineering has become manifold applied in large parts of Brazil in recent years. The first projects were realized in the region of Rio Grande do Sul within river stabilization works to protect agricultural land of small regional farmers. As result of research work the species Sebastiania schottiana and Phyllanthus sellowianus showed very adequate morpho-physiological properties and seem to be appropriate for the use in soil bioengineering. The aim of the present study was to examine a still unknown but crucial factor, the resistance of the above mentioned species against being pulled out. The pull out resistance is an indicator for the stability of the soil-root matrix and expresses the stabilizing effects of plants on soil. Furthermore it is an applicable index to compare the qualification of the species to be used in soil bioengineering works. Another objective was to investigate plant characteristics, which correlate to the pull out resistance of the investigated species, to be able to draft up efficient plant strategies for future restoration works on eroded river embankments. For the experiment a special apparatus was designed, which enables to implement a pull out process with a constant rate and generate a graph of the plants resistance force versus its displacement. P. sellowianus showed a significant higher resistance against being pulled out than S. schottiana. The analyses of root and shoot properties of P. sellowianus showed more favorable morpho-physiological properties in terms of pull out resistance, a bigger amount of biomass, both above and below ground and also a higher amount of anchorage. The Cross-Sectional-Areas (CSA) of the shoots showed in both species the strongest correlation of the investigated shoot and root properties with the maximum resistance against being pulled out. Thus it can be concluded that the CSA can be used as a value to assess the stabilization effects of the plants. The experiments showed that some root and shoot properties do have a great impact on the pullout strength and that P. sellowianus can be preferred for slope stabilization works as it exhibits outstanding resistance against being pulled out.

Identification of proteins associated with the yeast mitochondrial RNA polymerase by tandem affinity purification

PubMed Central

Markov, Dmitriy A; Savkina, Maria; Anikin, Michael; Del Campo, Mark; Ecker, Karen; Lambowitz, Alan M; De Gnore, Jon P; McAllister, William T

2009-01-01

The abundance of mitochondrial (mt) transcripts varies under different conditions, and is thought to depend upon rates of transcription initiation, transcription termination/attenuation and RNA processing/degradation. The requirement to maintain the balance between RNA synthesis and processing may involve coordination between these processes; however, little is known about factors that regulate the activity of mtRNA polymerase (mtRNAP). Recent attempts to identify mtRNAP–protein interactions in yeast by means of a generalized tandem affinity purification (TAP) protocol were not successful, most likely because they involved a C-terminal mtRNAP–TAP fusion (which is incompatible with mtRNAP function) and because of the use of whole-cell solubilization protocols that did not preserve the integrity of mt protein complexes. Based upon the structure of T7 RNAP (to which mtRNAPs show high sequence similarity), we identified positions in yeast mtRNAP that allow insertion of a small affinity tag, confirmed the mature N-terminus, constructed a functional N-terminal TAP–mtRNAP fusion, pulled down associated proteins, and identified them by LC–MS–MS. Among the proteins found in the pull-down were a DEAD-box protein (Mss116p) and an RNA-binding protein (Pet127p). Previous genetic experiments suggested a role for these proteins in linking transcription and RNA degradation, in that a defect in the mt degradadosome could be suppressed by overexpression of either of these proteins or, independently, by mutations in either mtRNAP or its initiation factor Mtf1p. Further, we found that Mss116p inhibits transcription by mtRNAP in vitro in a steady-state reaction. Our results support the hypothesis that Mss116p and Pet127p are involved in modulation of mtRNAP activity. Copyright © 2009 John Wiley & Sons, Ltd. PMID:19536766

Direct interaction between surface β1,4-galactosyltransferase 1 and epidermal growth factor receptor (EGFR) inhibits EGFR activation in hepatocellular carcinoma

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

Tang, Wenqing; Weng, Shuqiang; Zhang, Si

2013-05-10

Highlights: •β1,4GT1 interacts with EGFR both in vitro and in vivo. •β1,4GT1 co-localizes with EGFR on the cell surface. •β1,4GT1 inhibits {sup 125}I-EGF binding to EGFR. •β1,4GT1 inhibits EGF induced EGFR dimerization and phosphorylation. -- Abstract: Our previous studies showed that cell surface β1,4-galactosyltransferase 1 (β1,4GT1) negatively regulated cell survival through inhibition and modulation of the epidermal growth factor receptor (EGFR) signaling pathway in human hepatocellular carcinoma (HCC) SMMC-7721 cells. However, the underlying mechanism remains unclear. Here we demonstrated that β1,4-galactosyltransferase 1 (β1,4GT1) interacted with EGFR in vitro by GST pull-down analysis. Furthermore, we demonstrated that β1,4GT1 bound to EGFRmore » in vivo by co-immunoprecipitation and determined the co-localization of β1,4GT1 and EGFR on the cell surface via confocal laser scanning microscopy analysis. Finally, using {sup 125}I-EGF binding experiments and Western blot analysis, we found that overexpression of β1,4GT1 inhibited {sup 125}I-EGF binding to EGFR, and consequently reduced the levels of EGFR dimerization and phosphorylation. In contrast, RNAi-mediated knockdown of β1,4GT1 increased the levels of EGFR dimerization and phosphorylation. These data suggest that cell surface β1,4GT1 interacts with EGFR and inhibits EGFR activation.« less

S.E.E. Program Parents Manual: How to Raise a Child with Epilepsy. Part Two: Coping with Stigma

ERIC Educational Resources Information Center

Mittan, Robert J.

2005-01-01

Epilepsy is the most misunderstood of all neurological disorders known to man. Even though modern medicine (a very recent development in human history) learned that epilepsy was a common variation in biology, the roots laid down by centuries of misunderstanding have yet to be pulled from the society's social consciousness. While medicine and now…

Criterion 5: Maintenance of forest contributions to global carbon cycles

Treesearch

Stephen R. Shifley; Francisco X. Aguilar; Nianfu Song; Susan I. Stewart; David J. Nowak; Dale D. Gormanson; W. Keith Moser; Sherri Wormstead; Eric J. Greenfield

2012-01-01

Northern forests cover more than 42 percent of the region and are enormous reservoirs of carbon. Through photosynthesis, live trees emit oxygen in exchange for carbon dioxide they pull from the atmosphere. As a tree grows it stores carbon in wood above and below ground, and sequestered carbon comprises about half of its dry weight. Dead trees and down logs are also...

Theater in Physics Teacher Education

ERIC Educational Resources Information Center

van den Berg, Ed

2009-01-01

Ten years ago I sat down with the first batch of students in our science/math teacher education program in the Philippines, then third-year students, and asked them what they could do for the opening of the new science building. One of them pulled a stack of papers out of his bag and put it in front of me: a complete script for a science play!…

Arabidopsis DNA methyltransferase AtDNMT2 associates with histone deacetylase AtHD2s activity

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

Song, Yuan; Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3; Wu, Keqiang

2010-05-28

DNA methyltransferase2 (DNMT2) is always deemed to be enigmatic, because it contains highly conserved DNA methyltransferase motifs but lacks the DNA methylation catalytic capability. Here we show that Arabidopsis DNA methyltransferase2 (AtDNMT2) is localized in nucleus and associates with histone deacetylation. Bimolecular fluorescence complementation and pull-down assays show AtDNMT2 interacts with type-2 histone deacetylases (AtHD2s), a unique type of histone deacetylase family in plants. Through analyzing the expression of AtDNMT2: ss-glucuronidase (GUS) fusion protein, we demonstrate that AtDNMT2 has the ability to repress gene expression at transcription level. Meanwhile, the expression of AtDNMT2 gene is altered in athd2c mutant plants.more » We propose that AtDNMT2 possibly involves in the activity of histone deacetylation and plant epigenetic regulatory network.« less

DNA-binding activity of TNF-{alpha} inducing protein from Helicobacter pylori

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

Kuzuhara, T.; Suganuma, M.; Oka, K.

2007-11-03

Tumor necrosis factor-{alpha} (TNF-{alpha}) inducing protein (Tip{alpha}) is a carcinogenic factor secreted from Helicobacter pylori (H. pylori), mediated through both enhanced expression of TNF-{alpha} and chemokine genes and activation of nuclear factor-{kappa}B. Since Tip{alpha} enters gastric cancer cells, the Tip{alpha} binding molecules in the cells should be investigated. The direct DNA-binding activity of Tip{alpha} was observed by pull down assay using single- and double-stranded genomic DNA cellulose. The surface plasmon resonance assay, indicating an association between Tip{alpha} and DNA, revealed that the affinity of Tip{alpha} for (dGdC)10 is 2400 times stronger than that of del-Tip{alpha}, an inactive Tip{alpha}. This suggestsmore » a strong correlation between DNA-binding activity and carcinogenic activity of Tip{alpha}. And the DNA-binding activity of Tip{alpha} was first demonstrated with a molecule secreted from H. pylori.« less

The X-windows interactive navigation data editor

NASA Technical Reports Server (NTRS)

Rinker, G. C.

1992-01-01

A new computer program called the X-Windows Interactive Data Editor (XIDE) was developed and demonstrated as a prototype application for editing radio metric data in the orbit-determination process. The program runs on a variety of workstations and employs pull-down menus and graphical displays, which allow users to easily inspect and edit radio metric data in the orbit data files received from the Deep Space Network (DSN). The XIDE program is based on the Open Software Foundation OSF/Motif Graphical User Interface (GUI) and has proven to be an efficient tool for editing radio metric data in the navigation operations environment. It was adopted by the Magellan Navigation Team as their primary data-editing tool. Because the software was designed from the beginning to be portable, the prototype was successfully moved to new workstation environments. It was also itegrated into the design of the next-generation software tool for DSN multimission navigation interactive launch support.

Characterization of New Cationic N,N-Dimethyl[70]fulleropyrrolidinium Iodide Derivatives as Potent HIV-1 Maturation Inhibitors.

PubMed

Castro, Edison; Martinez, Zachary S; Seong, Chang-Soo; Cabrera-Espinoza, Andrea; Ruiz, Mauro; Hernandez Garcia, Andrea; Valdez, Federico; Llano, Manuel; Echegoyen, Luis

2016-12-22

HIV-1 maturation can be impaired by altering protease (PR) activity, the structure of the Gag-Pol substrate, or the molecular interactions of viral structural proteins. Here we report the synthesis and characterization of new cationic N,N-dimethyl[70]fulleropyrrolidinium iodide derivatives that inhibit more than 99% of HIV-1 infectivity at low micromolar concentrations. Analysis of the HIV-1 life cycle indicated that these compounds inhibit viral maturation by impairing Gag and Gag-Pol processing. Importantly, fullerene derivatives 2a-c did not inhibit in vitro PR activity and strongly interacted with HIV immature capsid protein in pull-down experiments. Furthermore, these compounds potently blocked infectivity of viruses harboring mutant PR that are resistant to multiple PR inhibitors or mutant Gag proteins that confer resistance to the maturation inhibitor Bevirimat. Collectively, our studies indicate fullerene derivatives 2a-c as potent and novel HIV-1 maturation inhibitors.

Push-pull tests for estimating effective porosity: expanded analytical solution and in situ application

NASA Astrophysics Data System (ADS)

Paradis, Charles J.; McKay, Larry D.; Perfect, Edmund; Istok, Jonathan D.; Hazen, Terry C.

2018-03-01

The analytical solution describing the one-dimensional displacement of the center of mass of a tracer during an injection, drift, and extraction test (push-pull test) was expanded to account for displacement during the injection phase. The solution was expanded to improve the in situ estimation of effective porosity. The truncated equation assumed displacement during the injection phase was negligible, which may theoretically lead to an underestimation of the true value of effective porosity. To experimentally compare the expanded and truncated equations, single-well push-pull tests were conducted across six test wells located in a shallow, unconfined aquifer comprised of unconsolidated and heterogeneous silty and clayey fill materials. The push-pull tests were conducted by injection of bromide tracer, followed by a non-pumping period, and subsequent extraction of groundwater. The values of effective porosity from the expanded equation (0.6-5.0%) were substantially greater than from the truncated equation (0.1-1.3%). The expanded and truncated equations were compared to data from previous push-pull studies in the literature and demonstrated that displacement during the injection phase may or may not be negligible, depending on the aquifer properties and the push-pull test parameters. The results presented here also demonstrated the spatial variability of effective porosity within a relatively small study site can be substantial, and the error-propagated uncertainty of effective porosity can be mitigated to a reasonable level (< ± 0.5%). The tests presented here are also the first that the authors are aware of that estimate, in situ, the effective porosity of fine-grained fill material.

Release mechanism of doxazosin from carrageenan matrix tablets: Effect of ionic strength and addition of sodium dodecyl sulphate.

PubMed

Kos, Petra; Pavli, Matej; Baumgartner, Saša; Kogej, Ksenija

2017-08-30

The polyelectrolyte matrix tablets loaded with an oppositely charged drug exhibit complex drug-release mechanisms. In this study, the release mechanism of a cationic drug doxazosin mesylate (DM) from matrix tablets based on an anionic polyelectrolyte λ-carrageenan (λ-CARR) is investigated. The drug release rates from λ-CARR matrices are correlated with binding results based on potentiometric measurements using the DM ion-sensitive membrane electrode and with molecular characteristics of the DM-λ-CARR-complex particles through hydrodynamic size measurements. Experiments are performed in solutions with different ionic strength and with the addition of an anionic surfactant sodium dodecyl sulphate (SDS). It is demonstrated that in addition to swelling and erosion of tablets, the release rates depend strongly on cooperative interactions between DM and λ-CARR. Addition of SDS at concentrations below its critical micelle concentration (CMC) slows down the DM release through hydrophobic binding of SDS to the DM-λ-CARR complex. On the contrary, at concentrations above the CMC SDS pulls DM from the complex by forming mixed micelles with it and thus accelerates the release. Results involving SDS show that the concentration of surfactants that are naturally present in gastrointestinal environment may have a great impact on the drug release process. Copyright © 2017 Elsevier B.V. All rights reserved.

Activation of orphan nuclear constitutive androstane receptor requires subnuclear targeting by peroxisome proliferator-activated receptor gamma coactivator-1 alpha. A possible link between xenobiotic response and nutritional state.

PubMed

Shiraki, Takuma; Sakai, Noriko; Kanaya, Eiko; Jingami, Hisato

2003-03-28

In contrast to the classical nuclear receptors, the constitutive androstane receptor (CAR) is transcriptionally active in the absence of ligand. In the course of searching for the mediator of CAR activation, we found that ligand-independent activation of CAR was achieved in cooperation with the peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1 alpha). PGC-1 beta, a PGC-1 alpha homologue, also activated CAR to less of an extent than PGC-1 alpha. Coexpression of the ligand-binding domain of a heterodimerization partner, retinoid X receptor alpha, enhanced the PGC-1 alpha-mediated activation of CAR, although it had a weak effect on the basal activity of CAR in the absence of PGC-1 alpha. Both the N-terminal region, with the LXXLL motif, and the C-terminal region, with a serine/arginine-rich domain (RS domain), in PGC-1 alpha were required for full activation of CAR. Pull-down experiments using recombinant proteins revealed that CAR directly interacted with both the LXXLL motif and the RS domain. Furthermore, we demonstrated that the RS domain of PGC-1 alpha was required for CAR localization at nuclear speckles. These results indicate that PGC-1 alpha mediates the ligand-independent activation of CAR by means of subnuclear targeting through the RS domain of PGC-1 alpha.

Evaluation of selected binding domains for the analysis of ubiquitinated proteomes

PubMed Central

Nakayasu, Ernesto S.; Ansong, Charles; Brown, Joseph N.; Yang, Feng; Lopez-Ferrer, Daniel; Qian, Wei-Jun; Smith, Richard D.; Adkins, Joshua N.

2013-01-01

Ubiquitination is an abundant post-translational modification that consists of covalent attachment of ubiquitin to lysine residues or the N-terminus of proteins. Mono and polyubiquitination have been shown to be involved in many critical eukaryotic cellular functions and are often disrupted by intracellular bacterial pathogens. Affinity enrichment of ubiquitinated proteins enables global analysis of this key modification. In this context, the use of ubiquitin-binding domains is a promising, but relatively unexplored alternative to more broadly used immunoaffinity or tagged affinity enrichment methods. In this study, we evaluated the application of eight ubiquitin-binding domains that have differing affinities for ubiquitination states. Small-scale proteomics analysis identified ∼200 ubiquitinated protein candidates per ubiquitin-binding domain pull-down experiment. Results from subsequent Western blot analyses that employed anti-ubiquitin or monoclonal antibodies against polyubiquitination at lysine 48 and 63 suggest that ubiquitin-binding domains from Dsk2 and ubiquilin-1 have the broadest specificity in that they captured most types of ubiquitination, whereas the binding domain from NBR1 was more selective to polyubiquitination. These data demonstrate that with optimized purification conditions, ubiquitin-binding domains can be an alternative tool for proteomic applications. This approach is especially promising for the analysis of tissues or cells resistant to transfection, of which the overexpression of tagged ubiquitin is a major hurdle. PMID:23649778

Differential lead component pulling as a possible mechanism of inside-out abrasion and conductor cable externalization.

PubMed

Lau, Ernest W

2013-09-01

Conductor cable externalization with protrusion (CCE*) is highly prevalent among the Riata 8F and ST 7F defibrillation (DF) leads and infrequently present in the QuickSite and the QuickFlex coronary sinus (CS) leads (St. Jude Medical, Sylmar, CA, USA). A model for CCE* based on differential lead component pulling and conjugate extension with reciprocal compression-bending was developed. Extension of a proximal lead body segment by pectoral or cardiac movements causes reciprocal compression-bending of a distal lead body segment mediated by inextensible conductor cables running down a lead body fixed at various points by fibrous adhesions. The "sawing" action of these cables under tension causes inside-out abrasion of insulation leading to CCE*. DF leads from different manufacturers and the QuickFlex and QuickFlex μ CS leads were subjected to simulated differential pulling. Restitution from differential pulling followed three patterns: complete, partial without escalation, and incomplete with escalation. Only the last pattern (only shown by the Riata 8F and ST 7F leads) was associated with an increased risk to CCE*. For CS leads, deformation concentrated on the more flexible segment when the lead body did not have a uniform construction. The Durata, Riata ST Optim, QuickFlex μ, and Quartet leads should be relatively immune to CCE*. The Durata leads are extremely resistant to longitudinal deformation and probably cause mediastinal displacement rather than differential pulling in response to pectoral movements in vivo. Implantation techniques and lead designs can be used to minimize the risk of CCE*. A bench test for CCE* can be constructed. ©2013, The Author. Journal compilation ©2013 Wiley Periodicals, Inc.

‘Natural experiment’ Demonstrates Top-Down Control of Spiders by Birds on a Landscape Level

PubMed Central

Rogers, Haldre; Hille Ris Lambers, Janneke; Miller, Ross; Tewksbury, Joshua J.

2012-01-01

The combination of small-scale manipulative experiments and large-scale natural experiments provides a powerful approach for demonstrating the importance of top-down trophic control on the ecosystem scale. The most compelling natural experiments have come from studies examining the landscape-scale loss of apex predators like sea otters, wolves, fish and land crabs. Birds are dominant apex predators in terrestrial systems around the world, yet all studies on their role as predators have come from small-scale experiments; the top-down impact of bird loss on their arthropod prey has yet to be examined at a landscape scale. Here, we use a unique natural experiment, the extirpation of insectivorous birds from nearly all forests on the island of Guam by the invasive brown tree snake, to produce the first assessment of the impacts of bird loss on their prey. We focused on spiders because experimental studies showed a consistent top-down effect of birds on spiders. We conducted spider web surveys in native forest on Guam and three nearby islands with healthy bird populations. Spider web densities on the island of Guam were 40 times greater than densities on islands with birds during the wet season, and 2.3 times greater during the dry season. These results confirm the general trend from manipulative experiments conducted in other systems however, the effect size was much greater in this natural experiment than in most manipulative experiments. In addition, bird loss appears to have removed the seasonality of spider webs and led to larger webs in at least one spider species in the forests of Guam than on nearby islands with birds. We discuss several possible mechanisms for the observed changes. Overall, our results suggest that effect sizes from smaller-scale experimental studies may significantly underestimate the impact of bird loss on spider density as demonstrated by this large-scale natural experiment. PMID:22970126

Scapula kinematics of pull-up techniques: Avoiding impingement risk with training changes.

PubMed

Prinold, Joe A I; Bull, Anthony M J

2016-08-01

Overhead athletic activities and scapula dyskinesia are linked with shoulder pathology; pull-ups are a common training method for some overhead sports. Different pull-up techniques exist: anecdotally some are easier to perform, and others linked to greater incidences of pathology. This study aims to quantify scapular kinematics and external forces for three pull-up techniques, thus discussing potential injury implications. An observational study was performed with eleven participants (age=26.8±2.4 years) who regularly perform pull-ups. The upward motions of three pull-up techniques were analysed: palms facing anterior, palms facing posterior and wide-grip. A skin-fixed scapula tracking technique with attached retro-reflective markers was used. High intra-participant repeatability was observed: mean coefficients of multiple correlations of 0.87-1.00 in humerothoracic rotations and 0.77-0.90 for scapulothoracic rotations. Standard deviations of hand force was low: <5% body weight. Significantly different patterns of humerothoracic, scapulothoracic and glenohumeral kinematics were observed between the pull-up techniques. The reverse technique has extreme glenohumeral internal-external rotation and large deviation from the scapula plane. The wide technique has a reduced range of pro/retraction in the same HT plane of elevation and 90° of arm abduction with 45° external rotation was observed. All these factors suggest increased sub-acromial impingement risk. The scapula tracking technique showed high repeatability. High arm elevation during pull-ups reduces sub-acromial space and increases pressure, increasing the risk of impingement injury. Wide and reverse pull-ups demonstrate kinematics patterns linked with increased impingement risk. Weight-assisted front pull-ups require further investigation and could be recommended for weaker participants. Copyright © 2015 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

Quantitative in-situ scanning electron microscope pull-out experiments and molecular dynamics simulations of carbon nanotubes embedded in palladium

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

Hartmann, S., E-mail: steffen.hartmann@etit.tu-chemnitz.de; Blaudeck, T.; Hermann, S.

2014-04-14

In this paper, we present our results of experimental and numerical pull-out tests on carbon nanotubes (CNTs) embedded in palladium. We prepared simple specimens by employing standard silicon wafers, physical vapor deposition of palladium and deposition of CNTs with a simple drop coating technique. An AFM cantilever with known stiffness connected to a nanomanipulation system was utilized inside a scanning electron microscope (SEM) as a force sensor to determine forces acting on a CNT during the pull-out process. SEM-images of the cantilever attached to a CNT have been evaluated for subsequent displacement steps with greyscale correlation to determine the cantilevermore » deflection. We compare the experimentally obtained pull-out forces with values of numerical investigations by means of molecular dynamics and give interpretations for deviations according to material impurities or defects and their influence on the pull-out data. We find a very good agreement of force data from simulation and experiment, which is 17 nN and in the range of 10–61 nN, respectively. Our findings contribute to the ongoing research of the mechanical characterization of CNT-metal interfaces. This is of significant interest for the design of future mechanical sensors utilizing the intrinsic piezoresistive effect of CNTs or other future devices incorporating CNT-metal interfaces.« less

Pressure-Distribution Measurements on O-2H Airplane in Flight

NASA Technical Reports Server (NTRS)

Pearson, H A

1937-01-01

Results are given of pressure-distribution measurements made over two different horizontal tail surfaces and the right wing cellule, including the slipstream area, of an observation-type biplane. Measurements were also taken of air speed, control-surface positions, control-stick forces, angular velocities, and accelerations during various abrupt maneuvers. These maneuvers consisted of push-downs and pull-ups from level flight, dive pull-outs, and aileron rolls with various thrust conditions. The results from the pressure-distribution measurements over the wing cellule are given on charts showing the variation of individual rib coefficients with wing coefficients; the data from the tail-surface pressure-distribution measurements are given mainly as total loads and moments. These data are supplemented by time histories of the measured quantities and isometric views of the rib pressure distributions occurring in abrupt maneuvers.

The Tablecloth Pull Revisited

ERIC Educational Resources Information Center

Vollmer, Michael; Mollmann, Klaus-Peter

2015-01-01

A very old and well-known magical trick is the so-called tablecloth pull. A table is covered with a tablecloth, on top of which are certain objects. The task is to remove the tablecloth while the objects--which must not be touched--stay on top of the table. This article describes the physics behind the experiment, and presents examples recorded…

Integrated IoT technology in industrial lasers for the improved user experience

NASA Astrophysics Data System (ADS)

Ding, Jianwu; Liu, Jinhui

2018-02-01

The end users' biggest concern for any industrial equipment is the reliability and the service down-time. This is especially true for industrial lasers as they are typically used in fully or semi- automated processes. Here we demonstrate how to use the integrated Internet of Things (IoT) technology in industrial lasers to address the reliability and the service down-time so to improve end users' experience.

Remote Control of Tissue Interactions via Engineered Photo-switchable Cell Surfaces

NASA Astrophysics Data System (ADS)

Luo, Wei; Pulsipher, Abigail; Dutta, Debjit; Lamb, Brian M.; Yousaf, Muhammad N.

2014-09-01

We report a general cell surface molecular engineering strategy via liposome fusion delivery to create a dual photo-active and bio-orthogonal cell surface for remote controlled spatial and temporal manipulation of microtissue assembly and disassembly. Cell surface tailoring of chemoselective functional groups was achieved by a liposome fusion delivery method and quantified by flow cytometry and characterized by a new cell surface lipid pull down mass spectrometry strategy. Dynamic co-culture spheroid tissue assembly in solution and co-culture tissue multilayer assembly on materials was demonstrated by an intercellular photo-oxime ligation that could be remotely cleaved and disassembled on demand. Spatial and temporal control of microtissue structures containing multiple cell types was demonstrated by the generation of patterned multilayers for controlling stem cell differentiation. Remote control of cell interactions via cell surface engineering that allows for real-time manipulation of tissue dynamics may provide tools with the scope to answer fundamental questions of cell communication and initiate new biotechnologies ranging from imaging probes to drug delivery vehicles to regenerative medicine, inexpensive bioreactor technology and tissue engineering therapies.

Localization of one-photon state in space and Einstein-Podolsky-Rosen paradox in spontaneous parametric down conversion

NASA Technical Reports Server (NTRS)

Penin, A. N.; Reutova, T. A.; Sergienko, A. V.

1992-01-01

An experiment on one-photon state localization in space using a correlation technique in Spontaneous Parametric Down Conversion (SPDC) process is discussed. Results of measurements demonstrate an idea of the Einstein-Podolsky-Rosen (EPR) paradox for coordinate and momentum variables of photon states. Results of the experiment can be explained with the help of an advanced wave technique. The experiment is based on the idea that two-photon states of optical electromagnetic fields arising in the nonlinear process of the spontaneous parametric down conversion (spontaneous parametric light scattering) can be explained by quantum mechanical theory with the help of a single wave function.

Localization of one-photon state in space and Einstein-Podolsky-Rosen paradox in spontaneous parametric down conversion

NASA Astrophysics Data System (ADS)

Penin, A. N.; Reutova, T. A.; Sergienko, A. V.

1992-02-01

An experiment on one-photon state localization in space using a correlation technique in Spontaneous Parametric Down Conversion (SPDC) process is discussed. Results of measurements demonstrate an idea of the Einstein-Podolsky-Rosen (EPR) paradox for coordinate and momentum variables of photon states. Results of the experiment can be explained with the help of an advanced wave technique. The experiment is based on the idea that two-photon states of optical electromagnetic fields arising in the nonlinear process of the spontaneous parametric down conversion (spontaneous parametric light scattering) can be explained by quantum mechanical theory with the help of a single wave function.

MoS2 Negative-Capacitance Field-Effect Transistors with Subthreshold Swing below the Physics Limit.

PubMed

Liu, Xingqiang; Liang, Renrong; Gao, Guoyun; Pan, Caofeng; Jiang, Chunsheng; Xu, Qian; Luo, Jun; Zou, Xuming; Yang, Zhenyu; Liao, Lei; Wang, Zhong Lin

2018-05-21

The Boltzmann distribution of electrons induced fundamental barrier prevents subthreshold swing (SS) from less than 60 mV dec -1 at room temperature, leading to high energy consumption of MOSFETs. Herein, it is demonstrated that an aggressive introduction of the negative capacitance (NC) effect of ferroelectrics can decisively break the fundamental limit governed by the "Boltzmann tyranny". Such MoS 2 negative-capacitance field-effect transistors (NC-FETs) with self-aligned top-gated geometry demonstrated here pull down the SS value to 42.5 mV dec -1 , and simultaneously achieve superior performance of a transconductance of 45.5 μS μm and an on/off ratio of 4 × 10 6 with channel length less than 100 nm. Furthermore, the inserted HfO 2 layer not only realizes a stable NC gate stack structure, but also prevents the ferroelectric P(VDF-TrFE) from fatigue with robust stability. Notably, the fabricated MoS 2 NC-FETs are distinctly different from traditional MOSFETs. The on-state current increases as the temperature decreases even down to 20 K, and the SS values exhibit nonlinear dependence with temperature due to the implementation of the ferroelectric gate stack. The NC-FETs enable fundamental applications through overcoming the Boltzmann limit in nanoelectronics and open up an avenue to low-power transistors needed for many exciting long-endurance portable consumer products. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Unfolding of Vortices into Topological Stripes in a Multiferroic Material

NASA Astrophysics Data System (ADS)

Wang, X.; Mostovoy, M.; Han, M. G.; Horibe, Y.; Aoki, T.; Zhu, Y.; Cheong, S.-W.

2014-06-01

Multiferroic hexagonal RMnO3 (R =rare earths) crystals exhibit dense networks of vortex lines at which six domain walls merge. While the domain walls can be readily moved with an applied electric field, the vortex cores so far have been impossible to control. Our experiments demonstrate that shear strain induces a Magnus-type force pulling vortices and antivortices in opposite directions and unfolding them into a topological stripe domain state. We discuss the analogy between this effect and the current-driven dynamics of vortices in superconductors and superfluids.

An "on-matrix" digestion procedure for AP-MS experiments dissects the interplay between complex-conserved and serotype-specific reactivities in Dengue virus-human plasma interactome.

PubMed

Ramos, Yassel; Huerta, Vivian; Martín, Dayron; Palomares, Sucel; Yero, Alexis; Pupo, Dianne; Gallien, Sebastien; Martín, Alejandro M; Pérez-Riverol, Yasset; Sarría, Mónica; Guirola, Osmany; Chinea, Glay; Domon, Bruno; González, Luis Javier

2017-07-13

The interactions between the four Dengue virus (DENV) serotypes and plasma proteins are crucial in the initial steps of viral infection to humans. Affinity purification combined with quantitative mass spectrometry analysis, has become one of the most powerful tools for the investigation on novel protein-protein interactions. Using this approach, we report here that a significant number of bait-interacting proteins do not dissociate under standard elution conditions, i.e. acid pH and chaotropic agents, and that this problem can be circumvented by using the "on-matrix" digestion procedure described here. This procedure enabled the identification of 16 human plasma proteins interacting with domain III from the envelope protein of DENV serotypes 1, 3 and 4 that would have not been detected otherwise and increased the known DIIIE interactors in human plasma to 59 proteins. Selected Reaction Monitoring analysis evidenced DENV interactome in human plasma is rather conserved although significant differences on the reactivity of viral serotypes with specific proteins do exist. A comparison between the serotype-dependent profile of reactivity and the conservation pattern of amino acid residues suggests an evolutionary selection of highly conserved interactions with the host and other interactions mediated for surface regions of higher variability. False negative results on the identification of interacting proteins in pull-down experiments compromise the subsequent interpretation of results and the formulation of a working hypothesis for the derived future work. In this study we demonstrate the presence of bait-interacting proteins reluctant to dissociate under elution conditions of acid pH and presence of chaotropics. We propose the direct proteolytic digestion of proteins while still bound to the affinity matrix ("on-matrix" digestion) and evaluate the impact of this methodology in the comparative study of the interactome of the four serotypes of Dengue virus mediated by the domain III of the viral envelope glycoprotein. Fifty nine proteins were identified as putative interaction partners of Dengue virus (IPs) either due to direct binding or by co-isolation with interacting proteins. Collectively the IPs identified from the pull-down with the recombinant domain III proteins representing the four viral serotypes, 29% were identified only after "on-matrix" digestion which demonstrate the usefulness of this method of recovering bait-bound proteins. Results highlight a particular importance of "on-matrix" digestion procedure for comparative studies where a stronger interaction with one of the interest baits could prevent a bound protein to elute under standard conditions thus leading to misinterpretation as absent in the interactome of this particular bait. The analysis of the Interaction Network indicates that Dengue virus interactome mediated by the domain III of the envelope protein is rather conserved in the viral complex suggesting a key role of these interactions for viral infection thus making candidates to explore for potential biomarkers of clinical outcome in DENV-caused disease. Interestingly, some particular IPs exhibit significant differences in the strength of the interaction with the viral serotypes representing interactions that involve more variable regions in the surface of the domain III. Since such variable regions are the consequence of the interaction with antibodies generated by human immune response; this result relates the interaction with proteins from human plasma with the interplay of the virus and the human immune system. Copyright © 2017 Elsevier B.V. All rights reserved.

8. VIEW SHOWING THE DEMOSSING OF GRAND CANAL LOCATION UNKNOWN. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. VIEW SHOWING THE DEMOSSING OF GRAND CANAL LOCATION UNKNOWN. AT TEAM OF HORSES ON OPPOSITE BANKS OF THE CANAL DRAG A CHAIN BETWEEN THEM ALONG THE BOTTOM OF THE CANAL, WHICH PULLS THE MOSS AND WEEDS LOOSE. THE PLANS THEN FLOAT DOWN THE CANAL AND ARE CAUGHT IN A SCREEN AND REMOVED. Photographer unknown, 1923 - Grand Canal, North side of Salt River, Tempe, Maricopa County, AZ

Breast Cancer Translational Research Center of Excellence

DTIC Science & Technology

2015-09-01

we will identify partner proteins by either Y2H or pull-down studies using Mass Spectrometry Analysis. Aim 2. Development of DNA aptamers against...CD44 that inhibit breast cancer invasion and metastasis. During the previous period, we developed novel and unique DNA aptamers that specifically...bound to exon v10 of CD44 using Systematic Evolution of Ligands by Exponential Enrichment (SELEX) systems. These DNA aptamers inhibited TN breast

Studying Protein-Protein Interactions by Biotin AP-Tagged Pulldown and LTQ-Orbitrap Mass Spectrometry.

PubMed

Xie, Zhongqiu; Jia, Yuemeng; Li, Hui

2017-01-01

The study of protein-protein interactions represents a key aspect of biological research. Identifying unknown protein binding partners using mass spectrometry (MS)-based proteomics has evolved into an indispensable strategy in drug discovery. The classic approach of immunoprecipitation with specific antibodies against the proteins of interest has limitations, such as the need for immunoprecipitation-qualified antibody. The biotin AP-tag pull-down system has the advantage of high specificity, ease of use, and no requirement for antibody. It is based on the high specificity, high affinity interaction between biotin and streptavidin. After pulldown, in-gel tryptic digestion and tandem mass spectrometry (MS/MS) analysis of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) protein bands can be performed. In this work, we provide protocols that can be used for the identification of proteins that interact with FOXM1, a protein that has recently emerged as a potential biomarker and drug target in oncotherapy, as an example. We focus on the pull-down procedure and assess the efficacy of the pulldown with known FOXM1 interactors such as β-catenin. We use a high performance LTQ Orbitrap MSn system that combines rapid LTQ ion trap data acquisition with high mass accuracy Orbitrap analysis to identify the interacting proteins.

Gluten-specific antibodies of celiac disease gut plasma cells recognize long proteolytic fragments that typically harbor T-cell epitopes.

PubMed

Dørum, Siri; Steinsbø, Øyvind; Bergseng, Elin; Arntzen, Magnus Ø; de Souza, Gustavo A; Sollid, Ludvig M

2016-05-05

This study aimed to identify proteolytic fragments of gluten proteins recognized by recombinant IgG1 monoclonal antibodies generated from single IgA plasma cells of celiac disease lesions. Peptides bound by monoclonal antibodies in complex gut-enzyme digests of gluten treated with the deamidating enzyme transglutaminase 2, were identified by mass spectrometry after antibody pull-down with protein G beads. The antibody bound peptides were long deamidated peptide fragments that contained the substrate recognition sequence of transglutaminase 2. Characteristically, the fragments contained epitopes with the sequence QPEQPFP and variants thereof in multiple copies, and they typically also harbored many different gluten T-cell epitopes. In the pull-down setting where antibodies were immobilized on a solid phase, peptide fragments with multivalent display of epitopes were targeted. This scenario resembles the situation of the B-cell receptor on the surface of B cells. Conceivably, B cells of celiac disease patients select gluten epitopes that are repeated multiple times in long peptide fragments generated by gut digestive enzymes. As the fragments also contain many different T-cell epitopes, this will lead to generation of strong antibody responses by effective presentation of several distinct T-cell epitopes and establishment of T-cell help to B cells.

Gluten-specific antibodies of celiac disease gut plasma cells recognize long proteolytic fragments that typically harbor T-cell epitopes

PubMed Central

Dørum, Siri; Steinsbø, Øyvind; Bergseng, Elin; Arntzen, Magnus Ø.; de Souza, Gustavo A.; Sollid, Ludvig M.

2016-01-01

This study aimed to identify proteolytic fragments of gluten proteins recognized by recombinant IgG1 monoclonal antibodies generated from single IgA plasma cells of celiac disease lesions. Peptides bound by monoclonal antibodies in complex gut-enzyme digests of gluten treated with the deamidating enzyme transglutaminase 2, were identified by mass spectrometry after antibody pull-down with protein G beads. The antibody bound peptides were long deamidated peptide fragments that contained the substrate recognition sequence of transglutaminase 2. Characteristically, the fragments contained epitopes with the sequence QPEQPFP and variants thereof in multiple copies, and they typically also harbored many different gluten T-cell epitopes. In the pull-down setting where antibodies were immobilized on a solid phase, peptide fragments with multivalent display of epitopes were targeted. This scenario resembles the situation of the B-cell receptor on the surface of B cells. Conceivably, B cells of celiac disease patients select gluten epitopes that are repeated multiple times in long peptide fragments generated by gut digestive enzymes. As the fragments also contain many different T-cell epitopes, this will lead to generation of strong antibody responses by effective presentation of several distinct T-cell epitopes and establishment of T-cell help to B cells. PMID:27146306

Directionally solidified Al2O3/GAP eutectic ceramics by micro-pulling-down method

NASA Astrophysics Data System (ADS)

Cao, Xue; Su, Haijun; Guo, Fengwei; Tan, Xi; Cao, Lamei

2016-11-01

We reported a novel route to prepare directionally solidified (DS) Al2O3/GAP eutectic ceramics by micro-pulling-down (μ-PD) method. The eutectic crystallizations, microstructure characters and evolutions, and their mechanical properties were investigated in detail. The results showed that the Al2O3/GAP eutectic composites can be successfully fabricated through μ-PD method, possessed smooth surface, full density and large crystal size (the maximal size: φ90 mm × 20 mm). At the process of Diameter, the as-solidified Al2O3/GAP eutectic presented a combination of "Chinese script" and elongated colony microstructure with complex regular structure. Inside the colonies, the rod-type or lamellar-type eutectic microstructures with ultra-fine GAP surrounded by the Al2O3 matrix were observed. At an appropriate solidificational rate, the binary eutectic exhibited a typical DS irregular eutectic structure of "chinese script" consisting of interpenetrating network of α-Al2O3 and GAP phases without any other phases. Therefore, the interphase spacing was refined to 1-2 µm and the irregular microstructure led to an outstanding vickers hardness of 17.04 GPa and fracture toughness of 6.3 MPa × m1/2 at room temperature.

Identification of DNA-binding proteins that interact with the 5'-flanking region of the human D-amino acid oxidase gene by pull-down assay coupled with two-dimensional gel electrophoresis and mass spectrometry.

PubMed

Tran, Diem Hong; Shishido, Yuji; Chung, Seong Pil; Trinh, Huong Thi Thanh; Yorita, Kazuko; Sakai, Takashi; Fukui, Kiyoshi

2015-12-10

D-Amino acid oxidase (DAO) is a flavoenzyme that metabolizes D-amino acids and is expected to be a promising therapeutic target of schizophrenia and glioblastoma. The study of DNA-binding proteins has yielded much information in the regulation of transcription and other biological processes. However, proteins interacting with DAO gene have not been elucidated. Our assessment of human DAO promoter activity using luciferase reporter system indicated the 5'-flanking region of this gene (-4289 bp from transcription initiation site) has a regulatory sequence for gene expression, which is regulated by multi-protein complexes interacting with this region. By using pull-down assay coupled with two-dimensional gel electrophoresis and mass spectrometry, we identified six proteins binding to the 5'-flanking region of the human DAO gene (zinc finger C2HC domain-containing protein 1A; histidine-tRNA ligase, cytoplasmic; molybdenum cofactor biosynthesis protein; 60S ribosomal protein L37; calponin-1; calmodulin binding protein and heterogeneous nuclear ribonucleoprotein A2/B1). These preliminary results will contribute to the advance in the understanding of the potential factors associated with the regulatory mechanism of DAO expression. Copyright © 2015 Elsevier B.V. All rights reserved.

Casingless down-hole for sealing an ablation volume and obtaining a sample for analysis

DOEpatents

Noble, D.T.; Braymen, S.D.; Anderson, M.S.

1996-10-01

A casing-less down hole sampling system for acquiring a subsurface sample for analysis using an inductively coupled plasma system is disclosed. The system includes a probe which is pushed into the formation to be analyzed using a hydraulic ram system. The probe includes a detachable tip member which has a soil point and a barb, with the soil point aiding the penetration of the earth, and the barb causing the tip member to disengage from the probe and remain in the formation when the probe is pulled up. The probe is forced into the formation to be tested, and then pulled up slightly, to disengage the tip member and expose a column of the subsurface formation to be tested. An instrumentation tube mounted in the probe is then extended outward from the probe to longitudinally extend into the exposed column. A balloon seal mounted on the end of the instrumentation tube allows the bottom of the column to be sealed. A source of laser radiation is emitted from the instrumentation tube to ablate a sample from the exposed column. The instrumentation tube can be rotated in the probe to sweep the laser source across the surface of the exposed column. An aerosol transport system carries the ablated sample from the probe to the surface for testing in an inductively coupled plasma system. By testing at various levels in the down-hole as the probe is extracted from the soil, a profile of the subsurface formation may be obtained. 9 figs.

Optical trapping, pulling, and Raman spectroscopy of airborne absorbing particles based on negative photophoretic force

NASA Astrophysics Data System (ADS)

Chen, Gui-hua; He, Lin; Wu, Mu-ying; Yang, Guang; Li, Y. Q.

2017-08-01

Optical pulling is the attraction of objects back to the light source by the use of optically induced "negative forces". The light-induced photophoretic force is generated by the momentum transfer between the heating particles and surrounding gas molecules and can be several orders of magnitude larger than the radiation force and gravitation force. Here, we demonstrate that micron-sized absorbing particles can be optically pulled and manipulated towards the light source over a long distance in air with a collimated Gaussian laser beam based on a negative photophoretic force. A variety of airborne absorbing particles can be pulled by this optical pipeline to the region where they are optically trapped with another focused laser beam and their chemical compositions are characterized with Raman spectroscopy. We found that micron-sized particles are pulled over a meter-scale distance in air with a pulling speed of 1-10 cm/s in the optical pulling pipeline and its speed can be controlled by changing the laser intensity. When an aerosol particle is optically trapped with a focused Gaussian beam, we measured its rotation motion around the laser propagation direction and measured its Raman spectroscopy for chemical identification by molecular fingerprints. The centripetal acceleration of the trapped particle as high as 20 times the gravitational acceleration was observed. Optical pulling over large distances with lasers in combination with Raman spectroscopy opens up potential applications for the collection and identification of atmospheric particles.

Push-pull tracer tests: Their information content and use for characterizing non-Fickian, mobile-immobile behavior: INFORMATION CONTENT OF PUSH-PULL TESTS

DOE PAGES

Hansen, Scott K.; Berkowitz, Brian; Vesselinov, Velimir V.; ...

2016-12-01

Path reversibility and radial symmetry are often assumed in push-pull tracer test analysis. In reality, heterogeneous flow fields mean that both assumptions are idealizations. In this paper, to understand their impact, we perform a parametric study which quantifies the scattering effects of ambient flow, local-scale dispersion, and velocity field heterogeneity on push-pull breakthrough curves and compares them to the effects of mobile-immobile mass transfer (MIMT) processes including sorption and diffusion into secondary porosity. We identify specific circumstances in which MIMT overwhelmingly determines the breakthrough curve, which may then be considered uninformative about drift and local-scale dispersion. Assuming path reversibility, wemore » develop a continuous-time-random-walk-based interpretation framework which is flow-field-agnostic and well suited to quantifying MIMT. Adopting this perspective, we show that the radial flow assumption is often harmless: to the extent that solute paths are reversible, the breakthrough curve is uninformative about velocity field heterogeneity. Our interpretation method determines a mapping function (i.e., subordinator) from travel time in the absence of MIMT to travel time in its presence. A mathematical theory allowing this function to be directly “plugged into” an existing Laplace-domain transport model to incorporate MIMT is presented and demonstrated. Algorithms implementing the calibration are presented and applied to interpretation of data from a push-pull test performed in a heterogeneous environment. A successful four-parameter fit is obtained, of comparable fidelity to one obtained using a million-node 3-D numerical model. In conclusion, we demonstrate analytically and numerically how push-pull tests quantifying MIMT are sensitive to remobilization, but not immobilization, kinetics.« less

Push-pull tracer tests: Their information content and use for characterizing non-Fickian, mobile-immobile behavior: INFORMATION CONTENT OF PUSH-PULL TESTS

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

Hansen, Scott K.; Berkowitz, Brian; Vesselinov, Velimir V.

Path reversibility and radial symmetry are often assumed in push-pull tracer test analysis. In reality, heterogeneous flow fields mean that both assumptions are idealizations. In this paper, to understand their impact, we perform a parametric study which quantifies the scattering effects of ambient flow, local-scale dispersion, and velocity field heterogeneity on push-pull breakthrough curves and compares them to the effects of mobile-immobile mass transfer (MIMT) processes including sorption and diffusion into secondary porosity. We identify specific circumstances in which MIMT overwhelmingly determines the breakthrough curve, which may then be considered uninformative about drift and local-scale dispersion. Assuming path reversibility, wemore » develop a continuous-time-random-walk-based interpretation framework which is flow-field-agnostic and well suited to quantifying MIMT. Adopting this perspective, we show that the radial flow assumption is often harmless: to the extent that solute paths are reversible, the breakthrough curve is uninformative about velocity field heterogeneity. Our interpretation method determines a mapping function (i.e., subordinator) from travel time in the absence of MIMT to travel time in its presence. A mathematical theory allowing this function to be directly “plugged into” an existing Laplace-domain transport model to incorporate MIMT is presented and demonstrated. Algorithms implementing the calibration are presented and applied to interpretation of data from a push-pull test performed in a heterogeneous environment. A successful four-parameter fit is obtained, of comparable fidelity to one obtained using a million-node 3-D numerical model. In conclusion, we demonstrate analytically and numerically how push-pull tests quantifying MIMT are sensitive to remobilization, but not immobilization, kinetics.« less

Fast Pulling of n-Type Si Ingots for Enhanced Si Solar Cell Production

NASA Astrophysics Data System (ADS)

Kim, Kwanghun; Park, Sanghyun; Park, Jaechang; Pang, Ilsun; Ryu, Sangwoo; Oh, Jihun

2018-07-01

Reducing the manufacturing costs of silicon substrates is an important issue in the silicon-based solar cell industry. In this study, we developed a high-throughput ingot growth method by accelerating the pulling speed in the Czochralski process. By controlling the heat flow of the ingot growth chamber and at the solid-liquid interfaces, the pulling speed of an ingot could be increased by 15% compared to the conventional method, while retaining high quality. The wafer obtained at a high pulling speed showed an enhanced minority carrier lifetime compared with conventional wafers, due to the vacancy passivation effect, and also demonstrated comparable bulk resistivity and impurities. The results in this work are expected to open a new way to enhance the productivity of Si wafers used for Si solar cells, and therefore, to reduce the overall manufacturing cost.

Fast Pulling of n-Type Si Ingots for Enhanced Si Solar Cell Production

NASA Astrophysics Data System (ADS)

Kim, Kwanghun; Park, Sanghyun; Park, Jaechang; Pang, Ilsun; Ryu, Sangwoo; Oh, Jihun

2018-03-01

Reducing the manufacturing costs of silicon substrates is an important issue in the silicon-based solar cell industry. In this study, we developed a high-throughput ingot growth method by accelerating the pulling speed in the Czochralski process. By controlling the heat flow of the ingot growth chamber and at the solid-liquid interfaces, the pulling speed of an ingot could be increased by 15% compared to the conventional method, while retaining high quality. The wafer obtained at a high pulling speed showed an enhanced minority carrier lifetime compared with conventional wafers, due to the vacancy passivation effect, and also demonstrated comparable bulk resistivity and impurities. The results in this work are expected to open a new way to enhance the productivity of Si wafers used for Si solar cells, and therefore, to reduce the overall manufacturing cost.

Free energy calculation of single molecular interaction using Jarzynski's identity method: the case of HIV-1 protease inhibitor system

NASA Astrophysics Data System (ADS)

Li, De-Chang; Ji, Bao-Hua

2012-06-01

Jarzynski' identity (JI) method was suggested a promising tool for reconstructing free energy landscape of biomolecular interactions in numerical simulations and experiments. However, JI method has not yet been well tested in complex systems such as ligand-receptor molecular pairs. In this paper, we applied a huge number of steered molecular dynamics (SMD) simulations to dissociate the protease of human immunodeficiency type I virus (HIV-1 protease) and its inhibitors. We showed that because of intrinsic complexity of the ligand-receptor system, the energy barrier predicted by JI method at high pulling rates is much higher than experimental results. However, with a slower pulling rate and fewer switch times of simulations, the predictions of JI method can approach to the experiments. These results suggested that the JI method is more appropriate for reconstructing free energy landscape using the data taken from experiments, since the pulling rates used in experiments are often much slower than those in SMD simulations. Furthermore, we showed that a higher loading stiffness can produce higher precision of calculation of energy landscape because it yields a lower mean value and narrower bandwidth of work distribution in SMD simulations.

Trunk response and stability in standing under sagittal-symmetric pull-push forces at different orientations, elevations and magnitudes.

PubMed

El Ouaaid, Z; Shirazi-Adl, A; Plamondon, A

2018-03-21

To reduce lifting and associated low back injuries, manual material handling operations often involve pulling-pushing of carts at different weights, orientations, and heights. The loads on spine and risk of injury however need to be investigated. The aim of this study was to evaluate muscle forces, spinal loads and trunk stability in pull-push tasks in sagittal-symmetric, static upright standing posture. Three hand-held load magnitudes (80, 120 and 160 N) at four elevations (0, 20, 40 and 60 cm to the L5-S1) and 24 force directions covering all pull/push orientations were considered. For this purpose, a musculoskeletal finite element model with kinematics measured earlier were used. Results demonstrated that peak spinal forces occur under inclined pull (lift) at upper elevations but inclined push at the lowermost one. Minimal spinal loads, on the other hand, occurred at and around vertical pull directions. Overall, spinal forces closely followed variations in the net external moment of pull-push forces at the L5-S1. Local lumbar muscles were most active in pulls while global extensor muscles in lifts. The trunk stability margin decreased with load elevation except at and around horizontal push; it peaked under pulls and reached minimum at vertical lifts. It also increased with antagonist activity in muscles and intra-abdominal pressure. Results provide insight into the marked effects of variation in the load orientation and elevation on muscle forces, spinal loads and trunk stability and hence offer help in rehabilitation, performance enhancement training and design of safer workplaces. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

Chen, L.; McGruer, N. E.; Adams, G. G.

We report the observation of two distinct modes of rate-dependent behavior during contact cycling tests. One is a higher pull-off force at low cycling rates and the other is a higher pull-off force at high cycling rates. Subsequent investigation of these contacts using scanning electron microscopy (SEM) demonstrates that these two rate-dependent modes can be related to brittle and ductile separation modes. The former behavior is indicative of brittle separation, whereas the latter accompanies ductile separation. Thus by monitoring the rate dependence of the pull-off force, the type of separation mode can be identified during cycling without interrupting the testmore » to perform SEM.« less

A demonstration of NIOSH push-pull ventilation criteria.

PubMed

Klein, M K

1987-03-01

This paper summarizes the results of a study performed on an actual chrome plating tank in order to validate criteria for push-pull ventilation systems developed by Huebener and Hughes at NIOSH. Validation of the criteria was made by taking area industrial hygiene samples for hexavalent and total chrome at ten locations around the plating tank. The sampling was performed during actual production runs or while the tank was operating with a dummy load. The sampling data are summarized. The data show that the push-pull system, operating at Huebener's criteria, could control emissions to below the current standards and guidelines. Conclusions and recommendations are included.

Identification of HYPK-Interacting Proteins Reveals Involvement of HYPK in Regulating Cell Growth, Cell Cycle, Unfolded Protein Response and Cell Death

PubMed Central

Choudhury, Kamalika Roy; Raychaudhuri, Swasti; Bhattacharyya, Nitai P.

2012-01-01

Huntingtin Yeast Two-Hybrid Protein K (HYPK) is an intrinsically unstructured huntingtin (HTT)-interacting protein with chaperone-like activity. To obtain more information about the function(s) of the protein, we identified 27 novel interacting partners of HYPK by pull-down assay coupled with mass spectrometry and, further, 9 proteins were identified by co-localization and co-immunoprecipitation (co-IP) assays. In neuronal cells, (EEF1A1 and HSPA1A), (HTT and LMNB2) and (TP53 and RELA) were identified in complex with HYPK in different experiments. Various Gene Ontology (GO) terms for biological processes, like protein folding (GO: 0006457), response to unfolded protein (GO: 0006986), cell cycle arrest (GO: 0007050), anti-apoptosis (GO: 0006916) and regulation of transcription (GO: 0006355) were significantly enriched with the HYPK-interacting proteins. Cell growth and the ability to refold heat-denatured reporter luciferase were decreased, but cytotoxicity was increased in neuronal cells where HYPK was knocked-down using HYPK antisense DNA construct. The proportion of cells in different phases of cell cycle was also altered in cells with reduced levels of HYPK. These results show that HYPK is involved in several biological processes, possibly through interaction with its partners. PMID:23272104

Free energy profiles from single-molecule pulling experiments.

PubMed

Hummer, Gerhard; Szabo, Attila

2010-12-14

Nonequilibrium pulling experiments provide detailed information about the thermodynamic and kinetic properties of molecules. We show that unperturbed free energy profiles as a function of molecular extension can be obtained rigorously from such experiments without using work-weighted position histograms. An inverse Weierstrass transform is used to relate the system free energy obtained from the Jarzynski equality directly to the underlying molecular free energy surface. An accurate approximation for the free energy surface is obtained by using the method of steepest descent to evaluate the inverse transform. The formalism is applied to simulated data obtained from a kinetic model of RNA folding, in which the dynamics consists of jumping between linker-dominated folded and unfolded free energy surfaces.

The effect of aspect ratio on adhesion and stiffness for soft elastic fibres

PubMed Central

Aksak, Burak; Hui, Chung-Yuen; Sitti, Metin

2011-01-01

The effect of aspect ratio on the pull-off stress and stiffness of soft elastic fibres is studied using elasticity and numerical analysis. The adhesive interface between a soft fibre and a smooth rigid surface is modelled using the Dugdale–Barenblatt model. Numerical simulations show that, while pull-off stress increases with decreasing aspect ratio, fibres get stiffer. Also, for sufficiently low aspect ratio fibres, failure occurs via the growth of internal cracks and pull-off stress approaches the intrinsic adhesive strength. Experiments carried out with various aspect ratio polyurethane elastomer fibres are consistent with the numerical simulations. PMID:21227962

FELERION: a new approach for leakage power reduction

NASA Astrophysics Data System (ADS)

R, Anjana; Somkuwar, Ajay

2014-12-01

The circuit proposed in this paper simultaneously reduces the sub threshold leakage power and saves the state of art aspect of the logic circuits. Sleep transistors and PMOS-only logic are used to further reduce the leakage power. Sleep transistors are used as the keepers to reduce the sub threshold leakage current providing the low resistance path to the output. PMOS-only logic is used between the pull up and pull down devices to mitigate the leakage power further. Our proposed fast efficient leakage reduction circuit not only reduces the leakage current but also reduces the power dissipation. Power and delay are analyzed at the 32 nm BSIM4 model for a chain of four inverters, NAND, NOR and ISCAS-85 c17 benchmark circuits using DSCH3 and the Microwind tool. The simulation results reveal that our proposed approach mitigates leakage power by 90%-94% as compared to the conventional approach.

Electrostatically operated optical microshutter array for a miniature integrated optical spectrometer

NASA Astrophysics Data System (ADS)

Ilias, Samir; Picard, Francis; Larouche, Carl; Kruzelecky, Roman; Jamroz, Wes

2017-11-01

16x1 programmable microshutter arrays allowing control of the light transmitted through a transparent substrate supporting the array were successfully fabricated using surface micromachining technology. Each microshutter is basically an electrostatic zipping actuator having a curved shape induced by a stress gradient through the actuator thickness. When a sufficient voltage is applied between the microshutter and the actuation electrode surrounding the associated microslit area, the generated electrostatic force pulls the actuator down to the substrate which closes the microslit. Opening the slit relies on the restoring force. High light transmission through the slit area is obtained with the actuator in the open position and excellent light blocking is observed when the shutter is closed. Static and dynamic responses of the device were determined. The pull-in voltage to close the microslit was about 110 V and the response times to close and open the microslit were about 2 ms and 7 ms, respectively.

MAPPER: A personal computer map projection tool

NASA Technical Reports Server (NTRS)

Bailey, Steven A.

1993-01-01

MAPPER is a set of software tools designed to let users create and manipulate map projections on a personal computer (PC). The capability exists to generate five popular map projections. These include azimuthal, cylindrical, mercator, lambert, and sinusoidal projections. Data for projections are contained in five coordinate databases at various resolutions. MAPPER is managed by a system of pull-down windows. This interface allows the user to intuitively create, view and export maps to other platforms.

"I Just Saw It as Something that Would Pull You Down, Rather than Lift You Up": Resilience in Never-Smokers with Mental Illness

ERIC Educational Resources Information Center

Lawn, Sharon; Hersh, Deborah; Ward, Paul R.; Tsourtos, George; Muller, Robert; Winefield, Anthony; Coveney, John

2011-01-01

Self ConceptWhy people smoke despite the health risks is an important public health question. Equally important is why and how some people resist smoking in spite of circumstances that clearly place them at high risk of becoming smokers. This study used in-depth interviews to explore the narratives of 12 people diagnosed with mental illness, who…

East Europe Report, Economic and Industrial Affairs, No. 2408.

DTIC Science & Technology

1983-06-08

these power plants ; —[the crisis resulting from] insufficient geological surveys (too few drill- ings, insufficient reserves’ analysis ), serious...which swallow the sardines of the local factories and plants . Regional banks pull their shutters down the moment a few ounces of gold raise or lower...ac- cordance with the multilateral program of R & D cooperation entitled "Diagnos- tics of WER Nuclear Power Plants ’ Operating State," a unique

A MEMS Multi-Cantilever Variable Capacitor On Metamaterial

DTIC Science & Technology

2009-03-26

tuning range [38]. 21 Bakri- Kassem and Mansour [39] have developed a parallel-plate variable capac- itor with carrier beams between the plates to...downwards, however, the carrier beams slightly bend down with the movable plate, still prevent- ing it from pulling-in. Bakri- Kassem and Mansour’s... Kassem and R. R. Mansour, “A high-tuning-range mems variable ca- pacitor using carrier beams,” Canadian Journal of Electrical and Computer En- gineering

Sensitivity of tumor cells towards CIGB-300 anticancer peptide relies on its nucleolar localization.

PubMed

Perera, Yasser; Costales, Heydi C; Diaz, Yakelin; Reyes, Osvaldo; Farina, Hernan G; Mendez, Lissandra; Gómez, Roberto E; Acevedo, Boris E; Gomez, Daniel E; Alonso, Daniel F; Perea, Silvio E

2012-04-01

CIGB-300 is a novel anticancer peptide that impairs the casein kinase 2-mediated phosphorylation by direct binding to the conserved phosphoacceptor site on their substrates. Previous findings indicated that CIGB-300 inhibits tumor cell proliferation in vitro and induces tumor growth delay in vivo in cancer animal models. Interestingly, we had previously demonstrated that the putative oncogene B23/nucleophosmin (NPM) is the major intracellular target for CIGB-300 in a sensitive human lung cancer cell line. However, the ability of this peptide to target B23/NPM in cancer cells with differential CIGB-300 response phenotype remained to be determined. Interestingly, in this work, we evidenced that CIGB-300's antiproliferative activity on tumor cells strongly correlates with its nucleolar localization, the main subcellular localization of the previously identified B23/NPM target. Likewise, using CIGB-300 equipotent doses (concentration that inhibits 50% of proliferation), we demonstrated that this peptide interacts and inhibits B23/NPM phosphorylation in different cancer cell lines as evidenced by in vivo pull-down and metabolic labeling experiments. Moreover, such inhibition was followed by a fast apoptosis on CIGB-300-treated cells and also an impairment of cell cycle progression mainly after 5 h of treatment. Altogether, our data not only validates B23/NPM as a main target for CIGB-300 in cancer cells but also provides the first experimental clues to explain their differential antiproliferative response. Importantly, our findings suggest that further improvements to this cell penetrating peptide-based drug should entail its more efficient intracellular delivery at such subcellular localization. Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.

Analytical solutions for efficient interpretation of single-well push-pull tracer tests

NASA Astrophysics Data System (ADS)

Huang, Junqi; Christ, John A.; Goltz, Mark N.

2010-08-01

Single-well push-pull tracer tests have been used to characterize the extent, fate, and transport of subsurface contamination. Analytical solutions provide one alternative for interpreting test results. In this work, an exact analytical solution to two-dimensional equations describing the governing processes acting on a dissolved compound during a modified push-pull test (advection, longitudinal and transverse dispersion, first-order decay, and rate-limited sorption/partitioning in steady, divergent, and convergent flow fields) is developed. The coupling of this solution with inverse modeling to estimate aquifer parameters provides an efficient methodology for subsurface characterization. Synthetic data for single-well push-pull tests are employed to demonstrate the utility of the solution for determining (1) estimates of aquifer longitudinal and transverse dispersivities, (2) sorption distribution coefficients and rate constants, and (3) non-aqueous phase liquid (NAPL) saturations. Employment of the solution to estimate NAPL saturations based on partitioning and non-partitioning tracers is designed to overcome limitations of previous efforts by including rate-limited mass transfer. This solution provides a new tool for use by practitioners when interpreting single-well push-pull test results.

High mobility group box-1 is phosphorylated by protein kinase C zeta and secreted in colon cancer cells

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

Lee, Hanna; Park, Minhee; Shin, Nara

2012-07-27

Highlights: Black-Right-Pointing-Pointer Specific enzyme for HMGB1 phosphorylation and its secretion is proposed. Black-Right-Pointing-Pointer Inhibition of PKC-{zeta} leads to significant reduction of the secreted HMGB1. Black-Right-Pointing-Pointer Phosphorylation of specific site of HMGB1 redirects its secretion in cancer cells. Black-Right-Pointing-Pointer Activation of PKC-{zeta} in cancers explains the enhanced HMGB1 secretion. -- Abstract: High mobility group box-1 (HMGB1), a nuclear protein, is overexpressed and secreted in cancer cells. Phosphorylation on two different nuclear localization signal regions are known to be important for the nuclear-to-cytoplasmic transport and secretion of HMGB1. However, little is known about the biochemical mechanism of HMGB1 modifications and its subsequentmore » secretion from cancer cells. To identify the specific enzyme and important sites for HMGB1 phosphorylation, we screened the protein kinase C (PKC) family in a colon cancer cell line (HCT116) for HMGB1 binding by pull-down experiments using a 3XFLAG-HMGB1 construct. Strong interactions between atypical PKCs (PKC-{zeta}, {lambda}, and {iota}) and cytoplasmic HMGB1 were observed in HCT116 cells. We further identified the most critical PKC isotype that regulates HMGB1 secretion is PKC-{zeta} by using PKC inhibitors and siRNA experiments. The serine residues at S39, S53 and S181 of HMGB1 were related to enhancing HMGB1 secretion. We also demonstrated overexpression and activation of PKC-{zeta} in colon cancer tissues. Our findings suggest that PKC-{zeta} is involved in the phosphorylation of HMGB1, and the phosphorylation of specific serine residues in the nuclear localization signal regions is related to enhanced HMGB1 secretion in colon cancer cells.« less

Motivation and Perception of Tourists as Push and Pull Factors to Visit National Park

NASA Astrophysics Data System (ADS)

Said, Jumrin; Maryono

2018-02-01

Push-pull theoretical framework is a popular theory to explain the reason why the tourists decide to visit the destination rather than other place, the kind of experience they want to get and the type of activity they want to do. In this paper, it is explained the motivation as push factors and the perception as pull factors of the tourist in deciding the destination based on previous literature and research using descriptive method. The framework asumed that tourists are motivated to fulfill their needs, including to reduce the psychological imbalance and to gain recognition of social status. National Park is one of destination based on nature or commonly knowns as ecotourism. In choosing the destination, the tourists tend to classify their alternative choice based on several criteria, such as the domination perception of tourist from one destination (pull factor), self motivation (push factor) and the available time and money (situational constraints).

Angle-Dependent Atomic Force Microscopy Single-Chain Pulling of Adsorbed Macromolecules from Planar Surfaces Unveils the Signature of an Adsorption-Desorption Transition.

PubMed

Grebíková, Lucie; Whittington, Stuart G; Vancso, Julius G

2018-05-23

The adsorption-desorption behavior of polymer chains is at the heart of macromolecular surface science and technology. With the current developments in atomic force microscopy (AFM), it has now become possible to address the desorption problem from the perspective of a single macromolecule. Here, we report on desorption of single polymer chains on planar surfaces by AFM-based single molecule force spectroscopy (SMFS) as a function of the pulling angle with respect to the surface-normal direction. SMFS experiments were performed in water with various substrates using different polymers covalently attached to the AFM probe tip. End-grafting at the AFM tip was achieved by surface-initiated polymerization using initiator functionalized tips. We found that the desorption force increases with a decreasing pulling angle, i.e., an enhanced adhesion of the polymer chain was observed. The magnitude of the desorption force shows a weak angular dependence at pulling angles close to the surface normal. A significant increase of the force is observed at shallower pulling from a certain pulling angle. This behavior carries the signature of an adsorption-desorption transition. The angular dependence of the normalized desorption force exhibits a universal behavior. We compared and interpreted our results using theoretical predictions for single-chain adsorption-desorption transitions.

Angle-Dependent Atomic Force Microscopy Single-Chain Pulling of Adsorbed Macromolecules from Planar Surfaces Unveils the Signature of an Adsorption–Desorption Transition

PubMed Central

2018-01-01

The adsorption–desorption behavior of polymer chains is at the heart of macromolecular surface science and technology. With the current developments in atomic force microscopy (AFM), it has now become possible to address the desorption problem from the perspective of a single macromolecule. Here, we report on desorption of single polymer chains on planar surfaces by AFM-based single molecule force spectroscopy (SMFS) as a function of the pulling angle with respect to the surface-normal direction. SMFS experiments were performed in water with various substrates using different polymers covalently attached to the AFM probe tip. End-grafting at the AFM tip was achieved by surface-initiated polymerization using initiator functionalized tips. We found that the desorption force increases with a decreasing pulling angle, i.e., an enhanced adhesion of the polymer chain was observed. The magnitude of the desorption force shows a weak angular dependence at pulling angles close to the surface normal. A significant increase of the force is observed at shallower pulling from a certain pulling angle. This behavior carries the signature of an adsorption–desorption transition. The angular dependence of the normalized desorption force exhibits a universal behavior. We compared and interpreted our results using theoretical predictions for single-chain adsorption–desorption transitions. PMID:29712430

Kinematics and kinetics of the bench-press and bench-pull exercises in a strength-trained sporting population.

PubMed

Pearson, Simon N; Cronin, John B; Hume, Patria A; Slyfield, David

2009-09-01

Understanding how loading affects power production in resistance training is a key step in identifying the most optimal way of training muscular power - an essential trait in most sporting movements. Twelve elite male sailors with extensive strength-training experience participated in a comparison of kinematics and kinetics from the upper body musculature, with upper body push (bench press) and pull (bench pull) movements performed across loads of 10-100% of one repetition maximum (1RM). 1RM strength and force were shown to be greater in the bench press, while velocity and power outputs were greater for the bench pull across the range of loads. While power output was at a similar level for the two movements at a low load (10% 1RM), significantly greater power outputs were observed for the bench pull in comparison to the bench press with increased load. Power output (Pmax) was maximized at higher relative loads for both mean and peak power in the bench pull (78.6 +/- 5.7% and 70.4 +/- 5.4% of 1RM) compared to the bench press (53.3 +/- 1.7% and 49.7 +/- 4.4% of 1RM). Findings can most likely be attributed to differences in muscle architecture, which may have training implications for these muscles.

The effect of complex rehabilitation training for 12 weeks on trunk muscle function and spine deformation of patients with SCI.

PubMed

Sung, Dong-Hun; Yoon, Seong-Deok; Park, Gi Duck

2015-03-01

[Purpose] It is important for patients with incomplete spinal cord injury (SCI) to strengthen their muscle strength and return to the work force one of the ultimate objectives of rehabilitation. This study reports how a single patient with SCI became stabilized in terms of abdominal muscles and back extension muscles, as well as returning the back to the neutral position from spinal deformation, as result of complex exercises performed for 12 weeks. [Subjects] The degree of damage of the subject was rated as C grade. The subject of this study had unstable posture due to paralysis in the lower extremities of the left side after removal of a malignant tumor by surgical operation, and tilting and torsion in the pelvis increased followed by increase of kyphosis in the thoracolumbar spine. The subject was more than two years since diagnosis of incomplete SCI after surgery. [Methods] Using isokinetic lumbar muscle strength measurement equipment, peak torque/weight, total work and average power in flexion and extension of the lumbar region were measured. A trunk measurement system (Formetric 4D, DIERS, Germany), which is a 3D image processing apparatus with high resolution for vertebrae, was used in order to measure 3D vertebrae and pelvis deformation as well as static balance abilities. As an exercise method, a foam roller was used to conduct fascia relaxation massage for warming-up, and postural kyphosis was changed into postural lordosis by lat pull-down using equipment, performed in 5 sets of 15 times preset at 60% intensity of 1RM 4 set of 10 crunch exercises per set using Togu's were done while sitting at the end of Balance pad, and 4 sets of 15 bridge exercises. [Results] All angular speed tests showed a gradual increase in muscle strength. Flexion and extension showed 10% and 3% improvements, respectively. The spine deformation test showed that isokinetic exercise and lat pull-down exercise for 12 weeks resulted in improved spinal shape. [Conclusion] In this study, core stability exercise for deep muscle training and lat pull-down exercise had positive effects on lower extremity muscle strength and the spinal shape of a patient with SCI.

Ka-band to L-band frequency down-conversion based on III-V-on-silicon photonic integrated circuits

NASA Astrophysics Data System (ADS)

Van Gasse, K.; Wang, Z.; Uvin, S.; De Deckere, B.; Mariën, J.; Thomassen, L.; Roelkens, G.

2017-12-01

In this work, we present the design, simulation and characterization of a frequency down-converter based on III-V-on-silicon photonic integrated circuit technology. We first demonstrate the concept using commercial discrete components, after which we demonstrate frequency conversion using an integrated mode-locked laser and integrated modulator. In our experiments, five channels in the Ka-band (27.5-30 GHz) with 500 MHz bandwidth are down-converted to the L-band (1.5 GHz). The breadboard demonstration shows a conversion efficiency of - 20 dB and a flat response over the 500 MHz bandwidth. The simulation of a fully integrated circuit indicates that a positive conversion gain can be obtained on a millimeter-sized photonic integrated circuit.

Human Body Mechanics of Pushing and Pulling: Analyzing the Factors of Task-related Strain on the Musculoskeletal System.

PubMed

Argubi-Wollesen, Andreas; Wollesen, Bettina; Leitner, Martin; Mattes, Klaus

2017-03-01

The purpose of this review is to name and describe the important factors of musculoskeletal strain originating from pushing and pulling tasks such as cart handling that are commonly found in industrial contexts. A literature database search was performed using the research platform Web of Science. For a study to be included in this review differences in measured or calculated strain had to be investigated with regard to: (1) cart weight/ load; (2) handle position and design; (3) exerted forces; (4) handling task (push and pull); or (5) task experience. Thirteen studies met the inclusion criteria and proved to be of adequate methodological quality by the standards of the Alberta Heritage Foundation for Medical Research. External load or cart weight proved to be the most influential factor of strain. The ideal handle positions ranged from hip to shoulder height and were dependent on the strain factor that was focused on as well as the handling task. Furthermore, task experience and subsequently handling technique were also key to reducing strain. Workplace settings that regularly involve pushing and pulling should be checked for potential improvements with regards to lower weight of the loaded handling device, handle design, and good practice guidelines to further reduce musculoskeletal disease prevalence.

Simulation of drop movement over an inclined surface using smoothed particle hydrodynamics.

PubMed

Das, Arup K; Das, Prasanta K

2009-10-06

Smoothed particle hydrodynamics (SPH) is used to numerically simulate the movement of drops down an inclined plane. Diffuse interfaces have been assumed for tracking the motion of the contact line. The asymmetric shape of the three-dimensional drop and the variation of contact angle along its periphery can be calculated using the simulation. During the motion of a liquid drop down an inclined plane, an internal circulation of liquid particles is observed due to gravitational pull which causes periodic change in the drop shape. The critical angle of inclination required for the inception of drop motion is also evaluated for different fluids as a function of drop volume. The numerical predictions exhibit a good agreement with the published experimental results.

On the pull-out of fibers with fractal-tree structure and the interference of strength and fracture toughness of composites

NASA Astrophysics Data System (ADS)

Fe, Shaoyun; Zhou, Benlian; Lung, Chiwei

1992-06-01

An approximate theory of pull-out of fiber with fractal-tree structure from a matrix is developed with the aim of quantifying the effects of the fractal-tree structure of the fiber. In the experimental investigation of the pull-out of the synthetic fiber with fractal-tree structure, it was generally observed that the force and energy of fiber pullout increase with the branching angle. The application of this theory to experiment is successful. The strength and fracture toughness of composites reinforced by this kind of fiber are inferred to be greater than those of composites reinforced by plane fibers.

On the automatic link between affect and tendencies to approach and avoid: Chen and Bargh (1999) revisited

PubMed Central

Rotteveel, Mark; Gierholz, Alexander; Koch, Gijs; van Aalst, Cherelle; Pinto, Yair; Matzke, Dora; Steingroever, Helen; Verhagen, Josine; Beek, Titia F.; Selker, Ravi; Sasiadek, Adam; Wagenmakers, Eric-Jan

2015-01-01

Within the literature on emotion and behavioral action, studies on approach-avoidance take up a prominent place. Several experimental paradigms feature successful conceptual replications but many original studies have not yet been replicated directly. We present such a direct replication attempt of two seminal experiments originally conducted by Chen and Bargh (1999). In their first experiment, participants affectively evaluated attitude objects by pulling or pushing a lever. Participants who had to pull the lever with positively valenced attitude objects and push the lever with negatively valenced attitude objects (i.e., congruent instruction) did so faster than participants who had to follow the reverse (i.e., incongruent) instruction. In Chen and Bargh's second experiment, the explicit evaluative instructions were absent and participants merely responded to the attitude objects by either always pushing or always pulling the lever. Similar results were obtained as in Experiment 1. Based on these findings, Chen and Bargh concluded that (1) attitude objects are evaluated automatically; and (2) attitude objects automatically trigger a behavioral tendency to approach or avoid. We attempted to replicate both experiments and failed to find the effects reported by Chen and Bargh as indicated by our pre-registered Bayesian data analyses; nevertheless, the evidence in favor of the null hypotheses was only anecdotal, and definitive conclusions await further study. PMID:25883572

ADVANCED DESIGNS OF MAGNETIC JACK-TYPE CONTROL ROD DRIVE

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

Young, J.N.

1959-11-01

The magnetic jack is a device for positioning the control rods In a nuclear reactor, especially in a reactor containing water under pressure. Magnetic actuation precludes the need for shaft seals and eliminates the problems associated with mechanisms operating in water. It consists of a pressure shell, four sets of external stationary magnet coils (hold, grip, lift, pull down), and one Internal moving part (ammature) that impants linear motion to a cluster of rods. (W.L.H.)

Targeting Common but Complex Proteoglycans on Breast Cancer Cells and Stem Cells Using Evolutionary Refined Malaria Proteins

DTIC Science & Technology

2014-09-01

chondroitin sulfate A proteoglycans present on all tested breast cancer cells and the vast majority of tested tissue biopsies. Using pull down assays...Invited, Daugaard. C) 2014. Gordon research conference, July 6-11; Targeting of cancer-specific chondroitin sulfate on circulating tumor cells using...now successfully identified a number of proteoglycans that interact with the recombinant malaria protein VAR2CSA when sulfated on carbon 4 of the CS

Social Awareness and Leader Influence: A Proposed Model and Training Intervention

DTIC Science & Technology

2007-07-01

lips pulled down • Lower lip pushed up • Corner of lips tightened and pressed • Soft volume • Slow pace • Monotone • Head tilted downward...an overall approach or strategy for the influence attempt (e.g., one strategy might be to rely on logical arguments to convince the target that it is...Behavior/ Influence Tactics Target’s Interpretation Target’s Reaction Influencer’s Revised Strategy Influence Goal Evaluation of the Situation

Military Review: The Professional Journal of the U.S. Army. Volume 89, Number 3, May-June 2009

DTIC Science & Technology

2009-06-01

information in other official U.S. Army publications. Authors are responsible for the accuracy and source documentation of material they provide. Military...Review reserves the right to edit material . Basis of official distribution is one per 10 officers for major commands, corps, divisions, major staff...officers pulled their handguns threatening to shoot any rangers who didn’t immediately drop down and take up defensive positions on a line to our

Responses to Disasters, Natural and Man-Made, and Interventions with Social Supports

DTIC Science & Technology

1994-08-01

million homes without power , destroyed 90,000 homes, left 250,000 homeless, and killed 43. Homestead Air Force Base and the surrounding communities of...engineering and military police secured the area and cleared roads of debris and downed power lines. On Friday August 28th, families were allowed to...12%) "* Cold/virus/infection (12%) "* Neck/back (.I%) "* Allergic reaction (03 %) "* Broken bones/pulled muscle (03%) "* Chest/joint pains (03

Direction-dependent force-induced dissociation dynamics of an entropic-driven lock-and-key assembly.

PubMed

Chen, Yen-Fu; Chen, Hsuan-Yi; Sheng, Yu-Jane; Tsao, Heng-Kwong

2017-09-01

The unbinding dynamics of a nanosized sphere-and-cavity assembly under the pulling of constant force and constant loading rate is explored by dissipative particle dynamics simulations. The formation of this matched lock-and-key pair in a polymer solution is driven by the depletion attraction. The two-dimensional free energy landscape U(x,z) associated with this assembly is constructed. Our results indicate that the unbinding pathway along the orientation of the assembly is unfavorable due to the relatively high energy barrier compared to that along the tortuous minimum path whose energy barrier is not high. It is also found that the dissociation rate depends on the direction of the external force (θ) with respect to the assembly orientation. The presence of the force component perpendicular to the assembly orientation can reduce the bond lifetime significantly by driving the key particle to approach the minimum path. Moreover, the dissociation dynamics can be facilitated even by a pushing force compared to the spontaneous dissociation (without forces). To elucidate the effective pathway under pulling, the escaping position is analyzed and its mean direction with respect to the assembly orientation rises generally with increasing θ, revealing that the presence of the force component along the minimum pathway is helpful. The importance of the direction of the external pulling has been demonstrated in our simple system. Therefore, this effect should be considered in more complicated unbinding experiments.

Elephants know when they need a helping trunk in a cooperative task

PubMed Central

Plotnik, Joshua M.; Lair, Richard; Suphachoksahakun, Wirot; de Waal, Frans B. M.

2011-01-01

Elephants are widely assumed to be among the most cognitively advanced animals, even though systematic evidence is lacking. This void in knowledge is mainly due to the danger and difficulty of submitting the largest land animal to behavioral experiments. In an attempt to change this situation, a classical 1930s cooperation paradigm commonly tested on monkeys and apes was modified by using a procedure originally designed for chimpanzees (Pan troglodytes) to measure the reactions of Asian elephants (Elephas maximus). This paradigm explores the cognition underlying coordination toward a shared goal. What do animals know or learn about the benefits of cooperation? Can they learn critical elements of a partner's role in cooperation? Whereas observations in nature suggest such understanding in nonhuman primates, experimental results have been mixed, and little evidence exists with regards to nonprimates. Here, we show that elephants can learn to coordinate with a partner in a task requiring two individuals to simultaneously pull two ends of the same rope to obtain a reward. Not only did the elephants act together, they inhibited the pulling response for up to 45 s if the arrival of a partner was delayed. They also grasped that there was no point to pulling if the partner lacked access to the rope. Such results have been interpreted as demonstrating an understanding of cooperation. Through convergent evolution, elephants may have reached a cooperative skill level on a par with that of chimpanzees. PMID:21383191

Direction-dependent force-induced dissociation dynamics of an entropic-driven lock-and-key assembly

NASA Astrophysics Data System (ADS)

Chen, Yen-Fu; Chen, Hsuan-Yi; Sheng, Yu-Jane; Tsao, Heng-Kwong

2017-09-01

The unbinding dynamics of a nanosized sphere-and-cavity assembly under the pulling of constant force and constant loading rate is explored by dissipative particle dynamics simulations. The formation of this matched lock-and-key pair in a polymer solution is driven by the depletion attraction. The two-dimensional free energy landscape U (x ,z ) associated with this assembly is constructed. Our results indicate that the unbinding pathway along the orientation of the assembly is unfavorable due to the relatively high energy barrier compared to that along the tortuous minimum path whose energy barrier is not high. It is also found that the dissociation rate depends on the direction of the external force (θ ) with respect to the assembly orientation. The presence of the force component perpendicular to the assembly orientation can reduce the bond lifetime significantly by driving the key particle to approach the minimum path. Moreover, the dissociation dynamics can be facilitated even by a pushing force compared to the spontaneous dissociation (without forces). To elucidate the effective pathway under pulling, the escaping position is analyzed and its mean direction with respect to the assembly orientation rises generally with increasing θ , revealing that the presence of the force component along the minimum pathway is helpful. The importance of the direction of the external pulling has been demonstrated in our simple system. Therefore, this effect should be considered in more complicated unbinding experiments.

Long non-coding RNA MIAT promotes growth and metastasis of colorectal cancer cells through regulation of miR-132/Derlin-1 pathway.

PubMed

Liu, Zhaoxia; Wang, Hai; Cai, Hongwei; Hong, Ye; Li, Yan; Su, Dongming; Fan, Zhining

2018-01-01

Recently, long non-coding RNA (lncRNA) MIAT has been demonstrated as an oncogenic gene in several types of cancer. However, the role and mechanism of MIAT in colorectal cancer (CRC) have not been investigated. Real-time PCR was used to measure MIAT expression in CRC tissues and cells. Small interfering RNA specific for MIAT (si-MIAT) was used to down-regulate MIAT expression in CRC cells. The interaction of MIAT and miR-132 was measured by RNA pull-down assay. The effect of si-MIAT on CRC cells apoptosis and metastasis were measured by flow cytometry assay, invasion and migration assay, respectively. In present study, we found that MIAT was highly expressed in CRC tissues and cells. MIAT knockdown inhibited proliferation, migration and invasion and enhanced apoptosis of CRC cells. Further, we demonstrated that MIAT acted as a competing endogenous RNA for miR-132, antagonized its functions, and resulted in the de-repression of its target gene Derlin-1, which acted as an oncogene in promoting growth and metastasis of CRC cells. In LOVO and SW480 cells with si-MIAT, miR-132 inhibitor resulted in an increase of cell proliferation, migration and invasion and a decrease of cell apoptosis, which was partially abolished by transfection of Derlin-1 shRNA. Our data indicated that highly expressed MIAT was an oncogenic lncRNA that promoted the growth and metastasis of CRC through miR-132/Derlin-1 axis.

Interaction between human BAP31 and respiratory syncytial virus small hydrophobic (SH) protein

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

Li, Yan; Jain, Neeraj; Limpanawat, Suweeraya

2015-08-15

The small hydrophobic (SH) protein is a short channel-forming polypeptide encoded by the human respiratory syncytial virus (hRSV). Deletion of SH protein leads to the viral attenuation in mice and primates, and delayed apoptosis in infected cells. We have used a membrane-based yeast two-hybrid system (MbY2H) and a library from human lung cDNA to detect proteins that bind SH protein. This led to the identification of a membrane protein, B-cell associated protein 31 (BAP31). Transfected SH protein co-localizes with transfected BAP31 in cells, and pulls down endogenous BAP31. Titration of purified C-terminal endodomain of BAP31 against isotopically labeled SH proteinmore » in detergent micelles suggests direct interaction between the two proteins. Given the key role of BAP31 in protein trafficking and its critical involvement in pro- and anti-apoptotic pathways, this novel interaction may constitute a potential drug target. - Highlights: • A yeast two-hybrid system (MbY2H) detected BAP31 as a binder of RSV SH protein. • Transfected SH and BAP31 co-localize in lung epithelial cells. • Endogenous BAP31 is pulled down by RSV SH protein. • BAP31 endodomain interacts with the N-terminal α-helix of SH protein in micelles. • This interaction is proposed to be a potential drug target.« less

Cingulin Contains Globular and Coiled-Coil Domains and Interacts with Zo-1, Zo-2, Zo-3, and Myosin

PubMed Central

Cordenonsi, Michelangelo; D'Atri, Fabio; Hammar, Eva; Parry, David A.D.; Kendrick-Jones, John; Shore, David; Citi, Sandra

1999-01-01

We characterized the sequence and protein interactions of cingulin, an M r 140–160-kD phosphoprotein localized on the cytoplasmic surface of epithelial tight junctions (TJ). The derived amino acid sequence of a full-length Xenopus laevis cingulin cDNA shows globular head (residues 1–439) and tail (1,326–1,368) domains and a central α-helical rod domain (440–1,325). Sequence analysis, electron microscopy, and pull-down assays indicate that the cingulin rod is responsible for the formation of coiled-coil parallel dimers, which can further aggregate through intermolecular interactions. Pull-down assays from epithelial, insect cell, and reticulocyte lysates show that an NH2-terminal fragment of cingulin (1–378) interacts in vitro with ZO-1 (K d ∼5 nM), ZO-2, ZO-3, myosin, and AF-6, but not with symplekin, and a COOH-terminal fragment (377–1,368) interacts with myosin and ZO-3. ZO-1 and ZO-2 immunoprecipitates contain cingulin, suggesting in vivo interactions. Full-length cingulin, but not NH2-terminal and COOH-terminal fragments, colocalizes with endogenous cingulin in transfected MDCK cells, indicating that sequences within both head and rod domains are required for TJ localization. We propose that cingulin is a functionally important component of TJ, linking the submembrane plaque domain of TJ to the actomyosin cytoskeleton. PMID:10613913

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

Mishra, Arjun K.; Agnihotri, Pragati; Srivastava, Vijay Kumar

Highlights: • L. donovani spermidine synthase and S-adenosylmethionine decarboxylase have been cloned and purified. • S-adenosylmethionine decarboxylase has autocatalytic property. • GST pull down assay shows the two proteins to form a metabolon. • Isothermal titration calorimetry shows that binding was exothermic having K{sub d} value of 0.4 μM. • Interaction confirmed by fluorescence spectroscopy and size exclusion chromatography. - Abstract: Polyamine biosynthesis pathway has long been considered an essential drug target for trypanosomatids including Leishmania. S-adenosylmethionine decarboxylase (AdoMetDc) and spermidine synthase (SpdSyn) are enzymes of this pathway that catalyze successive steps, with the product of the former, decarboxylated S-adenosylmethioninemore » (dcSAM), acting as an aminopropyl donor for the latter enzyme. Here we have explored the possibility of and identified the protein–protein interaction between SpdSyn and AdoMetDc. The protein–protein interaction has been identified using GST pull down assay. Isothermal titration calorimetry reveals that the interaction is thermodynamically favorable. Fluorescence spectroscopy studies also confirms the interaction, with SpdSyn exhibiting a change in tertiary structure with increasing concentrations of AdoMetDc. Size exclusion chromatography suggests the presence of the complex as a hetero-oligomer. Taken together, these results suggest that the enzymes indeed form a heteromer. Computational analyses suggest that this complex differs significantly from the corresponding human complex, implying that this complex could be a better therapeutic target than the individual enzymes.« less

RTVP-1 regulates glioma cell migration and invasion via interaction with N-WASP and hnRNPK

PubMed Central

Ziv-Av, Amotz; Giladi, Nissim David; Lee, Hae Kyung; Cazacu, Simona; Finniss, Susan; Xiang, Cunli; Pauker, Maor H.; Barda-Saad, Mira; Poisson, Laila; Brodie, Chaya

2015-01-01

Glioblastoma (GBM) are characterized by increased invasion into the surrounding normal brain tissue. RTVP-1 is highly expressed in GBM and regulates the migration and invasion of glioma cells. To further study RTVP-1 effects we performed a pull-down assay using His-tagged RTVP-1 followed by mass spectrometry and found that RTVP-1 was associated with the actin polymerization regulator, N-WASP. This association was further validated by co-immunoprecipitation and FRET analysis. We found that RTVP-1 increased cell spreading, migration and invasion and these effects were at least partly mediated by N-WASP. Another protein which was found by the pull-down assay to interact with RTVP-1 is hnRNPK. This protein has been recently reported to associate with and to inhibit the effect of N-WASP on cell spreading. hnRNPK decreased cell migration, spreading and invasion in glioma cells. Using co-immunoprecipitation we validated the interactions of hnRNPK with N-WASP and RTVP-1 in glioma cells. In addition, we found that overexpression of RTVP-1 decreased the association of N-WASP and hnRNPK. In summary, we report that RTVP-1 regulates glioma cell spreading, migration and invasion and that these effects are mediated via interaction with N-WASP and by interfering with the inhibitory effect of hnRNPK on the function of this protein. PMID:26305187

Novel protein interactions with an actin homolog (MreB) of Helicobacter pylori determined by bacterial two-hybrid system.

PubMed

Zepeda Gurrola, Reyna Cristina; Fu, Yajuan; Rodríguez Luna, Isabel Cristina; Benítez Cardoza, Claudia Guadalupe; López López, María de Jesús; López Vidal, Yolanda; Gutíerrez, Germán Rubén Aguilar; Rodríguez Pérez, Mario A; Guo, Xianwu

2017-08-01

The bacterium Helicobacter pylori infects more than 50% of the world population and causes several gastroduodenal diseases, including gastric cancer. Nevertheless, we still need to explore some protein interactions that may be involved in pathogenesis. MreB, an actin homolog, showed some special characteristics in previous studies, indicating that it could have different functions. Protein functions could be realized via protein-protein interactions. In the present study, the MreB protein from H. pylori 26695 fused with two tags 10×His and GST in tandem was overexpressed and purified from Escherchia coli. The purified recombinant protein was used to perform a pull-down assay with H. pylori 26695 cell lysate. The pulled-down proteins were identified by mass spectrometry (MALDI-TOF), in which the known important proteins related to morphogenesis were absent but several proteins related to pathogenesis process were observed. The bacterial two-hybrid system was further used to evaluate the protein interactions and showed that new interactions of MreB respectively with VacA, UreB, HydB, HylB and AddA were confirmed but the interaction MreB-MreC was not validated. These results indicated that the protein MreB in H. pylori has a distinct interactome, does not participate in cell morphogenesis via MreB-MreC but could be related to pathogenesis. Copyright © 2017 Elsevier GmbH. All rights reserved.

Peptide affinity analysis of proteins that bind to an unstructured NH2-terminal region of the osmoprotective transcription factor NFAT5

PubMed Central

DuMond, Jenna F.; Ramkissoon, Kevin; Zhang, Xue; Izumi, Yuichiro; Wang, Xujing; Eguchi, Koji; Gao, Shouguo; Mukoyama, Masashi; Ferraris, Joan D.

2016-01-01

NFAT5 is an osmoregulated transcription factor that particularly increases expression of genes involved in protection against hypertonicity. Transcription factors often contain unstructured regions that bind co-regulatory proteins that are crucial for their function. The NH2-terminal region of NFAT5 contains regions predicted to be intrinsically disordered. We used peptide aptamer-based affinity chromatography coupled with mass spectrometry to identify protein preys pulled down by one or more overlapping 20 amino acid peptide baits within a predicted NH2-terminal unstructured region of NFAT5. We identify a total of 467 unique protein preys that associate with at least one NH2-terminal peptide bait from NFAT5 in either cytoplasmic or nuclear extracts from HEK293 cells treated with elevated, normal, or reduced NaCl concentrations. Different sets of proteins are pulled down from nuclear vs. cytoplasmic extracts. We used GeneCards to ascertain known functions of the protein preys. The protein preys include many that were previously known, but also many novel ones. Consideration of the novel ones suggests many aspects of NFAT5 regulation, interaction and function that were not previously appreciated, for example, hypertonicity inhibits NFAT5 by sumoylating it and the NFAT5 protein preys include components of the CHTOP complex that desumoylate proteins, an action that should contribute to activation of NFAT5. PMID:26757802

HMSN/ACC truncation mutations disrupt brain-type creatine kinase-dependant activation of K+/Cl- co-transporter 3.

PubMed

Salin-Cantegrel, Adèle; Shekarabi, Masoud; Holbert, Sébastien; Dion, Patrick; Rochefort, Daniel; Laganière, Janet; Dacal, Sandra; Hince, Pascale; Karemera, Liliane; Gaspar, Claudia; Lapointe, Jean-Yves; Rouleau, Guy A

2008-09-01

The potassium-chloride co-transporter 3 (KCC3) is mutated in hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC); however, the molecular mechanisms of HMSN/ACC pathogenesis and the exact role of KCC3 in the development of the nervous system remain poorly understood. The functional regulation of this transporter by protein partners is also largely unknown. Using a yeast two-hybrid approach, we discovered that the C-terminal domain (CTD) of KCC3, which is lost in most HMSN/ACC-causing mutations, directly interacts with brain-specific creatine kinase (CK-B), an ATP-generating enzyme that is also a partner of KCC2. The interaction of KCC3 with CK-B was further confirmed by in vitro glutathione S-transferase pull-down assay, followed by sequencing of the pulled-down complexes. In transfected cultured cells, immunofluorescence labeling showed that CK-B co-localizes with wild-type KCC3, whereas the kinase fails to interact with the inactive truncated KCC3. Finally, CK-B's inhibition by DNFB results in reduction of activity of KCC3 in functional assays using Xenopus laevis oocytes. This physical and functional association between the co-transporter and CK-B is, therefore, the first protein-protein interaction identified to be potentially involved in the pathophysiology of HMSN/ACC.

In Planta Single-Molecule Pull-Down Reveals Tetrameric Stoichiometry of HD-ZIPIII:LITTLE ZIPPER Complexes.

PubMed

Husbands, Aman Y; Aggarwal, Vasudha; Ha, Taekjip; Timmermans, Marja C P

2016-08-01

Deciphering complex biological processes markedly benefits from approaches that directly assess the underlying biomolecular interactions. Most commonly used approaches to monitor protein-protein interactions typically provide nonquantitative readouts that lack statistical power and do not yield information on the heterogeneity or stoichiometry of protein complexes. Single-molecule pull-down (SiMPull) uses single-molecule fluorescence detection to mitigate these disadvantages and can quantitatively interrogate interactions between proteins and other compounds, such as nucleic acids, small molecule ligands, and lipids. Here, we establish SiMPull in plants using the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) and LITTLE ZIPPER (ZPR) interaction as proof-of-principle. Colocalization analysis of fluorophore-tagged HD-ZIPIII and ZPR proteins provides strong statistical evidence of complex formation. In addition, we use SiMPull to directly quantify YFP and mCherry maturation probabilities, showing these differ substantially from values obtained in mammalian systems. Leveraging these probabilities, in conjunction with fluorophore photobleaching assays on over 2000 individual complexes, we determined HD-ZIPIII:ZPR stoichiometry. Intriguingly, these complexes appear as heterotetramers, comprising two HD-ZIPIII and two ZPR molecules, rather than heterodimers as described in the current model. This surprising result raises new questions about the regulation of these key developmental factors and is illustrative of the unique contribution SiMPull is poised to make to in planta protein interaction studies. © 2016 American Society of Plant Biologists. All rights reserved.

Is RAF1 protein from Synechocystis sp. PCC 6803 really needed in the cyanobacterial Rubisco assembly process?

PubMed

Kolesinski, Piotr; Rydzy, Malgorzata; Szczepaniak, Andrzej

2017-05-01

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is responsible for carbon dioxide conversion during photosynthesis and, therefore, is the most important protein in biomass generation. Modifications of this biocatalyst toward improvements in its properties are hindered by the complicated and not yet fully understood assembly process required for the formation of active holoenzymes. An entire set of auxiliary factors, including chaperonin GroEL/GroES and assembly chaperones RbcX or Rubisco accumulation factor 1 (RAF1), is involved in the folding and subsequent assembly of Rubisco subunits. Recently, it has been shown that cyanobacterial RAF1 acts during the formation of the large Rubisco subunit (RbcL) dimer. However, both its physiological function and its necessity in the prokaryotic Rubisco formation process remain elusive. Here, we demonstrate that the Synechocystis sp. PCC 6803 strain with raf1 gene disruption shows the same growth rate as wild-type cells under standard conditions. Moreover, the Rubisco biosynthesis process seems to be unperturbed in mutant cells despite the absence of RbcL-RAF1 complexes. However, in the tested environmental conditions, sulfur starvation triggers the degradation of RbcL and subsequent proteolysis of other polypeptides in wild-type but not Δraf1 strains. Pull-down experiments also indicate that, apart from Rubisco, RAF1 co-purifies with phycocyanins. We postulate that RAF1 is not an obligatory factor in cyanobacterial Rubisco assembly, but rather participates in environmentally regulated Rubisco homeostasis.

PCAF interacts with tax and stimulates tax transactivation in a histone acetyltransferase-independent manner.

PubMed

Jiang, H; Lu, H; Schiltz, R L; Pise-Masison, C A; Ogryzko, V V; Nakatani, Y; Brady, J N

1999-12-01

Recent studies have shown that the p300/CREB binding protein (CBP)-associated factor (PCAF) is involved in transcriptional activation. PCAF activity has been shown strongly associated with histone acetyltransferase (HAT) activity. In this report, we present evidence for a HAT-independent transcription function that is activated in the presence of the human T-cell leukemia virus type 1 (HTLV-1) Tax protein. In vitro and in vivo GST-Tax pull-down and coimmunoprecipitation experiments demonstrate that there is a direct interaction between Tax and PCAF, independent of p300/CBP. PCAF can be recruited to the HTLV-1 Tax responsive element in the presence of Tax, and PCAF cooperates with Tax in vivo to activate transcription from the HTLV-1 LTR over 10-fold. Point mutations at Tax amino acid 318 (TaxS318A) or 319 to 320 (Tax M47), which have decreased or no activity on the HTLV-1 promoter, are defective for PCAF binding. Strikingly, the ability of PCAF to stimulate Tax transactivation is not solely dependent on the PCAF HAT domain. Two independent PCAF HAT mutants, which knock out acetyltransferase enzyme activity, activate Tax transactivation to approximately the same level as wild-type PCAF. In contrast, p300 stimulation of Tax transactivation is HAT dependent. These studies provide experimental evidence that PCAF contains a coactivator transcription function independent of the HAT activity on the viral long terminal repeat.

PCAF Interacts with Tax and Stimulates Tax Transactivation in a Histone Acetyltransferase-Independent Manner

PubMed Central

Jiang, Hua; Lu, Hanxin; Schiltz, R. Louis; Pise-Masison, Cynthia A.; Ogryzko, Vasily V.; Nakatani, Yoshihiro; Brady, John N.

1999-01-01

Recent studies have shown that the p300/CREB binding protein (CBP)-associated factor (PCAF) is involved in transcriptional activation. PCAF activity has been shown strongly associated with histone acetyltransferase (HAT) activity. In this report, we present evidence for a HAT-independent transcription function that is activated in the presence of the human T-cell leukemia virus type 1 (HTLV-1) Tax protein. In vitro and in vivo GST-Tax pull-down and coimmunoprecipitation experiments demonstrate that there is a direct interaction between Tax and PCAF, independent of p300/CBP. PCAF can be recruited to the HTLV-1 Tax responsive element in the presence of Tax, and PCAF cooperates with Tax in vivo to activate transcription from the HTLV-1 LTR over 10-fold. Point mutations at Tax amino acid 318 (TaxS318A) or 319 to 320 (Tax M47), which have decreased or no activity on the HTLV-1 promoter, are defective for PCAF binding. Strikingly, the ability of PCAF to stimulate Tax transactivation is not solely dependent on the PCAF HAT domain. Two independent PCAF HAT mutants, which knock out acetyltransferase enzyme activity, activate Tax transactivation to approximately the same level as wild-type PCAF. In contrast, p300 stimulation of Tax transactivation is HAT dependent. These studies provide experimental evidence that PCAF contains a coactivator transcription function independent of the HAT activity on the viral long terminal repeat. PMID:10567539

The human Kell blood group binds the erythroid 4.1R protein: new insights into the 4.1R-dependent red cell membrane complex

PubMed Central

Azouzi, Slim; Collec, Emmanuel; Mohandas, Narla; An, Xiuli; Colin, Yves; Le Van Kim, Caroline

2015-01-01

Summary Protein 4.1R plays an important role in maintaining the mechanical properties of the erythrocyte membrane. We analysed the expression of Kell blood group protein in erythrocytes from a patient with hereditary elliptocytosis associated with complete 4.1R deficiency (4.1(−) HE). Flow cytometry and Western blot analyses revealed a severe reduction of Kell. In vitro pull down and co-immunoprecipitation experiments from erythrocyte membranes showed a direct interaction between Kell and 4.1R. Using different recombinant domains of 4.1R and the cytoplasmic domain of Kell, we demonstrated that the R46R motif in the juxta-membrane region of Kell binds to lobe B of the 4.1R FERM domain. We also observed that 4.1R deficiency is associated with a reduction of XK and DARC (also termed ACKR1) proteins, the absence of the glycosylated form of the urea transporter B and a slight decrease of band 3. The functional alteration of the 4.1(−) HE erythrocyte membranes was also determined by measuring various transport activities. We documented a slower rate of HCO3−/Cl− exchange, but normal water and ammonia transport across erythrocyte membrane in the absence of 4.1. These findings provide novel insights into the structural organization of blood group antigen proteins into the 4.1R complex of the human red cell membrane. PMID:26455906

The mycobacterial antibiotic resistance determinant WhiB7 acts as a transcriptional activator by binding the primary sigma factor SigA (RpoV)

PubMed Central

Burian, Ján; Yim, Grace; Hsing, Michael; Axerio-Cilies, Peter; Cherkasov, Artem; Spiegelman, George B.; Thompson, Charles J.

2013-01-01

Tuberculosis therapeutic options are limited by the high intrinsic antibiotic resistance of Mycobacterium tuberculosis. The putative transcriptional regulator WhiB7 is crucial for the activation of systems that provide resistance to diverse antibiotic classes. Here, we used in vitro run-off, two-hybrid assays, as well as mutagenic, complementation and protein pull-down experiments, to characterize WhiB7 as an auto-regulatory, redox-sensitive transcriptional activator in Mycobacterium smegmatis. We provide the first direct biochemical proof that a WhiB protein promotes transcription and also demonstrate that this activity is sensitive to oxidation (diamide). Its partner protein for transcriptional activation was identified as SigA, the primary sigma factor subunit of RNA polymerase. Residues required for the interaction mapped to region 4 of SigA (including R515H) or adjacent domains of WhiB7 (including E63D). WhiB7’s ability to provide a specific spectrum of antibiotic-resistance was dependent on these residues as well as its C-terminal AT-hook module that binds to an AT-rich motif immediately upstream of the −35 hexamer recognized by SigA. These experimentally established constrains, combined with protein structure predictions, were used to generate a working model of the WhiB7–SigA-promoter complex. Inhibitors preventing WhiB7 interactions could allow the use of previously ineffective antibiotics for treatment of mycobacterial diseases. PMID:23990327

The mycobacterial antibiotic resistance determinant WhiB7 acts as a transcriptional activator by binding the primary sigma factor SigA (RpoV).

PubMed

Burian, Ján; Yim, Grace; Hsing, Michael; Axerio-Cilies, Peter; Cherkasov, Artem; Spiegelman, George B; Thompson, Charles J

2013-12-01

Tuberculosis therapeutic options are limited by the high intrinsic antibiotic resistance of Mycobacterium tuberculosis. The putative transcriptional regulator WhiB7 is crucial for the activation of systems that provide resistance to diverse antibiotic classes. Here, we used in vitro run-off, two-hybrid assays, as well as mutagenic, complementation and protein pull-down experiments, to characterize WhiB7 as an auto-regulatory, redox-sensitive transcriptional activator in Mycobacterium smegmatis. We provide the first direct biochemical proof that a WhiB protein promotes transcription and also demonstrate that this activity is sensitive to oxidation (diamide). Its partner protein for transcriptional activation was identified as SigA, the primary sigma factor subunit of RNA polymerase. Residues required for the interaction mapped to region 4 of SigA (including R515H) or adjacent domains of WhiB7 (including E63D). WhiB7's ability to provide a specific spectrum of antibiotic-resistance was dependent on these residues as well as its C-terminal AT-hook module that binds to an AT-rich motif immediately upstream of the -35 hexamer recognized by SigA. These experimentally established constrains, combined with protein structure predictions, were used to generate a working model of the WhiB7-SigA-promoter complex. Inhibitors preventing WhiB7 interactions could allow the use of previously ineffective antibiotics for treatment of mycobacterial diseases.

The role of transanal endorectal pull-through in the treatment of Hirschsprung's disease - a multicenter experience.

PubMed

Höllwarth, M E; Rivosecchi, M; Schleef, J; Deluggi, S; Fasching, G; Ceriati, E; Ciprandi, G; DePeppo, F

2002-09-01

The transanal approach (TAA) is a new technique for surgery of Hirschsprung's disease (HD) that was introduced by de la Torre in 1998. The purpose of this multicenter study, including experience from three Austrian and one Italian departments of peadiatric surgery, was to evaluate the role of this approach in HD in 18 children aged 1-72 months. In 14 children the TAA only was performed; in 3 an additional laparoscopy was performed and in 1 conversion to a laparotomy was necessary. One complication (abscess) occurred after laparoscopic-assisted pull-through. The postoperative recovery was rapid, no severe long-term problems were observed. The transanal pull-through technique is generally possible in most classic cases of HD with extension of the disease to the sigmoid colon. If necessary, it can be combined with laparoscopy. Our preliminary results show that the technique is safe, less invasive, and gives excellent cosmetic results, and allows rapid recovery. Long-term results are still pending.

Tug of war in motility assay experiments

NASA Astrophysics Data System (ADS)

Hexner, Daniel; Kafri, Yariv

2009-09-01

The dynamics of two groups of molecular motors pulling in opposite directions on a rigid filament is studied theoretically. To this end we first consider the behavior of one set of motors pulling in a single direction against an external force using a new mean-field approach. Based on these results we analyze a similar setup with two sets of motors pulling in opposite directions in a tug of war in the presence of an external force. In both cases we find that the interplay of fluid friction and protein friction leads to a complex phase diagram where the force-velocity relations can exhibit regions of bistability and spontaneous symmetry breaking. Finally, motivated by recent work, we turn to the case of motility assay experiments where motors bound to a surface push on a bundle of filaments. We find that, depending on the absence or the presence of bistability in the force-velocity curve at zero force, the bundle exhibits anomalous or biased diffusion on long-time and large-length scales.

Low Expression of lncRNA-GAS5 Is Implicated in Human Primary Varicose Great Saphenous Veins

PubMed Central

Yuan, Tian-You; Wang, Shi-Yi; Feng, Jing; Wang, Jing; Liu, Yuan; Wu, Ya-Han; Ma, Xiu-E; Ge, Jin; Cui, Ying-Yu; Jiang, Xiao-Yan

2015-01-01

The cellular mechanisms of primary varicose great saphenous veins (GSVs) involve inflammation, apoptosis, and proliferation of local cells and extracellular matrix degradation. Long non-coding RNAs (lncRNAs) play important roles in these cellular processes; however, which and how lncRNAs related to these mechanisms take effect on GSVs remain unclear. By screening lncRNAs that might experience changes in GSV varicosities, we selected the lower expressed lncRNA-GAS5 (growth arrest specific transcript 5) for functional assessments. Silencing of lncRNA-GAS5 promoted cell proliferation and migration, and cell cycle of the human saphenous vein smooth muscle cells (HSVSMCs), whereas overexpressing it inhibited these cellular behaviors and reduced apoptosis of HSVSMCs. RNA pull-down experiment revealed a direct bind of lncRNA-GAS5 to a Ca2+-dependent RNA-binding protein, Annexin A2. Further experiments showed that silencing of Annexin A2 reduced the HSVSMCs proliferation and vice versa. In the context of lncRNA-GAS5 knockdown, silencing of Annexin A2 reduced the proliferation of HSVSMCs while overexpression of Annexin A2 increased the proliferation. Thus, the low expression of lncRNA-GAS5 may facilitate HSVSMCs proliferation and migration through Annexin A2 and thereby the pathogenesis of GSV varicosities. PMID:25806802

Stars Can't Spin Out of Control (Artist's Animation)

NASA Technical Reports Server (NTRS)

2006-01-01

[figure removed for brevity, see original site] Click on the image for QuickTime Movie of Stars Can't Spin Out of Control

This artist's animation demonstrates how a dusty planet-forming disk can slow down a whirling young star, essentially saving the star from spinning itself to death. Evidence for this phenomenon comes from NASA's Spitzer Space Telescope.

The movie begins by showing a developing star (red ball). The star is basically a giant ball of gas that is collapsing onto itself. As it shrinks, it spins faster and faster, like a skater folding in his or her arms. The green lines represent magnetic fields.

As gravity continues to pull matter inward, the star spins so fast, it starts to flatten out. The same principle applies to the planet Saturn, whose spin has caused it to be slightly squashed or oblate.

A forming star can theoretically whip around fast enough to overcome gravity and flatten itself into a state where it can no longer become a full-fledged star. But stars don't spin out of control, possibly because swirling disks of dust slow them down. Such disks can be found orbiting young stars, and are filled with dust that might ultimately stick together to form planets.

The second half of the animation demonstrates how a disk is thought to keep its star's speed in check. A developing star is shown twirling inside its disk. As it turns, its magnetic fields pass through the disk and get bogged down like a spoon in molasses. This locks the star's rotation to the slower-turning disk, so the star, while continuing to shrink, does not spin faster.

Spitzer found evidence for star-slowing disks in a survey of nearly 500 forming stars in the Orion nebula. It observed that slowly spinning stars are five times more likely to host disks than rapidly spinning stars.

Communication and self-esteem in adults with Down syndrome.

PubMed

Jackson, Claire; Cavenagh, Penny; Clibbens, John

2014-01-01

It is estimated that around 50-90% of people with learning disabilities experience difficulties in communicating. Previous research has linked communication difficulties and self-esteem in other populations, yet this relationship has not previously been investigated for people with Down syndrome. To explore the relationship between communication and self-esteem in adults with Down syndrome. This research also aimed to offer an empowering and inclusive opportunity for adults with learning disabilities to be involved in research. Recruitment took place via a charity established for people with Down syndrome. Twelve adults with Down syndrome and an adult they nominated were approached. A qualitative methodology was selected to explore communication and self-esteem. A thematic analysis focused on the qualitative data collected via semi-structured interviews. Major themes within the data revealed that a relationship between communication and self-esteem may exist for adults with Down syndrome. This study demonstrated that people with Down syndrome may experience low self-esteem that may be linked to communication. The clinical implications of this are considered and areas for future research are identified. © 2014 Royal College of Speech and Language Therapists.

SONAR Discovers RNA-Binding Proteins from Analysis of Large-Scale Protein-Protein Interactomes.

PubMed

Brannan, Kristopher W; Jin, Wenhao; Huelga, Stephanie C; Banks, Charles A S; Gilmore, Joshua M; Florens, Laurence; Washburn, Michael P; Van Nostrand, Eric L; Pratt, Gabriel A; Schwinn, Marie K; Daniels, Danette L; Yeo, Gene W

2016-10-20

RNA metabolism is controlled by an expanding, yet incomplete, catalog of RNA-binding proteins (RBPs), many of which lack characterized RNA binding domains. Approaches to expand the RBP repertoire to discover non-canonical RBPs are currently needed. Here, HaloTag fusion pull down of 12 nuclear and cytoplasmic RBPs followed by quantitative mass spectrometry (MS) demonstrates that proteins interacting with multiple RBPs in an RNA-dependent manner are enriched for RBPs. This motivated SONAR, a computational approach that predicts RNA binding activity by analyzing large-scale affinity precipitation-MS protein-protein interactomes. Without relying on sequence or structure information, SONAR identifies 1,923 human, 489 fly, and 745 yeast RBPs, including over 100 human candidate RBPs that contain zinc finger domains. Enhanced CLIP confirms RNA binding activity and identifies transcriptome-wide RNA binding sites for SONAR-predicted RBPs, revealing unexpected RNA binding activity for disease-relevant proteins and DNA binding proteins. Copyright © 2016 Elsevier Inc. All rights reserved.

Retraction Assembly for Space Shuttle Extended Nose Landing Gear

NASA Technical Reports Server (NTRS)

Files, Bradley S.; Nicholson, Leonard S. (Technical Monitor)

2000-01-01

As part of a project to encourage the use of shape memory alloy actuators for space actuators, this mechanism uses a nitinol ribbon to provide the necessary motion to help retract the proposed extended nose landing gear (ENLG) for the space shuttle. Initial proof-of-concept design of the ENLG did not include the ability to retract the gear automatically. One proposed actuator for this purpose was designed at Johnson Space Center and uses resistive heating to rotate the ribbon around a cylinder. This rotation then allows the assembly to pull down a wedge that is used to hold the landing gear strut in place, thus returning the landing gear to its previous height before extension. The presentation will follow the design of this assembly from working with the nitinol ribbon to providing mechanical connections and allowing minimal friction for motion of three wraps around a cylinder. Also to be presented is preliminary work on design of a shape memory alloy gripper, a design project to demonstrate uses of NiTi.

sAPP modulates iron efflux from brain microvascular endothelial cells by stabilizing the ferrous iron exporter ferroportin

PubMed Central

McCarthy, Ryan C; Park, Yun-Hee; Kosman, Daniel J

2014-01-01

A sequence within the E2 domain of soluble amyloid precursor protein (sAPP) stimulates iron efflux. This activity has been attributed to a ferroxidase activity suggested for this motif. We demonstrate that the stimulation of efflux supported by this peptide and by sAPPα is due to their stabilization of the ferrous iron exporter, ferroportin (Fpn), in the plasma membrane of human brain microvascular endothelial cells (hBMVEC). The peptide does not bind ferric iron explaining why it does not and thermodynamically cannot promote ferrous iron autoxidation. This peptide specifically pulls Fpn down from the plasma membrane of hBMVEC; based on these results, FTP, for ferroportin-targeting peptide, correctly identifies the function of this peptide. The data suggest that in stabilizing Fpn via the targeting due to the FTP sequence, sAPP will increase the flux of iron into the cerebral interstitium. This inference correlates with the observation of significant iron deposition in the amyloid plaques characteristic of Alzheimer’s disease. PMID:24867889

Crystal Growth and Scintillation Properties of Ho-Doped Lu3Al5O12 Single Crystals

NASA Astrophysics Data System (ADS)

Sugiyama, Makoto; Yanagida, Takayuki; Fujimoto, Yutaka; Totsuka, Daisuke; Yokota, Yuui; Kurosawa, Shunsuke; Futami, Yoshisuke; Yoshikawa, Akira

2012-10-01

The crystals of 0.1, 0.5, 1, and 3% Ho doped Lu3Al5O12 (Ho:LuAG) grown by the micro-pulling-down method were examined for their scintillation properties. At wavelengths longer than 300 nm, Ho:LuAG crystals demonstrated around 60% transparency with many absorption peaks attributed to Ho3+ 4f10-4 f10 transitions. When excited by 241Am α-ray to obtain radio luminescence spectra, broad host emission and four sharp Ho3+ 4f10-4 f10 emission peaks were detected in the visible region. Light yields and decay time profiles of the samples irradiated by 137Cs γ-ray were measured using photomultiplier tubes R7600 (Hamamatsu). Ho 0.5%:LuAG showed the highest light yield of 3100 ±310 photons/MeV among the present samples. The decay time profiles were well reproduced by two components exponential approximation consisting of 0.5-1 μs and 3-6 μs.

Cdc45-induced loading of human RPA onto single-stranded DNA

PubMed Central

Tessmer, Ingrid; Prus, Piotr; Schlott, Bernhard; Pospiech, Helmut

2017-01-01

Abstract Cell division cycle protein 45 (Cdc45) is an essential component of the eukaryotic replicative DNA helicase. We found that human Cdc45 forms a complex with the single-stranded DNA (ssDNA) binding protein RPA. Moreover, it actively loads RPA onto nascent ssDNA. Pull-down assays and surface plasmon resonance studies revealed that Cdc45-bound RPA complexed with ssDNA in the 8–10 nucleotide binding mode, but dissociated when RPA covered a 30-mer. Real-time analysis of RPA-ssDNA binding demonstrated that Cdc45 catalytically loaded RPA onto ssDNA. This placement reaction required physical contacts of Cdc45 with the RPA70A subdomain. Our results imply that Cdc45 controlled stabilization of the 8-nt RPA binding mode, the subsequent RPA transition into 30-mer mode and facilitated an ordered binding to ssDNA. We propose that a Cdc45-mediated loading guarantees a seamless deposition of RPA on newly emerging ssDNA at the nascent replication fork. PMID:28100698

CD6 and Linker of Activated T Cells are Potential Interaction Partners for T Cell-Specific Adaptor Protein.

PubMed

Hem, C D; Ekornhol, M; Granum, S; Sundvold-Gjerstad, V; Spurkland, A

2017-02-01

The T cell-specific adaptor protein (TSAd) contains several protein interaction domains, and is merging as a modulator of T cell activation. Several interaction partners for the TSAd proline-rich region and phosphotyrosines have been identified, including the Src and Tec family kinases lymphocyte-specific protein tyrosine kinase and interleukin 2-inducible T cell kinase. Via its Src homology 2 (SH2) domain, TSAd may thus function as a link between these enzymes and other signalling molecules. However, few binding partners to the TSAd SH2 domain in T cells are hitherto known. Through the use of in silico ligand prediction, peptide spot arrays, pull-down and immunoprecipitation experiments, we here report novel interactions between the TSAd SH2 domain and CD6 phosphotyrosine (pTyr) 629 and linker of activated T cells (LAT) pTyr 171 , pTyr 191 and pTyr 226 . © 2016 The Foundation for the Scandinavian Journal of Immunology.

Dimerization deficiency of enigmatic retinitis pigmentosa-linked rhodopsin mutants

PubMed Central

Ploier, Birgit; Caro, Lydia N.; Morizumi, Takefumi; Pandey, Kalpana; Pearring, Jillian N.; Goren, Michael A.; Finnemann, Silvia C.; Graumann, Johannes; Arshavsky, Vadim Y.; Dittman, Jeremy S.; Ernst, Oliver P.; Menon, Anant K.

2016-01-01

Retinitis pigmentosa (RP) is a blinding disease often associated with mutations in rhodopsin, a light-sensing G protein-coupled receptor and phospholipid scramblase. Most RP-associated mutations affect rhodopsin's activity or transport to disc membranes. Intriguingly, some mutations produce apparently normal rhodopsins that nevertheless cause disease. Here we show that three such enigmatic mutations—F45L, V209M and F220C—yield fully functional visual pigments that bind the 11-cis retinal chromophore, activate the G protein transducin, traffic to the light-sensitive photoreceptor compartment and scramble phospholipids. However, tests of scramblase activity show that unlike wild-type rhodopsin that functionally reconstitutes into liposomes as dimers or multimers, F45L, V209M and F220C rhodopsins behave as monomers. This result was confirmed in pull-down experiments. Our data suggest that the photoreceptor pathology associated with expression of these enigmatic RP-associated pigments arises from their unexpected inability to dimerize via transmembrane helices 1 and 5. PMID:27694816

Investigation of reliability attributes and accelerated stress factors of terrestrial solar cells. First annual report

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

Prince, J.L.; Lathrop, J.W.

1979-05-01

The results of accelerated stress testing of four different types of silicon terrestrial solar cells are discussed. The accelerated stress tests used included bias-temperature tests, bias-temperature-humidity tests, thermal cycle and thermal shock tests, and power cycle tests. Characterization of the cells was performed before stress testing and at periodic down-times, using electrical measurement, visual inspection, and metal adherence pull tests. Electrical parameters measured included short-circuit current, I/sub sc/, open circuit voltage, V/sub oc/, and output power, voltage, and current at the maximum power point, P/sub m/, V/sub m/, and I/sub m/ respectively. Incorporated in the report are the distributions ofmore » the prestress electrical data for all cell types. Data was also obtained on cell series and shunt resistance. Significant differences in the response to the various stress tests was observed between cell types. On the basis of the experience gained in this research work, a suggested Reliability Qualification Test Schedule was developed.« less

Serum Amyloid P Component (SAP) Interactome in Human Plasma Containing Physiological Calcium Levels.

PubMed

Poulsen, Ebbe Toftgaard; Pedersen, Kata Wolff; Marzeda, Anna Maria; Enghild, Jan J

2017-02-14

The pentraxin serum amyloid P component (SAP) is secreted by the liver and found in plasma at a concentration of approximately 30 mg/L. SAP is a 25 kDa homopentamer known to bind both protein and nonprotein ligands, all in a calcium-dependent manner. The function of SAP is unclear but likely involves the humoral innate immune system spanning the complement system, inflammation, and coagulation. Also, SAP is known to bind to the generic structure of amyloid deposits and possibly to protect them against proteolysis. In this study, we have characterized the SAP interactome in human plasma containing the physiological Ca 2+ concentration using SAP affinity pull-down and co-immunoprecipitation experiments followed by mass spectrometry analyses. The analyses resulted in the identification of 33 proteins, of which 24 were direct or indirect interaction partners not previously reported. The SAP interactome can be divided into categories that include apolipoproteins, the complement system, coagulation, and proteolytic regulation.

Dimerization deficiency of enigmatic retinitis pigmentosa-linked rhodopsin mutants

NASA Astrophysics Data System (ADS)

Ploier, Birgit; Caro, Lydia N.; Morizumi, Takefumi; Pandey, Kalpana; Pearring, Jillian N.; Goren, Michael A.; Finnemann, Silvia C.; Graumann, Johannes; Arshavsky, Vadim Y.; Dittman, Jeremy S.; Ernst, Oliver P.; Menon, Anant K.

2016-10-01

Retinitis pigmentosa (RP) is a blinding disease often associated with mutations in rhodopsin, a light-sensing G protein-coupled receptor and phospholipid scramblase. Most RP-associated mutations affect rhodopsin's activity or transport to disc membranes. Intriguingly, some mutations produce apparently normal rhodopsins that nevertheless cause disease. Here we show that three such enigmatic mutations--F45L, V209M and F220C--yield fully functional visual pigments that bind the 11-cis retinal chromophore, activate the G protein transducin, traffic to the light-sensitive photoreceptor compartment and scramble phospholipids. However, tests of scramblase activity show that unlike wild-type rhodopsin that functionally reconstitutes into liposomes as dimers or multimers, F45L, V209M and F220C rhodopsins behave as monomers. This result was confirmed in pull-down experiments. Our data suggest that the photoreceptor pathology associated with expression of these enigmatic RP-associated pigments arises from their unexpected inability to dimerize via transmembrane helices 1 and 5.

Nuclear import of Nkx2-2 is mediated by multiple pathways

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

Lin, Wenbo; Xu, PengPeng; Guo, YingYing

Nkx2-2 homeoprotein is essential for the development of the central nervous system and pancreas. Although the nuclear localization signals of Nkx2-2 have been identified, the responsible transport receptor is still unknown. Here, we demonstrate that imp α1 not only interacts with Nkx2-2 but also transports it into the nucleus in vitro by acting together with imp β1. However, the nuclear import of Nkx2-2 in cells was not inhibited in response to knockdown expression of endogenous imp β1 or over-expression of Bimax2. Furthermore, imp β1 and imp 13, but not imp 4, directly interact with Nkx2-2 and are capable of transporting Nkx2-2more » in an in vitro import assay. By GST pull-down assay, we demonstrate that mutation of NLS1 or NLS2 has no effect on interaction with imp α1 or imp 13, but significantly reduced binding to imp β1. Thus, the nuclear import of Nkx2-2 is mediated not only by the classical import pathway but also directly by imp β1 or imp 13.« less

Increased levels of the long noncoding RNA, HOXA-AS3, promote proliferation of A549 cells.

PubMed

Zhang, Hongyue; Liu, Ying; Yan, Lixin; Zhang, Min; Yu, Xiufeng; Du, Wei; Wang, Siqi; Li, Qiaozhi; Chen, He; Zhang, Yafeng; Sun, Hanliang; Tang, Zhidong; Zhu, Daling

2018-06-13

Many long noncoding RNAs (lncRNAs) have been identified as powerful regulators of lung adenocarcinoma (LAD). However, the role of HOXA-AS3, a novel lncRNA, in LAD is largely unknown. In this study, we showed that HOXA-AS3 was significantly upregulated in LAD tissues and A549 cells. After knockdown of HOXA-AS3, cell proliferation, migration, and invasion were inhibited. Xenografts derived from A549 cells transfected with shRNA/HOXA-AS3 had significantly lower tumor weights and smaller tumor volumes. We also demonstrated that HOXA-AS3 increased HOXA6 mRNA stability by forming an RNA duplex. In addition, HOXA6 promoted cell proliferation, migration, and invasion in vitro. Using a RNA pull-down assay, we found that HOXA-AS3 bonded with NF110, which regulated the cell localization of HOXA-AS3. Moreover, histone acetylation was involved in upregulation of HOXA-AS3. These results demonstrate that HOXA-AS3 was activated in LAD and supported cancer cell progression. Therefore, inhibition of HOXA-AS3 could be an effective targeted therapy for patients with LAD.

Insights about transport mechanisms and fracture flow channeling from multi-scale observations of tracer dispersion in shallow fractured crystalline rock.

PubMed

Guihéneuf, N; Bour, O; Boisson, A; Le Borgne, T; Becker, M W; Nigon, B; Wajiduddin, M; Ahmed, S; Maréchal, J-C

2017-11-01

In fractured media, solute transport is controlled by advection in open and connected fractures and by matrix diffusion that may be enhanced by chemical weathering of the fracture walls. These phenomena may lead to non-Fickian dispersion characterized by early tracer arrival time, late-time tailing on the breakthrough curves and potential scale effect on transport processes. Here we investigate the scale dependency of these processes by analyzing a series of convergent and push-pull tracer experiments with distance of investigation ranging from 4m to 41m in shallow fractured granite. The small and intermediate distances convergent experiments display a non-Fickian tailing, characterized by a -2 power law slope. However, the largest distance experiment does not display a clear power law behavior and indicates possibly two main pathways. The push-pull experiments show breakthrough curve tailing decreases as the volume of investigation increases, with a power law slope ranging from -3 to -2.3 from the smallest to the largest volume. The multipath model developed by Becker and Shapiro (2003) is used here to evaluate the hypothesis of the independence of flow pathways. The multipath model is found to explain the convergent data, when increasing local dispersivity and reducing the number of pathways with distance which suggest a transition from non-Fickian to Fickian transport at fracture scale. However, this model predicts an increase of tailing with push-pull distance, while the experiments show the opposite trend. This inconsistency may suggest the activation of cross channel mass transfer at larger volume of investigation, which leads to non-reversible heterogeneous advection with scale. This transition from independent channels to connected channels when the volume of investigation increases suggest that both convergent and push-pull breakthrough curves can inform the existence of characteristic length scales. Copyright © 2017 Elsevier B.V. All rights reserved.

Insights about transport mechanisms and fracture flow channeling from multi-scale observations of tracer dispersion in shallow fractured crystalline rock

NASA Astrophysics Data System (ADS)

Guihéneuf, N.; Bour, O.; Boisson, A.; Le Borgne, T.; Becker, M. W.; Nigon, B.; Wajiduddin, M.; Ahmed, S.; Maréchal, J.-C.

2017-11-01

In fractured media, solute transport is controlled by advection in open and connected fractures and by matrix diffusion that may be enhanced by chemical weathering of the fracture walls. These phenomena may lead to non-Fickian dispersion characterized by early tracer arrival time, late-time tailing on the breakthrough curves and potential scale effect on transport processes. Here we investigate the scale dependency of these processes by analyzing a series of convergent and push-pull tracer experiments with distance of investigation ranging from 4 m to 41 m in shallow fractured granite. The small and intermediate distances convergent experiments display a non-Fickian tailing, characterized by a -2 power law slope. However, the largest distance experiment does not display a clear power law behavior and indicates possibly two main pathways. The push-pull experiments show breakthrough curve tailing decreases as the volume of investigation increases, with a power law slope ranging from - 3 to - 2.3 from the smallest to the largest volume. The multipath model developed by Becker and Shapiro (2003) is used here to evaluate the hypothesis of the independence of flow pathways. The multipath model is found to explain the convergent data, when increasing local dispersivity and reducing the number of pathways with distance which suggest a transition from non-Fickian to Fickian transport at fracture scale. However, this model predicts an increase of tailing with push-pull distance, while the experiments show the opposite trend. This inconsistency may suggest the activation of cross channel mass transfer at larger volume of investigation, which leads to non-reversible heterogeneous advection with scale. This transition from independent channels to connected channels when the volume of investigation increases suggest that both convergent and push-pull breakthrough curves can inform the existence of characteristic length scales.

Investigation of Structures of Microwave Microelectromechanical-System Switches by Taguchi Method

NASA Astrophysics Data System (ADS)

Lai, Yeong-Lin; Lin, Chien-Hung

2007-10-01

The optimal design of microwave microelectromechanical-system (MEMS) switches by the Taguchi method is presented. The structures of the switches are analyzed and optimized in terms of the effective stiffness constant, the maximum von Mises stress, and the natural frequency in order to improve the reliability and the performance of the MEMS switches. There are four factors, each of which has three levels in the Taguchi method for the MEMS switches. An L9(34) orthogonal array is used for the matrix experiments. The characteristics of the experiments are studied by the finite-element method and the analytical method. The responses of the signal-to-noise (S/N) ratios of the characteristics of the switches are investigated. The statistical analysis of variance (ANOVA) is used to interpret the experimental results and decide the significant factors. The final optimum setting, A1B3C1D2, predicts that the effective stiffness constant is 1.06 N/m, the maximum von Mises stress is 76.9 MPa, and the natural frequency is 29.331 kHz. The corresponding switching time is 34 μs, and the pull-down voltage is 9.8 V.

Rotational Energy.

ERIC Educational Resources Information Center

Lockett, Keith

1988-01-01

Demonstrates several objects rolling down a slope to explain the energy transition among potential energy, translational kinetic energy, and rotational kinetic energy. Contains a problem from Galileo's rolling ball experiment. (YP)

Anomalous transport in fracture networks: field scale experiments and modelling

NASA Astrophysics Data System (ADS)

Kang, P. K.; Le Borgne, T.; Bour, O.; Dentz, M.; Juanes, R.

2012-12-01

Anomalous transport is widely observed in different settings and scales of transport through porous and fractured geologic media. A common signature of anomalous transport is the late-time power law tailing in breakthrough curves (BTCs) during tracer tests. Various conceptual models of anomalous transport have been proposed, including multirate mass transfer, continuous time random walk, and stream tube models. Since different conceptual models can produce equally good fits to a single BTC, tracer test interpretation has been plagued with ambiguity. Here, we propose to resolve such ambiguity by analyzing BTCs obtained from both convergent and push-pull flow configurations at two different fracture planes. We conducted field tracer tests in a fractured granite formation close to Ploemeur, France. We observe that BTC tailing depends on the flow configuration and the injection fracture. Specifically the tailing disappears under push-pull geometry, and when we injected at a fracture with high flux (Figure 1). This indicates that for this fractured granite, BTC tailing is controlled by heterogeneous advection and not by matrix diffusion. To explain the change in tailing behavior for different flow configurations, we employ a simple lattice network model with heterogeneous conductivity distribution. The model assigns random conductivities to the fractures and solves the Darcy equation for an incompressible fluid, enforcing mass conservation at fracture intersections. The mass conservation constraint yields a correlated random flow through the fracture system. We investigate whether BTC tailing can be explained by the spatial distribution of preferential flow paths and stagnation zones, which is controlled by the conductivity variance and correlation length. By combining the results from the field tests and numerical modeling, we show that the reversibility of spreading is a key mechanism that needs to be captured. We also demonstrate the dominant role of the injection fracture on the tailing behavior: where we inject makes the difference in the tailing. Blue line is a BTC with injection into a slow velocity zone under convergent flow configuration. The late-time tailing observed for the convergent test diminished for push-pull experiment performed in the same zone(red line). Black line is a BTC with injection into a high velocity zone under convergent flow configuration. Insets: illustration of convergent and push-pull tracer tests using a double packer system.

GREAT I. PPIWG (Public Participation and Information Work Group) Executive Board Position Paper. Appendicies.

DTIC Science & Technology

1978-08-01

houses, parking and picnic areas could be maintained. To my knowledge there has been no response nor are such facilities mentioned in the status report. 8...requires constant speed, so it is improbable that a boat pulling a skier will slow down when passing a fisherman. It should be remembered that the...Page 2 * April 27, 1978 6) We feel that the Computerized Inventory Analysis project (page 102) has been oversold. Such a tool, while quite useful

BRSCW Reference Set Application: Karen Abbott -University of Arkansas (2014) — EDRN Public Portal

Cancer.gov

Our earlier glycoproteomic studies have identified bisecting glycoslyation and core fucosylation changes on particular glycoproteins in endometrioid ovarian cancer tissues and plasma (Abbott et al, 2010, Proteomics). We have validated that these glycan changes occur on the same glycoproteins in serous ovarian cancer plasma using a lectin-pull down western blot assays. We would like to used pooled reference samples to develop a sensitive magnetic bead-based assay to detect these glycoproteins with bisecting and core fucosylation changes.

Analysis of the interaction between membrane proteins and soluble binding partners by surface plasmon resonance.

PubMed

Wu, Zht Cheng; de Keyzer, Jeanine; Kusters, Ilja; Driessen, Arnold J M

2013-01-01

The interaction between membrane proteins and their (protein) ligands is conventionally investigated by nonequilibrium methods such as co-sedimentation or pull-down assays. Surface Plasmon Resonance can be used to monitor such binding events in real-time using isolated membranes immobilized to a surface providing insights in the kinetics of binding under equilibrium conditions. This application provides a fast, automated way to detect interacting species and to determine the kinetics and affinity (Kd) of the interaction.

Advancing radiation balanced lasers (RBLs) in rare-earth (RE)-doped solids

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

Hehlen, Markus Peter

2016-11-21

These slides cover the following topics: Mid-IR lasers in crystals using two-tone RBL (Single-dopant two-tone RBLs: Tm 3+, Er 3+, and Co-doped two-tone RBLs: (Yb 3+, Nd 3+) and (Ho 3+, Tm 3+); Advanced approaches to RBL crystals (Precursor purification, Micro-pulling-down crystal growth, and Bridgman crystal growth); Advanced approaches to RBL fibers (Materials for RBL glass fibers, Micro-structured fibers for RBL, and Fiber preform synthesis); and finally objectives.

[Neuroanatomy of Isolated Body Lateropulsion].

PubMed

Nakazato, Yoshihiko; Tamura, Naotoshi; Ikeda, Kei; Tanaka, Ai; Yamamoto, Toshimasa

2016-03-01

Axial body lateropulsion, a phenomenon where the body is pulled toward the side of the lesion, with tendency of falling down, is the well-known transient feature of lateral medullary syndrome. In some cases, axial body lateropulsion occurs without vestibular and cerebellar symptoms (isolated body lateropulsion:[iBL]). Patients with iBL have a lesion located in the spinocerebellar tract, descending lateral vestibulospinal tract, vestibulo-thalamic pathway, dentatorubrothalamic pathway, or thalamocortical fascicle. This review deals with the anatomic basis and clinical significance of iBL.

Role of GGAP/PIKE-A in prostate cancer progression

DTIC Science & Technology

2009-05-01

linking alterations of GGAP2 activity to neo- plastic transformation. The GGAP2 locus at 12q13.3 is amplified in glioblastoma cell lines, primary glioma ...prostate cancer tissue lysates . Protein (1.5 mg of total) was incubated with anti-GGAP2 rabbit antibody and protein A agarose beads for 4 h before the...agarose pull-down assay. Purified proteins or cell lysates expressing wild-type and mutant GGAP2 were equilibrated in GTP binding buffer [20 mmol/L Tris

Floating baffle to improve efficiency of liquid transfer from tanks

NASA Technical Reports Server (NTRS)

Howard, F. S. (Inventor)

1973-01-01

A floating baffle is described which rides up and down on a vertical shaft over a drain in a tank as the liquid level within the tank varies. When the baffle is in the raised position, the liquid is allowed to flow out of the drain at an unrestricted rate. When the baffle is in the lowered position, pull-through of air or gas that is above the liquid is presented, which would interfere and reduce the flow of liquid from the tank.

In Situ Protein Binding Assay Using Fc-Fusion Proteins.

PubMed

Padmanabhan, Nirmala; Siddiqui, Tabrez J

2017-01-01

This protocol describes an in situ protein-protein interaction assay between tagged recombinant proteins and cell-surface expressed synaptic proteins. The assay is arguably more sensitive than other traditional protein binding assays such as co-immunoprecipitation and pull-downs and provides a visual readout for binding. This assay has been widely used to determine the dissociation constant of binding of trans-synaptic adhesion proteins. The step-wise description in the protocol should facilitate the adoption of this method in other laboratories.

Proceedings of Seminar on Air Antitank Warfare Held at Springfield, Virginia on 25-26 May 1978

DTIC Science & Technology

1979-05-01

there is strategy . You have to get inside your adversary’s organization—learn his strengths, weaknesses, movement, and intentions. In other words...doing it maybe on a larger scale instead of down at the battalion or platoon levels . Finally, our third basis of attack is army. You should still do...all these things at the army level so that you can literally shatter them and pull them apart. Finally he brings up the notion of attacking cities

Spatial Repellency and the Field Evaluation of a Push-Pull Strategy for the Control of Malaria Vectors in Northern Belize, Central America

DTIC Science & Technology

2014-09-18

monitoring of knocked down mosquitoes. To control for residual chemical contamination from repellent treatments, all huts and interception traps were...to discontinue any ongoing trial if the institution is found to have contravened any of the above conditions. 7. The applicant shall cover food ...albopictus (Skuse) from Selangor, Malaysia . Trap Biomed 30:220-30 31. Cherington E, Ek E, Cho P, Burgess F, Hernandez B, et al. 2010. Forest Cover and

Basic Lessons in ORA and AutoMap 2012

DTIC Science & Technology

2012-06-11

boy named Dave. He has 2 balls. 1 ball is red. 1 ball is blue. milkAndCookies.txt: Dave wants milk and cookies. He drives to the store. He then buys... milk and cookies. 2. Create Concept List From the Pull Down Menu select Generate => Concept List => Concept List (per text). Navigate to where you...the thesaurus. Using the ThesauriContentOnly option You create a Meta-Network (Carley, 200) with the one-grams dog, cow , and farm. If you are going

Forming a Better Joint Team: Understanding Service Culture Impact on the Effectiveness of Senior Military Leaders

DTIC Science & Technology

2015-04-13

arrived, and Blue was now starting to understand fully the implications of the order. The JOC was a hub of activity. Staff officers ranging from O...if we divert the hogs down to Kandahar, that pulls them off station here and what happens if we get a troops in contact around Kabul or start ...between them. This paper’s goal is to do just that-- start examining the uniqueness of the service cultures and speculate, sometimes based on

Orienting attention to locations in mental representations

PubMed Central

Astle, Duncan Edward; Summerfield, Jennifer; Griffin, Ivan; Nobre, Anna Christina

2014-01-01

Many cognitive processes depend on our ability to hold information in mind, often well beyond the offset of the original sensory input. The capacity of this ‘visual short-term memory’ (VSTM) is limited to around three to four items. Recent research has demonstrated that the content of VSTM can be modulated by top-down attentional biases. This has been demonstrated using retrodictive spatial cues, termed ‘retro-cues’, which orient participants’ attention to spatial locations within VSTM. In the current paper, we tested whether the use of these cues is modulated by memory load and cue delay. There are a number of important conclusions: i) top-down biases can operate upon very brief iconic traces as well as older VSTM representations (Experiment 1); ii) when operating within capacity, subjects use the cue to prioritize where they initiate their memory search, rather than to discard un-cued items (Experiments 2 and 3); iii) when capacity is exceeded there is little benefit to top-down biasing relative to a neutral condition, however, unattended items are lost, with there being a substantial cost of invalid spatial cueing (Experiment 3); iv) these costs and benefits of orienting spatial attention differ across iconic memory and VSTM representations when VSTM capacity is exceeded (Experiment 4). PMID:21972046

Work volume and strength training responses to resistive exercise improve with periodic heat extraction from the palm.

PubMed

Grahn, Dennis A; Cao, Vinh H; Nguyen, Christopher M; Liu, Mengyuan T; Heller, H Craig

2012-09-01

Body core cooling via the palm of a hand increases work volume during resistive exercise. We asked: (a) "Is there a correlation between elevated core temperatures and fatigue onset during resistive exercise?" and (b) "Does palm cooling between sets of resistive exercise affect strength and work volume training responses?" Core temperature was manipulated by 30-45 minutes of fixed load and duration treadmill exercise in the heat with or without palm cooling. Work volume was then assessed by 4 sets of fixed load bench press exercises. Core temperatures were reduced and work volumes increased after palm cooling (Control: Tes = 39.0 ± 0.1° C, 36 ± 7 reps vs. Cooling: Tes = 38.4 ± 0.2° C, 42 ± 7 reps, mean ± SD, n = 8, p < 0.001). In separate experiments, the impact of palm cooling on work volume and strength training responses were assessed. The participants completed biweekly bench press or pull-up exercises for multiple successive weeks. Palm cooling was applied for 3 minutes between sets of exercise. Over 3 weeks of bench press training, palm cooling increased work volume by 40% (vs. 13% with no treatment; n = 8, p < 0.05). Over 6 weeks of pull-up training, palm cooling increased work volume by 144% in pull-up experienced subjects (vs. 5% over 2 weeks with no treatment; n = 7, p < 0.001) and by 80% in pull-up naïve subjects (vs. 20% with no treatment; n = 11, p < 0.01). Strength (1 repetition maximum) increased 22% over 10 weeks of pyramid bench press training (4 weeks with no treatment followed by 6 weeks with palm cooling; n = 10, p < 0.001). These results verify previous observations about the effects of palm cooling on work volume, demonstrate a link between core temperature and fatigue onset during resistive exercise, and suggest a novel means for improving strength and work volume training responses.

Metamorphic density controls on early-stage subduction dynamics

NASA Astrophysics Data System (ADS)

Duesterhoeft, Erik; Oberhänsli, Roland; Bousquet, Romain

2013-04-01

Subduction is primarily driven by the densification of the downgoing oceanic slab, due to dynamic P-T-fields in subduction zones. It is crucial to unravel slab densification induced by metamorphic reactions to understand the influence on plate dynamics. By analyzing the density and metamorphic structure of subduction zones, we may gain knowledge about the driving, metamorphic processes in a subduction zone like the eclogitization (i.e., the transformation of a MORB to an eclogite), the breakdown of hydrous minerals and the release of fluid or the generation of partial melts. We have therefore developed a 2D subduction zone model down to 250 km that is based on thermodynamic equilibrium assemblage computations. Our model computes the "metamorphic density" of rocks as a function of pressure, temperature and chemical composition using the Theriak-Domino software package at different time stages. We have used this model to investigate how the hydration, dehydration, partial melting and fractionation processes of rocks all influence the metamorphic density and greatly depend on the temperature field within subduction systems. These processes are commonly neglected by other approaches (e.g., gravitational or thermomechanical in nature) reproducing the density distribution within this tectonic setting. The process of eclogitization is assumed as being important to subduction dynamics, based on the very high density (3.6 g/cm3) of eclogitic rocks. The eclogitization in a MORB-type crust is possible only if the rock reaches the garnet phase stability field. This process is primarily temperature driven. Our model demonstrates that the initiation of eclogitization of the slab is not the only significant process that makes the descending slab denser and is responsible for the slab pull force. Indeed, our results show that the densification of the downgoing lithospheric mantle (due to an increase of pressure) starts in the early subduction stage and makes a significant contribution to the slab pull, where eclogitization does not occur. Thus, the lithospheric mantle acts as additional ballast below the sinking slab shortly after the initiation of subduction. Our calculation shows that the dogma of eclogitized basaltic, oceanic crust as the driving force of slab pull is overestimated during the early stage of subduction. These results improve our understanding of the force budget for slab pull during the intial and early stage of subduction. Therefore, the complex metamorphic structure of a slab and mantle wedge has an important impact on the development and dynamics of subduction zones. Further Reading: Duesterhoeft, Oberhänsli & Bousquet (2013), submitted to Earth and Planetary Science Letters

The photon: Experimental emphasis on its wave-particle duality

NASA Technical Reports Server (NTRS)

Shih, Yan-Hua; Sergienko, A. V.; Rubin, Morton H.; Kiess, Thomas E.; Alley, Carroll O.

1994-01-01

Two types of Einstein-Podolsky-Rosen experiments were demonstrated recently in our laboratory. It is interesting to see that in an interference experiment (wave-like experiment) the photon exhibits its particle property, and in a beam-splitting experiment (particle-like experiment) the photon exhibits its wave property. The two-photon states are produced from Type 1 and Type 2 optical spontaneous parametric down conversion, respectively.

Tool use and the effect of action on the imagination.

PubMed

Schwartz, D L; Holton, D L

2000-11-01

Three studies examined the claim that hand movements can facilitate imagery for object rotations but that this facilitation depends on people's model of the situation. In Experiment 1, physically turning a block without vision reduced mental rotation times compared with imagining the same rotation without bodily movement. In Experiment 2, pulling a string from a spool facilitated participants' mental rotation of an object sitting on the spool. In Experiment 3, depending on participants' model of the spool, the exact same pulling movement facilitated or interfered with the exact same imagery transformation. Results of Experiments 2 and 3 indicate that the geometric characteristics of an action do not specify the trajectory of an imagery transformation. Instead, they point to people's ability to model the tools that mediate between motor activity and its environmental consequences and to transfer tool knowledge to a new situation.

Myogenic origin of the hypotension induced by rapid changes in posture in awake dogs following autonomic blockade

PubMed Central

Wong, Brett J.; Sheriff, Don D.

2008-01-01

The “push-pull” effect denotes the reduced tolerance to +Gz (hypergravity) when +Gz stress is preceded by exposure to hypogravity, i.e., fractional, zero, or negative Gz. The purpose of this study was to test the hypothesis that an exaggerated, myogenically mediated rise in leg vascular conductance contributes to the push-pull effect, using heart level arterial blood pressure as a measure of G tolerance. The approach was to impose control (30 s of 30° head-up tilt) and push-pull (30 s of 30° head-up tilt immediately preceded by 10 s of −15° head-down tilt) gravitational stress after administration of hexamethonium (5 mg/kg) to inhibit autonomic ganglionic neurotransmission in seven dogs. Cardiac output or thigh level arterial pressure (myogenic stimulus) was maintained constant by computer-controlled ventricular pacing. The animals were sedated with acepromazine and lightly restrained in lateral recumbency on a tilt table. Following the onset of head-up tilt, the magnitude of the fall in heart level arterial pressure from baseline was −11.6 ± 2.9 and −17.1 ± 2.2 mmHg for the control and push-pull trials, respectively (P < 0.05), when cardiac output was maintained constant. Over 40% of the exaggerated fall in heart level arterial pressure was attributable to an exaggerated rise in hindlimb vascular conductance (P < 0.05). Maintaining thigh level arterial pressure constant abolished the exaggerated rise in hindlimb blood flow. Thus a push-pull effect largely attributable to a myogenically induced rise in leg vascular conductance occurs when autonomic function is inhibited. PMID:18927267

MBD2 is a critical component of a methyl cytosine-binding protein complex isolated from primary erythroid cells

PubMed Central

Kransdorf, Evan P.; Wang, Shou Zhen; Zhu, Sheng Zu; Langston, Timothy B.; Rupon, Jeremy W.; Ginder, Gordon D.

2006-01-01

The chicken embryonic β-type globin gene, ρ, is a member of a small group of vertebrate genes whose developmentally regulated expression is mediated by DNA methylation. Previously, we have shown that a methyl cytosine-binding complex binds to the methylated ρ-globin gene in vitro. We have now chromatographically purified and characterized this complex from adult chicken primary erythroid cells. Four components of the MeCP1 transcriptional repression complex were identified: MBD2, RBAP48, HDAC2, and MTA1. These 4 proteins, as well as the zinc-finger protein p66 and the chromatin remodeling factor Mi2, were found to coelute by gel-filtration analysis and pull-down assays. We conclude that these 6 proteins are components of the MeCPC. In adult erythrocytes, significant enrichment for MBD2 is seen at the inactive ρ-globin gene by chromatin immunoprecipitation assay, whereas no enrichment is observed at the active βA-globin gene, demonstrating MBD2 binds to the methylated and transcriptionally silent ρ-globin gene in vivo. Knock-down of MBD2 resulted in up-regulation of a methylated ρ-gene construct in mouse erythroleukemic (MEL)-ρ cells. These results represent the first purification of a MeCP1-like complex from a primary cell source and provide support for a role for MBD2 in developmental gene regulation. PMID:16778143

VEGF controls lung Th2 inflammation via the miR-1-Mpl (myeloproliferative leukemia virus oncogene)-P-selectin axis.

PubMed

Takyar, Seyedtaghi; Vasavada, Hema; Zhang, Jian-ge; Ahangari, Farida; Niu, Naiqian; Liu, Qing; Lee, Chun Geun; Cohn, Lauren; Elias, Jack A

2013-09-23

Asthma, the prototypic Th2-mediated inflammatory disorder of the lung, is an emergent disease worldwide. Vascular endothelial growth factor (VEGF) is a critical regulator of pulmonary Th2 inflammation, but the underlying mechanism and the roles of microRNAs (miRNAs) in this process have not been defined. Here we show that lung-specific overexpression of VEGF decreases miR-1 expression in the lung, most prominently in the endothelium, and a similar down-regulation occurs in lung endothelium in Th2 inflammation models. Intranasal delivery of miR-1 inhibited inflammatory responses to ovalbumin, house dust mite, and IL-13 overexpression. Blocking VEGF inhibited Th2-mediated lung inflammation, and this was restored by antagonizing miR-1. Using mRNA arrays, Argonaute pull-down assays, luciferase expression assays, and mutational analysis, we identified Mpl as a direct target of miR-1 and showed that VEGF controls the expression of endothelial Mpl during Th2 inflammation via the regulation of miR-1. In vivo knockdown of Mpl inhibited Th2 inflammation and indirectly inhibited the expression of P-selectin in lung endothelium. These experiments define a novel VEGF-miR-1-Mpl-P-selectin effector pathway in lung Th2 inflammation and herald the utility of miR-1 and Mpl as potential therapeutic targets for asthma.

VEGF controls lung Th2 inflammation via the miR-1–Mpl (myeloproliferative leukemia virus oncogene)–P-selectin axis

PubMed Central

Vasavada, Hema; Zhang, Jian-ge; Ahangari, Farida; Niu, Naiqian; Liu, Qing; Lee, Chun Geun; Cohn, Lauren

2013-01-01

Asthma, the prototypic Th2-mediated inflammatory disorder of the lung, is an emergent disease worldwide. Vascular endothelial growth factor (VEGF) is a critical regulator of pulmonary Th2 inflammation, but the underlying mechanism and the roles of microRNAs (miRNAs) in this process have not been defined. Here we show that lung-specific overexpression of VEGF decreases miR-1 expression in the lung, most prominently in the endothelium, and a similar down-regulation occurs in lung endothelium in Th2 inflammation models. Intranasal delivery of miR-1 inhibited inflammatory responses to ovalbumin, house dust mite, and IL-13 overexpression. Blocking VEGF inhibited Th2-mediated lung inflammation, and this was restored by antagonizing miR-1. Using mRNA arrays, Argonaute pull-down assays, luciferase expression assays, and mutational analysis, we identified Mpl as a direct target of miR-1 and showed that VEGF controls the expression of endothelial Mpl during Th2 inflammation via the regulation of miR-1. In vivo knockdown of Mpl inhibited Th2 inflammation and indirectly inhibited the expression of P-selectin in lung endothelium. These experiments define a novel VEGF–miR-1–Mpl–P-selectin effector pathway in lung Th2 inflammation and herald the utility of miR-1 and Mpl as potential therapeutic targets for asthma. PMID:24043765

Interpreting tracer breakthrough tailing from different forced-gradient tracer experiment configurations in fractured bedrock

USGS Publications Warehouse

Becker, M.W.; Shapiro, A.M.

2003-01-01

Conceptual and mathematical models are presented that explain tracer breakthrough tailing in the absence of significant matrix diffusion. Model predictions are compared to field results from radially convergent, weak-dipole, and push-pull tracer experiments conducted in a saturated crystalline bedrock. The models are based upon the assumption that flow is highly channelized, that the mass of tracer in a channel is proportional to the cube of the mean channel aperture, and the mean transport time in the channel is related to the square of the mean channel aperture. These models predict the consistent -2 straight line power law slope observed in breakthrough from radially convergent and weak-dipole tracer experiments and the variable straight line power law slope observed in push-pull tracer experiments with varying injection volumes. The power law breakthrough slope is predicted in the absence of matrix diffusion. A comparison of tracer experiments in which the flow field was reversed to those in which it was not indicates that the apparent dispersion in the breakthrough curve is partially reversible. We hypothesize that the observed breakthrough tailing is due to a combination of local hydrodynamic dispersion, which always increases in the direction of fluid velocity, and heterogeneous advection, which is partially reversed when the flow field is reversed. In spite of our attempt to account for heterogeneous advection using a multipath approach, a much smaller estimate of hydrodynamic dispersivity was obtained from push-pull experiments than from radially convergent or weak dipole experiments. These results suggest that although we can explain breakthrough tailing as an advective phenomenon, we cannot ignore the relationship between hydrodynamic dispersion and flow field geometry at this site. The design of the tracer experiment can severely impact the estimation of hydrodynamic dispersion and matrix diffusion in highly heterogeneous geologic media.

The importance of an underestimated grazer under climate change: how crab density, consumer competition, and physical stress affect salt marsh resilience.

PubMed

Angelini, Christine; van Montfrans, Schuyler G; Hensel, Marc J S; He, Qiang; Silliman, Brian R

2018-05-01

Climate change and consumer outbreaks are driving ecosystem collapse worldwide. Although much research has demonstrated that these factors can interact, how heterogeneity in top-down control intensity and physical forcing modulates ecosystem resilience to climate stress remains poorly understood. Here, we explore whether the nocturnal herbivorous crab Sesarma reticulatum can control spatially dominant cordgrass (Spartina alterniflora) growth and how its top-down effects vary with crab density, drought stress, and large-scale disturbance in southeastern US salt marshes. In multiple field experiments and surveys, we show that Sesarma depresses cordgrass growth and that its effects increase in a saturating manner with increasing crab density, such that the highest naturally occurring densities of this consumer can trigger local cordgrass die-off. This top-down effect of Sesarma is similar in magnitude to what is thought to be the dominant grazer in the system, the marsh periwinkle snail Littoraria irrorata. In a drought stress by Sesarma density experiment, we further show that salinity stress and intensive crab herbivory additively suppress cordgrass drought resistance. After drought subsides, surveys and experiments reveal that Sesarma also stifles cordgrass re-growth into existing die-off areas. Together, these results show that multiple grazers powerfully regulate the productivity and drought resilience of these intertidal grasslands and that heterogeneity in physical stress and consumer density can dictate when and where top-down forcing is important. More generally, this work provides a rare, experimental demonstration of the critical role top-down control can play across the initiation and recovery stages of ecosystem die-off.

Interactive Data Exploration with Smart Drill-Down

PubMed Central

Joglekar, Manas; Garcia-Molina, Hector; Parameswaran, Aditya

2017-01-01

We present smart drill-down, an operator for interactively exploring a relational table to discover and summarize “interesting” groups of tuples. Each group of tuples is described by a rule. For instance, the rule (a, b, ⋆, 1000) tells us that there are a thousand tuples with value a in the first column and b in the second column (and any value in the third column). Smart drill-down presents an analyst with a list of rules that together describe interesting aspects of the table. The analyst can tailor the definition of interesting, and can interactively apply smart drill-down on an existing rule to explore that part of the table. We demonstrate that the underlying optimization problems are NP-Hard, and describe an algorithm for finding the approximately optimal list of rules to display when the user uses a smart drill-down, and a dynamic sampling scheme for efficiently interacting with large tables. Finally, we perform experiments on real datasets on our experimental prototype to demonstrate the usefulness of smart drill-down and study the performance of our algorithms. PMID:28210096

Fiber glass pulling. [in space

NASA Technical Reports Server (NTRS)

Workman, Gary L.

1987-01-01

Experiments were conducted to determine the viability of performing containerless glass fiber pulling in space. The optical transmission properties and glass-forming capabilities of the heavy metal fluorides are reviewed and the acoustic characteristics required for a molten glass levitation system are examined. The design limitations of, and necessary modifications to the acoustic levitation furnace used in the experiments are discussed in detail. Acoustic levitator force measurements were performed and a thermal map of the furnace was generated from thermocouple data. It was determined that the thermal capability of the furnace was inadequate to melt a glass sample in the center. The substitution of a 10 KW carbon monoxide laser for the original furnace heating elements resulted in improved melt heating.

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

Kaminski, Michael

The Irreversible Wash Aid Additive process has been under development by the U.S. Environmental Protection Agency (EPA) and Argonne National Laboratory (Argonne). This process for radioactive cesium mitigation consists of a solution to wash down contaminated structures, roadways, and vehicles and a sequestering agent to bind the radionuclides from the wash water and render them environmentally immobile. The purpose of this process is to restore functionality to basic services and immediately reduce the consequences of a radiologically-contaminated urban environment. Research and development have resulted in a down-selection of technologies for integration and demonstration at the pilot-scale level as part ofmore » the Wide Area Recovery and Resiliency Program (WARRP) under the Department of Homeland Security and the Denver Urban Area Security Initiative. As part of developing the methods for performing a pilot-scale demonstration at the WARRP conference in Denver in 2012, Argonne conducted small-scale field experiments at Separmatic Systems. The main purpose of these experiments was to refine the wash water collection and separations systems and demonstrate key unit operations to help in planning for the large scale demonstration in Denver. Since the purpose of these tests was to demonstrate the operations of the system, we used no radioactive materials. After a brief set of experiments with the LAKOS unit to familiarize ourselves with its operation, two experiments were completed on two separate dates with the Separmatic systems.« less

Vertical electrostatic actuator with extended digital range via tailored topology

NASA Astrophysics Data System (ADS)

Zhang, Yanhang; Dunn, Martin L.

2002-07-01

We describe the design, fabrication, and testing of an electrostatic vertical actuator that exhibits a range of motion that covers the entire initial gap between the actuator and substrate and provides controllable digital output motion. This is obtained by spatially tailoring the electrode arrangement and the stiffness characteristics of the microstructure to control the voltage-deflection characteristics. The concept is based on the electrostatic pull down of bimaterial beams, via a series of electrodes attached to the beams by flexures with tailored stiffness characteristics. The range of travel of the actuator is defined by the post-release deformed shape of the bilayer beams, and can be controlled by a post-release heat-treat process combined with a tailored actuator topology (material distribution and geometry, including spatial geometrical patterning of the individual layers of the bilayer beams). Not only does this allow an increase in the range of travel to cover the entire initial gap, but it also permits digital control of the tip of the actuator which can be designed to yield linear displacement - pull in step characteristics. We fabricated these actuators using the MUMPs surface micromachining process, and packaged them in-house. We measured, using an interferometric microscope, full field deformed shapes of the actuator at each pull in step. The measurements compare well with companion simulation results, both qualitatively and quantitatively.

What is known about the patient's experience of medical tourism? A scoping review.

PubMed

Crooks, Valorie A; Kingsbury, Paul; Snyder, Jeremy; Johnston, Rory

2010-09-08

Medical tourism is understood as travel abroad with the intention of obtaining non-emergency medical services. This practice is the subject of increasing interest, but little is known about its scope. A comprehensive scoping review of published academic articles, media sources, and grey literature reports was performed to answer the question: what is known about the patient's experience of medical tourism? The review was accomplished in three steps: (1) identifying the question and relevant literature; (2) selecting the literature; (3) charting, collating, and summarizing the information. Overall themes were identified from this process. 291 sources were identified for review from the databases searched, the majority of which were media pieces (n = 176). A further 57 sources were included for review after hand searching reference lists. Of the 348 sources that were gathered, 216 were ultimately included in this scoping review. Only a small minority of sources reported on empirical studies that involved the collection of primary data (n = 5). The four themes identified via the review were: (1) decision-making (e.g., push and pull factors that operate to shape patients' decisions); (2) motivations (e.g., procedure-, cost-, and travel-based factors motivating patients to seek care abroad); (3) risks (e.g., health and travel risks); and (4) first-hand accounts (e.g., patients' experiential accounts of having gone abroad for medical care). These themes represent the most discussed issues about the patient's experience of medical tourism in the English-language academic, media, and grey literatures. This review demonstrates the need for additional research on numerous issues, including: (1) understanding how multiple information sources are consulted and evaluated by patients before deciding upon medical tourism; (2) examining how patients understand the risks of care abroad; (3) gathering patients' prospective and retrospective accounts; and (4) the push and pull factors, as well as the motives of patients to participate in medical tourism. The findings from this scoping review and the knowledge gaps it uncovered also demonstrate that there is great potential for new contributions to our understanding of the patient's experience of medical tourism.

What is known about the patient's experience of medical tourism? A scoping review

PubMed Central

2010-01-01

Background Medical tourism is understood as travel abroad with the intention of obtaining non-emergency medical services. This practice is the subject of increasing interest, but little is known about its scope. Methods A comprehensive scoping review of published academic articles, media sources, and grey literature reports was performed to answer the question: what is known about the patient's experience of medical tourism? The review was accomplished in three steps: (1) identifying the question and relevant literature; (2) selecting the literature; (3) charting, collating, and summarizing the information. Overall themes were identified from this process. Results 291 sources were identified for review from the databases searched, the majority of which were media pieces (n = 176). A further 57 sources were included for review after hand searching reference lists. Of the 348 sources that were gathered, 216 were ultimately included in this scoping review. Only a small minority of sources reported on empirical studies that involved the collection of primary data (n = 5). The four themes identified via the review were: (1) decision-making (e.g., push and pull factors that operate to shape patients' decisions); (2) motivations (e.g., procedure-, cost-, and travel-based factors motivating patients to seek care abroad); (3) risks (e.g., health and travel risks); and (4) first-hand accounts (e.g., patients' experiential accounts of having gone abroad for medical care). These themes represent the most discussed issues about the patient's experience of medical tourism in the English-language academic, media, and grey literatures. Conclusions This review demonstrates the need for additional research on numerous issues, including: (1) understanding how multiple information sources are consulted and evaluated by patients before deciding upon medical tourism; (2) examining how patients understand the risks of care abroad; (3) gathering patients' prospective and retrospective accounts; and (4) the push and pull factors, as well as the motives of patients to participate in medical tourism. The findings from this scoping review and the knowledge gaps it uncovered also demonstrate that there is great potential for new contributions to our understanding of the patient's experience of medical tourism. PMID:20825667

Passing crisis and emergency risk communications: the effects of communication channel, information type, and repetition.

PubMed

Edworthy, Judy; Hellier, Elizabeth; Newbold, Lex; Titchener, Kirsteen

2015-05-01

Three experiments explore several factors which influence information transmission when warning messages are passed from person to person. In Experiment 1, messages were passed down chains of participants using five different modes of communication. Written communication channels resulted in more accurate message transmission than verbal. In addition, some elements of the message endured further down the chain than others. Experiment 2 largely replicated these effects and also demonstrated that simple repetition of a message eliminated differences between written and spoken communication. In a final field experiment, chains of participants passed information however they wanted to, with the proviso that half of the chains could not use telephones. Here, the lack of ability to use a telephone did not affect accuracy, but did slow down the speed of transmission from the recipient of the message to the last person in the chain. Implications of the findings for crisis and emergency risk communication are discussed. Copyright © 2015 Elsevier Ltd and The Ergonomics Society. All rights reserved.

Gravity: Simple Experiments for Young Scientists.

ERIC Educational Resources Information Center

White, Larry

This book contains 12 simple experiments through which students can learn about gravity and its implications. Some of the topics included are weight, weightlessness, artificial gravity, the pull of gravity on different shapes, center of gravity, the universal law of gravity, and balancing. Experiments include: finding the balancing point; weighing…

The Dual Estrogen Receptor α Inhibitory Effects of the Tissue-Selective Estrogen Complex for Endometrial and Breast Safety

PubMed Central

Han, Sang Jun; Begum, Khurshida; Foulds, Charles E.; Hamilton, Ross A.; Bailey, Suzanna; Malovannaya, Anna; Chan, Doug; Qin, Jun

2016-01-01

The conjugated estrogen/bazedoxifene tissue-selective estrogen complex (TSEC) is designed to minimize the undesirable effects of estrogen in the uterus and breast tissues and to allow the beneficial effects of estrogen in other estrogen-target tissues, such as the bone and brain. However, the molecular mechanism underlying endometrial and breast safety during TSEC use is not fully understood. Estrogen receptor α (ERα)–estrogen response element (ERE)–DNA pull-down assays using HeLa nuclear extracts followed by mass spectrometry–immunoblotting analyses revealed that, upon TSEC treatment, ERα interacted with transcriptional repressors rather than coactivators. Therefore, the TSEC-mediated recruitment of transcriptional repressors suppresses ERα-mediated transcription in the breast and uterus. In addition, TSEC treatment also degraded ERα protein in uterine tissue and breast cancer cells, but not in bone cells. Interestingly, ERα-ERE-DNA pull-down assays also revealed that, upon TSEC treatment, ERα interacted with the F-box protein 45 (FBXO45) E3 ubiquitin ligase. The loss-of- and gain-of-FBXO45 function analyses indicated that FBXO45 is involved in TSEC-mediated degradation of the ERα protein in endometrial and breast cells. In preclinical studies, these synergistic effects of TSEC on ERα inhibition also suppressed the estrogen-dependent progression of endometriosis. Therefore, the endometrial and breast safety effects of TSEC are associated with synergy between the selective recruitment of transcriptional repressors to ERα and FBXO45-mediated degradation of the ERα protein. PMID:26487511

A new mutation identified in SPATA16 in two globozoospermic patients.

PubMed

ElInati, Elias; Fossard, Camille; Okutman, Ozlem; Ghédir, Houda; Ibala-Romdhane, Samira; Ray, Pierre F; Saad, Ali; Hennebicq, Sylvianne; Viville, Stéphane

2016-06-01

The aim of this study is to identify potential genes involved in human globozoopsermia. Nineteen globozoospermic patients (previously screened for DPY19L2 mutations with no causative mutation) were recruited in this study and screened for mutations in genes implicated in human globozoospermia SPATA16 and PICK1. Using the candidate gene approach and the determination of Spata16 partners by Glutathione S-transferase (GST) pull-down four genes were also selected and screened for mutations. We identified a novel mutation of SPATA16: deletion of 22.6 Kb encompassing the first coding exon in two unrelated Tunisian patients who presented the same deletion breakpoints. The two patients shared the same haplotype, suggesting a possible ancestral founder effect for this new deletion. Four genes were selected using the candidate gene approach and the GST pull-down (GOPC, PICK1, AGFG1 and IRGC) and were screened for mutation, but no variation was identified. The present study confirms the pathogenicity of the SPATA16 mutations. The fact that no variation was detected in the coding sequence of AFGF1, GOPC, PICK1 and IRGC does not mean that they are not involved in human globozoospermia. A larger globozoospermic cohort must be studied in order to accelerate the process of identifying new genes involved in such phenotypes. Until sufficient numbers of patients have been screened, AFGF1, GOPC, PICK1 and IRGC should still be considered as candidate genes.

Colloidal core-seeded semiconductor nanorods as fluorescent labels for in-vitro diagnostics (Conference Presentation)

NASA Astrophysics Data System (ADS)

Chan, YinThai

2016-03-01

Colloidal semiconductor nanocrystals are ideal fluorophores for clinical diagnostics, therapeutics, and highly sensitive biochip applications due to their high photostability, size-tunable color of emission and flexible surface chemistry. The relatively recent development of core-seeded semiconductor nanorods showed that the presence of a rod-like shell can confer even more advantageous physicochemical properties than their spherical counterparts, such as large multi-photon absorption cross-sections and facet-specific chemistry that can be exploited to deposit secondary nanoparticles. It may be envisaged that these highly fluorescent nanorods can be integrated with large scale integrated (LSI) microfluidic systems that allow miniaturization and integration of multiple biochemical processes in a single device at the nanoliter scale, resulting in a highly sensitive and automated detection platform. In this talk, I will describe a LSI microfluidic device that integrates RNA extraction, reverse transcription to cDNA, amplification and target pull-down to detect histidine decarboxylase (HDC) gene directly from human white blood cells samples. When anisotropic colloidal semiconductor nanorods (NRs) were used as the fluorescent readout, the detection limit was found to be 0.4 ng of total RNA, which was much lower than that obtained using spherical quantum dots (QDs) or organic dyes. This was attributed to the large action cross-section of NRs and their high probability of target capture in a pull-down detection scheme. The combination of large scale integrated microfluidics with highly fluorescent semiconductor NRs may find widespread utility in point-of-care devices and multi-target diagnostics.

Interaction between Na-K-ATPase and Bcl-2 proteins BclXL and Bak.

PubMed

Lauf, Peter K; Alqahtani, Tariq; Flues, Karin; Meller, Jaroslaw; Adragna, Norma C

2015-01-01

In silico analysis predicts interaction between Na-K-ATPase (NKA) and Bcl-2 protein canonical BH3- and BH1-like motifs, consistent with NKA inhibition by the benzo-phenanthridine alkaloid chelerythrine, a BH3 mimetic, in fetal human lens epithelial cells (FHLCs) (Lauf PK, Heiny J, Meller J, Lepera MA, Koikov L, Alter GM, Brown TL, Adragna NC. Cell Physiol Biochem 31: 257-276, 2013). This report establishes proof of concept: coimmunoprecipitation and immunocolocalization showed unequivocal and direct physical interaction between NKA and Bcl-2 proteins. Specifically, NKA antibodies (ABs) coimmunoprecipitated BclXL (B-cell lymphoma extra large) and BAK (Bcl-2 antagonist killer) proteins in FHLCs and A549 lung cancer cells. In contrast, both anti-Bcl-2 ABs failed to pull down NKA. Notably, the molecular mass of BAK1 proteins pulled down by NKA and BclXL ABs appeared to be some 4-kDa larger than found in input monomers. In silico analysis predicts these higher molecular mass BAK1 proteins as alternative splicing variants, encoding 42 amino acid (aa) larger proteins than the known 211-aa long canonical BAK1 protein. These BAK1 variants may constitute a pool separate from that forming mitochondrial pores by specifically interacting with NKA and BclXL proteins. We propose a NKA-Bcl-2 protein ternary complex supporting our hypothesis for a special sensor role of NKA in Bcl-2 protein control of cell survival and apoptosis. Copyright © 2015 the American Physiological Society.

Phosphoproteomics and Bioinformatics Analyses of Spinal Cord Proteins in Rats with Morphine Tolerance

PubMed Central

Liaw, Wen-Jinn; Tsao, Cheng-Ming; Huang, Go-Shine; Wu, Chin-Chen; Ho, Shung-Tai; Wang, Jhi-Joung; Tao, Yuan-Xiang; Shui, Hao-Ai

2014-01-01

Introduction Morphine is the most effective pain-relieving drug, but it can cause unwanted side effects. Direct neuraxial administration of morphine to spinal cord not only can provide effective, reliable pain relief but also can prevent the development of supraspinal side effects. However, repeated neuraxial administration of morphine may still lead to morphine tolerance. Methods To better understand the mechanism that causes morphine tolerance, we induced tolerance in rats at the spinal cord level by giving them twice-daily injections of morphine (20 µg/10 µL) for 4 days. We confirmed tolerance by measuring paw withdrawal latencies and maximal possible analgesic effect of morphine on day 5. We then carried out phosphoproteomic analysis to investigate the global phosphorylation of spinal proteins associated with morphine tolerance. Finally, pull-down assays were used to identify phosphorylated types and sites of 14-3-3 proteins, and bioinformatics was applied to predict biological networks impacted by the morphine-regulated proteins. Results Our proteomics data showed that repeated morphine treatment altered phosphorylation of 10 proteins in the spinal cord. Pull-down assays identified 2 serine/threonine phosphorylated sites in 14-3-3 proteins. Bioinformatics further revealed that morphine impacted on cytoskeletal reorganization, neuroplasticity, protein folding and modulation, signal transduction and biomolecular metabolism. Conclusions Repeated morphine administration may affect multiple biological networks by altering protein phosphorylation. These data may provide insight into the mechanism that underlies the development of morphine tolerance. PMID:24392096

Identification of Karyopherin α1 and α7 Interacting Proteins in Porcine Tissue

PubMed Central

Park, Ki-Eun; Inerowicz, H. Dorota; Wang, Xin; Li, Yanfang; Koser, Stephanie; Cabot, Ryan A.

2012-01-01

Specialized trafficking systems in eukaryotic cells serve a critical role in partitioning intracellular proteins between the nucleus and cytoplasm. Cytoplasmic proteins (including chromatin remodeling enzymes and transcription factors) must gain access to the nucleus to exert their functions to properly program fundamental cellular events ranging from cell cycle progression to gene transcription. Knowing that nuclear import mediated by members of the karyopherin α family of transport receptors plays a critical role in regulating development and differentiation, we wanted to determine the identity of proteins that are trafficked by this karyopherin α pathway. To this end, we performed a GST pull-down assay using porcine orthologs of karyopherin α1 (KPNA1) and karyopherin α7 (KPNA7) and prey protein derived from porcine fibroblast cells and used a liquid chromatography and tandem mass spectrometry (LC-MS/MS) approach to determine the identity of KPNA1 and KPNA7 interacting proteins. Our screen revealed that the proteins that interact with KPNA1 and KPNA7 are generally nuclear proteins that possess nuclear localization signals. We further validated two candidate proteins from this screen and showed that they are able to be imported into the nucleus in vivo and also interact with members of the karyopherin α family of proteins in vitro. Our results also reveal the utility of using a GST pull-down approach coupled with LC-MS/MS to screen for protein interaction partners in a non-traditional model system. PMID:22720010

Association between anthropometry and upper-body strength qualities with sprint paddling performance in competitive wave surfers.

PubMed

Sheppard, Jeremy M; McNamara, Phil; Osborne, Mark; Andrews, Mark; Oliveira Borges, Thiago; Walshe, Phil; Chapman, Dale W

2012-12-01

This study aimed to evaluate the potential association with anthropometry and upper-body pulling strength with sprint kinematics of competitive surfers. Ten competitive male surfers (23.9 ± 6.8 years, 177.0 ± 6.5 cm, 72.2 ± 2.4 kg) were assessed for stature, mass, arm span, ∑ 7 site skinfold thickness, pronated pull-up strength, and sprint paddling performance from a stationary start to 15 m. Pearson correlation analysis, and independent t-tests were used to compare potential differences between the slower and faster group of sprint paddlers. Strong associations were found between relative (total kilograms lifted per athlete mass) upper-body pulling strength and sprint paddling time to 5, 10, and 15 m, and peak sprint paddling velocity (r = 0.94, 0.93, 0.88, 0.66, respectively, p < 0.05) and relative upper-body pulling strength was found to be superior (p < 0.05) in the faster group, with large effect (d = 1.88). The results of this study demonstrate a strong association between relative upper-body pulling strength and sprint paddling ability in surfers. Strength and conditioning coaches working with competitive surfers should implement strength training with surfers, including an emphasis on developing relative strength, because this may have a strong influence on sprint paddling performance.

Super-active shape memory alloy composites

NASA Astrophysics Data System (ADS)

Barrett, Ronald M.; Gross, R. Steven

1995-05-01

A new type of very low stiffness super-active composite material is presented. This laminate uses shape-memory alloy (SMA) filaments which are embedded within a low Durometer silicone matrix. The purpose is to develop an active composite in which the local strains within the SMA actuator material will be approximately 1% while the laminate strains will be at least an order of magnitude larger. This type of laminate will be useful for biomimetic, biomedical, surgical and prosthetic applications in which the very high actuator strength of conventional SMA filaments is too great for biological tissues. A modified form of moment and force-balance analysis is used to model the performance of the super-active shape-memory alloy composite (SASMAC). The analytical models are used to predict the performance of a SASMAC pull-pull actuator which uses 10 mil diameter Tinel alloy K actuators embedded in a 0.10' thick, 25 Durometer silicon matrix. The results of testing demonstrate that the laminate is capable of straining up to 10% with theory and experiment in good agreement. Fatigue testing was conducted on the actuator for 1,000 cycles. Because the local strains within the SMA were kept to less than 1%, the element showed no degradation in performance.

Super-active shape-memory alloy composites

NASA Astrophysics Data System (ADS)

Barrett, Ron; Gross, R. Steven

1996-06-01

A new type of very-low-stiffness super-active composite material is presented. This laminate uses shape-memory alloy (SMA) filaments which are embedded within a low-hardness silicone matrix. The purpose is to develop an active composite in which the local strains within the SMA actuator material will be approximately 1%, while the laminate strains will be at least an order of magnitude larger. This type of laminate will be useful for biomimetic, biomedical, surgical and prosthetic applications in which the very high stiffness and actuation strength of conventional SMA filaments are too great for biological tissues. A modified form of moment and force-balance analysis is used to model the performance of the super-active shape-memory alloy composite (SASMAC). The analytical models are used to predict the performance of a SASMAC pull - pull actuator which uses 10 mil diameter Tinel alloy K actuators embedded in a 0.10" thick, 25 Durometer silicone matrix. The results of testing demonstrate that the laminate is capable of straining up to 10% with theory and experiment in good agreement. Fatigue testing was conducted on the actuator for 1 000 cycles. Because the local strains within the SMA were kept to less than 1%, the element showed no degradation in performance.

Exploring a QoS Driven Scheduling Approach for Peer-to-Peer Live Streaming Systems with Network Coding

PubMed Central

Cui, Laizhong; Lu, Nan; Chen, Fu

2014-01-01

Most large-scale peer-to-peer (P2P) live streaming systems use mesh to organize peers and leverage pull scheduling to transmit packets for providing robustness in dynamic environment. The pull scheduling brings large packet delay. Network coding makes the push scheduling feasible in mesh P2P live streaming and improves the efficiency. However, it may also introduce some extra delays and coding computational overhead. To improve the packet delay, streaming quality, and coding overhead, in this paper are as follows. we propose a QoS driven push scheduling approach. The main contributions of this paper are: (i) We introduce a new network coding method to increase the content diversity and reduce the complexity of scheduling; (ii) we formulate the push scheduling as an optimization problem and transform it to a min-cost flow problem for solving it in polynomial time; (iii) we propose a push scheduling algorithm to reduce the coding overhead and do extensive experiments to validate the effectiveness of our approach. Compared with previous approaches, the simulation results demonstrate that packet delay, continuity index, and coding ratio of our system can be significantly improved, especially in dynamic environments. PMID:25114968

Symptom accommodation, trichotillomania-by-proxy, and interpersonal functioning in trichotillomania (hair-pulling disorder).

PubMed

Falkenstein, Martha J; Haaga, David A F

2016-02-01

This study investigated relationship functioning in trichotillomania (TTM) as well as specific interpersonal behaviors that have received little attention in TTM research, including by-proxy pulling, symptom accommodation, and self-disclosure. The objective was to contribute data for future development of components of treatment that focus on interpersonal functioning. Data were collected through survey about relationships and related difficulties among adults who endorsed criteria consistent with DSM-5 criteria for TTM (n=670). Consistent with our hypotheses, TTM symptom severity was correlated negatively with relationship satisfaction and perceived social support, positively with perceived criticism, perceived risk in intimacy, and social interaction anxiety, though these correlations were small (absolute values r=.08 to .17). Approximately one-quarter of survey respondents had not told their closest friend about their trichotillomania, and one-fifth had not told their spouse or long-term romantic partner. TTM-by-proxy urges were reported by 54% of participants, and 37% of participants reported having actually pulled hair from other people, with the most common proxies specified as significant others (51%), parents (13%), friends (8%), siblings (8%), children (7%) and pets (5%). Higher levels of TTM-by-proxy urges were associated with "focused" pulling (d=.37) and perfectionistic thinking (d=.16 to .20), yet current by-proxy urges were not associated with, functional impairment. A small minority of individuals (7%) reported having asked other people to pull hair for them (78% of these requests were granted); there was increased endorsement of "focused" pulling among these individuals. The people who participants asked to pull hairs for them included significant others (66%), mothers (20%), siblings (11%), friends (9%) and one's children (9%). More than one-third of respondents had pulled hair from others, 7% had asked others to pull their hair, and sizable minorities kept TTM secret from their closest friends or even spouse/partners. Clinical levels of social interaction anxiety were endorsed by 51% of the sample. Understanding these interpersonal experiences more fully could improve our understanding of relationship functioning in TTM and guide efforts to individualize treatment for adults with TTM. Published by Elsevier Inc.

OSBP-related protein 8 (ORP8) interacts with Homo sapiens sperm associated antigen 5 (SPAG5) and mediates oxysterol interference of HepG2 cell cycle

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

Zhong, Wenbin; Zhou, You; Li, Jiwei

We earlier identified OSBP-related protein 8 (ORP8) as an endoplasmic reticulum/nuclear envelope oxysterol-binding protein implicated in cellular lipid homeostasis, migration, and organization of the microtubule cytoskeleton. Here, a yeast two-hybrid screen identified Homo sapiens sperm associated antigen 5 (SPAG5)/Astrin as interaction partner of ORP8. The putative interaction was further confirmed by pull-down and co-immunoprecipitation assays. ORP8 did not colocalize with kinetochore-associated SPAG5 in mitotic HepG2 or HuH7 cells, but overexpressed ORP8 was capable of recruiting SPAG5 onto endoplasmic reticulum membranes in interphase cells. In our experiments, 25-hydroxycholesterol (25OHC) retarded the HepG2 cell cycle, causing accumulation in G2/M phase; ORP8 overexpressionmore » resulted in the same phenotype. Importantly, ORP8 knock-down dramatically inhibited the oxysterol effect on HepG2 cell cycle, suggesting a mediating role of ORP8. Furthermore, knock-down of SPAG5 significantly reduced the effects of both ORP8 overexpression and 25OHC on the cell cycle, placing SPAG5 downstream of the two cell-cycle interfering factors. Taken together, the present results suggest that ORP8 may via SPAG5 mediate oxysterol interference of the HepG2 cell cycle. - Highlights: • The oxysterol-binding protein ORP8 was found to interact with the mitotic regulator SPAG5/Astrin. • Treatment of HepG2 cells with 25-hydroxycholesterol caused cell cycle retardation in G2/M. • ORP8 overexpression caused a similar G2/M accumulation, and ORP8 knock-down reversed the 25-hydroxycholesterol effect. • Reduction of cellular of SPAG5/Astrin reversed the cell cycle effects of both 25-hydroxycholesterol and ORP8 overexpression. • Our results suggest that ORP8 mediates via SPAG5/Astrin the oxysterol interference of HepG2 cell cycle.« less

Analysis of the STAT3 interactome using in-situ biotinylation and SILAC.

PubMed

Blumert, Conny; Kalkhof, Stefan; Brocke-Heidrich, Katja; Kohajda, Tibor; von Bergen, Martin; Horn, Friedemann

2013-12-06

Signal transducer and activator of transcription 3 (STAT3) is activated by a variety of cytokines and growth factors. To generate a comprehensive data set of proteins interacting specifically with STAT3, we applied stable isotope labeling with amino acids in cell culture (SILAC). For high-affinity pull-down using streptavidin, we fused STAT3 with a short peptide tag allowing biotinylation in situ (bio-tag), which did not affect STAT3 functions. By this approach, 3642 coprecipitated proteins were detected in human embryonic kidney-293 cells. Filtering using statistical and functional criteria finally extracted 136 proteins as putative interaction partners of STAT3. Both, a physical interaction network analysis and the enrichment of known and predicted interaction partners suggested that our filtering criteria successfully enriched true STAT3 interactors. Our approach identified numerous novel interactors, including ones previously predicted to associate with STAT3. By reciprocal coprecipitation, we were able to verify the physical association between STAT3 and selected interactors, including the novel interaction with TOX4, a member of the TOX high mobility group box family. Applying the same method, we next investigated the activation-dependency of the STAT3 interactome. Again, we identified both known and novel interactions. Thus, our approach allows to study protein-protein interaction effectively and comprehensively. The location, activity, function, degradation, and synthesis of proteins are significantly regulated by interactions of proteins with other proteins, biopolymers and small molecules. Thus, the comprehensive characterization of interactions of proteins in a given proteome is the next milestone on the path to understanding the biochemistry of the cell. In order to generate a comprehensive interactome dataset of proteins specifically interacting with a selected bait protein, we fused our bait protein STAT3 with a short peptide tag allowing biotinylation in situ (bio-tag). This bio-tag allows an affinity pull-down using streptavidin but affected neither the activation of STAT3 by tyrosine phosphorylation nor its transactivating potential. We combined SILAC for accurate relative protein quantification, subcellular fractionation to increase the coverage of interacting proteins, high-affinity pull-down and a stringent filtering method to successfully analyze the interactome of STAT3. With our approach we confirmed several already known and identified numerous novel STAT3 interactors. The approach applied provides a rapid and effective method, which is broadly applicable for studying protein-protein interactions and their dependency on post-translational modifications. © 2013. Published by Elsevier B.V. All rights reserved.

Measuring Input Thresholds on an Existing Board

NASA Technical Reports Server (NTRS)

Kuperman, Igor; Gutrich, Daniel G.; Berkun, Andrew C.

2011-01-01

A critical PECL (positive emitter-coupled logic) interface to Xilinx interface needed to be changed on an existing flight board. The new Xilinx input interface used a CMOS (complementary metal-oxide semiconductor) type of input, and the driver could meet its thresholds typically, but not in worst-case, according to the data sheet. The previous interface had been based on comparison with an external reference, but the CMOS input is based on comparison with an internal divider from the power supply. A way to measure what the exact input threshold was for this device for 64 inputs on a flight board was needed. The measurement technique allowed an accurate measurement of the voltage required to switch a Xilinx input from high to low for each of the 64 lines, while only probing two of them. Directly driving an external voltage was considered too risky, and tests done on any other unit could not be used to qualify the flight board. The two lines directly probed gave an absolute voltage threshold calibration, while data collected on the remaining 62 lines without probing gave relative measurements that could be used to identify any outliers. The PECL interface was forced to a long-period square wave by driving a saturated square wave into the ADC (analog to digital converter). The active pull-down circuit was turned off, causing each line to rise rapidly and fall slowly according to the input s weak pull-down circuitry. The fall time shows up as a change in the pulse width of the signal ready by the Xilinx. This change in pulse width is a function of capacitance, pulldown current, and input threshold. Capacitance was known from the different trace lengths, plus a gate input capacitance, which is the same for all inputs. The pull-down current is the same for all inputs including the two that are probed directly. The data was combined, and the Excel solver tool was used to find input thresholds for the 62 lines. This was repeated over different supply voltages and temperatures to show that the interface had voltage margin under all worst case conditions. Gate input thresholds are normally measured at the manufacturer when the device is on a chip tester. A key function of this machine was duplicated on an existing flight board with no modifications to the nets to be tested, with the exception of changes in the FPGA program.

Distinct regions of triadin are required for targeting and retention at the junctional domain of the sarcoplasmic reticulum.

PubMed

Rossi, Daniela; Bencini, Cristina; Maritati, Marina; Benini, Francesca; Lorenzini, Stefania; Pierantozzi, Enrico; Scarcella, Angela Maria; Paolini, Cecilia; Protasi, Feliciano; Sorrentino, Vincenzo

2014-03-01

Ca2+ release, which is necessary for muscle contraction, occurs at the j-SR (junctional domain of the sarcoplasmic reticulum). It requires the assembly of a large multiprotein complex containing the RyR (ryanodine receptor) and additional proteins, including triadin and calsequestrin. The signals which drive these proteins to the j-SR and how they assemble to form this multiprotein complex are poorly understood. To address aspects of these questions we studied the localization, dynamic properties and molecular interactions of triadin. We identified three regions, named TR1 (targeting region 1), TR2 and TR3, that contribute to the localization of triadin at the j-SR. FRAP experiments showed that triadin is stably associated with the j-SR and that this association is mediated by TR3. Protein pull-down experiments indicated that TR3 contains binding sites for calsequestrin-1 and that triadin clustering can be enhanced by binding to calsequestrin-1. These findings were confirmed by FRET experiments. Interestingly, the stable association of triadin to the j-SR was significantly decreased in myotubes from calsequestrin-1 knockout mice. Taken together, these results identify three regions in triadin that mediate targeting to the j-SR and reveal a role for calsequestrin-1 in promoting the stable association of triadin to the multiprotein complex associated with RyR.

February 2017 - NIF Highlights

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

Fournier, K. B.

2017-03-13

February was a very productive month with only 20 shot days on the calendar. There were 41 target shots performed for the HED, ICF, and the Discovery Science (DS) program. The DS program had a week dedicated to their experiments that was extraordinarily fruitful: 14 target shots were performed for five independent teams, each of whom had a unique experimental platform to field. The teams and the facility worked extraordinarily well to pull off this feat! Additionally, the facility developed high-energy laser operations on a demonstration quad to investigate taking NIF to a new level of performance, and the ICFmore » program demonstrated a 40% increase in the yield from a capsule that had a new, 5-μm-diameter fill tube that apparently mitigates some of the issues that have affected implosions to date. Details follow below.« less

A push-pull system to reduce house entry of malaria mosquitoes

PubMed Central

2014-01-01

Background Mosquitoes are the dominant vectors of pathogens that cause infectious diseases such as malaria, dengue, yellow fever and filariasis. Current vector control strategies often rely on the use of pyrethroids against which mosquitoes are increasingly developing resistance. Here, a push-pull system is presented, that operates by the simultaneous use of repellent and attractive volatile odorants. Method/Results Experiments were carried out in a semi-field set-up: a traditional house which was constructed inside a screenhouse. The release of different repellent compounds, para-menthane-3,8-diol (PMD), catnip oil e.o. and delta-undecalactone, from the four corners of the house resulted in significant reductions of 45% to 81.5% in house entry of host-seeking malaria mosquitoes. The highest reductions in house entry (up to 95.5%), were achieved by simultaneously repelling mosquitoes from the house (push) and removing them from the experimental set-up using attractant-baited traps (pull). Conclusions The outcome of this study suggests that a push-pull system based on attractive and repellent volatiles may successfully be employed to target mosquito vectors of human disease. Reductions in house entry of malaria vectors, of the magnitude that was achieved in these experiments, would likely affect malaria transmission. The repellents used are non-toxic and can be used safely in a human environment. Delta-undecalactone is a novel repellent that showed higher effectiveness than the established repellent PMD. These results encourage further development of the system for practical implementation in the field. PMID:24674451

Wt1a, Foxc1a, and the Notch mediator Rbpj Physically Interact and Regulate the Formation of Podocytes in Zebrafish

PubMed Central

O’Brien, Lori L.; Grimaldi, Michael; Kostun, Zachary; Wingert, Rebecca A.; Selleck, Rori; Davidson, Alan J.

2011-01-01

Podocytes help form the glomerular blood filtration barrier in the kidney and their injury or loss leads to renal disease. The Wilms’ tumor suppressor-1 (Wt1) and the FoxC1/2 transcription factors, as well as Notch signaling, have been implicated as important regulators of podocyte fate. It is not known whether these factors work in parallel or sequentially on different gene targets, or as higher-order transcriptional complexes on common genes. Here, we use the zebrafish to demonstrate that embryos treated with morpholinos against wt1a, foxc1a, or the Notch transcriptional mediator rbpj develop fewer podocytes, as determined by wt1b, hey1 and nephrin expression, while embryos deficient in any two of these factors completely lack podocytes. From GST-pull-downs and co-immunoprecipitation experiments we show that Wt1a, Foxc1a, and Rbpj can physically interact with each other, whereas only Rbpj binds to the Notch intracellular domain (NICD). In transactivation assays, combinations of Wt1, FoxC1/2, and NICD synergistically induce the Hey1 promoter, and have additive or repressive effects on the Podocalyxin promoter, depending on dosage. Taken together, these data suggest that Wt1, FoxC1/2, and Notch signaling converge on common target genes where they physically interact to regulate a podocyte-specific gene program. These findings further our understanding of the transcriptional circuitry responsible for podocyte formation and differentiation during kidney development. PMID:21871448

Identification of a functional interaction between Kv4.3 channels and c-Src tyrosine kinase.

PubMed

Gomes, Pedro; Saito, Tomoaki; Del Corsso, Cris; Alioua, Abderrahmane; Eghbali, Mansoureh; Toro, Ligia; Stefani, Enrico

2008-10-01

Voltage-gated K(+) (Kv) channels are key determinants of cardiac and neuronal excitability. A substantial body of evidence has accumulated in support of a role for Src family tyrosine kinases in the regulation of Kv channels. In this study, we examined the possibility that c-Src tyrosine kinase participates in the modulation of the transient voltage-dependent K(+) channel Kv4.3. Supporting a mechanistic link between Kv4.3 and c-Src, confocal microscopy analysis of HEK293 cells stably transfected with Kv4.3 showed high degree of co-localization of the two proteins at the plasma membrane. Our results further demonstrate an association between Kv4.3 and c-Src by co-immunoprecipitation and GST pull-down assays, this interaction being mediated by the SH2 and SH3 domains of c-Src. Furthermore, we show that Kv4.3 is tyrosine phosphorylated under basal conditions. The functional relevance of the observed interaction between Kv4.3 and c-Src was established in patch-clamp experiments, where application of the Src inhibitor PP2 caused a decrease in Kv4.3 peak current amplitude, but not the inactive structural analogue PP3. Conversely, intracellular application of recombinant c-Src kinase or the protein tyrosine phosphatase inhibitor bpV(phen) increased Kv4.3 peak current amplitude. In conclusion, our findings provide evidence that c-Src-induced Kv4.3 channel activation involves their association in a macromolecular complex and suggest a role for c-Src-Kv4.3 pathway in regulating cardiac and neuronal excitability.

New Partners in Regulation of Gene Expression: The Enhancer of Trithorax and Polycomb Corto Interacts with Methylated Ribosomal Protein L12 Via Its Chromodomain

PubMed Central

Coléno-Costes, Anne; Jang, Suk Min; de Vanssay, Augustin; Rougeot, Julien; Bouceba, Tahar; Randsholt, Neel B.; Gibert, Jean-Michel; Le Crom, Stéphane; Mouchel-Vielh, Emmanuèle

2012-01-01

Chromodomains are found in many regulators of chromatin structure, and most of them recognize methylated lysines on histones. Here, we investigate the role of the Drosophila melanogaster protein Corto's chromodomain. The Enhancer of Trithorax and Polycomb Corto is involved in both silencing and activation of gene expression. Over-expression of the Corto chromodomain (CortoCD) in transgenic flies shows that it is a chromatin-targeting module, critical for Corto function. Unexpectedly, mass spectrometry analysis reveals that polypeptides pulled down by CortoCD from nuclear extracts correspond to ribosomal proteins. Furthermore, real-time interaction analyses demonstrate that CortoCD binds with high affinity RPL12 tri-methylated on lysine 3. Corto and RPL12 co-localize with active epigenetic marks on polytene chromosomes, suggesting that both are involved in fine-tuning transcription of genes in open chromatin. RNA–seq based transcriptomes of wing imaginal discs over-expressing either CortoCD or RPL12 reveal that both factors deregulate large sets of common genes, which are enriched in heat-response and ribosomal protein genes, suggesting that they could be implicated in dynamic coordination of ribosome biogenesis. Chromatin immunoprecipitation experiments show that Corto and RPL12 bind hsp70 and are similarly recruited on gene body after heat shock. Hence, Corto and RPL12 could be involved together in regulation of gene transcription. We discuss whether pseudo-ribosomal complexes composed of various ribosomal proteins might participate in regulation of gene expression in connection with chromatin regulators. PMID:23071455

Association of Mumps Virus V Protein with RACK1 Results in Dissociation of STAT-1 from the Alpha Interferon Receptor Complex

PubMed Central

Kubota, Toru; Yokosawa, Noriko; Yokota, Shin-ichi; Fujii, Nobuhiro

2002-01-01

It has been reported that mumps virus protein V or the C-terminal Cys-rich region of protein V (Vsp) is associated with blocking of the interferon (IFN) signal transduction pathway through a decrease in STAT-1 production. The intracellular target of the V protein was investigated by using a two-hybrid screening system with Vsp as bait. Full-length V protein and Vsp were able to bind to RACK1, and the interaction did not require two WD domains, WD1 and WD2, in RACK1. A significant interaction between V protein and RACK1 was also demonstrated in cells persistently infected with mumps virus (FLMT cells), and the formation of the complex was not affected by treatment with IFN. On the other hand, in uninfected cells, STAT-1 was associated with the long form of the β subunit of the alpha IFN receptor, and this association was mediated by the function of RACK1 as an adaptor protein. Immunoprecipitation and glutathione S-transferase pull-down experiments revealed that the association of RACK1 or mumps virus V protein with the IFN receptor was undetectable in mumps virus-infected cells. Furthermore, RACK1 interacted with mumps virus V protein with a higher affinity than STAT-1 did. Therefore, it is suggested that mumps virus V protein has the ability to interact strongly with RACK1 and consequently to bring about the disruption of the complex formed from STAT-1, RACK1, and the IFN receptor. PMID:12438593

Optical tweezers reveal force plateau and internal friction in PEG-induced DNA condensation.

PubMed

Ojala, Heikki; Ziedaite, Gabija; Wallin, Anders E; Bamford, Dennis H; Hæggström, Edward

2014-03-01

The simplified artificial environments in which highly complex biological systems are studied do not represent the crowded, dense, salty, and dynamic environment inside the living cell. Consequently, it is important to investigate the effect of crowding agents on DNA. We used a dual-trap optical tweezers instrument to perform force spectroscopy experiments at pull speeds ranging from 0.3 to 270 μm/s on single dsDNA molecules in the presence of poly(ethylene glycol) (PEG) and monovalent salt. PEG of sizes 1,500 and 4,000 Da condensed DNA, and force-extension data contained a force plateau at approximately 1 pN. The level of the force plateau increased with increasing pull speed. During slow pulling the dissipated work increased linearly with pull speed. The calculated friction coefficient did not depend on amount of DNA incorporated in the condensate, indicating internal friction is independent of the condensate size. PEG300 had no effect on the dsDNA force-extension curve. The force plateau implies that condensation induced by crowding agents resembles condensation induced by multivalent cations.

Force probe simulations of a reversibly rebinding system: Impact of pulling device stiffness

NASA Astrophysics Data System (ADS)

Jaschonek, Stefan; Diezemann, Gregor

2017-03-01

We present a detailed study of the parameter dependence of force probe molecular dynamics (FPMD) simulations. Using a well studied calix[4]arene catenane dimer as a model system, we systematically vary the pulling velocity and the stiffness of the applied external potential. This allows us to investigate how the results of pulling simulations operating in the constant velocity mode (force-ramp mode) depend on the details of the simulation setup. The system studied has the further advantage of showing reversible rebinding meaning that we can monitor the opening and the rebinding transition. Many models designed to extract kinetic information from rupture force distributions work in the limit of soft springs and all quantities are found to depend solely on the so-called loading rate, the product of spring stiffness and pulling velocity. This approximation is known to break down when stiff springs are used, a situation often encountered in molecular simulations. We find that while some quantities only depend on the loading rate, others show an explicit dependence on the spring constant used in the FPMD simulation. In particular, the force versus extension curves show an almost stiffness independent rupture force but the force jump after the rupture transition does depend roughly linearly on the value of the stiffness. The kinetic rates determined from the rupture force distributions show a dependence on the stiffness that can be understood in terms of the corresponding dependence of the characteristic forces alone. These dependencies can be understood qualitatively in terms of a harmonic model for the molecular free energy landscape. It appears that the pulling velocities employed are so large that the crossover from activated dynamics to diffusive dynamics takes place on the time scale of our simulations. We determine the effective distance of the free energy minima of the closed and the open configurations of the system from the barrier via an analysis of the hydrogen-bond network with results in accord with earlier simulations. We find that the system is quite brittle in the force regime monitored in the sense that the barrier is located near to the closed state.

Addressing challenging behaviour in children with Down syndrome: the use of applied behaviour analysis for assessment and intervention.

PubMed

Feeley, Kathleen M; Jones, Emily A

2006-09-01

Children with Down syndrome are at an increased risk for engaging in challenging behaviour that may be part of a behavioural phenotype characteristic of Down syndrome. The methodology of applied behaviour analysis has been demonstrated effective with a wide range of challenging behaviours, across various disabilities. Applications to children with Down syndrome and the examination of behaviourally based strategies to specifically address the unique characteristics of children with Down syndrome are limited. However, there are several studies in which a subset of the participants did have Down syndrome. A handful of these studies are reviewed within the context of functional behaviour assessment and Positive Behavioural Supports. Drawing from these studies and the behavioural literature, as well as the authors' clinical experience and research, suggestions regarding early intervention for challenging behaviour with children with Down syndrome are provided.

Scalability of Robotic Controllers: Effects of Progressive Autonomy on Intelligence, Surveillance, and Reconnaissance Robotic Tasks

DTIC Science & Technology

2012-09-01

away from the MOCU. The semi-autonomous mode was preferred over the teleoperated mode for multitasking , maintaining SA, avoiding obstacles, and...0 23 Software with icons 0 0 0 0 2 25 Pull-down menu * 0 0 0 0 3 24 Graphics/drawing features in software packages* 3 8 1 4 3 8 Email 1 0 0 0 1...r. Navigate to the next waypoint or set of hash lines 5.27 5.08 6.25 s. Ability to multitask (operate/monitor robot and communicate on the radio

A personal computer-based, multitasking data acquisition system

NASA Technical Reports Server (NTRS)

Bailey, Steven A.

1990-01-01

A multitasking, data acquisition system was written to simultaneously collect meteorological radar and telemetry data from two sources. This system is based on the personal computer architecture. Data is collected via two asynchronous serial ports and is deposited to disk. The system is written in both the C programming language and assembler. It consists of three parts: a multitasking kernel for data collection, a shell with pull down windows as user interface, and a graphics processor for editing data and creating coded messages. An explanation of both system principles and program structure is presented.

Growth and scintillation properties of Eu doped BaCl2/LiF eutectic scintillator

NASA Astrophysics Data System (ADS)

Kamada, Kei; Hishinuma, Kosuke; Kurosawa, Shunsuke; Yamaji, Akihiro; Shoji, Yasuhiro; Pejchal, Jan; Yokota, Yuui; Ohashi, Yuji; Yoshikawa, Akira

2015-12-01

Eu doped BaCl2/LiF eutectics were grown by the micro-pulling down method and their directionally solidified eutectic (DSE) system has been investigated. The grown eutectic showed main phases of cubic LiF and orthorhombic BaCl2. In these eutectics, the 399 nm emission of Eu2+ 4f5d was obtained. It shows the intrinsic decay time of about 410 ns. The light yield of the 1-mm-thick eutectic showed 7000 ph/5.5 MeV alpha-ray.

Optimized Ce:LiCAF amplifier pumping configurations

NASA Astrophysics Data System (ADS)

Cadatal-Raduban, Marilou; Pham, Minh Hong; Gabayno, Jacque Lynn; Yamanoi, Kohei; Empizo, Melvin John F.; Shimizu, Toshihiko; Sarukura, Nobuhiko; Nguyen, Hung Dai; Yoshikawa, Akira; Fukuda, Tsuguo

2018-02-01

Two side-pumping schemes suitable for the development of an ultraviolet femtosecond amplifier system using a Ce3+:LiCaAlF6 crystal are reported. Firstly, a Bethune-type prismatic cell configuration that uniformly illuminates the four sides of a micro-pulling down method-grown crystal is used to amplify 290 nm, femtosecond pulses with no significant increase in pulse duration and B-integral. The second pumping scheme uses a two-side-pumped large crystal. These two side-pumping schemes can pave new possibilities for achieving high-energy ultraviolet femtosecond pulses.

A mix-and-measure assay for determining the activation status of endogenous Cdc42 in cytokine-stimulated macrophage cell lysates.

PubMed

Miskolci, Veronika; Spiering, Désirée; Cox, Dianne; Hodgson, Louis

2014-01-01

Cytokine stimulations of leukocytes many times result in transient activation of the p21 Rho family of small GTPases. The role of these molecules during cell migration and chemotaxis is well established. The traditional approach to study the activation dynamics of these proteins involves affinity pull-downs that are often cumbersome and prone to errors. Here, we describe a reagent and a method of simple "mix-and-measure" approach useful for determining the activation status of endogenous Cdc42 GTPase from cell lysates.

Accelerator and reactor complementarity in coherent neutrino-nucleus scattering

NASA Astrophysics Data System (ADS)

Dent, James B.; Dutta, Bhaskar; Liao, Shu; Newstead, Jayden L.; Strigari, Louis E.; Walker, Joel W.

2018-02-01

We study the complementarity between accelerator and reactor coherent elastic neutrino-nucleus elastic scattering (CE ν NS ) experiments for constraining new physics in the form of nonstandard neutrino interactions (NSI). First, considering just data from the recent observation by the Coherent experiment, we explore interpretive degeneracies that emerge when activating either two or four unknown NSI parameters. Next, we demonstrate that simultaneous treatment of reactor and accelerator experiments, each employing at least two distinct target materials, can break a degeneracy between up and down flavor-diagonal NSI terms that survives analysis of neutrino oscillation experiments. Considering four flavor-diagonal (e e /μ μ ) up- and down-type NSI parameters, we find that all terms can be measured with high local precision (to a width as small as ˜5 % in Fermi units) by next-generation experiments, although discrete reflection ambiguities persist.

Upper body push and pull strength ratio in recreationally active adults.

PubMed

Negrete, Rodney J; Hanney, William J; Pabian, Patrick; Kolber, Morey J

2013-04-01

Agonist to antagonist strength data is commonly analyzed due to its association with injury and performance. The purpose of this study was to examine the agonist to antagonist ratio of upper body strength using two simple field tests (timed push up/timed modified pull up) in recreationally active adults and to establish the basis for reference standards. One hundred eighty (180) healthy recreationally active adults (111 females and 69 males, aged 18-45 years) performed two tests of upper body strength in random order: 1. Push-ups completed during 3 sets of 15 seconds with a 45 second rest period between each set and 2. Modified pull-ups completed during 3 sets of 15 seconds with a 45 second rest period between each set. The push-up to modified pull-up ratio for the males was 1.57:1, whereas females demonstrated a ratio of 2.72:1. The results suggest that for our group of healthy recreationally active subjects, the upper body "pushing" musculature is approximately 1.5-2.7 times stronger than the musculature involved for pulling. In this study, these recreationally active adults displayed greater strength during the timed push-ups than the modified pull-ups. The relationship of these imbalances to one's performance and or injury risk requires further investigation. The reference values, however, may serve the basis for future comparison and prospective investigations. The field tests in this study can be easily implemented by clinicians and an agonist/antagonist ratio can be determined and compared to our findings. 2b.

Uncovering a role for the tail of the Dictyostelium discoideum SadA protein in cell-substrate adhesion.

PubMed

Kowal, Anthony S; Chisholm, Rex L

2011-05-01

Previous work from our laboratory showed that the Dictyostelium discoideum SadA protein plays a central role in cell-substrate adhesion. SadA null cells exhibit a loss of adhesion, a disrupted actin cytoskeleton, and a cytokinesis defect. How SadA mediates these phenotypes is unknown. This work addresses the mechanism of SadA function, demonstrating an important role for the C-terminal cytoplasmic tail in SadA function. We found that a SadA tailless mutant was unable to rescue the sadA adhesion deficiency, and overexpression of the SadA tail domain reduced adhesion in wild-type cells. We also show that SadA is closely associated with the actin cytoskeleton. Mutagenesis studies suggested that four serine residues in the tail, S924/S925 and S940/S941, may regulate association of SadA with the actin cytoskeleton. Glutathione S-transferase pull-down assays identified at least one likely interaction partner of the SadA tail, cortexillin I, a known actin bundling protein. Thus, our data demonstrate an important role for the carboxy-terminal cytoplasmic tail in SadA function and strongly suggest that a phosphorylation event in this tail regulates an interaction with cortexillin I. Based on our data, we propose a model for the function of SadA.

Structural and calorimetric studies demonstrate that the hepatocyte nuclear factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence.

PubMed

Wiedmann, Mareike M; Aibara, Shintaro; Spring, David R; Stewart, Murray; Brenton, James D

2016-09-01

The transcription factor hepatocyte nuclear factor 1β (HNF1β) is ubiquitously overexpressed in ovarian clear cell carcinoma (CCC) and is a potential therapeutic target. To explore potential approaches that block HNF1β transcription we have identified and characterised extensively the nuclear localisation signal (NLS) for HNF1β and its interactions with the nuclear protein import receptor, Importin-α. Pull-down assays demonstrated that the DNA binding domain of HNF1β interacted with a spectrum of Importin-α isoforms and deletion constructs tagged with eGFP confirmed that the HNF1β (229)KKMRRNR(235) sequence was essential for nuclear localisation. We further characterised the interaction between the NLS and Importin-α using complementary biophysical techniques and have determined the 2.4Å resolution crystal structure of the HNF1β NLS peptide bound to Importin-α. The functional, biochemical, and structural characterisation of the nuclear localisation signal present on HNF1β and its interaction with the nuclear import protein Importin-α provide the basis for the development of compounds targeting transcription factor HNF1β via its nuclear import pathway. Copyright © 2016. Published by Elsevier Inc.

Cell-to-cell movement of Alfalfa mosaic virus can be mediated by the movement proteins of Ilar-, bromo-, cucumo-, tobamo- and comoviruses and does not require virion formation.

PubMed

Sánchez-Navarro, Jesús A; Carmen Herranz, María; Pallás, Vicente

2006-03-01

RNA 3 of Alfalfa mosaic virus (AMV) encodes the movement protein (MP) and coat protein (CP). Chimeric RNA 3 with the AMV MP gene replaced by the corresponding MP gene of Prunus necrotic ringspot virus, Brome mosaic virus, Cucumber mosaic virus or Cowpea mosaic virus efficiently moved from cell-to-cell only when the expressed MP was extended at its C-terminus with the C-terminal 44 amino acids of AMV MP. MP of Tobacco mosaic virus supported the movement of the chimeric RNA 3 whether or not the MP was extended with the C-terminal AMV MP sequence. The replacement of the CP gene in RNA 3 by a mutant gene encoding a CP defective in virion formation did not affect cell-to-cell transport of the chimera's with a functional MP. A GST pull-down technique was used to demonstrate for the first time that the C-terminal 44 amino acids of the MP of a virus belonging to the family Bromoviridae interact specifically with AMV virus particles. Together, these results demonstrate that AMV RNA 3 can be transported from cell-to-cell by both tubule-forming and non-tubule-forming MPs if a specific MP-CP interaction occurs.

Addressing Production System Failures Using Multi-agent Control

NASA Astrophysics Data System (ADS)

Gautam, Rajesh; Miyashita, Kazuo

Output in high-volume production facilities is limited by bottleneck machines. We propose a control mechanism by modeling workstations as agents that pull jobs from other agents based on their current WIP level and requirements. During failures, when flows of some jobs are disrupted, the agents pull alternative jobs to maintain utilization of their capacity at a high level. In this paper, we empirically demonstrate that the proposed mechanism can react to failures more appropriately than other control mechanisms using a benchmark problem of a semiconductor manufacturing process.

γ-Adducin Stimulates the Thiazide-sensitive NaCl Cotransporter

PubMed Central

Dimke, Henrik; San-Cristobal, Pedro; de Graaf, Mark; Lenders, Jacques W.; Deinum, Jaap; Hoenderop, Joost G.J.

2011-01-01

The thiazide-sensitive NaCl cotransporter (NCC) plays a key role in renal salt reabsorption and the determination of systemic BP, but the molecular mechanisms governing the regulation of NCC are not completely understood. Here, through pull-down experiments coupled to mass spectrometry, we found that γ-adducin interacts with the NCC transporter. γ-Adducin colocalized with NCC to the distal convoluted tubule. 22Na+ uptake experiments in the Xenopus laevis oocyte showed that γ-adducin stimulated NCC activity in a dose-dependent manner, an effect that occurred upstream from With No Lysine (WNK) 4 kinase. The binding site of γ-adducin mapped to the N terminus of NCC and encompassed three previously reported phosphorylation sites. Supporting this site of interaction, competition with the N-terminal domain of NCC abolished the stimulatory effect of γ-adducin on the transporter. γ-Adducin failed to increase NCC activity when these phosphorylation sites were constitutively inactive or active. In addition, γ-adducin bound only to the dephosphorylated N terminus of NCC. Taken together, our observations suggest that γ-adducin dynamically regulates NCC, likely by amending the phosphorylation state, and consequently the activity, of the transporter. These data suggest that γ-adducin may influence BP homeostasis by modulating renal NaCl transport. PMID:21164023

Seafloor Construction Experiment, SEACON II An Instrumented Tri-Moor for Evaluating Undersea Cable Structure Technology

DTIC Science & Technology

1976-12-01

ik’sigi. of undcruater cattle arrays wt’uld opertioal onsraitsbe the primari goali Arr. consitruction technolog% deseclopmcnt %%A% a %ccont!aro goal...weight of 12,500 pounds. struction mooring anchor was pulled out while load The anchor is composed of a 7-foot by 8-foot by and displacement were...out of the bottom. In contrast, anchor AI pulled out to 27,000 pounds for the construction moor anchor. at a load of 3,500 pounds although a 10,000

The temperature and tension characteristics of the FBGs embedded in the polythene sheath of an optical cable

NASA Astrophysics Data System (ADS)

Chen, Guanghui; Zhao, Ming; Sha, Jianbo; Zhang, Jun; Wu, Bingyan; Lin, Chen; Zhang, Mingliang; Gao, Kan

2015-10-01

The five of FBG were embedded in the PE sheath of a tether optical cable, which has about 18mm diameter and 7000mm length. The temperature and tension characteristics of the FBGs embedded in the polythene (PE) sheath had been demonstrated quantitatively. The Bragg wavelength of the embedded FBG shift linearly with the change of pulling force loaded on the tether optical cable and its tension sensitivity is about 3.75 pm/kg. The results of temperature experiment suggest the embedded FBG have been sensitized by PE material, so that its temperature sensitivity increase from 9.37pm/°C to about 12.51pm/°C.

Mechanical modeling and characteristic study for the adhesive contact of elastic layered media

NASA Astrophysics Data System (ADS)

Zhang, Yuyan; Wang, Xiaoli; Tu, Qiaoan; Sun, Jianjun; Ma, Chenbo

2017-11-01

This paper investigates the adhesive contact between a smooth rigid sphere and a smooth elastic layered medium with different layer thicknesses, layer-to-substrate elastic modulus ratios and adhesion energy ratios. A numerical model is established by combining elastic responses of the contact system and an equation of equivalent adhesive contact pressure which is derived based on the Hamaker summation method and the Lennard-Jones intermolecular potential law. Simulation results for hard layer cases demonstrate that variation trends of the pull-off force with the layer thickness and elastic modulus ratio are complex. On one hand, when the elastic modulus ratio increases, the pull-off force decreases at smaller layer thicknesses, decreases at first and then increases at middle layer thicknesses, while increases monotonously at larger layer thicknesses. On the other hand, the pull-off force decreases at first and then increases with the increase in the layer thickness. Furthermore, a critical layer thickness above which the introduction of hard layer cannot reduce adhesion and an optimum layer thickness under which the pull-off force reaches a minimum are found. Both the critical and optimum layer thicknesses become larger with an increase in the Tabor parameter, while they tend to decrease with the increase in the elastic modulus ratio. In addition, the pull-off force increases sublinearly with the adhesion energy ratio if the layer thickness and elastic modulus ratio are fixed.

Designing for diffusion: how can we increase uptake of cancer communication innovations?

PubMed

Dearing, James W; Kreuter, Matthew W

2010-12-01

The best innovations in cancer communication do not necessarily achieve uptake by researchers, public health and clinical practitioners, and policy makers. This paper describes design activities that can be applied and combined for the purpose of spreading effective cancer communication innovations. A previously developed Push-Pull-Infrastructure Model is used to organize and highlight the types of activities that can be deployed during the design phase of innovations. Scientific literature about the diffusion of innovations, knowledge utilization, marketing, public health, and our experiences in working to spread effective practices, programs, and policies are used for this purpose. Attempts to broaden the reach, quicken the uptake, and facilitate the use of cancer communication innovations can apply design activities to increase the likelihood of diffusion. Some simple design activities hold considerable promise for improving dissemination and subsequent diffusion. Augmenting current dissemination practices with evidence-based concepts from diffusion science, marketing science, and knowledge utilization hold promise for improving results by eliciting greater market pull. Inventors and change agencies seeking to spread cancer communication innovations can experience more success by explicit consideration of design activities that reflect an expanded version of the Push-Pull-Infrastructure Model. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

The Origin of Monsoon Onset. Part 2; Rotational ITCZ Attractors

NASA Technical Reports Server (NTRS)

Chao, Winston C.; Chen, Baode; Einaudi, Franco (Technical Monitor)

2001-01-01

Through various specially designed numerical experiments with an aqua-planet general circulation model and theoretical arguments. Chao showed the existence of multiple quasi-equilibria of the intertropical convergence zone (ITCZ). He also showed that monsoon onset could be interpreted as an abrupt transition between the quasi-equilibria of the ITCZ. He further showed that the origin of these quasi-equilibria is related to two different types of attraction pulling the ITCZ in opposite directions. One type of attraction on the ITCZ is due to earth's rotation, which pulls the ITCZ toward the equator or two equatorial latitudes symmetric with respect to the equator depending on the choice of convection scheme, and the other due to the peak of the sea surface temperature (SST, which is given in the experiments a Gaussian profile in latitude and is uniform in longitude), which pulls the ITCZ toward a latitude just poleward of the SST peak. The strength of the attraction due to the earth's rotation has a highly nonlinear dependence on the latitude and that due to the SST peak has a linear (at least in a relative sense) dependence on the latitude.

Designing for Diffusion: How Can We Increase Uptake of Cancer Communication Innovations?

PubMed Central

Dearing, James W.; Kreuter, Matthew W.

2010-01-01

Objective The best innovations in cancer communication do not necessarily achieve uptake by researchers, public health and clinical practitioners, and policy makers. This paper describes design activities that can be applied and combined for the purpose of spreading effective cancer communication innovations. Methods A previously developed Push-Pull-Infrastructure Model is used to organize and highlight the types of activities that can be deployed during the design phase of innovations. Scientific literature about the diffusion of innovations, knowledge utilization, marketing, public health, and our experiences in working to spread effective practices, programs, and policies are used for this purpose. Results Attempts to broaden the reach, quicken the uptake, and facilitate the use of cancer communication innovations can apply design activities to increase the likelihood of diffusion. Some simple design activities hold considerable promise for improving dissemination and subsequent diffusion. Conclusion Augmenting current dissemination practices with evidence-based concepts from diffusion science, marketing science, and knowledge utilization hold promise for improving results by eliciting greater market pull. Practice Implications Inventors and change agencies seeking to spread cancer communication innovations can experience more success by explicit consideration of design activities that reflect an expanded version of the Push-Pull-Infrastructure Model. PMID:21067884

A Single-Cell Biochemistry Approach Reveals PAR Complex Dynamics during Cell Polarization.

PubMed

Dickinson, Daniel J; Schwager, Francoise; Pintard, Lionel; Gotta, Monica; Goldstein, Bob

2017-08-21

Regulated protein-protein interactions are critical for cell signaling, differentiation, and development. For the study of dynamic regulation of protein interactions in vivo, there is a need for techniques that can yield time-resolved information and probe multiple protein binding partners simultaneously, using small amounts of starting material. Here we describe a single-cell protein interaction assay. Single-cell lysates are generated at defined time points and analyzed using single-molecule pull-down, yielding information about dynamic protein complex regulation in vivo. We established the utility of this approach by studying PAR polarity proteins, which mediate polarization of many animal cell types. We uncovered striking regulation of PAR complex composition and stoichiometry during Caenorhabditis elegans zygote polarization, which takes place in less than 20 min. PAR complex dynamics are linked to the cell cycle by Polo-like kinase 1 and govern the movement of PAR proteins to establish polarity. Our results demonstrate an approach to study dynamic biochemical events in vivo. Copyright © 2017 Elsevier Inc. All rights reserved.

SRC-2 orchestrates polygenic inputs for fine-tuning glucose homeostasis

PubMed Central

Fleet, Tiffany; Zhang, Bin; Lin, Fumin; Zhu, Bokai; Dasgupta, Subhamoy; Stashi, Erin; Tackett, Bryan; Thevananther, Sundararajah; Rajapakshe, Kimal I.; Gonzales, Naomi; Dean, Adam; Mao, Jianqiang; Timchenko, Nikolai; Malovannaya, Anna; Qin, Jun; Coarfa, Cristian; DeMayo, Francesco; Dacso, Clifford C.; Foulds, Charles E.; O’Malley, Bert W.; York, Brian

2015-01-01

Despite extensive efforts to understand the monogenic contributions to perturbed glucose homeostasis, the complexity of genetic events that fractionally contribute to the spectrum of this pathology remain poorly understood. Proper maintenance of glucose homeostasis is the central feature of a constellation of comorbidities that define the metabolic syndrome. The ability of the liver to balance carbohydrate uptake and release during the feeding-to-fasting transition is essential to the regulation of peripheral glucose availability. The liver coordinates the expression of gene programs that control glucose absorption, storage, and secretion. Herein, we demonstrate that Steroid Receptor Coactivator 2 (SRC-2) orchestrates a hierarchy of nutritionally responsive transcriptional complexes to precisely modulate plasma glucose availability. Using DNA pull-down technology coupled with mass spectrometry, we have identified SRC-2 as an indispensable integrator of transcriptional complexes that control the rate-limiting steps of hepatic glucose release and accretion. Collectively, these findings position SRC-2 as a major regulator of polygenic inputs to metabolic gene regulation and perhaps identify a previously unappreciated model that helps to explain the clinical spectrum of glucose dysregulation. PMID:26487680

Self-Phase-Matched Second-Harmonic and White-Light Generation in a Biaxial Zinc Tungstate Single Crystal

PubMed Central

Osewski, Pawel; Belardini, Alessandro; Petronijevic, Emilija; Centini, Marco; Leahu, Grigore; Diduszko, Ryszard; Pawlak, Dorota A.; Sibilia, Concita

2017-01-01

Second-order nonlinear optical materials are used to generate new frequencies by exploiting second-harmonic generation (SHG), a phenomenon where a nonlinear material generates light at double the optical frequency of the input beam. Maximum SHG is achieved when the pump and the generated waves are in phase, for example through birefringence in uniaxial crystals. However, applying these materials usually requires a complicated cutting procedure to yield a crystal with a particular orientation. Here we demonstrate the first example of phase matching under the normal incidence of SHG in a biaxial monoclinic single crystal of zinc tungstate. The crystal was grown by the micro-pulling-down method with the (102) plane perpendicular to the growth direction. Additionally, at the same time white light was generated as a result of stimulated Raman scattering and multiphoton luminescence induced by higher-order effects such as three-photon luminescence enhanced by cascaded third-harmonic generation. The annealed crystal offers SHG intensities approximately four times larger than the as grown one; optimized growth and annealing conditions may lead to much higher SHG intensities. PMID:28338074

Cellular Hsp27 interacts with classical swine fever virus NS5A protein and negatively regulates viral replication by the NF-κB signaling pathway.

PubMed

Ling, Shifeng; Luo, Mingyang; Jiang, Shengnan; Liu, Jiayu; Ding, Chunying; Zhang, Qinghuan; Guo, Huancheng; Gong, Wenjie; Tu, Changchun; Sun, Jinfu

2018-05-01

Classical swine fever virus (CSFV) nonstructural protein NS5A is a multifunctional protein functioning in regulation of viral genome replication, protein translation and assembly by interaction with viral or host proteins. Here, heat shock protein 27 (Hsp27) has been identified as a novel binding partner of NS5A by using His tag "pull down" coupled with shotgun LC-MS/MS, with interaction of both proteins further confirmed by co-immunoprecipitation and laser confocal assays. In PK-15 cells, silencing of Hsp27 expression by siRNA enhanced CSFV replication, and upregulation of Hsp27 inhibited viral proliferation. Additionally, we have shown that overexpression of Hsp27 increased NF-κB signaling induced by TNFα. Blocking NF-κB signaling in PK-15 cells overexpressing Hsp27 by ammonium pyrrolidinedithiocarbamate (PDTC) eliminated the inhibition of CSFV replication by Hsp27. These findings clearly demonstrate that the inhibition of CSFV replication by Hsp27 is mediated via the NF-κB signaling pathway. Copyright © 2018 Elsevier Inc. All rights reserved.

Self-Phase-Matched Second-Harmonic and White-Light Generation in a Biaxial Zinc Tungstate Single Crystal

NASA Astrophysics Data System (ADS)

Osewski, Pawel; Belardini, Alessandro; Petronijevic, Emilija; Centini, Marco; Leahu, Grigore; Diduszko, Ryszard; Pawlak, Dorota A.; Sibilia, Concita

2017-03-01

Second-order nonlinear optical materials are used to generate new frequencies by exploiting second-harmonic generation (SHG), a phenomenon where a nonlinear material generates light at double the optical frequency of the input beam. Maximum SHG is achieved when the pump and the generated waves are in phase, for example through birefringence in uniaxial crystals. However, applying these materials usually requires a complicated cutting procedure to yield a crystal with a particular orientation. Here we demonstrate the first example of phase matching under the normal incidence of SHG in a biaxial monoclinic single crystal of zinc tungstate. The crystal was grown by the micro-pulling-down method with the (102) plane perpendicular to the growth direction. Additionally, at the same time white light was generated as a result of stimulated Raman scattering and multiphoton luminescence induced by higher-order effects such as three-photon luminescence enhanced by cascaded third-harmonic generation. The annealed crystal offers SHG intensities approximately four times larger than the as grown one; optimized growth and annealing conditions may lead to much higher SHG intensities.

Cdc45-induced loading of human RPA onto single-stranded DNA.

PubMed

Szambowska, Anna; Tessmer, Ingrid; Prus, Piotr; Schlott, Bernhard; Pospiech, Helmut; Grosse, Frank

2017-04-07

Cell division cycle protein 45 (Cdc45) is an essential component of the eukaryotic replicative DNA helicase. We found that human Cdc45 forms a complex with the single-stranded DNA (ssDNA) binding protein RPA. Moreover, it actively loads RPA onto nascent ssDNA. Pull-down assays and surface plasmon resonance studies revealed that Cdc45-bound RPA complexed with ssDNA in the 8-10 nucleotide binding mode, but dissociated when RPA covered a 30-mer. Real-time analysis of RPA-ssDNA binding demonstrated that Cdc45 catalytically loaded RPA onto ssDNA. This placement reaction required physical contacts of Cdc45 with the RPA70A subdomain. Our results imply that Cdc45 controlled stabilization of the 8-nt RPA binding mode, the subsequent RPA transition into 30-mer mode and facilitated an ordered binding to ssDNA. We propose that a Cdc45-mediated loading guarantees a seamless deposition of RPA on newly emerging ssDNA at the nascent replication fork. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Simple methods for the 3' biotinylation of RNA.

PubMed

Moritz, Bodo; Wahle, Elmar

2014-03-01

Biotinylation of RNA allows its tight coupling to streptavidin and is thus useful for many types of experiments, e.g., pull-downs. Here we describe three simple techniques for biotinylating the 3' ends of RNA molecules generated by chemical or enzymatic synthesis. First, extension with either the Schizosaccharomyces pombe noncanonical poly(A) polymerase Cid1 or Escherichia coli poly(A) polymerase and N6-biotin-ATP is simple, efficient, and generally applicable independently of the 3'-end sequences of the RNA molecule to be labeled. However, depending on the enzyme and the reaction conditions, several or many biotinylated nucleotides are incorporated. Second, conditions are reported under which splint-dependent ligation by T4 DNA ligase can be used to join biotinylated and, presumably, other chemically modified DNA oligonucleotides to RNA 3' ends even if these are heterogeneous as is typical for products of enzymatic synthesis. Third, we describe the use of 29 DNA polymerase for a template-directed fill-in reaction that uses biotin-dUTP and, thanks to the enzyme's proofreading activity, can cope with more extended 3' heterogeneities.

Mapping transcription factor interactome networks using HaloTag protein arrays.

PubMed

Yazaki, Junshi; Galli, Mary; Kim, Alice Y; Nito, Kazumasa; Aleman, Fernando; Chang, Katherine N; Carvunis, Anne-Ruxandra; Quan, Rosa; Nguyen, Hien; Song, Liang; Alvarez, José M; Huang, Shao-Shan Carol; Chen, Huaming; Ramachandran, Niroshan; Altmann, Stefan; Gutiérrez, Rodrigo A; Hill, David E; Schroeder, Julian I; Chory, Joanne; LaBaer, Joshua; Vidal, Marc; Braun, Pascal; Ecker, Joseph R

2016-07-19

Protein microarrays enable investigation of diverse biochemical properties for thousands of proteins in a single experiment, an unparalleled capacity. Using a high-density system called HaloTag nucleic acid programmable protein array (HaloTag-NAPPA), we created high-density protein arrays comprising 12,000 Arabidopsis ORFs. We used these arrays to query protein-protein interactions for a set of 38 transcription factors and transcriptional regulators (TFs) that function in diverse plant hormone regulatory pathways. The resulting transcription factor interactome network, TF-NAPPA, contains thousands of novel interactions. Validation in a benchmarked in vitro pull-down assay revealed that a random subset of TF-NAPPA validated at the same rate of 64% as a positive reference set of literature-curated interactions. Moreover, using a bimolecular fluorescence complementation (BiFC) assay, we confirmed in planta several interactions of biological interest and determined the interaction localizations for seven pairs. The application of HaloTag-NAPPA technology to plant hormone signaling pathways allowed the identification of many novel transcription factor-protein interactions and led to the development of a proteome-wide plant hormone TF interactome network.

A mammalian germ cell-specific RNA-binding protein interacts with ubiquitously expressed proteins involved in splice site selection

NASA Astrophysics Data System (ADS)

Elliott, David J.; Bourgeois, Cyril F.; Klink, Albrecht; Stévenin, James; Cooke, Howard J.

2000-05-01

RNA-binding motif (RBM) genes are found on all mammalian Y chromosomes and are implicated in spermatogenesis. Within human germ cells, RBM protein shows a similar nuclear distribution to components of the pre-mRNA splicing machinery. To address the function of RBM, we have used protein-protein interaction assays to test for possible physical interactions between these proteins. We find that RBM protein directly interacts with members of the SR family of splicing factors and, in addition, strongly interacts with itself. We have mapped the protein domains responsible for mediating these interactions and expressed the mouse RBM interaction region as a bacterial fusion protein. This fusion protein can pull-down several functionally active SR protein species from cell extracts. Depletion and add-back experiments indicate that these SR proteins are the only splicing factors bound by RBM which are required for the splicing of a panel of pre-mRNAs. Our results suggest that RBM protein is an evolutionarily conserved mammalian splicing regulator which operates as a germ cell-specific cofactor for more ubiquitously expressed pre-mRNA splicing activators.

Preparative two-step purification of recombinant H1.0 linker histone and its domains.

PubMed

Ivic, Nives; Bilokapic, Silvija; Halic, Mario

2017-01-01

H1 linker histones are small basic proteins that have a key role in the formation and maintenance of higher-order chromatin structures. Additionally, many examples have shown that linker histones play an important role in gene regulation, modulated by their various subtypes and posttranslational modifications. Obtaining high amounts of very pure linker histones, especially for efficient antibody production, remains a demanding and challenging procedure. Here we present an easy and fast method to purify human linker histone H1.0 overexpressed in Escherichia coli, as well as its domains: N-terminal/globular domain and C-terminal intrinsically disordered domain. This purification protocol relies on a simple affinity chromatography step followed by cation exchange due to the highly basic properties of histone proteins. Therefore, this protocol can also be applied to other linker histones. Highly pure proteins in amounts sufficient for most biochemical experiments can be obtained. The functional quality of purified H1.0 histone and its domains has been confirmed by pull-down, gel-mobility shift assays and the nuclear import assay.

Targeting the r(CGG) repeats that cause FXTAS with modularly assembled small molecules and oligonucleotides.

PubMed

Tran, Tuan; Childs-Disney, Jessica L; Liu, Biao; Guan, Lirui; Rzuczek, Suzanne; Disney, Matthew D

2014-04-18

We designed small molecules that bind the structure of the RNA that causes fragile X-associated tremor ataxia syndrome (FXTAS), an incurable neuromuscular disease. FXTAS is caused by an expanded r(CGG) repeat (r(CGG)(exp)) that inactivates a protein regulator of alternative pre-mRNA splicing. Our designed compounds modulate r(CGG)(exp) toxicity in cellular models of FXTAS, and pull-down experiments confirm that they bind r(CGG)(exp) in vivo. Importantly, compound binding does not affect translation of the downstream open reading frame (ORF). We compared molecular recognition properties of our optimal compound to oligonucleotides. Studies show that r(CGG)(exp)'s self-structure is a significant energetic barrier for oligonucleotide binding. A fully modified 2'-OMethyl phosphorothioate is incapable of completely reversing an FXTAS-associated splicing defect and inhibits translation of the downstream ORF, which could have deleterious effects. Taken together, these studies suggest that a small molecule that recognizes structure may be more well suited for targeting highly structured RNAs that require strand invasion by a complementary oligonucleotide.

Targeting the r(CGG) Repeats That Cause FXTAS with Modularly Assembled Small Molecules and Oligonucleotides

PubMed Central

2015-01-01

We designed small molecules that bind the structure of the RNA that causes fragile X-associated tremor ataxia syndrome (FXTAS), an incurable neuromuscular disease. FXTAS is caused by an expanded r(CGG) repeat (r(CGG)exp) that inactivates a protein regulator of alternative pre-mRNA splicing. Our designed compounds modulate r(CGG)exp toxicity in cellular models of FXTAS, and pull-down experiments confirm that they bind r(CGG)expin vivo. Importantly, compound binding does not affect translation of the downstream open reading frame (ORF). We compared molecular recognition properties of our optimal compound to oligonucleotides. Studies show that r(CGG)exp’s self-structure is a significant energetic barrier for oligonucleotide binding. A fully modified 2′-OMethyl phosphorothioate is incapable of completely reversing an FXTAS-associated splicing defect and inhibits translation of the downstream ORF, which could have deleterious effects. Taken together, these studies suggest that a small molecule that recognizes structure may be more well suited for targeting highly structured RNAs that require strand invasion by a complementary oligonucleotide. PMID:24506227

Identification of a new Mpl-interacting protein, Atp5d.

PubMed

Liu, Hongyan; Zhao, Zhenhu; Zhong, Yuxu; Shan, Yajun; Sun, Xiaohong; Mao, Bingzhi; Cong, Yuwen

2014-06-01

Thrombopoietin (TPO) can regulate hematopoiesis and megakaryopoiesis via activation of its receptor, c-Mpl, and multiple downstream signal transduction pathways. Using the cytoplasmic domain of Mpl as bait, we performed yeast two-hybrid screening, and found that the protein Atp5d might associate with Mpl. Atp5d is known as the δ subunit of mitochondrial ATP synthase, but little is known about the function of dissociative Atp5d. The interaction between Mpl and Atp5d was confirmed by the yeast two-hybrid system, mammalian two-hybrid assay, pull-down experiment, and co-immunoprecipitation study in vivo and in vitro. An additional immunofluorescence assay showed that the two proteins can colocalize along the plasma membrane in the cytoplasm. Using the yeast two-hybrid system, we tested a series of cytoplasmic truncated mutations for their ability to bind Atp5d and found an association between Atp5d and the Aa98-113 domain of Mpl. The dissociation of Atp5d from Mpl after TPO stimulation suggests that Atp5d may be a new component of TPO signaling.

Interaction of the SPG21 protein ACP33/maspardin with the aldehyde dehydrogenase ALDH16A1

PubMed Central

2017-01-01

Mast syndrome (SPG21) is an autosomal-recessive complicated form of hereditary spastic paraplegia characterized by dementia, thin corpus callosum, white matter abnormalities, and cerebellar and extrapyramidal signs in addition to spastic paraparesis. A nucleotide insertion resulting in premature truncation of the SPG21 gene product acidic cluster protein 33 (ACP33)/maspardin underlies this disorder, likely causing loss of protein function. However, little is known about the function of maspardin. Here, we report that maspardin localizes prominently to cytoplasm as well as to membranes, possibly at trans-Golgi network/late endosomal compartments. Immunoprecipitation of maspardin with identification of coprecipitating proteins by mass spectrometry revealed the aldehyde dehydrogenase ALDH16A1 as an interacting protein. This interaction was confirmed using overexpressed proteins as well as by fusion protein pull down experiments, and these proteins colocalized in cells. Further studies of the function of ALDH16A1 and the role of the maspardin–ALDH16A1 interaction in neuronal cells may clarify the cellular pathogenesis of Mast syndrome. PMID:19184135

Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin β1

PubMed Central

Klockenbusch, Cordula; Kast, Juergen

2010-01-01

Formaldehyde cross-linking of protein complexes combined with immunoprecipitation and mass spectrometry analysis is a promising technique for analysing protein-protein interactions, including those of transient nature. Here we used integrin β1 as a model to describe the application of formaldehyde cross-linking in detail, particularly focusing on the optimal parameters for cross-linking, the detection of formaldehyde cross-linked complexes, the utility of antibodies, and the identification of binding partners. Integrin β1 was found in a high molecular weight complex after formaldehyde cross-linking. Eight different anti-integrin β1 antibodies were used for pull-down experiments and no loss in precipitation efficiency after cross-linking was observed. However, two of the antibodies could not precipitate the complex, probably due to hidden epitopes. Formaldehyde cross-linked complexes, precipitated from Jurkat cells or human platelets and analyzed by mass spectrometry, were found to be composed of integrin β1, α4 and α6 or β1, α6, α2, and α5, respectively. PMID:20634879

Puncture-and-Pull Biomechanics in the Teeth of Predatory Coelurosaurian Dinosaurs.

PubMed

Torices, Angelica; Wilkinson, Ryan; Arbour, Victoria M; Ruiz-Omeñaca, Jose Ignacio; Currie, Philip J

2018-05-07

The teeth of putatively carnivorous dinosaurs are often blade-shaped with well-defined serrated cutting edges (Figure 1). These ziphodont teeth are often easily differentiated based on the morphology and density of the denticles [1, 2]. A tearing function has been proposed for theropod denticles in general [3], but the functional significance of denticle phenotypic variation has received less attention. In particular, the unusual hooked denticles found in troodontids suggest a different feeding strategy or diet compared to other small theropods. We used a two-pronged approach to investigate the function of denticle shape variation across theropods with both congruent body shapes and sizes (e.g., dromaeosaurids versus troodontids) and highly disparate body shapes and sizes (e.g., troodontids versus tyrannosaurids), using microwear and finite element analyses (Figure 1). We found that many toothed coelurosaurian theropods employed a puncture-and-pull feeding movement, in which parallel scratches form while biting down into prey and oblique scratches form as the head is pulled backward with the jaws closed. In finite element simulations, theropod teeth had the lowest stresses when bite forces were aligned with the oblique family of microwear scratches. Different denticle morphologies performed differently under a variety of simulated biting angles: Dromaeosaurus and Saurornitholestes were well-adapted for handling struggling prey, whereas troodontid teeth were more likely to fail at non-optimal bite angles. Troodontids may have favored softer, smaller, or immobile prey. Copyright © 2018 Elsevier Ltd. All rights reserved.

Interface modification during oxidation of a glass-ceramic matrix/SiC fibre composite

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

Daniel, A.M.; Martin-Meizoso, A.; Plucknett, K.P.

Oxidation heat treatments between 375{degrees}C and 600{degrees}C for 100 hours in air, have been performed on the calcium aluminosilicate glass-ceramic matrix/SiC fibre reinforced composite CAS/Nicalon (manufactured by Corning, USA). Using a commercial nano-indentation system to perform fibre push-down tests, the fibre-matrix interfacial debond fracture surface energy (G{sub i}) and frictional shear stress ({tau}) have been determined. Modification of interface properties, compared to the as-fabricated material, was observed at heat treatment temperatures as low as 375{degrees}C, where a significant drop in G{sub i} and an increase in {tau} were recorded. With 450{degrees}C, 525{degrees}C and 600{degrees}C heat treatments, an increase in G{submore » i} but a dramatic increase in {tau} were recorded. Under four-point flexure testing, the as fabricated and the 375{degrees}C heat treated materials displayed tough, composite behaviour with extensive fibre pull out, but at {ge}450{degrees}C, brittle failure with minimal fibre pull out, was observed. This transition from tough mechanical response to one of brittleness is due to the large increase in {tau} reducing fibre pull out to a minimum and therefore reducing the total required work of fracture. The large increases in {tau} and G{sub i} have been attributed to the oxidative removal of the lubricating, carbon interface and the compressive residual stresses across the interface.« less

Interface modification during oxidation of a glass-ceramic matrix/SiC fibre composite

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

Daniel, A.M.; Martin-Meizoso, A.; Plucknett, K.P.

Oxidation heat treatments between 375{degrees}C and 600{degrees}C for 100 hours in air, have been performed on the calcium aluminosilicate glass-ceramic matrix/SiC fibre reinforced composite CAS/Nicalon (manufactured by Coming, USA). Using a commercial nano-indentation system to perform fibre push-down tests, the fibre-matrix interfacial debond fracture surface energy (G{sub i}) and frictional shear stress ({tau}) have been determined. Modification of interface properties, compared to the as fabricated material, was observed at heat treatment temperatures as low as 375{degrees}C, where a significant drop in G{sub i} and an increase in {tau} were recorded. With 450{degrees}C, 525{degrees}C and 600{degrees}C heat treatments, an increase inmore » G{sub i} but a dramatic increase in {tau} were recorded. Under four-point flexure testing, the as fabricated and the 375{degrees}C heat treated materials displayed tough, composite behaviour with extensive fibre pull out, but at {le}450{degrees}C, brittle failure with minimal fibre pull out, was observed. This transition from tough mechanical response to one of brittleness is due to the large increase in {tau} reducing fibre pull out to a minimum and therefore reducing the total required work of fracture. The large increases in {tau} and G{sub i} have been attributed to the oxidative removal of the lubricating, carbon interface and the compressive residual stresses across the interface.« less

Dual-Force Vaginoplasty for Treatment of Segmental Vaginal Aplasia.

PubMed

El Saman, Ali M; Farag, Mohamad A; Shazly, Sherif A; Noor, Mohamed; Ali, Mohammed K; Othman, Essam R; Khalifa, Mansour; Farghly, Tarek A; El Saman, Dina A

2017-05-01

Vaginal aplasia occurs in 1 in 5,000-10,000 female live births. In this report, we evaluated a novel dual-force vaginoplasty technique for treatment of 11 patients with segmental vaginal aplasia. The principle of the approach is to thin the atretic part between two counteracting forces. The instrument was inserted laparoscopically into the proximal hematocolpos. Two balloon catheters, one for drainage and one for traction, were threaded over the inserter. The traction catheter was then threaded over a silicon tube, leaving the balloon in the proximal portion of the vagina and connecting across the vaginal septum to a fenestrated Teflon olive, which was positioned against the distal surface of the vaginal septum. This created a dual "pushing and pulling" force across the septum, which, over 3-4 days, pulls the upper vaginal pouch down while the vaginal dimple is pushed up. The aplastic segment becomes thin and easy to dilate and permits achievement of vaginal patency. The drainage of the hematocolpos is predominantly through the balloon catheter so postoperative wound management is facilitated. Eleven menarchal girls were diagnosed with segmental vaginal aplasia. The dual-force vaginoplasty was performed on each and was tolerated well with no operative complications. They all reported establishment of the menstrual cycle and significant improvement of pain during follow-up. Creation of a dual pushing-pulling force on the atretic vaginal segment is a feasible short procedure for management of segmental vaginal aplasia.

Strength and power determinants of grinding performance in America's Cup sailors.

PubMed

Pearson, Simon N; Hume, Patria A; Cronin, John B; Slyfield, David

2009-09-01

The purpose grinding is a physically demanding component of America's Cup sailing that is important to overall team performance, but little research is available on the determinants of grinding performance. We examined the relationship between various measures of muscular performance and the performance of upper-body grinding. Eleven elite male America's Cup sailors (33.9 +/- 5.5 yr, 97.8 +/- 12.5 kg, 186.0 +/- 7.1 cm) who performed grinding as part of their on-board role with extensive strength training experience participated in this study. Muscular performance testing examined the force, velocity, and power capabilities of the upper-body musculature, with upper-body push (bench press) and pull (bench pull) movements performed across loads of 10-100% of 1 repetition maximum (1RM). Functional grinding performance was examined for both forward and backward grinding and at 2 different resistances (moderate = 48 N x m, heavy = 68 N x m) using a land-based ergometer. Bench press 1RM and maximum force capability were the measures demonstrating the strongest correlation with forward grinding performance (r = 0.88-0.99 and 0.87-0.99, respectively), with the relationship increasing with grinding load. For backward grinding, there was a very strong relationship with bench pull maximum power (r = 0.85-0.98) in addition to 1RM (r = 0.90-0.95) and maximum force (r = 0.87-0.95). It appears that although maximal strength is a crucial muscular performance characteristic for grinding performance in all conditions, for backward grinding, there is the additional need to focus on the development of speed strength/power to maximize performance gains. This information was used by the Emirates Team New Zealand physical conditioner to develop a conditioning intervention to help improve grinding performance.

Comprehensive identification of proteins binding to RNA G-quadruplex motifs in the 5' UTR of tumor-associated mRNAs.

PubMed

Serikawa, Tatsuo; Spanos, Christos; von Hacht, Annekathrin; Budisa, Nediljko; Rappsilber, Juri; Kurreck, Jens

2018-01-01

G-quadruplex structures in the 5' UTR of mRNAs are widely considered to suppress translation without affecting transcription. The current study describes the comprehensive analysis of proteins binding to four different G-quadruplex motifs located in mRNAs of the cancer-related genes Bcl-2, NRAS, MMP16, and ARPC2. Following metabolic labeling (Stable Isotope Labeling with Amino acids in Cell culture, SILAC) of proteins in the human cell line HEK293, G-quadruplex binding proteins were enriched by pull-down assays and identified by LC-orbitrap mass spectrometry. We found different patterns of interactions for the G-quadruplex motifs under investigation. While the G-quadruplexes in the mRNAs of NRAS and MMP16 specifically interacted with a small number of proteins, the Bcl-2 and ARPC2 G-quadruplexes exhibited a broad range of proteinaceous interaction partners with 99 and 82 candidate proteins identified in at least two replicates, respectively. The use of a control composed of samples from all G-quadruplex-forming sequences and their mutated controls ensured that the identified proteins are specific for RNA G-quadruplex structures and are not general RNA-binding proteins. Independent validation experiments based on pull-down assays and Western blotting confirmed the MS data. Among the interaction partners were many proteins known to bind to RNA, including multiple heterogenous nuclear ribonucleoproteins (hnRNPs). Several of the candidate proteins are likely to reflect stalling of the ribosome by RNA G-quadruplex structures. Interestingly, additional proteins were identified that have not previously been described to interact with RNA. Gene ontology analysis of the candidate proteins revealed that many interaction partners are known to be tumor related. The majority of the identified RNA G-quadruplex interacting proteins are thought to be involved in post-transcriptional processes, particularly in splicing. These findings indicate that protein-G-quadruplex interactions are not only important for the fine-tuning of translation but are also relevant to the regulation of mRNA maturation and may play an important role in tumor biology. Proteomic data are available via ProteomeXchange with identifier PXD005761. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Analysis of TID process, geometry, and bias condition dependence in 14-nm FinFETs and implications for RF and SRAM performance

DOE PAGES

King, M. P.; Wu, X.; Eller, Manfred; ...

2016-12-07

Here, total ionizing dose results are provided, showing the effects of different threshold adjust implant processes and irradiation bias conditions of 14-nm FinFETs. Minimal radiation-induced threshold voltage shift across a variety of transistor types is observed. Off-state leakage current of nMOSFET transistors exhibits a strong gate bias dependence, indicating electrostatic gate control of the sub-fin region and the corresponding parasitic conduction path are the largest concern for radiation hardness in FinFET technology. The high-Vth transistors exhibit the best irradiation performance across all bias conditions, showing a reasonably small change in off-state leakage current and Vth, while the low-Vth transistors exhibitmore » a larger change in off-state leakage current. The “worst-case” bias condition during irradiation for both pull-down and pass-gate nMOSFETs in static random access memory is determined to be the on-state (Vgs = Vdd). We find the nMOSFET pull-down and pass-gate transistors of the SRAM bit-cell show less radiation-induced degradation due to transistor geometry and channel doping differences than the low-Vth transistor. Near-threshold operation is presented as a methodology for reducing radiation-induced increases in off-state device leakage current. In a 14-nm FinFET technology, the modeling indicates devices with high channel stop doping show the most robust response to TID allowing stable operation of ring oscillators and the SRAM bit-cell with minimal shift in critical operating characteristics.« less

Analysis of TID process, geometry, and bias condition dependence in 14-nm FinFETs and implications for RF and SRAM performance

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

King, M. P.; Wu, X.; Eller, Manfred

Here, total ionizing dose results are provided, showing the effects of different threshold adjust implant processes and irradiation bias conditions of 14-nm FinFETs. Minimal radiation-induced threshold voltage shift across a variety of transistor types is observed. Off-state leakage current of nMOSFET transistors exhibits a strong gate bias dependence, indicating electrostatic gate control of the sub-fin region and the corresponding parasitic conduction path are the largest concern for radiation hardness in FinFET technology. The high-Vth transistors exhibit the best irradiation performance across all bias conditions, showing a reasonably small change in off-state leakage current and Vth, while the low-Vth transistors exhibitmore » a larger change in off-state leakage current. The “worst-case” bias condition during irradiation for both pull-down and pass-gate nMOSFETs in static random access memory is determined to be the on-state (Vgs = Vdd). We find the nMOSFET pull-down and pass-gate transistors of the SRAM bit-cell show less radiation-induced degradation due to transistor geometry and channel doping differences than the low-Vth transistor. Near-threshold operation is presented as a methodology for reducing radiation-induced increases in off-state device leakage current. In a 14-nm FinFET technology, the modeling indicates devices with high channel stop doping show the most robust response to TID allowing stable operation of ring oscillators and the SRAM bit-cell with minimal shift in critical operating characteristics.« less

Identification of Molecular Targets for 4,5-Dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) in Teleosts: New Insight into Mechanism of Toxicity.

PubMed

Chen, Lianguo; Au, Doris W T; Hu, Chenyan; Peterson, Drew R; Zhou, Bingsheng; Qian, Pei-Yuan

2017-02-07

Environmental pollutants are capable of concomitantly inducing diverse toxic effects. However, it is largely unknown which effects are directly induced and which effects are secondary, thus calling for definitive identification of the initiating molecular event for a pollutant to elucidate the mechanism of toxicity. In the present study, affinity pull-down assays were used to identify target proteins for 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT), a costal pollutant of emerging concern, in various tissues (e.g., brain, liver, plasma, and gonad) from marine medaka (Oryzias melastigma) and zebrafish (Danio rerio). Pull-down results showed that, in male and female brains from medaka and zebrafish, DCOIT had a consistently high affinity for G protein alpha subunits (Gα), suggesting the targeted effects of DCOIT on signaling transduction from G protein-coupled receptors (GPCRs) and an extrapolatable mode of action in teleost brains. Validation using recombinant proteins and molecular docking analysis confirmed that binding of DCOIT to Gα protein competitively inhibited its activation by substrate. Considering the involvement of GPCRs in the regulation of myriad biological processes, including the hypothalamus-pituitary-gonadal-liver axis, binding of DCOIT to upstream Gα proteins in the brain may provide a plausible explanation for the diversity of toxic effects resulting from DCOIT challenge, especially abnormal hormonal production through the mitogen-activated protein kinase pathway. A new mechanism of action based on GPCR signaling is thus hypothesized for endocrine disrupting chemicals and warrants further research to clearly elucidate the link between GPCR signaling and endocrine disruption.

Low power, scalable multichannel high voltage controller

DOEpatents

Stamps, James Frederick [Livermore, CA; Crocker, Robert Ward [Fremont, CA; Yee, Daniel Dadwa [Dublin, CA; Dils, David Wright [Fort Worth, TX

2006-03-14

A low voltage control circuit is provided for individually controlling high voltage power provided over bus lines to a multitude of interconnected loads. An example of a load is a drive for capillary channels in a microfluidic system. Control is distributed from a central high voltage circuit, rather than using a number of large expensive central high voltage circuits to enable reducing circuit size and cost. Voltage is distributed to each individual load and controlled using a number of high voltage controller channel switches connected to high voltage bus lines. The channel switches each include complementary pull up and pull down photo isolator relays with photo isolator switching controlled from the central high voltage circuit to provide a desired bus line voltage. Switching of the photo isolator relays is further controlled in each channel switch using feedback from a resistor divider circuit to maintain the bus voltage swing within desired limits. Current sensing is provided using a switched resistive load in each channel switch, with switching of the resistive loads controlled from the central high voltage circuit.

Low power, scalable multichannel high voltage controller

DOEpatents

Stamps, James Frederick [Livermore, CA; Crocker, Robert Ward [Fremont, CA; Yee, Daniel Dadwa [Dublin, CA; Dils, David Wright [Fort Worth, TX

2008-03-25

A low voltage control circuit is provided for individually controlling high voltage power provided over bus lines to a multitude of interconnected loads. An example of a load is a drive for capillary channels in a microfluidic system. Control is distributed from a central high voltage circuit, rather than using a number of large expensive central high voltage circuits to enable reducing circuit size and cost. Voltage is distributed to each individual load and controlled using a number of high voltage controller channel switches connected to high voltage bus lines. The channel switches each include complementary pull up and pull down photo isolator relays with photo isolator switching controlled from the central high voltage circuit to provide a desired bus line voltage. Switching of the photo isolator relays is further controlled in each channel switch using feedback from a resistor divider circuit to maintain the bus voltage swing within desired limits. Current sensing is provided using a switched resistive load in each channel switch, with switching of the resistive loads controlled from the central high voltage circuit.

a High-Level Technique for Estimation and Optimization of Leakage Power for Full Adder

NASA Astrophysics Data System (ADS)

Shrivas, Jayram; Akashe, Shyam; Tiwari, Nitesh

2013-06-01

Optimization of power is a very important issue in low-voltage and low-power application. In this paper, we have proposed power gating technique to reduce leakage current and leakage power of one-bit full adder. In this power gating technique, we use two sleep transistors i.e., PMOS and NMOS. PMOS sleep transistor is inserted between power supply and pull up network. And NMOS sleep transistor is inserted between pull down network and ground terminal. These sleep transistors (PMOS and NMOS) are turned on when the circuit is working in active mode. And sleep transistors (PMOS and NMOS) are turned off when circuit is working in standby mode. We have simulated one-bit full adder and compared with the power gating technique using cadence virtuoso tool in 45 nm technology at 0.7 V at 27°C. By applying this technique, we have reduced leakage current from 2.935 pA to 1.905 pA and leakage power from 25.04μw to 9.233μw. By using this technique, we have reduced leakage power up to 63.12%.

Metal-Ferroelectric-Semiconductor Field-Effect Transistor NAND Gate Switching Time Analysis

NASA Technical Reports Server (NTRS)

Phillips, Thomas A.; Macleod, Todd C.; Ho, Fat D.

2006-01-01

Previous research investigated the modeling of a N Wga te constructed of Metal-Ferroelectric- Semiconductor Field-Effect Transistors (MFSFETs) to obtain voltage transfer curves. The NAND gate was modeled using n-channel MFSFETs with positive polarization for the standard CMOS n-channel transistors and n-channel MFSFETs with negative polarization for the standard CMOS p-channel transistors. This paper investigates the MFSFET NAND gate switching time propagation delay, which is one of the other important parameters required to characterize the performance of a logic gate. Initially, the switching time of an inverter circuit was analyzed. The low-to-high and high-to-low propagation time delays were calculated. During the low-to-high transition, the negatively polarized transistor pulls up the output voltage, and during the high-to-low transition, the positively polarized transistor pulls down the output voltage. The MFSFETs were simulated by using a previously developed model which utilized a partitioned ferroelectric layer. Then the switching time of a 2-input NAND gate was analyzed similarly to the inverter gate. Extension of this technique to more complicated logic gates using MFSFETs will be studied.

Correlation of two bioadhesion assays: the everted sac technique and the CAHN microbalance.

PubMed

Santos, C A; Jacob, J S; Hertzog, B A; Freedman, B D; Press, D L; Harnpicharnchai, P; Mathiowitz, E

1999-08-27

This contribution correlates two in vitro methods utilized to determine bioadhesion. One method, the everted intestinal sac technique, is a passive test for bioadhesion involving several polymer microspheres and a section of everted intestinal tissue. The other method, the CAHN microbalance, employs a CAHN dynamic contact angle analyzer with modified software to record the tensile forces measured as a single polymer microsphere is pulled from intestinal tissue. This study demonstrates that CAHN and everted sac experiments yield similar results when used to quantify the bioadhesive nature of polymer microsphere systems. A polymer showing high adhesion in one method also demonstrates high bioadhesion in the other method; polymers that exhibit high fracture strength and tensile work measurements with the CAHN microbalance also yield high binding percentages with the everted sac method. The polymers tested and reported here are poly(caprolactone) and different copolymer ratios of poly(fumaric-co-sebacic anhydride). The results of this correlation demonstrate that each method alone is a valuable indicator of bioadhesion.

Autolytic activity of human calpain 7 is enhanced by ESCRT-III-related protein IST1 through MIT-MIM interaction.

PubMed

Osako, Yohei; Maemoto, Yuki; Tanaka, Ryohei; Suzuki, Hironori; Shibata, Hideki; Maki, Masatoshi

2010-11-01

Calpain 7, a mammalian ortholog of yeast Cpl1/Rim13 and fungal PalB, is an atypical calpain that lacks a penta-EF-hand domain. Previously, we reported that a region containing a tandem repeat of microtubule-interacting and transport (MIT) domains in calpain 7 interacts with a subset of endosomal sorting complex required for transport (ESCRT)-III-related proteins, suggesting involvement of calpain 7 in the ESCRT system. Although yeast and fungal calpains are thought to be involved in alkaline adaptation via limited proteolysis of specific transcription factors, proteolytic activity of calpain 7 has not been demonstrated yet. In this study, we investigated the interaction between calpain 7 and a newly reported ESCRT-III family member, increased sodium tolerance-1 (IST1), which possesses two different types of MIT-interacting motifs (MIM1 and MIM2). We found that glutathione-S-transferase (GST)-fused tandem MIT domains of calpain 7 (calpain 7MIT) pulled down FLAG-tagged IST1 expressed in HEK293T cells. Coimmunoprecipitation assays with various deletion or point mutants of epitope-tagged calpain 7 and IST1 revealed that both repetitive MIT domains and MIMs are required for efficient interaction. Direct MIT-MIM binding was confirmed by a pulldown experiment with GST-fused IST1 MIM and purified recombinant calpain 7MIT. Furthermore, we found that the GST-MIM protein enhances the autolysis of purified Strep-tagged monomeric green fluorescent protein (mGFP)-fused calpain 7 (mGFP-calpain 7-Strep). The autolysis was almost completely abolished by 10 mmN-ethylmaleimide but only partially inhibited by 1 mm leupeptin or E-64. The putative catalytic Cys290-substituted mutant (mGFP-calpain 7(C290S)-Strep) showed no autolytic activity. These results demonstrate for the first time that human calpain 7 is proteolytically active, and imply that calpain 7 is activated in the ESCRT system. © 2010 The Authors Journal compilation © 2010 FEBS.

LncRNA-FEZF1-AS1 promotes tumor proliferation and metastasis in colorectal cancer by regulating PKM2 signaling.

PubMed

Bian, Zehua; Zhang, Jiwei; Li, Min; Feng, Yuyang; Wang, Xue; Zhang, Jia; Yao, Surui; Jin, Guoying; Du, Jun; Han, Weifeng; Yin, Yuan; Huang, Shenglin; Fei, Bojian; Zou, Jian; Huang, Zhaohui

2018-06-18

Long non-coding RNAs (lncRNAs) play key roles in human cancers. Here, FEZF1-AS1, a highly overexpressed lncRNA in colorectal cancer (CRC), was identified by lncRNA microarrays. We aimed to explore the roles and possible molecular mechanisms of FEZF1-AS1 in CRC. LncRNA expression in CRC tissues was measured by lncRNA microarray and qRT-PCR. The functional roles of FEZF1-AS1 in CRC were demonstrated by a series of in vitro and in vivo experiments. RNA pull-down, RNA immunoprecipitation and luciferase analyses were used to demonstrate the potential mechanisms of FEZF1-AS1. We identified a series of differentially expressed lncRNAs in CRC using lncRNA microarrays, and revealed that FEZF1-AS1 is one of the most overexpressed. Further validation in two expanded CRC cohorts confirmed the upregulation of FEZF1-AS1 in CRC, and revealed that increased FEZF1-AS1 expression is associated with poor survival. Functional assays revealed that FEZF1-AS1 promotes CRC cell proliferation and metastasis. Mechanistically, FEZF1-AS1 could bind and increase the stability of the pyruvate kinase 2 (PKM2) protein, resulting in increased cytoplasmic and nuclear PKM2 levels. Increased cytoplasmic PKM2 promoted pyruvate kinase activity and lactate production (aerobic glycolysis), whereas FEZF1-AS1-induced nuclear PKM2 upregulation further activated STAT3 signaling. In addition, PKM2 was upregulated in CRC tissues and correlated with FEZF1-AS1 expression and patient survival. Together, these data provide mechanistic insights into the regulation of FEZF1-AS1 on both STAT3 signaling and glycolysis by binding PKM2 and increasing its stability. Copyright ©2018, American Association for Cancer Research.

Theoretical modeling and experimental study of dielectric loss of the multi-push-pull mode magnetoelectric laminate composites

NASA Astrophysics Data System (ADS)

Xu, Bingbing; Ma, Jiashuai; Fang, Cong; Yao, Meng; Di, Wenning; Li, Xiaobing; Luo, Haosu

2018-02-01

In this work, we establish a dielectric loss model for multi-push-pull mode ME laminate composites. It deduces that the total dielectric loss of the ME composites equals the linear average of the dielectric loss of piezoelectric plate and epoxy resin. But further analysis of this model has indicated that we can ignore the dielectric loss of epoxy resin. To verify this model, we use three kinds of epoxy resin with different dielectric loss to fabricate multi-push-pull mode PMNT/Metglas ME laminate composites respectively. It turns out that the different kinds of epoxy resin have little influence on the total dielectric loss, capacitance and piezoelectricity of the composites, which demonstrates that our model conforms to the practical case. Therefore, we can pay more attention to the mechanical properties of epoxy resin rather than its dielectric loss on fabricating the ME laminate composites.

High-Cycle, Push-Pull Fatigue Fracture Behavior of High-C, Si-Al-Rich Nanostructured Bainite Steel.

PubMed

Zhao, Jing; Ji, Honghong; Wang, Tiansheng

2017-12-29

The high-cycle, push-pull fatigue fracture behavior of high-C, Si-Al-rich nanostructured bainitic steel was studied through the measurement of fatigue limits, a morphology examination and phase composition analysis of the fatigue fracture surface, as well as fractography of the fatigue crack propagation. The results demonstrated that the push-pull fatigue limits at 10⁷ cycles were estimated as 710-889 MPa, for the samples isothermally transformed at the temperature range of 220-260 °C through data extrapolation, measured under the maximum cycle number of 10⁵. Both the interior inclusion and the sample surface constituted the fatigue crack origins. During the fatigue crack propagation, a high amount of secondary cracks were formed in almost parallel arrangements. The apparent plastic deformation occurred in the fracture surface layer, which induced approximately all retained austenite to transform into martensite.

High-Cycle, Push–Pull Fatigue Fracture Behavior of High-C, Si–Al-Rich Nanostructured Bainite Steel

PubMed Central

Zhao, Jing; Ji, Honghong

2017-01-01

The high-cycle, push–pull fatigue fracture behavior of high-C, Si–Al-rich nanostructured bainitic steel was studied through the measurement of fatigue limits, a morphology examination and phase composition analysis of the fatigue fracture surface, as well as fractography of the fatigue crack propagation. The results demonstrated that the push–pull fatigue limits at 107 cycles were estimated as 710–889 MPa, for the samples isothermally transformed at the temperature range of 220–260 °C through data extrapolation, measured under the maximum cycle number of 105. Both the interior inclusion and the sample surface constituted the fatigue crack origins. During the fatigue crack propagation, a high amount of secondary cracks were formed in almost parallel arrangements. The apparent plastic deformation occurred in the fracture surface layer, which induced approximately all retained austenite to transform into martensite. PMID:29286325

Deformation Behavior during Processing in Carbon Fiber Reinforced Plastics

NASA Astrophysics Data System (ADS)

Ogihara, Shinji; Kobayashi, Satoshi

In this study, we manufacture the device for measuring the friction between the prepreg curing process and subjected to pull-out tests with it The prepreg used in this study is a unidirectional carbon/epoxy, produced by TORAY designation of T700SC/2592.When creating specimens 4-ply prepregs are prepared and laminated. The 2-ply prepregs in the middle are shifted 50mm. In order to measure the friction between the prepreg during the cure process, we simulate the environment in the autoclave in the device, and we experiment in pull-out test. Test environment simulating temperature and pressure. The speed of displacement should be calculated by coefficient of thermal expansions (CTE). By calculation, 0.05mm/min gives the order of magnitude of displacement speed. In this study, 3 pull-out speeds are used: 0.01, 0.05 and 0.1mm/min. The specimen was heated by a couple of heaters, and we controlled the heaters with a temperature controller along the curing conditions of the prepreg. We put pressure using 4 bolts. Two strain gages were put on the bolt. We can understand the load applied to the specimen from the strain of the bolt. Pressure was adjusted the tightness of the bolt according to curing conditions. By using such a device, the pull-out test performed by tensile testing machine while adding temperature and pressure. During the 5 hours, we perform experiments while recording the load and stroke. The shear stress determined from the load and the stroke, and evaluated.

Basic in vitro experiment on the adhesive effects of sheet-type hemostatic agents used in combination with a liquid fibrin sealant.

PubMed

Ishii, Keiichi; Kawashima, Hideki; Hayama, Takuma; Mayabashira, Sumika; Oka, Shiro; Sugimoto, Toshikado

2011-11-01

Various hemostatic agents have been used quite effectively for hemostasis, as well as for providing effective adhesion during laparoscopic partial nephrectomies. In this study, we investigated the adhesiveness to the renal tissue of some sheet-type hemostatic agents used in combination with a liquid fibrin sealant. In Experiment A, component solutions of the fibrin glue (liquid fibrin sealant) were dripped onto a kite string placed annularly on a porcine kidney slice. Then, one of the sheet-type hemostats--namely, the collagen, gelatin, or cellulose hemostat--was placed on the slices, and a string scale was used to measure the force needed to pull the string apart vertically from the kidney slice. Twelve slices were used for each group, and the weight data were analyzed statistically. The tissue adhering to each sheet-type hemostatic agent was fixed in formalin and sliced and then examined by light microscopy after hematoxylin and eosin staining. In Experiment B, the solutions were dripped onto the sheet-type hemostatic agent placed first on the slice, and the force needed for pulling apart the hemostat sheet from the slice was similarly examined. The combination of fibrin glue plus a collagen hemostat was clearly superior in Experiment A, but the hemostat and renal tissue could be pulled apart more easily in Experiment B. These results showed that fibrin glue could not exert its expected adhesive effect unless it is used in combination with another hemostatic agent or is directly applied to renal tissue. It is important to obtain further comparative data among agents and select the appropriate agents, taking into consideration the type of surgery.

The Friction Force Determination of Large-Sized Composite Rods in Pultrusion

NASA Astrophysics Data System (ADS)

Grigoriev, S. N.; Krasnovskii, A. N.; Kazakov, I. A.

2014-08-01

Nowadays, the simple pull-force models of pultrusion process are not suitable for large sized rods because they are not considered a chemical shrinkage and thermal expansion acting in cured material inside the die. But the pulling force of the resin-impregnated fibers as they travels through the heated die is essential factor in the pultrusion process. In order to minimize the number of trial-and-error experiments a new mathematical approach to determine the frictional force is presented. The governing equations of the model are stated in general terms and various simplifications are implemented in order to obtain solutions without extensive numerical efforts. The influence of different pultrusion parameters on the frictional force value is investigated. The results obtained by the model can establish a foundation by which process control parameters are selected to achieve an appropriate pull-force and can be used for optimization pultrusion process.

Computational model of polarized actin cables and cytokinetic actin ring formation in budding yeast

PubMed Central

Tang, Haosu; Bidone, Tamara C.

2015-01-01

The budding yeast actin cables and contractile ring are important for polarized growth and division, revealing basic aspects of cytoskeletal function. To study these formin-nucleated structures, we built a 3D computational model with actin filaments represented as beads connected by springs. Polymerization by formins at the bud tip and bud neck, crosslinking, severing, and myosin pulling, are included. Parameter values were estimated from prior experiments. The model generates actin cable structures and dynamics similar to those of wild type and formin deletion mutant cells. Simulations with increased polymerization rate result in long, wavy cables. Simulated pulling by type V myosin stretches actin cables. Increasing the affinity of actin filaments for the bud neck together with reduced myosin V pulling promotes the formation of a bundle of antiparallel filaments at the bud neck, which we suggest as a model for the assembly of actin filaments to the contractile ring. PMID:26538307

Impacts of Experiential Learning Depth and Breadth on Student Outcomes

ERIC Educational Resources Information Center

Coker, Jeffrey Scott; Heiser, Evan; Taylor, Laura; Book, Connie

2017-01-01

This 5-year study of graduating seniors at Elon University (n = 2,058) evaluates the impacts of experiential learning depth (amount of time commitment) and breadth (number of different types of experiences) on student outcomes. Data on study abroad, undergraduate research, internships, service, and leadership experiences were pulled from…

Serial dependence promotes object stability during occlusion

PubMed Central

Liberman, Alina; Zhang, Kathy; Whitney, David

2016-01-01

Object identities somehow appear stable and continuous over time despite eye movements, disruptions in visibility, and constantly changing visual input. Recent results have demonstrated that the perception of orientation, numerosity, and facial identity is systematically biased (i.e., pulled) toward visual input from the recent past. The spatial region over which current orientations or face identities are pulled by previous orientations or identities, respectively, is known as the continuity field, which is temporally tuned over the past several seconds (Fischer & Whitney, 2014). This perceptual pull could contribute to the visual stability of objects over short time periods, but does it also address how perceptual stability occurs during visual discontinuities? Here, we tested whether the continuity field helps maintain perceived object identity during occlusion. Specifically, we found that the perception of an oriented Gabor that emerged from behind an occluder was significantly pulled toward the random (and unrelated) orientation of the Gabor that was seen entering the occluder. Importantly, this serial dependence was stronger for predictable, continuously moving trajectories, compared to unpredictable ones or static displacements. This result suggests that our visual system takes advantage of expectations about a stable world, helping to maintain perceived object continuity despite interrupted visibility. PMID:28006066

Tectonic evolution of the Salton Sea inferred from seismic reflection data

USGS Publications Warehouse

Brothers, D.S.; Driscoll, N.W.; Kent, G.M.; Harding, A.J.; Babcock, J.M.; Baskin, R.L.

2009-01-01

Oblique extension across strike-slip faults causes subsidence and leads to the formation of pull-apart basins such as the Salton Sea in southern California. The formation of these basins has generally been studied using laboratory experiments or numerical models. Here we combine seismic reflection data and geological observations from the Salton Sea to understand the evolution of this nascent pull-apart basin. Our data reveal the presence of a northeast-trending hinge zone that separates the sea into northern and southern sub-basins. Differential subsidence (10 mm yr 1) in the southern sub-basin suggests the existence of northwest-dipping basin-bounding faults near the southern shoreline, which may control the spatial distribution of young volcanism. Rotated and truncated strata north of the hinge zone suggest that the onset of extension associated with this pull-apart basin began after 0.5 million years ago. We suggest that slip is partitioned spatially and temporally into vertical and horizontal domains in the Salton Sea. In contrast to previous models based on historical seismicity patterns, the rapid subsidence and fault architecture that we document in the southern part of the sea are consistent with experimental models for pull-apart basins. ?? 2009 Macmillan Publishers Limited.

Task Differences and Prosociality; Investigating Pet Dogs' Prosocial Preferences in a Token Choice Paradigm.

PubMed

Dale, Rachel; Quervel-Chaumette, Mylène; Huber, Ludwig; Range, Friederike; Marshall-Pescini, Sarah

2016-01-01

Prosociality has received increasing interest by non-human animal researchers since the initial discoveries that suggested it is not a uniquely human trait. However, thus far studies, even within the same species, have not garnered conclusive results. A prominent suggestion for this disparity is the effect methodology can have on prosocial responses in animals. We recently found evidence of prosociality in domestic dogs towards familiar conspecifics using a bar-pulling paradigm, in which a subject could pull a rope to deliver food to its partner. Therefore, the current study aimed to assess whether dogs would show a similar response in a different paradigm, based on the token exchange task paradigm frequently used with primates. In this task, dogs had the option to touch a token with their nose that delivered a reward to an adjacent receiver enclosure, which contained a familiar conspecific, a stranger or no dog at all. Crucially, we also included a social facilitation control condition, whereby the partner (stranger/familiar) was present but unable to access the food. We found that the familiarity effect remained consistent across tasks, with dogs of both the bar-pulling and token choice experiments providing more food to familiar partners than in a non-social control and providing less food to stranger partners than this same control. However, in contrast to our previous bar-pulling experiment, we could not exclude social facilitation as an underlying motive in the current task. We propose this difference in results between tasks may be related to increased task complexity in the token choice paradigm, making it harder for dogs to discriminate between the test and social facilitation conditions. Overall our findings suggest that subtle methodological changes can have an impact on prosocial behaviours in dogs and highlights the importance of controlling for social facilitation effects in such experiments.

Sequence-dependent folding landscapes of adenine riboswitch aptamers.

PubMed

Lin, Jong-Chin; Hyeon, Changbong; Thirumalai, D

2014-04-14

Expression of a large fraction of genes in bacteria is controlled by riboswitches, which are found in the untranslated region of mRNA. Structurally riboswitches have a conserved aptamer domain to which a metabolite binds, resulting in a conformational change in the downstream expression platform. Prediction of the functions of riboswitches requires a quantitative description of the folding landscape so that the barriers and time scales for the conformational change in the switching region in the aptamer can be estimated. Using a combination of all atom molecular dynamics (MD) and coarse-grained model simulations we studied the response of adenine (A) binding add and pbuE A-riboswitches to mechanical force. The two riboswitches contain a structurally similar three-way junction formed by three paired helices, P1, P2, and P3, but carry out different functions. Using pulling simulations, with structures generated in MD simulations, we show that after P1 rips the dominant unfolding pathway in the add A-riboswitch is the rupture of P2 followed by unraveling of P3. In the pbuE A-riboswitch, after P1 unfolds P3 ruptures ahead of P2. The order of unfolding of the helices, which is in accord with single molecule pulling experiments, is determined by the relative stabilities of the individual helices. Our results show that the stability of isolated helices determines the order of assembly and response to force in these non-coding regions. We use the simulated free energy profile for the pbuE A-riboswitch to estimate the time scale for allosteric switching, which shows that this riboswitch is under kinetic control lending additional support to the conclusion based on single molecule pulling experiments. A consequence of the stability hypothesis is that a single point mutation (U28C) in the P2 helix of the add A-riboswitch, which increases the stability of P2, would make the folding landscapes of the two riboswitches similar. This prediction can be tested in single molecule pulling experiments.

Field demonstration of CO2 leakage detection and potential impacts on groundwater quality at Brackenridge Field Laboratory

NASA Astrophysics Data System (ADS)

Zou, Y.; Yang, C.; Guzman, N.; Delgado, J.; Mickler, P. J.; Horvoka, S.; Trevino, R.

2015-12-01

One concern related to GCS is possible risk of unintended CO2 leakage from the storage formations into overlying potable aquifers on underground sources of drinking water (USDW). Here we present a series of field tests conducted in an alluvial aquifer which is on a river terrace at The University of Texas Brackenridge Field Laboratory. Several shallow groundwater wells were completed to the limestone bedrock at a depth of 6 m and screened in the lower 3 m. Core sediments recovered from the shallow aquifer show that the sediments vary in grain size from clay-rich layers to coarse sandy gravels. Two main types of field tests were conducted at the BFL: single- (or double-) well push-pull test and pulse-like CO2 release test. A single- (or double-) well push-pull test includes three phases: the injection phase, the resting phase and pulling phase. During the injection phase, groundwater pumped from the shallow aquifer was stored in a tank, equilibrated with CO2 gasand then injected into the shallow aquifer to mimic CO2 leakage. During the resting phase, the groundwater charged with CO2 reacted with minerals in the aquifer sediments. During the pulling phase, groundwater was pumped from the injection well and groundwater samples were collected continuously for groundwater chemistry analysis. In such tests, large volume of groundwater which was charged with CO2 can be injected into the shallow aquifer and thus maximize contact of groundwater charged with CO2. Different than a single- (or double-) well push-pull test, a pulse-like CO2 release test for validating chemical sensors for CO2 leakage detection involves a CO2 release phase that CO2 gas was directly bubbled into the testing well and a post monitoring phase that groundwater chemistry was continuously monitored through sensors and/or grounder sampling. Results of the single- (or double-) well push-pull tests conducted in the shallow aquifer shows that the unintended CO2 leakage could lead to dissolution of carbonates and some silicates and mobilization of heavy metals from the aquifer sediments to groundwater, however, such mobilization posed no risks on groundwater quality at this site. The pulse-like tests have demonstrated it is plausible to use chemical sensors for CO2 leakage detection in groundwater.

Top-down contextual knowledge guides visual attention in infancy.

PubMed

Tummeltshammer, Kristen; Amso, Dima

2017-10-26

The visual context in which an object or face resides can provide useful top-down information for guiding attention orienting, object recognition, and visual search. Although infants have demonstrated sensitivity to covariation in spatial arrays, it is presently unclear whether they can use rapidly acquired contextual knowledge to guide attention during visual search. In this eye-tracking experiment, 6- and 10-month-old infants searched for a target face hidden among colorful distracter shapes. Targets appeared in Old or New visual contexts, depending on whether the visual search arrays (defined by the spatial configuration, shape and color of component items in the search display) were repeated or newly generated throughout the experiment. Targets in Old contexts appeared in the same location within the same configuration, such that context covaried with target location. Both 6- and 10-month-olds successfully distinguished between Old and New contexts, exhibiting faster search times, fewer looks at distracters, and more anticipation of targets when contexts repeated. This initial demonstration of contextual cueing effects in infants indicates that they can use top-down information to facilitate orienting during memory-guided visual search. © 2017 John Wiley & Sons Ltd.

The essence of conscious conflict: subjective effects of sustaining incompatible intentions.

PubMed

Morsella, Ezequiel; Gray, Jeremy R; Krieger, Stephen C; Bargh, John A

2009-10-01

Conflict constitutes one of the fundamental "tuggings and pullings" of the human experience. Yet, the link between the various kinds of conflict in the nervous system and subjective experience remains unexplained. The authors tested a hypothesis that predicts why both the "hot" conflicts involving self-control and motivation and the "cooler" response conflicts of the laboratory lead to changes in subjective experience. From this standpoint, these changes arise automatically from the activation of incompatible skeletomotor intentions, because the primary function of consciousness is to integrate such intentions for adaptive skeletal muscle output. Accordingly, the authors demonstrated for the first time that merely sustaining incompatible intentions (to move right and left) in a motionless state produces stronger subjective effects than sustaining compatible intentions. The results held equally strongly for two different effector systems involving skeletal muscle: arm movements and finger movements. In contrast, no such effects were found with conflict in a smooth muscle effector system. Together, these findings illuminate aspects of the nature of subjective experience and the role of incompatible intentions in affect and failures of self-control.

Searching for the Hebb effect in Down syndrome: evidence for a dissociation between verbal short-term memory and domain-general learning of serial order.

PubMed

Mosse, E K; Jarrold, C

2010-04-01

The Hebb effect is a form of repetition-driven long-term learning that is thought to provide an analogue for the processes involved in new word learning. Other evidence suggests that verbal short-term memory also constrains now vocabulary acquisition, but if the Hebb effect is independent of short-term memory, then it may be possible to demonstrate its preservation in a sample of individuals with Down syndrome, who typically show a verbal short-term memory deficit alongside surprising relative strengths in vocabulary. In two experiments, individuals both with and without Down syndrome (matched for receptive vocabulary) completed immediate serial recall tasks incorporating a Hebb repetition paradigm in either verbal or visuospatial conditions. Both groups demonstrated equivalent benefit from Hebb repetition, despite individuals with Down syndrome showing significantly lower verbal short-term memory spans. The resultant Hebb effect was equivalent across verbal and visuospatial domains. These studies suggest that the Hebb effect is essentially preserved within Down syndrome, implying that explicit verbal short-term memory is dissociable from potentially more implicit Hebb learning. The relative strength in receptive vocabulary observed in Down syndrome may therefore be supported by largely intact long-term as opposed to short-term serial order learning. This in turn may have implications for teaching methods and interventions that present new phonological material to individuals with Down syndrome.

Chimpanzee 'Ham' In Biopack Couch

NASA Technical Reports Server (NTRS)

1961-01-01

A three-year-old chimpanzee, named Ham, in the biopack couch for the MR-2 suborbital test flight. On January 31, 1961, a Mercury-Redstone launch from Cape Canaveral carried the chimpanzee 'Ham' over 640 kilometers down range in an arching trajectory that reached a peak of 254 kilometers above the Earth. The mission was successful and Ham performed his lever-pulling task well in response to the flashing light. NASA used chimpanzees and other primates to test the Mercury Capsule before launching the first American astronaut Alan Shepard in May 1961. The successful flight and recovery confirmed the soundness of the Mercury-Redstone systems.

PlateRunner: A Search Engine to Identify EMR Boilerplates.

PubMed

Divita, Guy; Workman, T Elizabeth; Carter, Marjorie E; Redd, Andrew; Samore, Matthew H; Gundlapalli, Adi V

2016-01-01

Medical text contains boilerplated content, an artifact of pull-down forms from EMRs. Boilerplated content is the source of challenges for concept extraction on clinical text. This paper introduces PlateRunner, a search engine on boilerplates from the US Department of Veterans Affairs (VA) EMR. Boilerplates containing concepts should be identified and reviewed to recognize challenging formats, identify high yield document titles, and fine tune section zoning. This search engine has the capability to filter negated and asserted concepts, save and search query results. This tool can save queries, search results, and documents found for later analysis.

Mercury Project

NASA Image and Video Library

1961-01-31

A three-year-old chimpanzee, named Ham, in the biopack couch for the MR-2 suborbital test flight. On January 31, 1961, a Mercury-Redstone launch from Cape Canaveral carried the chimpanzee "Ham" over 640 kilometers down range in an arching trajectory that reached a peak of 254 kilometers above the Earth. The mission was successful and Ham performed his lever-pulling task well in response to the flashing light. NASA used chimpanzees and other primates to test the Mercury Capsule before launching the first American astronaut Alan Shepard in May 1961. The successful flight and recovery confirmed the soundness of the Mercury-Redstone systems.

Mercury Project

NASA Image and Video Library

1961-01-01

A three-year-old chimpanzee, named Ham, in the biopack couch for the MR-2 suborbital test flight. On January 31, 1961, a Mercury-Redstone launch from Cape Canaveral carried the chimpanzee "Ham" over 640 kilometers down range in an arching trajectory that reached a peak of 254 kilometers above the Earth. The mission was successful and Ham performed his lever-pulling task well in response to the flashing light. NASA used chimpanzees and other primates to test the Mercury Capsule before launching the first American astronaut Alan Shepard in May 1961. The successful flight and recovery confirmed the soundness of the Mercury-Redstone systems.

The Role of Nanodiamonds in the Polishing Zone During Magnetorheological Finishing (MRF)

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

DeGroote, J.E.; Marino, A.E.; WIlson, J.P.

2008-01-07

In this work we discuss the role that nanodiamond abrasives play in magnetorheological finishing. We hypothesize that, as the nanodiamond MR fluid is introduced to the magnetic field, the micron sized spherical carbonyl iron (CI) particles are pulled down towards the rotating wheel, leaving a thin layer of nanodiamonds at the surface of the stiffened MR fluid ribbon. Our experimental results shown here support this hypothesis. We also show that surface roughness values inside MRF spots show a strong correlation with the near surface mechanical properties of the glass substrates and with drag force.

Fermi-surface-free superconductivity in underdoped (Bi,Pb)(Sr,La) 2CuO 6+δ (Bi2201)

DOE PAGES

Mistark, Peter; Hafiz, Hasnain; Markiewicz, Robert S.; ...

2015-06-18

Fermi-surface-free superconductivity arises when the superconducting order pulls down spectral weight from a band that is completely above the Fermi energy in the normal state. Here, we show that this can arise in hole-doped cuprates when a competing order causes a reconstruction of the Fermi surface. The change in Fermi surface topology is accompanied by a characteristic rise in the spectral weight. Finally, our results support the presence of a trisected superconducting dome, and suggest that superconductivity is responsible for stabilizing the (π,π) magnetic order at higher doping.

Photoinduced intramolecular charge transfer in a cross-conjugated push-pull enediyne: implications toward photoreaction.

PubMed

Singh, Anuja; Pati, Avik Kumar; Mishra, Ashok Kumar

2018-05-30

Push-pull organic fluorophores are important owing to their interesting optoelectronical properties. Here we report the photophysics of a new cross-conjugated push-pull enediynyl dye which belongs to an unexplored class of π-conjugated donor-acceptor systems. Two N,N-dimethylaniline moieties serve as donors and one pyrene ring functions as an acceptor via a common Y-shaped 'enediyne' bridge which facilitates the cross-electronic communication. The dye exhibits dual emission from locally excited (LE) and intramolecular charge transfer (ICT) states. While the LE emission is dominant in non-polar solvents, the ICT emission predominates in polar solvents. Time-resolved fluorescence decay experiments reveal a relatively shorter lifetime component (∼0.5-0.9 ns) belonging to an ICT state and a relatively longer lifetime species (∼1.6-2.8 ns) corresponding to the LE state. The strong ICT behavior of the dye is manifested through the huge red-shift (4166 cm-1) of the emission spectra from non-polar cyclohexane to polar N,N-dimethylformamide. In contrast to many small push-pull organic dyes, the LE and ICT states of the push-pull enediynyl dye follow the same excitation pathway. The dominant red-shifted ICT emission (∼550 nm) intensity of the dye in polar solvent decreases with a concomitant appearance of the blue-shifted LE emission (∼385 nm) upon prolonged exposure to photons. This opens up a new photophysical strategy of achieving high contrast two fluorescence color conversion from yellow to blue.

Urethral pull-through operation for the management of pelvic fracture urethral distraction defects.

PubMed

Yin, Lei; Li, Zhenhua; Kong, Chuize; Yu, Xiuyue; Zhu, Yuyan; Zhang, Yuxi; Jiang, Yuanjun

2011-10-01

To present our institutional experience in the management of pelvic fracture urethral distraction defects with urethral pull-through operation. Seventy-six patients (average age 34.5 years) with posterior urethral strictures caused by pelvic fracture urethral distraction defects underwent urethral pull-through operation at our department from July 1995 to September 2009. The estimated urethral stricture length was 2.0-3.5 cm (mean 2.5). Of these patients, 31 (41%) had undergone failed urethroplasty or urethrotomy after the initial management, and 5 (7%) had urethrorectal fistula. Urethral pull-through operation was performed 4-7 months (mean 4.9) after initial treatment or failed urethral reconstruction. The clinical outcome was considered a failure when any postoperative intervention was needed. Follow-up was 14-74 months (mean 42.5). The overall success rate was 89% (68/76). All treatment failures occurred within the first 6 months postoperatively. Failed repairs were successfully managed with internal urethrotomy in 1 patient, by urethral dilation in 6, and by another urethroplasty in 1. All patients were urinary-continent postoperatively. Of the potent patients, 2 (5%) became impotent after urethroplasty. There was no chordee, penile shortening, or urethral fistula recurrence. Urethral pull-through operation might be a less demanding and less time-consuming procedure. It does not increase the rate of impotence or incontinence and, with a high success rate, might serve as an alternative method for the management of pelvic fracture urethral distraction defects. Copyright © 2011 Elsevier Inc. All rights reserved.

Growth differentiation factor-15 (GDF-15) suppresses in vitro angiogenesis through a novel interaction with connective tissue growth factor (CCN2).

PubMed

Whitson, Ramon J; Lucia, Marshall Scott; Lambert, James R

2013-06-01

Growth differentiation factor-15 (GDF-15) and the CCN family member, connective tissue growth factor (CCN2), are associated with cardiac disease, inflammation, and cancer. The precise role and signaling mechanism for these factors in normal and diseased tissues remains elusive. Here we demonstrate an interaction between GDF-15 and CCN2 using yeast two-hybrid assays and have mapped the domain of interaction to the von Willebrand factor type C domain of CCN2. Biochemical pull down assays using secreted GDF-15 and His-tagged CCN2 produced in PC-3 prostate cancer cells confirmed a direct interaction between these proteins. To investigate the functional consequences of this interaction, in vitro angiogenesis assays were performed. We demonstrate that GDF-15 blocks CCN2-mediated tube formation in human umbilical vein endothelial (HUVEC) cells. To examine the molecular mechanism whereby GDF-15 inhibits CCN2-mediated angiogenesis, activation of αV β3 integrins and focal adhesion kinase (FAK) was examined. CCN2-mediated FAK activation was inhibited by GDF-15 and was accompanied by a decrease in αV β3 integrin clustering in HUVEC cells. These results demonstrate, for the first time, a novel signaling pathway for GDF-15 through interaction with the matricellular signaling molecule CCN2. Furthermore, antagonism of CCN2 mediated angiogenesis by GDF-15 may provide insight into the functional role of GDF-15 in disease states. Copyright © 2012 Wiley Periodicals, Inc.

RNA Helicase Associated with AU-rich Element (RHAU/DHX36) Interacts with the 3′-Tail of the Long Non-coding RNA BC200 (BCYRN1)*

PubMed Central

Booy, Evan P.; McRae, Ewan K. S.; Howard, Ryan; Deo, Soumya R.; Ariyo, Emmanuel O.; Dzananovic, Edis; Meier, Markus; Stetefeld, Jörg; McKenna, Sean A.

2016-01-01

RNA helicase associated with AU-rich element (RHAU) is an ATP-dependent RNA helicase that demonstrates high affinity for quadruplex structures in DNA and RNA. To elucidate the significance of these quadruplex-RHAU interactions, we have performed RNA co-immunoprecipitation screens to identify novel RNAs bound to RHAU and characterize their function. In the course of this study, we have identified the non-coding RNA BC200 (BCYRN1) as specifically enriched upon RHAU immunoprecipitation. Although BC200 does not adopt a quadruplex structure and does not bind the quadruplex-interacting motif of RHAU, it has direct affinity for RHAU in vitro. Specifically designed BC200 truncations and RNase footprinting assays demonstrate that RHAU binds to an adenosine-rich region near the 3′-end of the RNA. RHAU truncations support binding that is dependent upon a region within the C terminus and is specific to RHAU isoform 1. Tests performed to assess whether BC200 interferes with RHAU helicase activity have demonstrated the ability of BC200 to act as an acceptor of unwound quadruplexes via a cytosine-rich region near the 3′-end of the RNA. Furthermore, an interaction between BC200 and the quadruplex-containing telomerase RNA was confirmed by pull-down assays of the endogenous RNAs. This leads to the possibility that RHAU may direct BC200 to bind and exert regulatory functions at quadruplex-containing RNA or DNA sequences. PMID:26740632

Learning to perceive haptic distance-to-break in the presence of friction.

PubMed

Altenhoff, Bliss M; Pagano, Christopher C; Kil, Irfan; Burg, Timothy C

2017-02-01

Two experiments employed attunement and calibration training to investigate whether observers are able to identify material break points in compliant materials through haptic force application. The task required participants to attune to a recently identified haptic invariant, distance-to-break (DTB), rather than haptic stimulation not related to the invariant, including friction. In the first experiment participants probed simulated force-displacement relationships (materials) under 3 levels of friction with the aim of pushing as far as possible into the materials without breaking them. In a second experiment a different set of participants pulled on the materials. Results revealed that participants are sensitive to DTB for both pushing and pulling, even in the presence of varying levels of friction, and this sensitivity can be improved through training. The results suggest that the simultaneous presence of friction may assist participants in perceiving DTB. Potential applications include the development of haptic training programs for minimally invasive (laparoscopic) surgery to reduce accidental tissue damage. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

A microelectromechanical systems (MEMS) force-displacement transducer for sub-5 nm nanoindentation and adhesion measurements

NASA Astrophysics Data System (ADS)

Zhang, Youfeng; Oh, Yunje; Stauffer, Douglas; Polycarpou, Andreas A.

2018-04-01

We present a highly sensitive force-displacement transducer capable of performing ultra-shallow nanoindentation and adhesion measurements. The transducer utilizes electrostatic actuation and capacitive sensing combined with microelectromechanical fabrication technologies. Air indentation experiments report a root-mean-square (RMS) force resolution of 1.8 nN and an RMS displacement resolution of 0.019 nm. Nanoindentation experiments on a standard fused quartz sample report a practical RMS force resolution of 5 nN and an RMS displacement resolution of 0.05 nm at sub-10 nm indentation depths, indicating that the system has a very low system noise for indentation experiments. The high sensitivity and low noise enables the transducer to obtain high-resolution nanoindentation data at sub-5 nm contact depths. The sensitive force transducer is used to successfully perform nanoindentation measurements on a 14 nm thin film. Adhesion measurements were also performed, clearly capturing the pull-on and pull-off forces during approach and separation of two contacting surfaces.

Push-Pull Receptive Field Organization and Synaptic Depression: Mechanisms for Reliably Encoding Naturalistic Stimuli in V1

PubMed Central

Kremkow, Jens; Perrinet, Laurent U.; Monier, Cyril; Alonso, Jose-Manuel; Aertsen, Ad; Frégnac, Yves; Masson, Guillaume S.

2016-01-01

Neurons in the primary visual cortex are known for responding vigorously but with high variability to classical stimuli such as drifting bars or gratings. By contrast, natural scenes are encoded more efficiently by sparse and temporal precise spiking responses. We used a conductance-based model of the visual system in higher mammals to investigate how two specific features of the thalamo-cortical pathway, namely push-pull receptive field organization and fast synaptic depression, can contribute to this contextual reshaping of V1 responses. By comparing cortical dynamics evoked respectively by natural vs. artificial stimuli in a comprehensive parametric space analysis, we demonstrate that the reliability and sparseness of the spiking responses during natural vision is not a mere consequence of the increased bandwidth in the sensory input spectrum. Rather, it results from the combined impacts of fast synaptic depression and push-pull inhibition, the later acting for natural scenes as a form of “effective” feed-forward inhibition as demonstrated in other sensory systems. Thus, the combination of feedforward-like inhibition with fast thalamo-cortical synaptic depression by simple cells receiving a direct structured input from thalamus composes a generic computational mechanism for generating a sparse and reliable encoding of natural sensory events. PMID:27242445

Complex formation by the human Rad51B and Rad51C DNA repair proteins and their activities in vitro

NASA Technical Reports Server (NTRS)

Lio, Yi-Ching; Mazin, Alexander V.; Kowalczykowski, Stephen C.; Chen, David J.

2003-01-01

The human Rad51 protein is essential for DNA repair by homologous recombination. In addition to Rad51 protein, five paralogs have been identified: Rad51B/Rad51L1, Rad51C/Rad51L2, Rad51D/Rad51L3, XRCC2, and XRCC3. To further characterize a subset of these proteins, recombinant Rad51, Rad51B-(His)(6), and Rad51C proteins were individually expressed employing the baculovirus system, and each was purified from Sf9 insect cells. Evidence from nickel-nitrilotriacetic acid pull-down experiments demonstrates a highly stable Rad51B.Rad51C heterodimer, which interacts weakly with Rad51. Rad51B and Rad51C proteins were found to bind single- and double-stranded DNA and to preferentially bind 3'-end-tailed double-stranded DNA. The ability to bind DNA was elevated with mixed Rad51 and Rad51C, as well as with mixed Rad51B and Rad51C, compared with that of the individual protein. In addition, both Rad51B and Rad51C exhibit DNA-stimulated ATPase activity. Rad51C displays an ATP-independent apparent DNA strand exchange activity, whereas Rad51B shows no such activity; this apparent strand exchange ability results actually from a duplex DNA destabilization capability of Rad51C. By analogy to the yeast Rad55 and Rad57, our results suggest that Rad51B and Rad51C function through interactions with the human Rad51 recombinase and play a crucial role in the homologous recombinational repair pathway.

Mechanism of Adenomatous Polyposis Coli (APC)-mediated Blockage of Longpatch Base Excision Repair†

PubMed Central

Jaiswal, Aruna S.; Balusu, Ramesh; Armas, Melissa L.; Kundu, Chanakya N.; Narayan, Satya

2008-01-01

Recently, we found an interaction between adenomatous polyposis coli (APC) and DNA polymerase β (pol-β) and showed that APC blocks strand-displacement synthesis of long-patch base excision repair (LP-BER) however, the mechanism is not clear. Using an in vivo LP-BER assay system, we now show that the LP-BER is higher in APC−/− cells than in APC+/+ cells. In addition to pol-β, the pull-down experiments showed that the full-length APC also interacted with flap endonuclease 1 (Fen-1). To further characterize the interaction of APC with pol-β and Fen-1, we performed a domain-mapping of APC and found that both pol-β and Fen-1 interact with a 138-amino acids peptide from the APC at the DRI-domain. Our functional assays showed that APC blocks pol-β-mediated 1-nucleotide (1-nt) as well as strand-displacement synthesis of reduced abasic, nicked-, or 1-nt gapped-DNA substrates. Our further studies demonstrated that APC blocks 5′-flap endonuclease as well as 5′-3′ exonuclease activity of Fen-1 resulting in the blockage of LP-BER. From these results we concluded that APC can have three different effects in the LP-BER pathway. First, APC can block pol-β-mediated 1-nt incorporation and strand-displacement synthesis. Second, APC can block LP-BER by blocking coordinated formation and removal of the strand-displaced flap. Third, APC can block LP-BER by blocking “Hit and Run” synthesis. These studies will have important implications of APC in DNA damage-induced carcinogenesis and chemoprevention. PMID:17176113

Functional analysis of the glycogen binding subunit CG9238/Gbs-70E of protein phosphatase 1 in Drosophila melanogaster.

PubMed

Kerekes, Éva; Kókai, Endre; Páldy, Ferenc Sándor; Dombrádi, Viktor

2014-06-01

The product of the CG9238 gene that we termed glycogen binding subunit 70E (Gbs-70E) was characterized by biochemical and molecular genetics methods. The interaction between Gbs-70E and all catalytic subunits of protein phosphatase 1 (Pp1-87B, Pp1-9C, Pp1-96A and Pp1-13C) of Drosophila melanogaster was confirmed by pairwise yeast two-hybrid tests, co-immunoprecipitation and pull down experiments. The binding of Gbs-70E to glycogen was demonstrated by sedimentation analysis. With RT-PCR we found that the mRNAs coding for the longer Gbs-70E PB/PC protein were expressed in all developmental stages of the fruit flies while the mRNA for the shorter Gbs-70E PA was restricted to the eggs and the ovaries of the adult females. The development specific expression of the shorter splice variant was not conserved in different Drosophila species. The expression level of the gene was manipulated by P-element insertions and gene deletion to analyze the functions of the gene product. A small or moderate reduction in the gene expression resulted in no significant changes, however, a deletion mutant expressing very low level of the transcript lived shorter and exhibited reduced glycogen content in the imagos. In addition, the gene deletion decreased the fertility of the fruit flies. Our results prove that Gbs-70E functions as the glycogen binding subunit of protein phosphatase 1 that regulates glycogen content and plays a role in the development of eggs in D. melanogaster. Copyright © 2014 Elsevier Ltd. All rights reserved.

Crossing borders to bind proteins--a new concept in protein recognition based on the conjugation of small organic molecules or short peptides to polypeptides from a designed set.

PubMed

Baltzer, Lars

2011-06-01

A new concept for protein recognition and binding is highlighted. The conjugation of small organic molecules or short peptides to polypeptides from a designed set provides binder molecules that bind proteins with high affinities, and with selectivities that are equal to those of antibodies. The small organic molecules or peptides need to bind the protein targets but only with modest affinities and selectivities, because conjugation to the polypeptides results in molecules with dramatically improved binder performance. The polypeptides are selected from a set of only sixteen sequences designed to bind, in principle, any protein. The small number of polypeptides used to prepare high-affinity binders contrasts sharply with the huge libraries used in binder technologies based on selection or immunization. Also, unlike antibodies and engineered proteins, the polypeptides have unordered three-dimensional structures and adapt to the proteins to which they bind. Binder molecules for the C-reactive protein, human carbonic anhydrase II, acetylcholine esterase, thymidine kinase 1, phosphorylated proteins, the D-dimer, and a number of antibodies are used as examples to demonstrate that affinities are achieved that are higher than those of the small molecules or peptides by as much as four orders of magnitude. Evaluation by pull-down experiments and ELISA-based tests in human serum show selectivities to be equal to those of antibodies. Small organic molecules and peptides are readily available from pools of endogenous ligands, enzyme substrates, inhibitors or products, from screened small molecule libraries, from phage display, and from mRNA display. The technology is an alternative to established binder concepts for applications in drug development, diagnostics, medical imaging, and protein separation.

Direct methylation of FXR by Set7/9, a lysine methyltransferase, regulates the expression of FXR target genes

PubMed Central

Balasubramaniyan, Natarajan; Ananthanarayanan, Meena

2012-01-01

The farnesoid X receptor (FXR) is a ligand (bile acid)-dependent nuclear receptor that regulates target genes involved in every aspect of bile acid homeostasis. Upon binding of ligand, FXR recruits an array of coactivators and associated proteins, some of which have intrinsic enzymatic activity that modify histones or even components of the transcriptional complex. In this study, we show chromatin occupancy by the Set7/9 methyltransferase at the FXR response element (FXRE) and direct methylation of FXR in vivo and in vitro at lysine 206. siRNA depletion of Set7/9 in the Huh-7 liver cell line decreased endogenous mRNAs of the FXR target genes, the short heterodimer partner (SHP) and bile salt export pump (BSEP). Mutation of the methylation site at K206 of FXR to an arginine prevented methylation by Set7/9. A pan-methyllysine antibody recognized the wild-type FXR but not the K206R mutant form. An electromobility shift assay showed that methylation by Set7/9 enhanced binding of FXR/retinoic X receptor-α to the FXRE. Interaction between hinge domain of FXR (containing K206) and Set7/9 was confirmed by coimmunoprecipitation, GST pull down, and mammalian two-hybrid experiments. Set7/9 overexpression in Huh-7 cells significantly enhanced transactivation of the SHP and BSEP promoters in a ligand-dependent fashion by wild-type FXR but not the K206R mutant FXR. A Set7/9 mutant deficient in methyltransferase activity was also not effective in increasing transactivation of the BSEP promoter. These studies demonstrate that posttranslational methylation of FXR by Set7/9 contributes to the transcriptional activation of FXR-target genes. PMID:22345554

High Temperature Electrolysis for Hydrogen Production from Nuclear Energy – TechnologySummary

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

J. E. O'Brien; C. M. Stoots; J. S. Herring

2010-02-01

The Department of Energy, Office of Nuclear Energy, has requested that a Hydrogen Technology Down-Selection be performed to identify the hydrogen production technology that has the best potential for timely commercial demonstration and for ultimate deployment with the Next Generation Nuclear Plant (NGNP). An Independent Review Team has been assembled to execute the down-selection. This report has been prepared to provide the members of the Independent Review Team with detailed background information on the High Temperature Electrolysis (HTE) process, hardware, and state of the art. The Idaho National Laboratory has been serving as the lead lab for HTE research andmore » development under the Nuclear Hydrogen Initiative. The INL HTE program has included small-scale experiments, detailed computational modeling, system modeling, and technology demonstration. Aspects of all of these activities are included in this report. In terms of technology demonstration, the INL successfully completed a 1000-hour test of the HTE Integrated Laboratory Scale (ILS) technology demonstration experiment during the fall of 2008. The HTE ILS achieved a hydrogen production rate in excess of 5.7 Nm3/hr, with a power consumption of 18 kW. This hydrogen production rate is far larger than has been demonstrated by any of the thermochemical or hybrid processes to date.« less

Human Asymptomatic Epitope Peptide/CXCL10-Based Prime/Pull Vaccine Induces Herpes Simplex Virus-Specific Gamma Interferon-Positive CD107+ CD8+ T Cells That Infiltrate the Cornea and Trigeminal Ganglia of Humanized HLA Transgenic Rabbits and Protect against Ocular Herpes Challenge.

PubMed

Khan, Arif A; Srivastava, Ruchi; Vahed, Hawa; Roy, Soumyabrata; Walia, Sager S; Kim, Grace J; Fouladi, Mona A; Yamada, Taikun; Ly, Vincent T; Lam, Cynthia; Lou, Anthony; Nguyen, Vivianna; Boldbaatar, Undariya; Geertsema, Roger; Fraser, Nigel W; BenMohamed, Lbachir

2018-06-13

Herpes simplex virus 1 (HSV-1) is a prevalent human pathogen that infects the cornea causing potentially blinding herpetic disease. A clinical herpes vaccine is still lacking. In the present study, a novel prime/pull vaccine was tested in Human Leukocyte Antigen- (HLA-) transgenic rabbit model of ocular herpes (HLA Tg rabbit). Three asymptomatic (ASYMP) peptide epitopes were selected from the HSV-1 membrane glycoprotein C (UL44 400-408 ), the DNA replication binding helicase (UL9 196-204 ), and the tegument protein (UL25 572-580 ), all preferentially recognized by CD8 + T cells from "naturally protected" HSV-1-seropositive healthy ASYMP individuals (who never had recurrent corneal herpetic disease). HLA Tg rabbits were immunized with a mixture of these three ASYMP CD8 + T cell peptide epitopes (UL44 400-408 , UL9 196-204 and UL25 572-580 ), delivered subcutaneously with CpG 2007 adjuvant (prime). Fifteen days later, half of the rabbits received a topical ocular treatment with a recombinant neurotropic AAV8 vector, expressing the T cell-attracting CXCL10 chemokine (pull). The frequency, function of HSV-specific CD8 + T cells induced by the prime/pull vaccine were assessed in peripheral blood, cornea, and trigeminal ganglia (TG). Compared to peptides alone, the peptides/CXCL10 prime/pull vaccine generated frequent polyfunctional gamma interferon-positive (IFN-γ + ) CD107 + CD8 + T cells that infiltrated both the cornea and TG. CD8 + T cells mobilization into cornea and TG of prime/pull- vaccinated rabbits was associated with a significant reduction in corneal herpes infection and disease following an ocular HSV-1 challenge (McKrae). These findings draw attention to the novel prime/pull vaccine strategy to mobilize anti-viral CD8 + T cells into tissues protecting them against herpes infection and disease. IMPORTANCE There is an urgent need for a vaccine against widespread herpes simplex virus infections. The present study demonstrates that immunization of HLA transgenic rabbits with a peptide/CXCL10 prime/pull vaccine triggered mobilization of HSV-specific CD8 + T cells locally in the cornea and TG, the sites of acute and latent herpes infections. Mobilization of antiviral CD8 + T cells into cornea and TG of rabbits that received the prime/pull vaccine was associated with protection against ocular herpes infection and disease following an ocular HSV-1 challenge. These results highlight the importance of the prime/pull vaccine strategy to bolster the number and function of protective CD8 + T cells within infected tissues. Copyright © 2018 American Society for Microbiology.

Research on the improvement of traditional dial instrument precision based on C8051F020.

NASA Astrophysics Data System (ADS)

Sun, Guiling; Liu, Yi; Lu, Li

2006-11-01

Two essential parameters to weigh the quality of a reinforcing steel bar are the value of its bending force and the maximum pull it can withstand, in order to measure them with higher precision, it is significant to describe the changing tendency of force with time and displacement by drawing a real-time curve directly during the process examining the quality of a bar when the pull exerted is variable continuously. Using C8051F020 as the core component, this paper improves traditional dial instruments whose precision can only reach the second level. Adopting a high precision pulling/pressing force sensor, an amplifier, a two-order Butterworth low-pass filter and a 12-bit AD converter which is in the C8051F020, the first level of precision can be obtained. A rotary encoder is used to measure the length increment of the bar during the pulling process, based on an algorithm, a force-displacement (or time) curve which is quite important for operators to control the course of experiment can be displayed on the LCD. Meanwhile, real-time experimental data can be stored in local flash, or uploaded to PC by RS-485 and stored in the center database. A real-time clock is also adopted to mark the time of each experiment that is useful to index the data. The measure system we describe here is characterized by simple structure, high precision and stabilization, and convenience operation, can be used in other actual measure systems by only changing the front sensor, so it is of great value of application and popularization.

Pushing and Pulling Sara: A Case Study of the Contrasting Influences of High School and University Experiences on Engineering Agency, Identity, and Participation

ERIC Educational Resources Information Center

Godwin, Allison; Potvin, Geoff

2017-01-01

This manuscript reports a longitudinal case study of how one woman, Sara, who had previously considered dropping out of high school, authored strong mathematics and science identities and purposefully exhibited agency through her experiences in high school science. These experiences empowered her to choose an engineering major in college; however,…

Arm-Locking with the GRACE Follow-On Laser Ranging Instrument

NASA Technical Reports Server (NTRS)

Thorpe, James Ira; Mckenzie, Kirk

2016-01-01

Arm-locking is a technique for stabilizing the frequency of a laser in an inter-spacecraft interferometer by using the spacecraft separation as the frequency reference. A candidate technique for future space-based gravitational wave detectors such as the Laser Interferometer Space Antenna (LISA), arm-locking has been extensive studied in this context through analytic models, time-domain simulations, and hardware-in-the-loop laboratory demonstrations. In this paper we show the Laser Ranging Instrument flying aboard the upcoming Gravity Recovery and Climate Experiment Follow-On (GRACE-FO) mission provides an appropriate platform for an on-orbit demonstration of the arm-locking technique. We describe an arm-locking controller design for the GRACE-FO system and a series of time-domain simulations that demonstrate its feasibility. We conclude that it is possible to achieve laser frequency noise suppression of roughly two orders of magnitude around a Fourier frequency of 1Hz with conservative margins on the system's stability. We further demonstrate that `pulling' of the master laser frequency due to fluctuating Doppler shifts and lock acquisition transients is less than 100MHz over several GRACE-FO orbits. These findings motivate further study of the implementation of such a demonstration.

Arm locking with the GRACE follow-on laser ranging interferometer

NASA Astrophysics Data System (ADS)

Thorpe, James Ira; McKenzie, Kirk

2016-02-01

Arm locking is a technique for stabilizing the frequency of a laser in an interspacecraft interferometer by using the spacecraft separation as the frequency reference. A candidate technique for future space-based gravitational wave detectors such as the Laser Interferometer Space Antenna, arm locking has been extensive studied in this context through analytic models, time-domain simulations, and hardware-in-the-loop laboratory demonstrations. In this paper we show the laser ranging interferometer instrument flying aboard the upcoming Gravity Recovery and Climate Experiment follow-on (GRACE-FO) mission provides an appropriate platform for an on-orbit demonstration of the arm-locking technique. We describe an arm-locking controller design for the GRACE-FO system and a series of time-domain simulations that demonstrate its feasibility. We conclude that it is possible to achieve laser frequency noise suppression of roughly 2 orders of magnitude around a Fourier frequency of 1 Hz with conservative margins on the system's stability. We further demonstrate that "pulling" of the master laser frequency due to fluctuating Doppler shifts and lock acquisition transients is less than 100 MHz over several GRACE-FO orbits. These findings motivate further study of the implementation of such a demonstration.

Variability of strength measurement in postmenopausal women who are overweight or obese: a Monet study.

PubMed

St-Onge, Maxime; Mathieu, Marie-Eve; Tousignant, Benoit; Faraj, May; Lavoie, Jean-Marc

2009-12-01

The main objective of this study was to establish whether a stable measurement of strength could be obtained without prior exercise familiarization in postmenopausal women who were overweight or obese. A second objective was to evaluate the influence of physical activity on the variability of strength measurement. Thirty postmenopausal women (age: 57.9 yr; SD: 5 yr; body mass index: 31.0 kg/m2; SD: 4 kg/m2) underwent 3 strength testing sessions (48 hr apart) each including 3 exercises (leg press, chest press, and lat pull down). Energy expenditure was measured before the strength testing week with the doubly labelled water method over a 10-day period. Resting metabolic rate was measured by indirect calorimetry. Physical activity energy expenditure was calculated as follows: total energy expenditure x 0.9, minus the resting metabolic rate. Repeated analysis of variance and paired t-test were used to assess the difference and the reliability of the testing sequence. Results from leg press and chest press exercises indicated no significant difference among the 3 testing sessions. The lat pull down exercise was associated with a significant systematic bias between sessions 1 and 2 (mean difference: 1.4 kg; SD: 3 kg; 95% confidence intervals; 0.2-2.7 kg), but the difference disappeared at the third testing session (mean difference: 0.7 kg; SD: 3 kg; 95% confidence intervals; 0.5-2 kg). Physical activity did not influence the variability of the strength results. Overall, our results showed that a relatively stable strength measurement can be obtained within a maximum of 3 testing sessions without prior familiarization. In addition, physical activity did not influence strength testing in postmenopausal women who were overweight or obese.

Isolation of pulmonary artery smooth muscle cells from neonatal mice.

PubMed

Lee, Keng Jin; Czech, Lyubov; Waypa, Gregory B; Farrow, Kathryn N

2013-10-19

Pulmonary hypertension is a significant cause of morbidity and mortality in infants. Historically, there has been significant study of the signaling pathways involved in vascular smooth muscle contraction in PASMC from fetal sheep. While sheep make an excellent model of term pulmonary hypertension, they are very expensive and lack the advantage of genetic manipulation found in mice. Conversely, the inability to isolate PASMC from mice was a significant limitation of that system. Here we described the isolation of primary cultures of mouse PASMC from P7, P14, and P21 mice using a variation of the previously described technique of Marshall et al. that was previously used to isolate rat PASMC. These murine PASMC represent a novel tool for the study of signaling pathways in the neonatal period. Briefly, a slurry of 0.5% (w/v) agarose + 0.5% iron particles in M199 media is infused into the pulmonary vascular bed via the right ventricle (RV). The iron particles are 0.2 μM in diameter and cannot pass through the pulmonary capillary bed. Thus, the iron lodges in the small pulmonary arteries (PA). The lungs are inflated with agarose, removed and dissociated. The iron-containing vessels are pulled down with a magnet. After collagenase (80 U/ml) treatment and further dissociation, the vessels are put into a tissue culture dish in M199 media containing 20% fetal bovine serum (FBS), and antibiotics (M199 complete media) to allow cell migration onto the culture dish. This initial plate of cells is a 50-50 mixture of fibroblasts and PASMC. Thus, the pull down procedure is repeated multiple times to achieve a more pure PASMC population and remove any residual iron. Smooth muscle cell identity is confirmed by immunostaining for smooth muscle myosin and desmin.

Estrogen Receptor β and Its Domains Interact with Casein Kinase 2, Phosphokinase C, and N-Myristoylation Sites of Mitochondrial and Nuclear Proteins in Mouse Brain*

PubMed Central

Paramanik, Vijay; Thakur, Mahendra Kumar

2012-01-01

The localization of estrogen receptor (ER)β in mitochondria suggests ERβ-dependent regulation of genes, which is poorly understood. Here, we analyzed the ERβ interacting mitochondrial as well as nuclear proteins in mouse brain using pull-down assay and matrix-assisted laser desorption ionization mass spectroscopy (MALDI-MS). In the case of mitochondria, ERβ interacted with six proteins of 35–152 kDa, its transactivation domain (TAD) interacted with four proteins of 37–172 kDa, and ligand binding domain (LBD) interacted with six proteins of 37–161 kDa. On the other hand, in nuclei, ERβ interacted with seven proteins of 30–203 kDa, TAD with ten proteins of 31–160 kDa, and LBD with fourteen proteins of 42–179 kDa. For further identification, these proteins were cleaved by trypsin into peptides and analyzed by MALDI-MS using mascot search engine, immunoprecipitation, immunoblotting, and far-Western blotting. To find the consensus binding motifs in interacting proteins, their unique tryptic peptides were analyzed by the motif scan software. All the interacting proteins were found to contain casein kinase (CK) 2, phosphokinase (PK)C phosphorylation, and N-myristoylation sites. These were further confirmed by peptide pull-down assays using specific mutations in the interacting sites. Thus, the present findings provide evidence for the interaction of ERβ with specific mitochondrial and nuclear proteins through consensus CK2, PKC phosphorylation, and N-myristoylation sites, and may represent an essential step toward designing selective ER modulators for regulating estrogen-mediated signaling. PMID:22566700

Finding the Cell Center by a Balance of Dynein and Myosin Pulling and Microtubule Pushing: A Computational Study

PubMed Central

Zhu, Jie; Burakov, Anton; Rodionov, Vladimir

2010-01-01

The centrosome position in many types of interphase cells is actively maintained in the cell center. Our previous work indicated that the centrosome is kept at the center by pulling force generated by dynein and actin flow produced by myosin contraction and that an unidentified factor that depends on microtubule dynamics destabilizes position of the centrosome. Here, we use modeling to simulate the centrosome positioning based on the idea that the balance of three forces—dyneins pulling along microtubule length, myosin-powered centripetal drag, and microtubules pushing on organelles—is responsible for the centrosome displacement. By comparing numerical predictions with centrosome behavior in wild-type and perturbed interphase cells, we rule out several plausible hypotheses about the nature of the microtubule-based force. We conclude that strong dynein- and weaker myosin-generated forces pull the microtubules inward competing with microtubule plus-ends pushing the microtubule aster outward and that the balance of these forces positions the centrosome at the cell center. The model also predicts that kinesin action could be another outward-pushing force. Simulations demonstrate that the force-balance centering mechanism is robust yet versatile. We use the experimental observations to reverse engineer the characteristic forces and centrosome mobility. PMID:20980619

Finding the cell center by a balance of dynein and myosin pulling and microtubule pushing: a computational study.

PubMed

Zhu, Jie; Burakov, Anton; Rodionov, Vladimir; Mogilner, Alex

2010-12-01

The centrosome position in many types of interphase cells is actively maintained in the cell center. Our previous work indicated that the centrosome is kept at the center by pulling force generated by dynein and actin flow produced by myosin contraction and that an unidentified factor that depends on microtubule dynamics destabilizes position of the centrosome. Here, we use modeling to simulate the centrosome positioning based on the idea that the balance of three forces-dyneins pulling along microtubule length, myosin-powered centripetal drag, and microtubules pushing on organelles-is responsible for the centrosome displacement. By comparing numerical predictions with centrosome behavior in wild-type and perturbed interphase cells, we rule out several plausible hypotheses about the nature of the microtubule-based force. We conclude that strong dynein- and weaker myosin-generated forces pull the microtubules inward competing with microtubule plus-ends pushing the microtubule aster outward and that the balance of these forces positions the centrosome at the cell center. The model also predicts that kinesin action could be another outward-pushing force. Simulations demonstrate that the force-balance centering mechanism is robust yet versatile. We use the experimental observations to reverse engineer the characteristic forces and centrosome mobility.

Uncovering a Role for the Tail of the Dictyostelium discoideum SadA Protein in Cell-Substrate Adhesion ▿ †

PubMed Central

Kowal, Anthony S.; Chisholm, Rex L.

2011-01-01

Previous work from our laboratory showed that the Dictyostelium discoideum SadA protein plays a central role in cell-substrate adhesion. SadA null cells exhibit a loss of adhesion, a disrupted actin cytoskeleton, and a cytokinesis defect. How SadA mediates these phenotypes is unknown. This work addresses the mechanism of SadA function, demonstrating an important role for the C-terminal cytoplasmic tail in SadA function. We found that a SadA tailless mutant was unable to rescue the sadA adhesion deficiency, and overexpression of the SadA tail domain reduced adhesion in wild-type cells. We also show that SadA is closely associated with the actin cytoskeleton. Mutagenesis studies suggested that four serine residues in the tail, S924/S925 and S940/S941, may regulate association of SadA with the actin cytoskeleton. Glutathione S-transferase pull-down assays identified at least one likely interaction partner of the SadA tail, cortexillin I, a known actin bundling protein. Thus, our data demonstrate an important role for the carboxy-terminal cytoplasmic tail in SadA function and strongly suggest that a phosphorylation event in this tail regulates an interaction with cortexillin I. Based on our data, we propose a model for the function of SadA. PMID:21441344

Identification and Characterization of Roseltide, a Knottin-type Neutrophil Elastase Inhibitor Derived from Hibiscus sabdariffa

PubMed Central

Loo, Shining; Kam, Antony; Xiao, Tianshu; Nguyen, Giang K. T.; Liu, Chuan Fa; Tam, James P.

2016-01-01

Plant knottins are of therapeutic interest due to their high metabolic stability and inhibitory activity against proteinases involved in human diseases. The only knottin-type proteinase inhibitor against porcine pancreatic elastase was first identified from the squash family in 1989. Here, we report the identification and characterization of a knottin-type human neutrophil elastase inhibitor from Hibiscus sabdariffa of the Malvaceae family. Combining proteomic and transcriptomic methods, we identified a panel of novel cysteine-rich peptides, roseltides (rT1-rT8), which range from 27 to 39 residues with six conserved cysteine residues. The 27-residue roseltide rT1 contains a cysteine spacing and amino acid sequence that is different from the squash knottin-type elastase inhibitor. NMR analysis demonstrated that roseltide rT1 adopts a cystine-knot fold. Transcriptome analyses suggested that roseltides are bioprocessed by asparagine endopeptidases from a three-domain precursor. The cystine-knot structure of roseltide rT1 confers its high resistance against degradation by endopeptidases, 0.2 N HCl, and human serum. Roseltide rT1 was shown to inhibit human neutrophil elastase using enzymatic and pull-down assays. Additionally, roseltide rT1 ameliorates neutrophil elastase-stimulated cAMP accumulation in vitro. Taken together, our findings demonstrate that roseltide rT1 is a novel knottin-type neutrophil elastase inhibitor with therapeutic potential for neutrophil elastase associated diseases. PMID:27991569

The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA.

PubMed

Su'etsugu, Masayuki; Harada, Yuji; Keyamura, Kenji; Matsunaga, Chika; Kasho, Kazutoshi; Abe, Yoshito; Ueda, Tadashi; Katayama, Tsutomu

2013-12-01

DnaA activity for replication initiation of the Escherichia coli chromosome is negatively regulated by feedback from the DNA-loaded form of the replicase clamp. In this process, called RIDA (regulatory inactivation of DnaA), ATP-bound DnaA transiently assembles into a complex consisting of Hda and the DNA-clamp, which promotes inter-AAA+ domain association between Hda and DnaA and stimulates hydrolysis of DnaA-bound ATP, producing inactive ADP-DnaA. Using a truncated DnaA mutant, we previously demonstrated that the DnaA N-terminal domain is involved in RIDA. However, the precise role of the N-terminal domain in RIDA has remained largely unclear. Here, we used an in vitro reconstituted system to demonstrate that the Asn-44 residue in the N-terminal domain of DnaA is crucial for RIDA but not for replication initiation. Moreover, an assay termed PDAX (pull-down after cross-linking) revealed an unstable interaction between a DnaA-N44A mutant and Hda. In vivo, this mutant exhibited an increase in the cellular level of ATP-bound DnaA. These results establish a model in which interaction between DnaA Asn-44 and Hda stabilizes the association between the AAA+ domains of DnaA and Hda to facilitate DnaA-ATP hydrolysis during RIDA. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

CD147-induced cell proliferation is associated with Smad4 signal inhibition.

PubMed

Qin, Hui; Rasul, Azhar; Li, Xin; Masood, Muqaddas; Yang, Guang; Wang, Na; Wei, Wei; He, Xi; Watanabe, Nobumoto; Li, Jiang; Li, Xiaomeng

2017-09-15

CD147 is a multifunctional trans-membrane glycoprotein, which is highly expressed in many cancers. However, the mechanism by which CD147 modulates cell proliferation is not fully understood. The aim of this study is to investigate the role of CD147 in cell proliferation associated with the TGF-β/Smad4 signaling pathway. Here, we used cell viability and clone formation assays in LNCaP prostate cancer cells to demonstrate that CD147 promotes cell proliferation. The luciferase assay and western blotting show that silencing CD147 using shRNA enhances transcription and expression of p21 WAF1 . Using immunofluorescence and nuclear-cytoplasmic separation, we show that this is primarily attributed to transport of Smad4 from the cytoplasm to nucleus. Other assays (GST pull-down, co-immunoprecipitation and immunofluorescence) demonstrate that Smad4 is a new interaction partner of CD147, with the Smad4 MH2 domain and CD147 intracellular domain (CD147-ICD) being involved in the interaction. Furthermore, we report that a phosphoserine (pSer) in CD147 (pSer252) is responsible for this interaction and inhibition of the Smad4/p21 WAF1 signal that promotes cell proliferation. Our results provide a novel molecular mechanism for CD147-induced cell proliferation associated with Smad4 signal inhibition. Copyright © 2017. Published by Elsevier Inc.

Evaluation of a peridomestic mosquito trap for integration into an Aedes aegypti (Diptera: Culicidae) push-pull control strategy.

PubMed

Salazar, Ferdinand V; Achee, Nicole L; Grieco, John P; Prabaripai, Atchariya; Eisen, Lars; Shah, Pankhil; Chareonviriyaphap, Theeraphap

2012-06-01

We determined the feasibility of using the BG-Sentinel™ mosquito trap (BGS) as the pull component in a push-pull strategy to reduce indoor biting by Aedes aegypti. This included evaluating varying numbers of traps (1-4) and mosquito release numbers (10, 25, 50, 100, 150, 200, and 250) on recapture rates under screen house conditions. Based on these variations in trap and mosquito numbers, release intervals were rotated through a completely randomized design with environmental factors (temperature, relative humidity, and light intensity) and monitored throughout each experiment. Data from four sampling time points (05:30, 09:30, 13:30, and 17:30) indicate a recapture range among treatments of 66-98%. Furthermore, 2-3 traps were as effective in recapturing mosquitoes as 4 traps for all mosquito release numbers. Time trends indicate Day 1 (the day the mosquitoes were released) as the "impact period" for recapture with peak numbers of marked mosquitoes collected at 09:30 or 4 h post-release. Information from this study will be used to guide the configuration of the BGS trap component of a push-pull vector control strategy currently in the proof-of-concept stage of development in Thailand and Peru. © 2012 The Society for Vector Ecology.

Actin cable distribution and dynamics arising from cross-linking, motor pulling, and filament turnover

PubMed Central

Tang, Haosu; Laporte, Damien; Vavylonis, Dimitrios

2014-01-01

The growth of fission yeast relies on the polymerization of actin filaments nucleated by formin For3p, which localizes at tip cortical sites. These actin filaments bundle to form actin cables that span the cell and guide the movement of vesicles toward the cell tips. A big challenge is to develop a quantitative understanding of these cellular actin structures. We used computer simulations to study the spatial and dynamical properties of actin cables. We simulated individual actin filaments as semiflexible polymers in three dimensions composed of beads connected with springs. Polymerization out of For3p cortical sites, bundling by cross-linkers, pulling by type V myosin, and severing by cofilin are simulated as growth, cross-linking, pulling, and turnover of the semiflexible polymers. With the foregoing mechanisms, the model generates actin cable structures and dynamics similar to those observed in live-cell experiments. Our simulations reproduce the particular actin cable structures in myoVΔ cells and predict the effect of increased myosin V pulling. Increasing cross-linking parameters generates thicker actin cables. It also leads to antiparallel and parallel phases with straight or curved cables, consistent with observations of cells overexpressing α-actinin. Finally, the model predicts that clustering of formins at cell tips promotes actin cable formation. PMID:25103242

Quantitative modeling of forces in electromagnetic tweezers

NASA Astrophysics Data System (ADS)

Bijamov, Alex; Shubitidze, Fridon; Oliver, Piercen M.; Vezenov, Dmitri V.

2010-11-01

This paper discusses numerical simulations of the magnetic field produced by an electromagnet for generation of forces on superparamagnetic microspheres used in manipulation of single molecules or cells. Single molecule force spectroscopy based on magnetic tweezers can be used in applications that require parallel readout of biopolymer stretching or biomolecular binding. The magnetic tweezers exert forces on the surface-immobilized macromolecule by pulling a magnetic bead attached to the free end of the molecule in the direction of the field gradient. In a typical force spectroscopy experiment, the pulling forces can range between subpiconewton to tens of piconewtons. In order to effectively provide such forces, an understanding of the source of the magnetic field is required as the first step in the design of force spectroscopy systems. In this study, we use a numerical technique, the method of auxiliary sources, to investigate the influence of electromagnet geometry and material parameters of the magnetic core on the magnetic forces pulling the target beads in the area of interest. The close proximity of the area of interest to the magnet body results in deviations from intuitive relations between magnet size and pulling force, as well as in the force decay with distance. We discuss the benefits and drawbacks of various geometric modifications affecting the magnitude and spatial distribution of forces achievable with an electromagnet.

Thermoplastic pultrusion development and characterization of residual in pultruded composites with modeling and experiments

NASA Astrophysics Data System (ADS)

Jamiyanaa, Khongor

Pultrusion processing is a technique to make highly aligned fiber reinforced polymer composites. Thermoset pultrusion is a mature process and well established, while thermoplastic pultrusion in still in its infancy. Thermoplastic pultrusion has not been well established because thermoplastic resins are difficult to process due to their high viscosity. However, thermoplastic resins offer distinct advantages that make thermoplastic pultrusion worth exploring. The present work centers on developing a method to design and validate a die for a thermoplastic pultrusion system. Analytical models and various software tools were used to design a pultrusion die. Experimental measurements have been made to validate the models. One-dimensional transient heat transfer analysis was used to calculate the time required for pre-impregnated E-Glass/Polypropylene tapes to melt and consolidate into profiled shapes. Creo Element/Pro 1.0 was used to design the die, while ANSYS Work Bench 14.0 was used to conduct heat transfer analysis to understand the temperature profile of the pultrusion apparatus. Additionally Star-CCM+ was used to create a three-dimensional fluid flow model to capture the molten polymer flow inside the pultrusion die. The fluid model was used to understand the temperature of the flow and the force required to pull the material at any given temperature and line speed. A complete pultrusion apparatus including the die, heating unit, cooling unit, and the frame has been designed and manufactured as guided by the models, and pultruded profiles have been successfully produced. The results show that the analytical model and the fluid model show excellent correlation. The predicted and measured pulling forces are in agreement and show that the pull force increases as the pull speed increases. Furthermore, process induced residual stress and its influence on dimensional instability, such as bending or bowing, on pultruded composites was analyzed. The study indicated that unbalanced layup can produce asymmetrical residual stress through the thickness and causes the part to bow. Furthermore, the residual stress through the thickness was mapped with excellent accuracy. A design of experiments around the processing parameters indicated that increase in pull speed or decrease in die temperature increased the residual stress within the part.

New Insights to Clathrin and Adaptor Protein 2 for the Design and Development of Therapeutic Strategies

PubMed Central

Poulsen, Ebbe Toftgaard; Larsen, Agnete; Zollo, Alen; Jørgensen, Arne L.; Sanggaard, Kristian W.; Enghild, Jan J.; Matrone, Carmela

2015-01-01

The Amyloid Precursor Protein (APP) has been extensively studied for its role as the precursor of the β-amyloid protein (Aβ) in Alzheimer’s disease (AD). However, our understanding of the normal function of APP is still patchy. Emerging evidence indicates that a dysfunction in APP trafficking and degradation can be responsible for neuronal deficits and progressive degeneration in humans. We recently reported that the Y682 mutation in the 682YENPTY687 domain of APP affects its binding to specific adaptor proteins and leads to its anomalous trafficking, to defects in the autophagy machinery and to neuronal degeneration. In order to identify adaptors that influence APP function, we performed pull-down experiments followed by quantitative mass spectrometry (MS) on hippocampal tissue extracts of three month-old mice incubated with either the 682YENPTY687 peptide, its mutated form, 682GENPTY687 or its phosphorylated form, 682pYENPTY687. Our experiments resulted in the identification of two proteins involved in APP internalization and trafficking: Clathrin heavy chain (hc) and its Adaptor Protein 2 (AP-2). Overall our results consolidate and refine the importance of Y682 in APP normal functions from an animal model of premature aging and dementia. Additionally, they open the perspective to consider Clathrin hc and AP-2 as potential targets for the design and development of new therapeutic strategies. PMID:26690411

Drawbar Pull

DTIC Science & Technology

2017-01-26

Includes procedures for hard surface, soil , and water tests. Discusses vehicle preparation, instrumentation method of computing results, data reduction...and amphibious vehicles. 15. SUBJECT TERMS Bollard pull Soft- soil mobility Drawbar pull Vehicle, amphibious Drawbar horsepower Vehicle...4.3 Drawbar Pull in Soft Soil ................................................. 8 4.4 Amphibious Vehicle Tests (Drawbar Pull in Water and Bollard Pull

How graphene slides: measurement and theory of strain-dependent frictional forces between graphene and SiO2.

PubMed

Kitt, Alexander L; Qi, Zenan; Rémi, Sebastian; Park, Harold S; Swan, Anna K; Goldberg, Bennett B

2013-06-12

Strain, bending rigidity, and adhesion are interwoven in determining how graphene responds when pulled across a substrate. Using Raman spectroscopy of circular, graphene-sealed microchambers under variable external pressure, we demonstrate that graphene is not firmly anchored to the substrate when pulled. Instead, as the suspended graphene is pushed into the chamber under pressure, the supported graphene outside the microchamber is stretched and slides, pulling in an annulus. Analyzing Raman G band line scans with a continuum model extended to include sliding, we extract the pressure dependent sliding friction between the SiO2 substrate and mono-, bi-, and trilayer graphene. The sliding friction for trilayer graphene is directly proportional to the applied load, but the friction for monolayer and bilayer graphene is inversely proportional to the strain in the graphene, which is in violation of Amontons' law. We attribute this behavior to the high surface conformation enabled by the low bending rigidity and strong adhesion of few layer graphene.

Using Social Networks to Enhance Teaching and Learning Experiences in Higher Learning Institutions

ERIC Educational Resources Information Center

Balakrishnan, Vimala

2014-01-01

The paper first explores the factors that affect the use of social networks to enhance teaching and learning experiences among students and lecturers, using structured questionnaires prepared based on the Push-Pull-Mooring framework. A total of 455 students and lecturers from higher learning institutions in Malaysia participated in this study.…

Robust control of electrostatic torsional micromirrors using adaptive sliding-mode control

NASA Astrophysics Data System (ADS)

Sane, Harshad S.; Yazdi, Navid; Mastrangelo, Carlos H.

2005-01-01

This paper presents high-resolution control of torsional electrostatic micromirrors beyond their inherent pull-in instability using robust sliding-mode control (SMC). The objectives of this paper are two-fold - firstly, to demonstrate the applicability of SMC for MEMS devices; secondly - to present a modified SMC algorithm that yields improved control accuracy. SMC enables compact realization of a robust controller tolerant of device characteristic variations and nonlinearities. Robustness of the control loop is demonstrated through extensive simulations and measurements on MEMS with a wide range in their characteristics. Control of two-axis gimbaled micromirrors beyond their pull-in instability with overall 10-bit pointing accuracy is confirmed experimentally. In addition, this paper presents an analysis of the sources of errors in discrete-time implementation of the control algorithm. To minimize these errors, we present an adaptive version of the SMC algorithm that yields substantial performance improvement without considerably increasing implementation complexity.

A smart microelectromechanical sensor and switch triggered by gas

NASA Astrophysics Data System (ADS)

Bouchaala, Adam; Jaber, Nizar; Shekhah, Osama; Chernikova, Valeriya; Eddaoudi, Mohamed; Younis, Mohammad I.

2016-07-01

There is an increasing interest to realize smarter sensors and actuators that can deliver a multitude of sophisticated functionalities while being compact in size and of low cost. We report here combining both sensing and actuation on the same device based on a single microstructure. Specifically, we demonstrate a smart resonant gas (mass) sensor, which in addition to being capable of quantifying the amount of absorbed gas, can be autonomously triggered as an electrical switch upon exceeding a preset threshold of absorbed gas. Toward this, an electrostatically actuated polymer microbeam is fabricated and is then functionalized with a metal-organic framework, namely, HKUST-1. The microbeam is demonstrated to absorb vapors up to a certain threshold, after which is shown to collapse through the dynamic pull-in instability. Upon pull-in, the microstructure can be made to act as an electrical switch to achieve desirable actions, such as alarming.

Role of combined tactile and kinesthetic feedback in minimally invasive surgery.

PubMed

Lim, Soo-Chul; Lee, Hyung-Kew; Park, Joonah

2014-10-18

Haptic feedback is of critical importance in surgical tasks. However, conventional surgical robots do not provide haptic feedback to surgeons during surgery. Thus, in this study, a combined tactile and kinesthetic feedback system was developed to provide haptic feedback to surgeons during robotic surgery. To assess haptic feasibility, the effects of two types of haptic feedback were examined empirically - kinesthetic and tactile feedback - to measure object-pulling force with a telesurgery robotics system at two desired pulling forces (1 N and 2 N). Participants answered a set of questionnaires after experiments. The experimental results reveal reductions in force error (39.1% and 40.9%) when using haptic feedback during 1 N and 2 N pulling tasks. Moreover, survey analyses show the effectiveness of the haptic feedback during teleoperation. The combined tactile and kinesthetic feedback of the master device in robotic surgery improves the surgeon's ability to control the interaction force applied to the tissue. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

Microgravity

NASA Image and Video Library

1998-02-05

Sections of ZBLAN fibers pulled in a conventional 1-g process (left) and in experiments aboard NASA's KC-135 low-gravity aircraft. The rough surface of the 1-g fiber indicates surface defects that would scatter an optical signal and greatly degrade its quality. ZBLAN is part of the family of heavy-metal fluoride glasses (fluorine combined zirconium, barium, lanthanum, aluminum, and sodium). NASA is conducting research on pulling ZBLAN fibers in the low-g environment of space to prevent crystallization that limits ZBLAN's usefulness in optical fiber-based communications. ZBLAN is a heavy-metal fluoride glass that shows exceptional promise for high-throughput communications with infrared lasers. Photo credit: NASA/Marshall Space Flight Center

Design of a haptic device with grasp and push-pull force feedback for a master-slave surgical robot.

PubMed

Hu, Zhenkai; Yoon, Chae-Hyun; Park, Samuel Byeongjun; Jo, Yung-Ho

2016-07-01

We propose a portable haptic device providing grasp (kinesthetic) and push-pull (cutaneous) sensations for optical-motion-capture master interfaces. Although optical-motion-capture master interfaces for surgical robot systems can overcome the stiffness, friction, and coupling problems of mechanical master interfaces, it is difficult to add haptic feedback to an optical-motion-capture master interface without constraining the free motion of the operator's hands. Therefore, we utilized a Bowden cable-driven mechanism to provide the grasp and push-pull sensation while retaining the free hand motion of the optical-motion capture master interface. To evaluate the haptic device, we construct a 2-DOF force sensing/force feedback system. We compare the sensed force and the reproduced force of the haptic device. Finally, a needle insertion test was done to evaluate the performance of the haptic interface in the master-slave system. The results demonstrate that both the grasp force feedback and the push-pull force feedback provided by the haptic interface closely matched with the sensed forces of the slave robot. We successfully apply our haptic interface in the optical-motion-capture master-slave system. The results of the needle insertion test showed that our haptic feedback can provide more safety than merely visual observation. We develop a suitable haptic device to produce both kinesthetic grasp force feedback and cutaneous push-pull force feedback. Our future research will include further objective performance evaluations of the optical-motion-capture master-slave robot system with our haptic interface in surgical scenarios.

Ingrowing Hair

PubMed Central

Luo, Di-Qing; Liang, Yu-Hua; Li, Xi-Qing; Zhao, Yu-Kun; Wang, Fang; Sarkar, Rashmi

2016-01-01

Abstract Cutaneous pili migrans and creeping eruption caused by parasitic diseases may present as a moving linear lesion in skin. The former, caused by a hair shaft or fragment embedded in the superficial skin or middle dermis, is a rare condition characterized by creeping eruption with a black line observed at the advancing end. In exceptionally rare instance, the hair grows inside the skin and burrows in the uppermost dermis, such a condition has been called “ingrown hair.” We report a 30-year-old Chinese man, who was accustomed to pull or extrude the beard hairs, with 1-year history of slowly extending black linear eruption on his right chin. Cutaneous examination revealed a 4-cm long black linear lesion beneath the skin associated with edematous erythema around and folliculitis on both ends of the lesion. After treatment with topical mupirocin ointment, the erythema and folliculitis improved and 2 hairs of the beard with hair follicles were pulled out from the skin. Two weeks later, another similar black line about 1 cm in length in the skin presented on the prior lesional area, which was pulled out by a shallow incision of the skin and was also demonstrated as a beard hair with hair follicle. The patient was diagnosed as “ingrowing hair” with multiple recurrences. The lesions recovered after the beard hairs were pulled out. No recurrence occurred in a year of follow-up. We suggest that “ingrowing hair” is better than “ingrown hair” to describe such a condition. Pulling out the involved hair and correcting the bad practice are its optimal management strategies. PMID:27175694

Ribbon curling

NASA Astrophysics Data System (ADS)

Juel, Anne; Prior, Chris; Moussou, Julien; Chakrabarti, Buddhapriya; Jensen, Oliver

The procedure of curling a ribbon by running it over a sharp blade is commonly used when wrapping presents. Despite its ubiquity, a quantitative explanation of this everyday phenomenon is still lacking. We address this using experiment and theory, examining the dependence of ribbon curvature on blade curvature, the longitudinal load imposed on the ribbon and the speed of pulling. Experiments in which a ribbon is drawn steadily over a blade under a fixed load show that the ribbon curvature is generated over a restricted range of loads, the curvature/load relationship can be non-monotonic, and faster pulling (under a constant imposed load) results in less tightly curled ribbons. We develop a theoretical model that captures these features, building on the concept that the ribbon under the imposed deformation undergoes differential plastic stretching across its thickness, resulting in a permanently curved shape. The model identifies factors that optimize curling and clarifies the physical mechanisms underlying the ribbon's nonlinear response to an apparently simple deformation.

NFIL3 suppresses hypoxia-induced apoptotic cell death by targeting the insulin-like growth factor 2 receptor.

PubMed

Lin, Kuan-Ho; Kuo, Chia-Hua; Kuo, Wei-Wen; Ho, Tsung-Jung; Pai, Peiying; Chen, Wei-Kung; Pan, Lung-Fa; Wang, Chien-Cheng; Padma, V Vijaya; Huang, Chih-Yang

2015-06-01

The insulin-like growth factor-II/mannose 6-phosphate receptor (IGF2R) over-expression correlates with heart disease progression. The IGF2R is not only an IGF2 clearance receptor, but it also triggers signal transduction, resulting in cardiac hypertrophy, apoptosis and fibrosis. The present study investigated the nuclear factor IL-3 (NFIL3), a transcription factor of the basic leucine zipper superfamily, and its potential pro-survival effects in cardiomyocytes. NFIL3 might play a key role in heart development and act as a survival factor in the heart, but the regulatory mechanisms are still unclear. IGF2 and IGF2R protein expression were highly increased in rat hearts subjected to hemorrhagic shock. IGF2R protein expression was also up-regulated in H9c2 cells exposed to hypoxia. Over-expression of NFIL3 in H9c2 cardiomyoblast cells inhibited the induction of hypoxia-induced apoptosis and down-regulated IGF2R expression levels. Gel shift assay, double-stranded DNA pull-down assay and chromatin immune-precipitation analyses indicated that NFIL3 binds directly to the IGF2R promoter region. Using a luciferase assay, we further observed NFIL3 repress IGF2R gene promoter activity. Our results demonstrate that NFIL3 is an important negative transcription factor, which through binding to the promoter of IGF2R, suppresses the apoptosis induced by IGF2R signaling in H9c2 cardiomyoblast cells under hypoxic conditions. © 2015 Wiley Periodicals, Inc.

Comparison of the Hang High-Pull and Loaded Jump Squat for the Development of Vertical Jump and Isometric Force-Time Characteristics.

PubMed

Oranchuk, Dustin J; Robinson, Tracey L; Switaj, Zachary J; Drinkwater, Eric J

2017-04-15

Weightlifting movements have high skill demands and require expert coaching. Loaded jumps have a comparably lower skill demand, but may be similarly effective for improving explosive performance. The purpose of this study was to compare vertical jump performance, isometric force, and rate of force development (RFD) following a ten-week intervention employing the hang high-pull (hang-pull) or trap-bar jump squat (jump-squat). Eighteen NCAA Division II swimmers (8 males, 10 females) with at least one year of resistance training experience volunteered to participate. Testing included the squat jump (SJ), countermovement jump (CMJ) and the isometric mid-thigh pull (IMTP). Vertical ground reaction forces were analyzed to obtain jump height and relative peak power. Relative peak force, peak RFD and relative force at five time bands were obtained from the IMTP. Subjects were randomly assigned to either a hang-pull (n = 9) or jump-squat (n = 9) training group and completed a ten-week, volume-equated, periodized training program. While there was a significant main effect of training for both groups, no statistically significant between-group differences were found (p ≥ 0.17) for any of the dependent variables. However, medium effect sizes in favor of the jump-squat training group were seen in SJ height (d = 0.56) and SJ peak power (d = 0.69). Loaded jumps seem equally effective as weightlifting derivatives for improving lower-body power in experienced athletes. Since loaded jumps require less skill and less coaching expertise than weightlifting, loaded jumps should be considered where coaching complex movements is difficult.

Higher order balance control: Distinct effects between cognitive task and manual steadiness constraint on automatic postural responses.

PubMed

Coelho, Daniel Boari; Bourlinova, Catarina; Teixeira, Luis Augusto

2016-12-01

In the present experiment, we aimed to evaluate the interactive effect of performing a cognitive task simultaneously with a manual task requiring either high or low steadiness on APRs. Young volunteers performed the task of recovering upright balance following a mechanical perturbation provoked by unanticipatedly releasing a load pulling the participant's body backwards. The postural task was performed while holding a cylinder steadily on a tray. One group performed that task under high (cylinder' round side down) and another one under low (cylinder' flat side down) manual steadiness constraint. Those tasks were evaluated in the conditions of performing concurrently a cognitive numeric subtraction task and under no cognitive task. Analysis showed that performance of the cognitive task led to increased body and tray displacement, associated with higher displacement at the hip and upper trunk, and lower magnitude of activation of the GM muscle in response to the perturbation. Conversely, high manual steadiness constraint led to reduced tray velocity in association with lower values of trunk displacement, and decreased rotation amplitude at the ankle and hip joints. We found no interactions between the effects of the cognitive and manual tasks on APRs, suggesting that they were processed in parallel in the generation of responses for balance recovery. Modulation of postural responses from the manual and cognitive tasks indicates participation of higher order neural structures in the generation of APRs, with postural responses being affected by multiple mental processes occurring in parallel. Copyright © 2016 Elsevier B.V. All rights reserved.

Differential Regulation of the Serotonin Transporter by Vesicle-Associated Membrane Protein 2 in Cells of Neuronal versus Non-Neuronal Origin

PubMed Central

Müller, Heidi Kaastrup; Kragballe, Marie; Fjorback, Anja Winther; Wiborg, Ove

2014-01-01

The serotonin transporter (SERT) is a key regulator of serotonergic signalling as it mediates the re-uptake of synaptic serotonin into nerve terminals, thereby terminating or modulating its signal. It is well-known that SERT regulation is a dynamic process orchestrated by a wide array of proteins and mechanisms. However, molecular details on possible coordinated regulation of SERT activity and 5-HT release are incomplete. Here, we report that vesicle-associated membrane protein 2 (VAMP2), a SNARE protein that mediates vesicle fusion with the plasma membrane, interacts with SERT. This was documented in vitro, through GST pull-down assays, by co-immunoprecipitation experiments on heterologous cells and rat hippocampal synaptosomes, and with FRET analysis in live transfected HEK-293 MSR cells. The related isoforms VAMP1 and VAMP3 also physically interact with SERT. However, comparison of the three VAMP isoforms shows that only VAMP2 possesses a functionally distinct role in relation to SERT. VAMP2 influences 5-HT uptake, cell surface expression and the delivery rate of SERT to the plasma membrane differentially in HEK-293 MSR and PC12 cells. Moreover, siRNA-mediated knock-down of endogenous VAMP2 reduces 5-HT uptake in CAD cells stably expressing low levels of heterologous SERT. Deletion and mutant analysis suggest a role for the isoform specific C-terminal domain of VAMP2 in regulating SERT function. Our data identify a novel interaction between SERT and a synaptic vesicle protein and support a link between 5-HT release and re-uptake. PMID:24878716

Teaching Basic Science Environmentally, Concept: Water that Comes Down as Rain Is Used Over and Over Again.

ERIC Educational Resources Information Center

Busch, Phyllis S.

1985-01-01

Provides directions for basic science experiments which demonstrate the rain cycle, fundamentals of cloud formation, and testing for the presence of acidity in local rainwater. Describes materials required, step-by-step instructions, and discussion topics. (NEC)

Particle sensor array

NASA Technical Reports Server (NTRS)

Buehler, Martin G. (Inventor); Blaes, Brent R. (Inventor); Lieneweg, Udo (Inventor)

1994-01-01

A particle sensor array which in a preferred embodiment comprises a static random access memory having a plurality of ion-sensitive memory cells, each such cell comprising at least one pull-down field effect transistor having a sensitive drain surface area (such as by bloating) and at least one pull-up field effect transistor having a source connected to an offset voltage. The sensitive drain surface area and the offset voltage are selected for memory cell upset by incident ions such as alpha-particles. The static random access memory of the present invention provides a means for selectively biasing the memory cells into the same state in which each of the sensitive drain surface areas is reverse biased and then selectively reducing the reversed bias on these sensitive drain surface areas for increasing the upset sensitivity of the cells to ions. The resulting selectively sensitive memory cells can be used in a number of applications. By way of example, the present invention can be used for measuring the linear energy transfer of ion particles, as well as a device for assessing the resistance of CMOS latches to Cosmic Ray induced single event upsets. The sensor of the present invention can also be used to determine the uniformity of an ion beam.

Addressable microshutter array for a high-performance infrared miniature dispersive spectrometer

NASA Astrophysics Data System (ADS)

Ilias, S.; Picard, F.; Larouche, C.; Kruzelecky, R.; Jamroz, W.

2009-02-01

Programmable microshutter arrays were designed to improve the attainable signal to noise ratio (SNR) of a miniature dispersive spectrometer developed for space applications. Integration of a microshutter array to this instrument provides advantages such as the addition of a binary coded optical input operation mode for the miniature spectrometer which results in SNR benefits without spectral resolution loss. These arrays were successfully fabricated using surface micromachining technology. Each microshutter is basically an electrostatic zipping actuator having a curved shape. Applying critical voltage to one microshutter pulls the actuator down to the substrate and closes the associated slit. Opening of the microslits relies on the restoring force generated within the actuated zippers. High light transmission is obtained with the actuator in the open position and excellent light blocking is observed when the shutter is closed. The pull-in voltage to close the microslits was about 110 V and the response times to close and open the microslits were about 2 ms and 7 ms, respectively. Selected array dies were mounted in modified off-the-shelf ceramic packages and electrically connected to package pins. The packages were hermetically sealed with AR coated sapphire windows. This last packaging step was performed in a dry nitrogen controlled atmosphere.

Programmable optical microshutter arrays for large aspect ratio microslits

NASA Astrophysics Data System (ADS)

Ilias, S.; Picard, F.; Larouche, C.; Kruzelecky, R.; Jamroz, W.; Le Noc, L.; Topart, P.

2008-06-01

Design, fabrication and characterization of a 16x1 programmable microshutter array are described. Each shutter controls the light transmitted through a microslit defined on the transparent substrate supporting the array. Two approaches were considered for the shutter array implementation: sweeping blades and zipping actuators. Simulation results and fabrication constraints led to the selection of the zipping actuators. The device was fabricated using a surface micromachining process. Each microshutter is basically an electrostatic zipping actuator having a curved shape induced by a stress gradient throughout the actuator thickness. When a sufficient voltage is applied between the microshutter and an actuation electrode surrounding the microslit area, the generated electrostatic force pulls the actuator down to the substrate which closes the microslit. Opening the slit relies on the restoring force due to the actuator deformation. Microshutter arrays were fabricated successfully. High light transmission through the slit area is obtained with the actuator in the open position and excellent light blocking is observed when the shutter is closed. Static and dynamic responses of the device were determined. A pull-in voltage of about 110 V closes the microslit and the response times to close and open the microslit are about 2 and 7 ms, respectively.

A focus group study of predictors of relapse in electronic gaming machine problem gambling, part 2: factors that 'pull' the gambler away from relapse.

PubMed

Oakes, J; Pols, R; Battersby, M; Lawn, S; Pulvirenti, M; Smith, D

2012-09-01

This study aimed to develop an empirically based description of relapse in Electronic Gaming Machine (EGM) problem gambling (PG) by describing the processes and factors that 'pull' the problem gambler away from relapse contrasted with the 'push' towards relapse. These conceptualisations describe two opposing, interacting emotional processes occurring within the problem gambler during any relapse episode. Each relapse episode comprises a complex set of psychological and social behaviours where many factors interact sequentially and simultaneously within the problem gambler to produce a series of mental and behaviour events that end (1) with relapse where 'push' overcomes 'pull' or (2) continued abstinence where 'pull' overcomes 'push'. Four focus groups comprising thirty participants who were EGM problem gamblers, gamblers' significant others, therapists and counsellors described their experiences and understanding of relapse. The groups were recorded, recordings were then transcribed and analysed using thematic textual analysis. It was established that vigilance, motivation to commit to change, positive social support, cognitive strategies such as remembering past gambling harms or distraction techniques to avoid thinking about gambling to enable gamblers to manage the urge to gamble and urge extinction were key factors that protected against relapse. Three complementary theories emerged from the analysis. Firstly, a process of reappraisal of personal gambling behaviour pulls the gambler away from relapse. This results in a commitment to change that develops over time and affects but is independent of each episode of relapse. Secondly, relapse may be halted by interacting factors that 'pull' the problem gambler away from the sequence of mental and behavioural events, which follow the triggering of the urge and cognitions to gamble. Thirdly, urge extinction and apparent 'cure' is possible for EGM gambling. This study provides a qualitative, empirical model for understanding protective factors against gambling relapse.

How to inhibit a distractor location? Statistical learning versus active, top-down suppression.

PubMed

Wang, Benchi; Theeuwes, Jan

2018-05-01

Recently, Wang and Theeuwes (Journal of Experimental Psychology: Human Perception and Performance, 44(1), 13-17, 2018a) demonstrated the role of lingering selection biases in an additional singleton search task in which the distractor singleton appeared much more often in one location than in all other locations. For this location, there was less capture and selection efficiency was reduced. It was argued that statistical learning induces plasticity within the spatial priority map such that particular locations that are high likely to contain a distractor are suppressed relative to all other locations. The current study replicated these findings regarding statistical learning (Experiment 1) and investigated whether similar effects can be obtained by cueing the distractor location in a top-down way on a trial-by-trial basis. The results show that top-down cueing of the distractor location with long (1,500 ms; Experiment 2) and short stimulus-onset symmetries (SOAs) (600 ms; Experiment 3) does not result in suppression: The amount of capture nor the efficiency of selection was affected by the cue. If anything, we found an attentional benefit (instead of the suppression) for the short SOA. We argue that through statistical learning, weights within the attentional priority map are changed such that one location containing a salient distractor is suppressed relative to all other locations. Our cueing experiments show that this effect cannot be accomplished by active, top-down suppression. Consequences for recent theories of distractor suppression are discussed.

Functional and Structural Properties of a Novel Protein and Virulence Factor (Protein sHIP) in Streptococcus pyogenes *

PubMed Central

Wisniewska, Magdalena; Happonen, Lotta; Kahn, Fredrik; Varjosalo, Markku; Malmström, Lars; Rosenberger, George; Karlsson, Christofer; Cazzamali, Giuseppe; Pozdnyakova, Irina; Frick, Inga-Maria; Björck, Lars; Streicher, Werner; Malmström, Johan; Wikström, Mats

2014-01-01

Streptococcus pyogenes is a significant bacterial pathogen in the human population. The importance of virulence factors for the survival and colonization of S. pyogenes is well established, and many of these factors are exposed to the extracellular environment, enabling bacterial interactions with the host. In the present study, we quantitatively analyzed and compared S. pyogenes proteins in the growth medium of a strain that is virulent to mice with a non-virulent strain. Particularly, one of these proteins was present at significantly higher levels in stationary growth medium from the virulent strain. We determined the three-dimensional structure of the protein that showed a unique tetrameric organization composed of four helix-loop-helix motifs. Affinity pull-down mass spectrometry analysis in human plasma demonstrated that the protein interacts with histidine-rich glycoprotein (HRG), and the name sHIP (streptococcal histidine-rich glycoprotein-interacting protein) is therefore proposed. HRG has antibacterial activity, and when challenged by HRG, sHIP was found to rescue S. pyogenes bacteria. This and the finding that patients with invasive S. pyogenes infection respond with antibody production against sHIP suggest a role for the protein in S. pyogenes pathogenesis. PMID:24825900

Characterisation of chicken TES and its role in cell spreading and motility.

PubMed

Griffith, Elen; Coutts, Amanda S; Black, Donald M

2004-03-01

Previously we identified TES as a candidate tumour suppressor gene that is located at human chromosome 7q31.1. More recently, we and others have shown TES to encode a novel LIM domain protein that localises to focal adhesions. Here, we present the cloning and functional analysis of the chicken orthologue of TES, cTES. The TES proteins are highly conserved between chicken and human, showing 89% identity at the amino acid level. We show that the cTES protein localised at focal adhesions, actin stress fibres, and sites of cell-cell contact, and GST-cTES can pull-down zyxin and actin. To investigate a functional role for cTES, we looked at the effect of its overexpression on cell spreading and cell motility. Cells overexpressing cTES showed increased cell spreading on fibronectin, and decreased cell motility, compared to RCAS vector transfected control cells. The data from our studies with cTES support our previous findings with human TES and further implicate TES as a member of a complex of proteins that function together to regulate cell adhesion and additionally demonstrate a role for TES in cell motility. Copyright 2004 Wiley-Liss, Inc.

Arf6 regulates EGF-induced internalization of E-cadherin in breast cancer cells.

PubMed

Xu, Rui; Zhang, Yujie; Gu, Luo; Zheng, Jianchao; Cui, Jie; Dong, Jing; Du, Jun

2015-01-01

E-cadherin internalization facilitates dissolution of adherens junctions and promotes tumor cell epithelial-mesenchymal transition (EMT) and migration. Our previous results have shown that Arf6 exerts pro-migratory action in breast cancer cells after EGF stimulation. Despite the fact that EGF signaling stimulates EMT of breast cancer cells, the effect of Arf6 on internalization of E-cadherin of breast cancer cells under EGF treatment remains to be determined. Here, we showed that EGF dose-dependently stimulated E-cadherin internalization by MCF-7 cells with the maximal effect at 50 ng/ml. Meanwhile, EGF treatment markedly increased Arf6 activation. Arf6 was involved in complexes of E-cadherin, and more E-cadherin was pulled down with Arf6 when the activity of the latter was increased. Immunoblotting and immunofluorescence assays showed that transfection breast cancer cells with Arf6-T27N or Arf6 siRNA suppressed EGF-induced E-cadherin internalization. Taken together, our study demonstrated that Arf6 activation plays a potential role in EGF-induced E-cadherin internalization, providing new mechanism underlying the effect of Arf6 on promoting breast cancer cell metastasis.