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Sample records for primed vesicle state

  1. Synaptobrevin N-terminally bound to syntaxin–SNAP-25 defines the primed vesicle state in regulated exocytosis

    PubMed Central

    Walter, Alexander M.; Wiederhold, Katrin; Bruns, Dieter; Fasshauer, Dirk

    2010-01-01

    Rapid neurotransmitter release depends on the ability to arrest the SNAP receptor (SNARE)–dependent exocytosis pathway at an intermediate “cocked” state, from which fusion can be triggered by Ca2+. It is not clear whether this state includes assembly of synaptobrevin (the vesicle membrane SNARE) to the syntaxin–SNAP-25 (target membrane SNAREs) acceptor complex or whether the reaction is arrested upstream of that step. In this study, by a combination of in vitro biophysical measurements and time-resolved exocytosis measurements in adrenal chromaffin cells, we find that mutations of the N-terminal interaction layers of the SNARE bundle inhibit assembly in vitro and vesicle priming in vivo without detectable changes in triggering speed or fusion pore properties. In contrast, mutations in the last C-terminal layer decrease triggering speed and fusion pore duration. Between the two domains, we identify a region exquisitely sensitive to mutation, possibly constituting a switch. Our data are consistent with a model in which the N terminus of the SNARE complex assembles during vesicle priming, followed by Ca2+-triggered C-terminal assembly and membrane fusion. PMID:20142423

  2. Variable priming of a docked synaptic vesicle

    PubMed Central

    Jung, Jae Hoon; Szule, Joseph A.; Marshall, Robert M.; McMahan, Uel J.

    2016-01-01

    The priming of a docked synaptic vesicle determines the probability of its membrane (VM) fusing with the presynaptic membrane (PM) when a nerve impulse arrives. To gain insight into the nature of priming, we searched by electron tomography for structural relationships correlated with fusion probability at active zones of axon terminals at frog neuromuscular junctions. For terminals fixed at rest, the contact area between the VM of docked vesicles and PM varied >10-fold with a normal distribution. There was no merging of the membranes. For terminals fixed during repetitive evoked synaptic transmission, the normal distribution of contact areas was shifted to the left, due in part to a decreased number of large contact areas, and there was a subpopulation of large contact areas where the membranes were hemifused, an intermediate preceding complete fusion. Thus, fusion probability of a docked vesicle is related to the extent of its VM–PM contact area. For terminals fixed 1 h after activity, the distribution of contact areas recovered to that at rest, indicating the extent of a VM–PM contact area is dynamic and in equilibrium. The extent of VM–PM contact areas in resting terminals correlated with eccentricity in vesicle shape caused by force toward the PM and with shortness of active zone material macromolecules linking vesicles to PM components, some thought to include Ca2+ channels. We propose that priming is a variable continuum of events imposing variable fusion probability on each vesicle and is regulated by force-generating shortening of active zone material macromolecules in dynamic equilibrium. PMID:26858418

  3. The Vesicle Priming Factor CAPS Functions as a Homodimer via C2 Domain Interactions to Promote Regulated Vesicle Exocytosis*

    PubMed Central

    Petrie, Matt; Esquibel, Joseph; Maciuba, Stephanie; Takahashi, Hirohide

    2016-01-01

    Neurotransmitters and peptide hormones are secreted by regulated vesicle exocytosis. CAPS (also known as CADPS) is a 145-kDa cytosolic and peripheral membrane protein required for vesicle docking and priming steps that precede Ca2+-triggered vesicle exocytosis. CAPS binds phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and SNARE proteins and is proposed to promote SNARE protein complex assembly for vesicle docking and priming. We characterized purified soluble CAPS as mainly monomer in equilibrium with small amounts of dimer. However, the active form of CAPS bound to PC12 cell membranes or to liposomes containing PI(4,5)P2 and Q-SNARE proteins was mainly dimer. CAPS dimer formation required its C2 domain based on mutation or deletion studies. Moreover, C2 domain mutations or deletions resulted in a loss of CAPS function in regulated vesicle exocytosis, indicating that dimerization is essential for CAPS function. Comparison of the CAPS C2 domain to a structurally defined Munc13-1 C2A domain dimer revealed conserved residues involved in CAPS dimerization. We conclude that CAPS functions as a C2 domain-mediated dimer in regulated vesicle exocytosis. The unique tandem C2-PH domain of CAPS may serve as a PI(4,5)P2-triggered switch for dimerization. CAPS dimerization may be coupled to oligomeric SNARE complex assembly for vesicle docking and priming. PMID:27528604

  4. Secretory vesicle priming by CAPS is independent of its SNARE-binding MUN domain.

    PubMed

    Nguyen Truong, Cuc Quynh; Nestvogel, Dennis; Ratai, Olga; Schirra, Claudia; Stevens, David R; Brose, Nils; Rhee, JeongSeop; Rettig, Jens

    2014-11-06

    Priming of secretory vesicles is a prerequisite for their Ca(2+)-dependent fusion with the plasma membrane. The key vesicle priming proteins, Munc13s and CAPSs, are thought to mediate vesicle priming by regulating the conformation of the t-SNARE syntaxin, thereby facilitating SNARE complex assembly. Munc13s execute their priming function through their MUN domain. Given that the MUN domain of Ca(2+)-dependent activator protein for secretion (CAPS) also binds syntaxin, it was assumed that CAPSs prime vesicles through the same mechanism as Munc13s. We studied naturally occurring splice variants of CAPS2 in CAPS1/CAPS2-deficient cells and found that CAPS2 primes vesicles independently of its MUN domain. Instead, the pleckstrin homology domain of CAPS2 seemingly is essential for its priming function. Our findings indicate a priming mode for secretory vesicles. This process apparently requires membrane phospholipids, does not involve the binding or direct conformational regulation of syntaxin by MUN domains of CAPSs, and is therefore not redundant with Munc13 action.

  5. CAPS-1 and CAPS-2 are essential synaptic vesicle priming proteins.

    PubMed

    Jockusch, Wolf J; Speidel, Dina; Sigler, Albrecht; Sørensen, Jakob B; Varoqueaux, Frederique; Rhee, Jeong-Seop; Brose, Nils

    2007-11-16

    Before transmitter-filled synaptic vesicles can fuse with the plasma membrane upon stimulation they have to be primed to fusion competence. The regulation of this priming process controls the strength and plasticity of synaptic transmission between neurons, which in turn determines many complex brain functions. We show that CAPS-1 and CAPS-2 are essential components of the synaptic vesicle priming machinery. CAPS-deficient neurons contain no or very few fusion competent synaptic vesicles, which causes a selective impairment of fast phasic transmitter release. Increases in the intracellular Ca(2+) levels can transiently revert this defect. Our findings demonstrate that CAPS proteins generate and maintain a highly fusion competent synaptic vesicle pool that supports phasic Ca(2+) triggered release of transmitters.

  6. Friends and Foes in Synaptic Transmission—the Role of Tomosyn in Vesicle Priming

    PubMed Central

    Ashery, Uri; Bielopolski, Noa; Barak, Boaz; Yizhar, Ofer

    2009-01-01

    Priming is the process by which vesicles become available for fusion at nerve terminals and it is modulated by numerous proteins and second messengers. One of the prominent members of this diverse family is Tomosyn. Tomosyn has been identified as a Syntaxin-binding protein; it inhibits vesicle priming, but its mode of action is not fully understood. Tomosyn's inhibitory activity depends on its N-terminal WD40-repeat domain and is regulated by the binding of its SNARE-motif to Syntaxin. The present review describes new physiological information on Tomosyn's function and addresses possible interpretations of these results in the framework of the recently described crystal structure of the yeast Tomosyn homolog Sro7. We also present possible molecular scenarios for vesicle priming and the involvement of Tomosyn in these processes. PMID:19307030

  7. Two distinct secretory vesicle-priming steps in adrenal chromaffin cells.

    PubMed

    Liu, Yuanyuan; Schirra, Claudia; Edelmann, Ludwig; Matti, Ulf; Rhee, JeongSeop; Hof, Detlef; Bruns, Dieter; Brose, Nils; Rieger, Heiko; Stevens, David R; Rettig, Jens

    2010-09-20

    Priming of large dense-core vesicles (LDCVs) is a Ca(2+)-dependent step by which LDCVs enter a release-ready pool, involving the formation of the soluble N-ethyl-maleimide sensitive fusion protein attachment protein (SNAP) receptor complex consisting of syntaxin, SNAP-25, and synaptobrevin. Using mice lacking both isoforms of the calcium-dependent activator protein for secretion (CAPS), we show that LDCV priming in adrenal chromaffin cells entails two distinct steps. CAPS is required for priming of the readily releasable LDCV pool and sustained secretion in the continued presence of high Ca(2+) concentrations. Either CAPS1 or CAPS2 can rescue secretion in cells lacking both CAPS isoforms. Furthermore, the deficit in the readily releasable LDCV pool resulting from CAPS deletion is reversed by a constitutively open form of syntaxin but not by Munc13-1, a priming protein that facilitates the conversion of syntaxin to the open conformation. Our data indicate that CAPS functions downstream of Munc13s but also interacts functionally with Munc13s in the LDCV-priming process.

  8. Acute destruction of the synaptic ribbon reveals a role for the ribbon in vesicle priming

    PubMed Central

    Snellman, Josefin; Mehta, Bhupesh; Babai, Norbert; Bartoletti, Theodore M.; Akmentin, Wendy; Francis, Adam; Matthews, Gary; Thoreson, Wallace; Zenisek, David

    2011-01-01

    In vision, balance, and hearing, sensory receptor cells translate sensory stimuli into electrical signals whose amplitude is graded with stimulus intensity. The output synapses of these sensory neurons must provide fast signaling to follow rapidly changing stimuli, while also transmitting graded information covering a wide range of stimulus intensity and sustained for long time periods. To meet these demands, specialized machinery for transmitter release—the synaptic ribbon—has evolved at the synaptic outputs of these neurons. Here we show that acute disruption of synaptic ribbons by photodamage to the ribbon dramatically reduces both sustained and transient components of neurotransmitter release in mouse bipolar cells and salamander cones, without affecting the ultrastructure of the ribbon or its ability to localize synaptic vesicles to the active zone. Our results indicate that ribbons mediate slow as well as fast signaling at sensory synapses, and support an additional role for the synaptic ribbon in priming vesicles for exocytosis at active zones. PMID:21785435

  9. Extracellular Vesicles from Metastatic Rat Prostate Tumors Prime the Normal Prostate Tissue to Facilitate Tumor Growth

    PubMed Central

    Halin Bergström, Sofia; Hägglöf, Christina; Thysell, Elin; Bergh, Anders; Wikström, Pernilla; Lundholm, Marie

    2016-01-01

    Accumulating data indicates that tumor-derived extracellular vesicles (EVs) are responsible for tumor-promoting effects. However, if tumor EVs also prepare the tumor-bearing organ for subsequent tumor growth, and if this effect is different in low and high malignant tumors is not thoroughly explored. Here we used orthotopic rat Dunning R-3327 prostate tumors to compare the role of EVs from fast growing and metastatic MatLyLu (MLL) tumors with EVs from more indolent and non-metastatic Dunning G (G) tumors. Prostate tissue pre-conditioned with MLL-EVs in vivo facilitated G tumor establishment compared to G-EVs. MLL-EVs increased prostate epithelial proliferation and macrophage infiltration into the prostate compared to G-EVs. Both types of EVs increased macrophage endocytosis and the mRNA expression of genes associated with M2 polarization in vitro, with MLL-EVs giving the most pronounced effects. MLL-EVs also altered the mRNA expression of growth factors and cytokines in primary rat prostate fibroblasts compared to G-EVs, suggesting fibroblast activation. Our findings propose that EVs from metastatic tumors have the ability to prime the prostate tissue and enhance tumor growth to a higher extent than EVs from non-metastatic tumors. Identifying these differences could lead to novel therapeutic targets and potential prognostic markers for prostate cancer. PMID:27550147

  10. Vesicle release probability and pre-primed pool at glutamatergic synapses in area CA1 of the rat neonatal hippocampus

    PubMed Central

    Hanse, Eric; Gustafsson, Bengt

    2001-01-01

    Factors determining the release probability were examined using whole-cell patch-clamp recording and minimal stimulation (10 impulses, 50 Hz) of individual glutamatergic synapses, containing single release sites, on pyramidal neurones in the CA1 region of hippocampal slices from neonatal rats. Release probability following the first action potential in the burst (P1) varied among the synapses from 0 to 0.87 (mean ± s.d.; 0.35 ± 0.28, n = 52) and the average release during the burst (burst pool) varied from 0.4 to 4.1 events (1.7 ± 0.85, n = 52). Heterogeneity in P1 did not co-vary with that of the burst pool. By selecting burst trials during which only one release event occurred, the vesicle release probability (Pves) at a release site could be determined. It was found to vary considerably among the synapses, from 0.04 to 0.94 (0.43 ± 0.28, n = 43). This heterogeneity correlated significantly with that of P1 such that more than half of the variation of P1 could be explained by a variation in Pves. The average number of vesicles directly available for release at the onset of the burst (the pre-primed pool) was estimated as the cumulative release up to that point in the burst where a second release event did not produce higher initial release probability than that found in trials where only one vesicle was released. The average pre-primed pool varied among the synapses from 0.4 to 2.1 (1.03 ± 0.42, n = 43). It co-varied significantly with that of P1 such that it could explain the remaining variation in P1. The difference between the burst pool and the pre-primed pool suggests the presence of a fast (< 100 ms), activity-dependent priming of vesicles. Some synapses (9/52) did not show any initial release (P1 = 0), but release occurred later during the burst (‘low frequency mute synapses’). Their behaviour was explained by an absence of a pre-primed pool. PMID:11230520

  11. Chlorophyll a triplet-state ESR in frozen phosphatidylcholine vesicles

    SciTech Connect

    Hiromitsu, I.; Kevan, L.

    1988-05-19

    Photoexcited chlorophyll a (Chla) triplet state in rapidly frozen egg phosphatidylcholine (EPC) vesicles is investigated at 77 K by electron spin resonance (ESR) spectroscopy using light intensity modulation. The electron spin polarization (ESP) intensity is stronger for 0.2 mM Chla than for 1.0 mM Chla. The absolute values of the zero field splitting parameter, D, are 283 (+/-1) x 10/sup -4/ and 276 (+/-2) x 10/sup -4/ cm/sup -1/, and the average depopulation rates of the triplet state are 0.671 +/- 0.052 and 1.054 +/- 0.036 ms/sup -1/ for 0.2 mM Chla and 1.0 mM Chla, respectively. This difference can be consistently attributed to faster triplet-state migration between adjacent Chla's at the higher 1.0 mM Chla concentration. A characteristic migration time of 2.6 ms is obtained. The ESP pattern of the Chla triplet state in the frozen EPC vesicles resembles that in polycrystals more than that in glasses. This suggests that the local environment around Chla in the vesicles is more structured than in glasses.

  12. Prime Contract Awards by State. Fiscal Year 1993

    DTIC Science & Technology

    1993-01-01

    additional prior year of civil function procurement award information. Table 6 -- A new table displaying county data within each state by DoD...Value of DoD Procurement Awards, by State and County , According to Defense Component: FY 1993 ......... ................ . .. T INTRODUCTION Prime...year a new table, Table 6, has been added. Table 6displays county data within each state by DoD Component (including Civil Function) for the current

  13. Pseudomonas aeruginosa Outer Membrane Vesicles Triggered by Human Mucosal Fluid and Lysozyme Can Prime Host Tissue Surfaces for Bacterial Adhesion

    PubMed Central

    Metruccio, Matteo M. E.; Evans, David J.; Gabriel, Manal M.; Kadurugamuwa, Jagath L.; Fleiszig, Suzanne M. J.

    2016-01-01

    Pseudomonas aeruginosa is a leading cause of human morbidity and mortality that often targets epithelial surfaces. Host immunocompromise, or the presence of indwelling medical devices, including contact lenses, can predispose to infection. While medical devices are known to accumulate bacterial biofilms, it is not well understood why resistant epithelial surfaces become susceptible to P. aeruginosa. Many bacteria, including P. aeruginosa, release outer membrane vesicles (OMVs) in response to stress that can fuse with host cells to alter their function. Here, we tested the hypothesis that mucosal fluid can trigger OMV release to compromise an epithelial barrier. This was tested using tear fluid and corneal epithelial cells in vitro and in vivo. After 1 h both human tear fluid, and the tear component lysozyme, greatly enhanced OMV release from P. aeruginosa strain PAO1 compared to phosphate buffered saline (PBS) controls (∼100-fold). Transmission electron microscopy (TEM) and SDS-PAGE showed tear fluid and lysozyme-induced OMVs were similar in size and protein composition, but differed from biofilm-harvested OMVs, the latter smaller with fewer proteins. Lysozyme-induced OMVs were cytotoxic to human corneal epithelial cells in vitro and murine corneal epithelium in vivo. OMV exposure in vivo enhanced Ly6G/C expression at the corneal surface, suggesting myeloid cell recruitment, and primed the cornea for bacterial adhesion (∼4-fold, P < 0.01). Sonication disrupted OMVs retained cytotoxic activity, but did not promote adhesion, suggesting the latter required OMV-mediated events beyond cell killing. These data suggest that mucosal fluid induced P. aeruginosa OMVs could contribute to loss of epithelial barrier function during medical device-related infections. PMID:27375592

  14. ELKS1 localizes the synaptic vesicle priming protein bMunc13-2 to a specific subset of active zones.

    PubMed

    Kawabe, Hiroshi; Mitkovski, Miso; Kaeser, Pascal S; Hirrlinger, Johannes; Opazo, Felipe; Nestvogel, Dennis; Kalla, Stefan; Fejtova, Anna; Verrier, Sophie E; Bungers, Simon R; Cooper, Benjamin H; Varoqueaux, Frederique; Wang, Yun; Nehring, Ralf B; Gundelfinger, Eckart D; Rosenmund, Christian; Rizzoli, Silvio O; Südhof, Thomas C; Rhee, Jeong-Seop; Brose, Nils

    2017-03-06

    Presynaptic active zones (AZs) are unique subcellular structures at neuronal synapses, which contain a network of specific proteins that control synaptic vesicle (SV) tethering, priming, and fusion. Munc13s are core AZ proteins with an essential function in SV priming. In hippocampal neurons, two different Munc13s-Munc13-1 and bMunc13-2-mediate opposite forms of presynaptic short-term plasticity and thus differentially affect neuronal network characteristics. We found that most presynapses of cortical and hippocampal neurons contain only Munc13-1, whereas ∼10% contain both Munc13-1 and bMunc13-2. Whereas the presynaptic recruitment and activation of Munc13-1 depends on Rab3-interacting proteins (RIMs), we demonstrate here that bMunc13-2 is recruited to synapses by the AZ protein ELKS1, but not ELKS2, and that this recruitment determines basal SV priming and short-term plasticity. Thus, synapse-specific interactions of different Munc13 isoforms with ELKS1 or RIMs are key determinants of the molecular and functional heterogeneity of presynaptic AZs.

  15. Triplet states of oxygen vacancy defects in α-quartz: Center \\bf{\\text{E}^{\\prime\\prime}_{9}}

    NASA Astrophysics Data System (ADS)

    Mashkovtsev, R. I.; Pan, Y.

    2014-08-01

    The paramagnetic E^{\\prime\\prime}_{9} center in triplet state occurring in electron-irradiated, synthetic α-quartz has been investigated by using electron paramagnetic resonance (EPR) spectroscopy. The primary spin Hamiltonian parameter matrices g, D and A (hyperfine interactions for five 29Si nuclei) have now been determined. The principal values and principal directions of D and A matrices allow us to suggest the structural model for this stable triplet defect. The E^{\\prime\\prime}_{9} center involves the two unpaired electrons located in the orbitals of two silicon atoms next to one oxygen vacancy each. Firm correlations between the spin Hamiltonian matrix principal axes and crystallographic directions have been attained.

  16. A Sequential Vesicle Pool Model with a Single Release Sensor and a Ca2+-Dependent Priming Catalyst Effectively Explains Ca2+-Dependent Properties of Neurosecretion

    PubMed Central

    Walter, Alexander M.; Pinheiro, Paulo S.; Verhage, Matthijs; Sørensen, Jakob B.

    2013-01-01

    Neurotransmitter release depends on the fusion of secretory vesicles with the plasma membrane and the release of their contents. The final fusion step displays higher-order Ca2+ dependence, but also upstream steps depend on Ca2+. After deletion of the Ca2+ sensor for fast release – synaptotagmin-1 – slower Ca2+-dependent release components persist. These findings have provoked working models involving parallel releasable vesicle pools (Parallel Pool Models, PPM) driven by alternative Ca2+ sensors for release, but no slow release sensor acting on a parallel vesicle pool has been identified. We here propose a Sequential Pool Model (SPM), assuming a novel Ca2+-dependent action: a Ca2+-dependent catalyst that accelerates both forward and reverse priming reactions. While both models account for fast fusion from the Readily-Releasable Pool (RRP) under control of synaptotagmin-1, the origins of slow release differ. In the SPM the slow release component is attributed to the Ca2+-dependent refilling of the RRP from a Non-Releasable upstream Pool (NRP), whereas the PPM attributes slow release to a separate slowly-releasable vesicle pool. Using numerical integration we compared model predictions to data from mouse chromaffin cells. Like the PPM, the SPM explains biphasic release, Ca2+-dependence and pool sizes in mouse chromaffin cells. In addition, the SPM accounts for the rapid recovery of the fast component after strong stimulation, where the PPM fails. The SPM also predicts the simultaneous changes in release rate and amplitude seen when mutating the SNARE-complex. Finally, it can account for the loss of fast- and the persistence of slow release in the synaptotagmin-1 knockout by assuming that the RRP is depleted, leading to slow and Ca2+-dependent fusion from the NRP. We conclude that the elusive ‘alternative Ca2+ sensor’ for slow release might be the upstream priming catalyst, and that a sequential model effectively explains Ca2+-dependent properties of

  17. α-Synuclein Populates Both Elongated and Broken Helix States on Small Unilamellar Vesicles

    PubMed Central

    Lokappa, Sowmya Bekshe; Ulmer, Tobias S.

    2011-01-01

    The misfolding of the protein α-synuclein (αS) has been implicated in the molecular chain of events leading to Parkinson disease. Physiologically, αS undergoes a transition from a random coil to helical conformation upon encountering synaptic vesicle membranes. On analogous small unilamellar vesicles (SUVs), the conformation of αS is dominated by a single elongated αS helix. However, alternative broken helix states have been postulated, mandating experimental clarification. Here, the upper limit for the free energy difference between elongated and broken helix conformations on SUVs resembling synaptic vesicles was determined to be 1.2 ± 0.4 kcal/mol, which amounts to a population ratio of 7.6:1 between both states (12% broken helices). In response to helix breaks at different positions, αS rearranged in an opportunistic manner, thereby minimizing helix abrogations to as little as one to two turns. Enthalpy and entropy measurements of gel state SUV-αS interactions indicated that broken helix states retain the ability to relieve membrane-packing stress. Thus, broken helix states are a distinct physiological feature of the vesicle-bound αS state, making it a “checkered” protein of multiple parallel conformations. A continuous interconversion between structural states may contribute to pathological αS misfolding. PMID:21524999

  18. Guilt by dissociation: guilt primes augment the relationship between dissociative tendencies and state dissociation.

    PubMed

    Rugens, Alex; Terhune, Devin Blair

    2013-03-30

    We examined the influence of guilt on the relationship between dissociative tendencies and state dissociation during mirror-gazing in a non-clinical sample. Dissociative tendencies correlated with state dissociation following guilt primes, but not after negative or neutral primes. This suggests that guilt augments the relationship between dissociative tendencies and state dissociation.

  19. The Munc18-1 domain 3a hinge-loop controls syntaxin-1A nanodomain assembly and engagement with the SNARE complex during secretory vesicle priming

    PubMed Central

    Kasula, Ravikiran; Bademosi, Adekunle T.; Harper, Callista B.; Morrow, Isabel C.; Hosy, Eric; Collins, Brett M.; Choquet, Daniel

    2016-01-01

    Munc18-1 and syntaxin-1A control SNARE-dependent neuroexocytosis and are organized in nanodomains on the plasma membrane of neurons and neurosecretory cells. Deciphering the intra- and intermolecular steps via which they prepare secretory vesicles (SVs) for fusion is key to understanding neuronal and hormonal communication. Here, we demonstrate that expression of a priming-deficient mutant lacking 17 residues of the domain 3a hinge-loop (Munc18-1Δ317-333) in PC12 cells engineered to knockdown Munc18-1/2 markedly prolonged SV docking. Single-molecule analysis revealed nonhomogeneous diffusion of Munc18-1 and syntaxin-1A in and out of partially overlapping nanodomains. Whereas Munc18-1WT mobility increased in response to stimulation, syntaxin-1A became less mobile. These Munc18-1 and syntaxin-1A diffusional switches were blocked by the expression of Munc18-1Δ317-333, suggesting that a conformational change in the Munc18-1 hinge-loop controls syntaxin-1A and subsequent SNARE complex assembly. Accordingly, syntaxin-1A confinement was prevented by expression of botulinum neurotoxin type E. The Munc18-1 domain 3a hinge-loop therefore controls syntaxin-1A engagement into SNARE complex formation during priming. PMID:27646276

  20. Seed priming: state of the art and new perspectives.

    PubMed

    Paparella, S; Araújo, S S; Rossi, G; Wijayasinghe, M; Carbonera, D; Balestrazzi, Alma

    2015-08-01

    Priming applied to commercial seed lots is widely used by seed technologists to enhance seed vigour in terms of germination potential and increased stress tolerance. Priming can be also valuable to seed bank operators who need improved protocols of ex situ conservation of germplasm collections (crop and native species). Depending on plant species, seed morphology and physiology, different priming treatments can be applied, all of them triggering the so-called 'pre-germinative metabolism'. This physiological process takes place during early seed imbibition and includes the seed repair response (activation of DNA repair pathways and antioxidant mechanisms), essential to preserve genome integrity, ensuring proper germination and seedling development. The review provides an overview of priming technology, describing the range of physical-chemical and biological treatments currently available. Optimised priming protocols can be designed using the 'hydrotime concept' analysis which provides the theoretical bases for assessing the relationship between water potential and germination rate. Despite the efforts so far reported to further improve seed priming, novel ideas and cutting-edge investigations need to be brought into this technological sector of agri-seed industry. Multidisciplinary translational research combining digital, bioinformatic and molecular tools will significantly contribute to expand the range of priming applications to other relevant commercial sectors, e.g. the native seed market.

  1. Adhesion energy can regulate vesicle fusion and stabilize partially fused states.

    PubMed

    Long, Rong; Hui, Chung-Yuen; Jagota, Anand; Bykhovskaia, Maria

    2012-07-07

    Release of neurotransmitters from nerve terminals occurs by fusion of synaptic vesicles with the plasma membrane, and this process is highly regulated. Although major molecular components that control docking and fusion of vesicles to the synaptic membrane have been identified, the detailed mechanics of this process is not yet understood. We have developed a mathematical model that predicts how adhesion forces imposed by docking and fusion molecular machinery would affect the fusion process. We have computed the membrane stress that is produced by adhesion-driven vesicle bending and find that it is compressive. Further, our computations of the membrane curvature predict that strong adhesion can create a metastable state with a partially opened pore that would correspond to the 'kiss and run' release mode. Our model predicts that the larger the vesicle size, the more likely the metastable state with a transiently opened pore. These results contribute to understanding the mechanics of the fusion process, including possible clamping of the fusion by increasing molecular adhesion, and a balance between 'kiss and run' and full collapse fusion modes.

  2. [Medical doctors as the state presidents and prime ministers--a biographical analysis].

    PubMed

    Lass, Piotr; Szarszewski, Adam; Gaworska-Krzemińska, Aleksandra; Sławek, Jarosław

    2012-01-01

    The authors overviewed the biographies of 29 medical doctors who became the heads of the state or the prime ministers of their countries. Most of them ruled in a countries of fresh or unstable democracies, most often in Asia, Africa and Latin America, three of them were bloody dictators. With the exemptions of Georges Clemenceau and Sun-Yat-Sen they were not famous as historical figures, although some were good administrators like the prime minister of Norway, Gro Harlem Brundtland, Malayan prime minister Mahatir bin Mohamad, Brasilian and Chilean presidents, Juscelino Kubitschek and Veronica Bachelet. Regarding specialisation was mostly general medics or they specialised in public health.

  3. Qualitative changes in the proteome of extracellular vesicles accompanying cancer cell transition to mesenchymal state.

    PubMed

    Garnier, Delphine; Magnus, Nathalie; Meehan, Brian; Kislinger, Thomas; Rak, Janusz

    2013-10-15

    Transitions of the cancer cell phenotype between epithelial and mesenchymal states (EMT) are likely to alter the patterns of intercellular communication. In this regard we have previously documented that EMT-like changes trigger quantitative rearrangements in exosomal vesicle emission in A431 cancer cells driven by oncogenic epidermal growth factor receptor (EGFR). Here we report that extracellular vesicles (EVs) produced by these cancer cells in their epithelial and mesenchymal states exhibit profound qualitative differences in their proteome. Thus, induction of the EMT-like state through blockade of E-cadherin and EGFR stimulation provoked a mesenchymal shift in cellular morphology and enrichment in the CD44-high/CD24-low immunophenotype, often linked to cellular stemness. This change also resulted in reprogramming of the EV-related proteome (distinct from that of corresponding cells), which contained 30 unique protein signals, and revealed enrichment in pathways related to cellular growth, cell-to-cell signaling, and cell movement. Some of the most prominent EV-related proteins were validated, including integrin α2 and tetraspanin CD9. We propose that changes in cellular differentiation status translate into unique qualitative rearrangements in the cargo of EVs, a process that may have implications for intercellular communication and could serve as source of new biomarkers to detect EMT-like processes in cancer.

  4. Vesicle Pools: Lessons from Adrenal Chromaffin Cells

    PubMed Central

    Stevens, David R.; Schirra, Claudia; Becherer, Ute; Rettig, Jens

    2011-01-01

    The adrenal chromaffin cell serves as a model system to study fast Ca2+-dependent exocytosis. Membrane capacitance measurements in combination with Ca2+ uncaging offers a temporal resolution in the millisecond range and reveals that catecholamine release occurs in three distinct phases. Release of a readily releasable (RRP) and a slowly releasable (SRP) pool are followed by sustained release, due to maturation, and release of vesicles which were not release-ready at the start of the stimulus. Trains of depolarizations, a more physiological stimulus, induce release from a small immediately releasable pool of vesicles residing adjacent to calcium channels, as well as from the RRP. The SRP is poorly activated by depolarization. A sequential model, in which non-releasable docked vesicles are primed to a slowly releasable state, and then further mature to the readily releasable state, has been proposed. The docked state, dependent on membrane proximity, requires SNAP-25, synaptotagmin, and syntaxin. The ablation or modification of SNAP-25 and syntaxin, components of the SNARE complex, as well as of synaptotagmin, the calcium sensor, and modulators such complexins and Snapin alter the properties and/or magnitudes of different phases of release, and in particular can ablate the RRP. These results indicate that the composition of the SNARE complex and its interaction with modulatory molecules drives priming and provides a molecular basis for different pools of releasable vesicles. PMID:21423410

  5. Subdiffractional tracking of internalized molecules reveals heterogeneous motion states of synaptic vesicles.

    PubMed

    Joensuu, Merja; Padmanabhan, Pranesh; Durisic, Nela; Bademosi, Adekunle T D; Cooper-Williams, Elizabeth; Morrow, Isabel C; Harper, Callista B; Jung, WooRam; Parton, Robert G; Goodhill, Geoffrey J; Papadopulos, Andreas; Meunier, Frédéric A

    2016-10-24

    Our understanding of endocytic pathway dynamics is severely restricted by the diffraction limit of light microscopy. To address this, we implemented a novel technique based on the subdiffractional tracking of internalized molecules (sdTIM). This allowed us to image anti-green fluorescent protein Atto647N-tagged nanobodies trapped in synaptic vesicles (SVs) from live hippocampal nerve terminals expressing vesicle-associated membrane protein 2 (VAMP2)-pHluorin with 36-nm localization precision. Our results showed that, once internalized, VAMP2-pHluorin/Atto647N-tagged nanobodies exhibited a markedly lower mobility than on the plasma membrane, an effect that was reversed upon restimulation in presynapses but not in neighboring axons. Using Bayesian model selection applied to hidden Markov modeling, we found that SVs oscillated between diffusive states or a combination of diffusive and transport states with opposite directionality. Importantly, SVs exhibiting diffusive motion were relatively less likely to switch to the transport motion. These results highlight the potential of the sdTIM technique to provide new insights into the dynamics of endocytic pathways in a wide variety of cellular settings.

  6. Dollar Summary of Prime Contract Awards by State, County, Contractor, and Place. Part 3. (Aitkin, Minnesota-Yamhill, Oregon)

    DTIC Science & Technology

    1989-01-01

    by State, County , Contractor, and Place. Part 3(Aitkin, Hinnosota-Yardhill, Oregon) 6. AUTHOR(S) 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) B...TITLE AND DESCRIPTION ESTIMATED AVAILABILITY DATE ST25. Dollar Summary of Prime Contract Awards by State, County , Contractor, and P!ace (Previous Title...Prime Contract Awards Over $25,000 by State, County , Contractor, and Place) March 15 This tabulation shows the net value of DoD prime contract

  7. Translation-priming effects on tip-of-the-tongue states

    PubMed Central

    Gollan, Tamar H.; Ferreira, Victor S.; Cera, Cynthia; Flett, Susanna

    2013-01-01

    Bilinguals experience more tip-of-the-tongue (TOT) states than monolinguals, but it is not known if this is caused in part by access of representations from both of bilinguals’ languages, or dual-language activation. In two translation priming experiments, bilinguals were given three Spanish primes and produced either semantically (Experiment 1) or phonologically related Spanish words (Experiment 2) to each. They then named a picture in English. On critical trials, one of the primes was the Spanish translation of the English picture name. Translation primes significantly increased TOTs regardless of task, and also speeded correct retrievals but only with the semantic task. In both experiments translation-primed TOTs were significantly more likely to resolve spontaneously. These results illustrate an effect of non-dominant language activation on dominant-language retrieval, as well as imply that TOTs can arise during (not after) lexical retrieval, at a level of processing where translation equivalent lexical representations normally interact (possibly competing for selection, or mutually activating each other, or both depending on the locus of retrieval failure). PMID:24644375

  8. Additive effects on the energy barrier for synaptic vesicle fusion cause supralinear effects on the vesicle fusion rate.

    PubMed

    Schotten, Sebastiaan; Meijer, Marieke; Walter, Alexander Matthias; Huson, Vincent; Mamer, Lauren; Kalogreades, Lawrence; ter Veer, Mirelle; Ruiter, Marvin; Brose, Nils; Rosenmund, Christian; Sørensen, Jakob Balslev; Verhage, Matthijs; Cornelisse, Lennart Niels

    2015-04-14

    The energy required to fuse synaptic vesicles with the plasma membrane ('activation energy') is considered a major determinant in synaptic efficacy. From reaction rate theory, we predict that a class of modulations exists, which utilize linear modulation of the energy barrier for fusion to achieve supralinear effects on the fusion rate. To test this prediction experimentally, we developed a method to assess the number of releasable vesicles, rate constants for vesicle priming, unpriming, and fusion, and the activation energy for fusion by fitting a vesicle state model to synaptic responses induced by hypertonic solutions. We show that complexinI/II deficiency or phorbol ester stimulation indeed affects responses to hypertonic solution in a supralinear manner. An additive vs multiplicative relationship between activation energy and fusion rate provides a novel explanation for previously observed non-linear effects of genetic/pharmacological perturbations on synaptic transmission and a novel interpretation of the cooperative nature of Ca(2+)-dependent release.

  9. Prime Knowledge about Primes

    ERIC Educational Resources Information Center

    Eisenberg, Theodore

    2007-01-01

    Several proofs demonstrating that there are infinitely many primes, different types of primes, tests of primality, pseudo primes, prime number generators and open questions about primes are discussed in Section 1. Some of these notions are elaborated upon in Section 2, with discussions of the Riemann zeta function and how algorithmic complexity…

  10. Vesicle Photonics

    SciTech Connect

    Vasdekis, Andreas E.; Scott, E. A.; Roke, Sylvie; Hubbell, J. A.; Psaltis, D.

    2013-04-03

    Thin membranes, under appropriate boundary conditions, can self-assemble into vesicles, nanoscale bubbles that encapsulate and hence protect or transport molecular payloads. In this paper, we review the types and applications of light fields interacting with vesicles. By encapsulating light-emitting molecules (e.g. dyes, fluorescent proteins, or quantum dots), vesicles can act as particles and imaging agents. Vesicle imaging can take place also under second harmonic generation from vesicle membrane, as well as employing mass spectrometry. Light fields can also be employed to transport vesicles using optical tweezers (photon momentum) or directly pertrurbe the stability of vesicles and hence trigger the delivery of the encapsulated payload (photon energy).

  11. Information on State versus Local Administration of CETA Prime Sponsors in Michigan. Report to the Chairman, Committee on Education and Labor, House of Representatives.

    ERIC Educational Resources Information Center

    General Accounting Office, Washington, DC. Div. of Human Resources.

    State administration of the Comprehensive Employment and Training Act (CETA) Balance of State (BOS) programs was compared with administration of CETA programs by local (regular) prime sponsors in Michigan to gain insight on the potential impact of increased state control. BOS prime sponsors spent less than regular prime sponsors for administration…

  12. Superpriming of synaptic vesicles after their recruitment to the readily releasable pool

    PubMed Central

    Lee, Jae Sung; Ho, Won-Kyung; Neher, Erwin; Lee, Suk-Ho

    2013-01-01

    Recruitment of release-competent vesicles during sustained synaptic activity is one of the major factors governing short-term plasticity. During bursts of synaptic activity, vesicles are recruited to a fast-releasing pool from a reluctant vesicle pool through an actin-dependent mechanism. We now show that newly recruited vesicles in the fast-releasing pool do not respond at full speed to a strong Ca2+ stimulus, but require approximately 4 s to mature to a “superprimed” state. Superpriming was found to be altered by agents that modulate the function of unc13 homolog proteins (Munc13s), but not by calmodulin inhibitors or actin-disrupting agents. These findings indicate that recruitment and superpriming of vesicles are regulated by separate mechanisms, which require integrity of the cytoskeleton and activation of Munc13s, respectively. We propose that refilling of the fast-releasing vesicle pool proceeds in two steps, rapid actin-dependent “positional priming,” which brings vesicles closer to Ca2+ sources, followed by slower superpriming, which enhances the Ca2+ sensitivity of primed vesicles. PMID:23980146

  13. Characterization and genetic manipulation of primed stem cells into a functional naïve state with ESRRB

    PubMed Central

    Rossello, Ricardo Antonio; Pfenning, Andreas; Howard, Jason T; Hochgeschwender, Ute

    2016-01-01

    AIM To identify differences between primed mouse embryonic stem cells (ESCs) and fully functional naive ESCs; to manipulate primed cells into a naive state. METHODS We have cultured 3 lines of cells from different mouse strains that have been shown to be naive or primed as determined by generating germline-transmitting chimeras. Cells were put through a battery of tests to measure the different features. RNA from cells was analyzed using microarrays, to determine a priority list of the differentially expressed genes. These were later validated by quantificational real-time polymerase chain reaction. Viral cassettes were created to induce expression of differentially expressed genes in the primed cells through lentiviral transduction. Primed reprogrammed cells were subjected to in-vivo incorporation studies. RESULTS Most results show that both primed and naive cells have similar features (morphology, proliferation rates, stem cell genes expressed). However, there were some genes that were differentially expressed in the naïve cells relative to the primed cells. Key upregulated genes in naïve cells include ESRRB, ERAS, ATRX, RNF17, KLF-5, and MYC. After over-expressing some of these genes the primed cells were able to incorporate into embryos in-vivo, re-acquiring a feature previously absent in these cells. CONCLUSION Although there are no notable phenotypic differences, there are key differences in gene expression between these naïve and primed stem cells. These differences can be overcome through overexpression. PMID:27822342

  14. Graph states of prime-power dimension from generalized CNOT quantum circuit

    PubMed Central

    Chen, Lin; Zhou, D. L.

    2016-01-01

    We construct multipartite graph states whose dimension is the power of a prime number. This is realized by the finite field, as well as the generalized controlled-NOT quantum circuit acting on two qudits. We propose the standard form of graph states up to local unitary transformations and particle permutations. The form greatly simplifies the classification of graph states as we illustrate up to five qudits. We also show that some graph states are multipartite maximally entangled states in the sense that any bipartition of the system produces a bipartite maximally entangled state. We further prove that 4-partite maximally entangled states exist when the dimension is an odd number at least three or a multiple of four. PMID:27272401

  15. Graph states of prime-power dimension from generalized CNOT quantum circuit.

    PubMed

    Chen, Lin; Zhou, D L

    2016-06-07

    We construct multipartite graph states whose dimension is the power of a prime number. This is realized by the finite field, as well as the generalized controlled-NOT quantum circuit acting on two qudits. We propose the standard form of graph states up to local unitary transformations and particle permutations. The form greatly simplifies the classification of graph states as we illustrate up to five qudits. We also show that some graph states are multipartite maximally entangled states in the sense that any bipartition of the system produces a bipartite maximally entangled state. We further prove that 4-partite maximally entangled states exist when the dimension is an odd number at least three or a multiple of four.

  16. Profiling of Altered Metabolomic States in Nicotiana tabacum Cells Induced by Priming Agents

    PubMed Central

    Mhlongo, Msizi I.; Steenkamp, Paul A.; Piater, Lizelle A.; Madala, Ntakadzeni E.; Dubery, Ian A.

    2016-01-01

    Metabolomics has developed into a valuable tool for advancing our understanding of plant metabolism. Plant innate immune defenses can be activated and enhanced so that, subsequent to being pre-sensitized, plants are able to launch a stronger and faster defense response upon exposure to pathogenic microorganisms, a phenomenon known as priming. Here, three contrasting chemical activators, namely acibenzolar-S-methyl, azelaic acid and riboflavin, were used to induce a primed state in Nicotiana tabacum cells. Identified biomarkers were then compared to responses induced by three phytohormones—abscisic acid, methyljasmonate, and salicylic acid. Altered metabolomes were studied using a metabolite fingerprinting approach based on liquid chromatography and mass spectrometry. Multivariate data models indicated that these inducers cause time-dependent metabolic perturbations in the cultured cells and revealed biomarkers of which the levels are affected by these agents. A total of 34 metabolites were annotated from the mass spectral data and online databases. Venn diagrams were used to identify common biomarkers as well as those unique to a specific agent. Results implicate 20 cinnamic acid derivatives conjugated to (i) quinic acid (chlorogenic acids), (ii) tyramine, (iii) polyamines, or (iv) glucose as discriminatory biomarkers of priming in tobacco cells. Functional roles for most of these metabolites in plant defense responses could thus be proposed. Metabolites induced by the activators belong to the early phenylpropanoid pathway, which indicates that different stimuli can activate similar pathways but with different metabolite fingerprints. Possible linkages to phytohormone-dependent pathways at a metabolomic level were indicated in the case of cells treated with salicylic acid and methyljasmonate. The results contribute to a better understanding of the priming phenomenon and advance our knowledge of cinnamic acid derivatives as versatile defense metabolites. PMID

  17. Gas vesicles.

    PubMed Central

    Walsby, A E

    1994-01-01

    The gas vesicle is a hollow structure made of protein. It usually has the form of a cylindrical tube closed by conical end caps. Gas vesicles occur in five phyla of the Bacteria and two groups of the Archaea, but they are mostly restricted to planktonic microorganisms, in which they provide buoyancy. By regulating their relative gas vesicle content aquatic microbes are able to perform vertical migrations. In slowly growing organisms such movements are made more efficiently than by swimming with flagella. The gas vesicle is impermeable to liquid water, but it is highly permeable to gases and is normally filled with air. It is a rigid structure of low compressibility, but it collapses flat under a certain critical pressure and buoyancy is then lost. Gas vesicles in different organisms vary in width, from 45 to > 200 nm; in accordance with engineering principles the narrower ones are stronger (have higher critical pressures) than wide ones, but they contain less gas space per wall volume and are therefore less efficient at providing buoyancy. A survey of gas-vacuolate cyanobacteria reveals that there has been natural selection for gas vesicles of the maximum width permitted by the pressure encountered in the natural environment, which is mainly determined by cell turgor pressure and water depth. Gas vesicle width is genetically determined, perhaps through the amino acid sequence of one of the constituent proteins. Up to 14 genes have been implicated in gas vesicle production, but so far the products of only two have been shown to be present in the gas vesicle: GvpA makes the ribs that form the structure, and GvpC binds to the outside of the ribs and stiffens the structure against collapse. The evolution of the gas vesicle is discussed in relation to the homologies of these proteins. Images PMID:8177173

  18. Ground- and excited-state tautomerism in 2-(3{prime}-hydroxy-2{prime}-pyridyl)benzimidazole

    SciTech Connect

    Prieto, F.R.; Rodriguez, M.C.R.; Gonzalez, M.M.; Fernandez, M.A.R.

    1994-09-01

    Ground-state HPyBI is determined to have keto-enol equilibrium in water, and the enol form predominates in nonaqueous solutions. The keto form is the only excited form in all the solvents considered. Ultrafast intramolecular proton transfer creates the enol form from the keto form. 47 refs., 6 figs., 3 tabs.

  19. Studying calcium triggered vesicle fusion in a single vesicle-vesicle content/lipid mixing system

    PubMed Central

    Kyoung, Minjoung; Zhang, Yunxiang; Diao, Jiajie; Chu, Steven; Brunger, Axel T.

    2013-01-01

    This Protocol describes a single vesicle-vesicle microscopy system to study Ca2+-triggered vesicle fusion. Donor vesicles contain reconstituted synaptobrevin and synaptotagmin-1. Acceptor vesicles contain reconstituted syntaxin and SNAP-25, and are tethered to a PEG-coated glass surface. Donor vesicles are mixed with the tethered acceptor vesicles and incubated for several minutes at zero Ca2+-concentration, resulting in a collection of single interacting vesicle pairs. The donor vesicles also contain two spectrally distinct fluorophores that allow simultaneous monitoring of temporal changes of the content and membrane. Upon Ca2+-injection into the sample chamber, our system therefore differentiates between hemifusion and complete fusion of interacting vesicle pairs and determines the temporal sequence of these events on a sub-hundred millisecond timescale. Other factors, such as complexin, can be easily added. Our system is unique by monitoring both content and lipid mixing, and by starting from a metastable state of interacting vesicle pairs prior to Ca2+-injection. PMID:23222454

  20. Disruption of the autoinhibited state primes the E3 ligase parkin for activation and catalysis.

    PubMed

    Kumar, Atul; Aguirre, Jacob D; Condos, Tara E C; Martinez-Torres, R Julio; Chaugule, Viduth K; Toth, Rachel; Sundaramoorthy, Ramasubramanian; Mercier, Pascal; Knebel, Axel; Spratt, Donald E; Barber, Kathryn R; Shaw, Gary S; Walden, Helen

    2015-10-14

    The PARK2 gene is mutated in 50% of autosomal recessive juvenile parkinsonism (ARJP) cases. It encodes parkin, an E3 ubiquitin ligase of the RBR family. Parkin exists in an autoinhibited state that is activated by phosphorylation of its N-terminal ubiquitin-like (Ubl) domain and binding of phosphoubiquitin. We describe the 1.8 Å crystal structure of human parkin in its fully inhibited state and identify the key interfaces to maintain parkin inhibition. We identify the phosphoubiquitin-binding interface, provide a model for the phosphoubiquitin-parkin complex and show how phosphorylation of the Ubl domain primes parkin for optimal phosphoubiquitin binding. Furthermore, we demonstrate that the addition of phosphoubiquitin leads to displacement of the Ubl domain through loss of structure, unveiling a ubiquitin-binding site used by the E2~Ub conjugate, thus leading to active parkin. We find the role of the Ubl domain is to prevent parkin activity in the absence of the phosphorylation signals, and propose a model for parkin inhibition, optimization for phosphoubiquitin recruitment, release of inhibition by the Ubl domain and engagement with an E2~Ub conjugate. Taken together, this model provides a mechanistic framework for activating parkin.

  1. Rapid Detection of an ABT-737-Sensitive Primed for Death State in Cells Using Microplate-Based Respirometry

    PubMed Central

    Clerc, Pascaline; Carey, Gregory B.; Mehrabian, Zara; Wei, Michael; Hwang, Hyehyun; Girnun, Geoffrey D.; Chen, Hegang; Martin, Stuart S.; Polster, Brian M.

    2012-01-01

    Cells that exhibit an absolute dependence on the anti-apoptotic BCL-2 protein for survival are termed “primed for death” and are killed by the BCL-2 antagonist ABT-737. Many cancers exhibit a primed phenotype, including some that are resistant to conventional chemotherapy due to high BCL-2 expression. We show here that 1) stable BCL-2 overexpression alone can induce a primed for death state and 2) that an ABT-737-induced loss of functional cytochrome c from the electron transport chain causes a reduction in maximal respiration that is readily detectable by microplate-based respirometry. Stable BCL-2 overexpression sensitized non-tumorigenic MCF10A mammary epithelial cells to ABT-737-induced caspase-dependent apoptosis. Mitochondria within permeabilized BCL-2 overexpressing cells were selectively vulnerable to ABT-737-induced cytochrome c release compared to those from control-transfected cells, consistent with a primed state. ABT-737 treatment caused a dose-dependent impairment of maximal O2 consumption in MCF10A BCL-2 overexpressing cells but not in control-transfected cells or in immortalized mouse embryonic fibroblasts lacking both BAX and BAK. This impairment was rescued by delivering exogenous cytochrome c to mitochondria via saponin-mediated plasma membrane permeabilization. An ABT-737-induced reduction in maximal O2 consumption was also detectable in SP53, JeKo-1, and WEHI-231 B-cell lymphoma cell lines, with sensitivity correlating with BCL-2:MCL-1 ratio and with susceptibility (SP53 and JeKo-1) or resistance (WEHI-231) to ABT-737-induced apoptosis. Multiplexing respirometry assays to ELISA-based determination of cytochrome c redistribution confirmed that respiratory inhibition was associated with cytochrome c release. In summary, cell-based respiration assays were able to rapidly identify a primed for death state in cells with either artificially overexpressed or high endogenous BCL-2. Rapid detection of a primed for death state in individual cancers

  2. Word encoding during sleep is suggested by correlations between word-evoked up-states and post-sleep semantic priming.

    PubMed

    Ruch, Simon; Koenig, Thomas; Mathis, Johannes; Roth, Corinne; Henke, Katharina

    2014-01-01

    To test whether humans can encode words during sleep we played everyday words to men while they were napping and assessed priming from sleep-played words following waking. Words were presented during non-rapid eye movement (NREM) sleep. Priming was assessed using a semantic and a perceptual priming test. These tests measured differences in the processing of words that had been or had not been played during sleep. Synonyms to sleep-played words were the targets in the semantic priming test that tapped the meaning of sleep-played words. All men responded to sleep-played words by producing up-states in their electroencephalogram. Up-states are NREM sleep-specific phases of briefly increased neuronal excitability. The word-evoked up-states might have promoted word processing during sleep. Yet, the mean performance in the priming tests administered following sleep was at chance level, which suggests that participants as a group failed to show priming following sleep. However, performance in the two priming tests was positively correlated to each other and to the magnitude of the word-evoked up-states. Hence, the larger a participant's word-evoked up-states, the larger his perceptual and semantic priming. Those participants who scored high on all variables must have encoded words during sleep. We conclude that some humans are able to encode words during sleep, but more research is needed to pin down the factors that modulate this ability.

  3. Word encoding during sleep is suggested by correlations between word-evoked up-states and post-sleep semantic priming

    PubMed Central

    Ruch, Simon; Koenig, Thomas; Mathis, Johannes; Roth, Corinne; Henke, Katharina

    2014-01-01

    To test whether humans can encode words during sleep we played everyday words to men while they were napping and assessed priming from sleep-played words following waking. Words were presented during non-rapid eye movement (NREM) sleep. Priming was assessed using a semantic and a perceptual priming test. These tests measured differences in the processing of words that had been or had not been played during sleep. Synonyms to sleep-played words were the targets in the semantic priming test that tapped the meaning of sleep-played words. All men responded to sleep-played words by producing up-states in their electroencephalogram. Up-states are NREM sleep-specific phases of briefly increased neuronal excitability. The word-evoked up-states might have promoted word processing during sleep. Yet, the mean performance in the priming tests administered following sleep was at chance level, which suggests that participants as a group failed to show priming following sleep. However, performance in the two priming tests was positively correlated to each other and to the magnitude of the word-evoked up-states. Hence, the larger a participant's word-evoked up-states, the larger his perceptual and semantic priming. Those participants who scored high on all variables must have encoded words during sleep. We conclude that some humans are able to encode words during sleep, but more research is needed to pin down the factors that modulate this ability. PMID:25452740

  4. CAPS1 stabilizes the state of readily releasable synaptic vesicles to fusion competence at CA3–CA1 synapses in adult hippocampus

    PubMed Central

    Shinoda, Yo; Ishii, Chiaki; Fukazawa, Yugo; Sadakata, Tetsushi; Ishii, Yuki; Sano, Yoshitake; Iwasato, Takuji; Itohara, Shigeyoshi; Furuichi, Teiichi

    2016-01-01

    Calcium-dependent activator protein for secretion 1 (CAPS1) regulates exocytosis of dense-core vesicles in neuroendocrine cells and of synaptic vesicles in neurons. However, the synaptic function of CAPS1 in the mature brain is unclear because Caps1 knockout (KO) results in neonatal death. Here, using forebrain-specific Caps1 conditional KO (cKO) mice, we demonstrate, for the first time, a critical role of CAPS1 in adult synapses. The amplitude of synaptic transmission at CA3–CA1 synapses was strongly reduced, and paired-pulse facilitation was significantly increased, in acute hippocampal slices from cKO mice compared with control mice, suggesting a perturbation in presynaptic function. Morphological analysis revealed an accumulation of synaptic vesicles in the presynapse without any overall morphological change. Interestingly, however, the percentage of docked vesicles was markedly decreased in the Caps1 cKO. Taken together, our findings suggest that CAPS1 stabilizes the state of readily releasable synaptic vesicles, thereby enhancing neurotransmitter release at hippocampal synapses. PMID:27545744

  5. Cell-Type-Specific Chromatin States Differentially Prime Squamous Cell Carcinoma Tumor-Initiating Cells for Epithelial to Mesenchymal Transition.

    PubMed

    Latil, Mathilde; Nassar, Dany; Beck, Benjamin; Boumahdi, Soufiane; Wang, Li; Brisebarre, Audrey; Dubois, Christine; Nkusi, Erwin; Lenglez, Sandrine; Checinska, Agnieszka; Vercauteren Drubbel, Alizée; Devos, Michael; Declercq, Wim; Yi, Rui; Blanpain, Cédric

    2017-02-02

    Epithelial to mesenchymal transition (EMT) in cancer cells has been associated with metastasis, stemness, and resistance to therapy. Some tumors undergo EMT while others do not, which may reflect intrinsic properties of their cell of origin. However, this possibility is largely unexplored. By targeting the same oncogenic mutations to discrete skin compartments, we show that cell-type-specific chromatin and transcriptional states differentially prime tumors to EMT. Squamous cell carcinomas (SCCs) derived from interfollicular epidermis (IFE) are generally well differentiated, while hair follicle (HF) stem cell-derived SCCs frequently exhibit EMT, efficiently form secondary tumors, and possess increased metastatic potential. Transcriptional and epigenomic profiling revealed that IFE and HF tumor-initiating cells possess distinct chromatin landscapes and gene regulatory networks associated with tumorigenesis and EMT that correlate with accessibility of key epithelial and EMT transcription factor binding sites. These findings highlight the importance of chromatin states and transcriptional priming in dictating tumor phenotypes and EMT.

  6. How state counter-industry campaigns help prime perceptions of tobacco industry practices to promote reductions in youth smoking

    PubMed Central

    Hersey, J; Niederdeppe, J; Ng, S; Mowery, P; Farrelly, M; Messeri, P

    2005-01-01

    Methods: Rates of youth smoking were compared in three groups of states: (1) those with long funded counter-industry campaigns (California, Florida, and Massachusetts); (2) states with more recently funded counter-industry media campaigns (Indiana, Minnesota, Mississippi, and New Jersey); and (3) other states. An analysis was performed for a series of national telephone surveys of 12–17 year olds between 1999 and 2002, controlling for differences in demographic background, the price of cigarettes, and exposure to the national truth® campaign. Results: Between 1999 and 2002, rates of current smoking and established smoking decreased significantly faster in states with established or more newly funded counter-industry campaigns than in other states. State counter-industry campaigns appear to prime, or make more salient, negative perceptions about tobacco industry practices. Conclusion: Results highlight the value of continued state counter-industry campaigns. PMID:16319360

  7. Double resonance spectroscopy of the D {}^1 \\Pi _u^+ and B^{\\prime \\prime }\\barB^1\\Sigma ^+_u states near the third dissociation threshold of H2

    NASA Astrophysics Data System (ADS)

    Ekey, R. C.; Cordova, A. E.; Duan, W.; Chartrand, A. M.; McCormack, E. F.

    2013-12-01

    Double-resonance laser spectroscopy via the E,F {}^1 \\Sigma _g^+, v^{\\prime }=6, J^{\\prime } state was used to probe the energy region below the third dissociation limit of molecular hydrogen. Resonantly enhanced multi-photon ionization spectra were recorded by detecting ion production as a function of energy using a time-of-flight mass spectrometer. Energies and line widths for the v = 14-17 levels of the D{}^1 \\Pi _u^+ state of H2 are reported and compared to experimental data obtained by using VUV synchrotron light excitation (Dickenson et al 2010 J. Chem. Phys. 133 144317) and fully ab initio non-adiabatic calculations of D {}^1 \\Pi _u^+ state energies and line widths (Glass-Maujean et al 2012 Phys. Rev. A 86 052507). Several high vibrational levels of the B^{\\prime \\prime }\\bar{B}^1\\Sigma ^+_u state were also observed in this region. Term energies and rotational constants for the v = 67-69 vibrational levels are reported and compared to highly accurate ro-vibrational energy level predictions from fully ab initio non-adiabatic calculations of the first six {}^1 \\Sigma _u^+ levels of H2 (Wolniewicz et al 2006 J. Mol. Spectrosc. 238 118). While additional observed transitions can be assigned to other states, several unassigned features in the spectra highlight the need for a fully integrated theoretical treatment of dissociation and ionization to understand the complex pattern of highly vibrationally excited states expected in this region.

  8. Additive effects on the energy barrier for synaptic vesicle fusion cause supralinear effects on the vesicle fusion rate

    PubMed Central

    Schotten, Sebastiaan; Meijer, Marieke; Walter, Alexander Matthias; Huson, Vincent; Mamer, Lauren; Kalogreades, Lawrence; ter Veer, Mirelle; Ruiter, Marvin; Brose, Nils; Rosenmund, Christian

    2015-01-01

    The energy required to fuse synaptic vesicles with the plasma membrane (‘activation energy’) is considered a major determinant in synaptic efficacy. From reaction rate theory, we predict that a class of modulations exists, which utilize linear modulation of the energy barrier for fusion to achieve supralinear effects on the fusion rate. To test this prediction experimentally, we developed a method to assess the number of releasable vesicles, rate constants for vesicle priming, unpriming, and fusion, and the activation energy for fusion by fitting a vesicle state model to synaptic responses induced by hypertonic solutions. We show that complexinI/II deficiency or phorbol ester stimulation indeed affects responses to hypertonic solution in a supralinear manner. An additive vs multiplicative relationship between activation energy and fusion rate provides a novel explanation for previously observed non-linear effects of genetic/pharmacological perturbations on synaptic transmission and a novel interpretation of the cooperative nature of Ca2+-dependent release. DOI: http://dx.doi.org/10.7554/eLife.05531.001 PMID:25871846

  9. The low-energy, charge-transfer excited states of 4-amino-4-prime-nitrodiphenyl sulfide

    NASA Technical Reports Server (NTRS)

    O'Connor, Donald B.; Scott, Gary W.; Tran, Kim; Coulter, Daniel R.; Miskowski, Vincent M.; Stiegman, Albert E.; Wnek, Gary E.

    1992-01-01

    Absorption and emission spectra of 4-amino-4-prime-nitrodiphenyl sulfide in polar and nonpolar solvents were used to characterize and assign the low-energy excited states of the molecule. Fluorescence-excitation anisotropy spectra and fluorescence quantum yields were also used to characterize the photophysics of these states. The lowest-energy fluorescent singlet state was determined to be an intramolecular charge transfer (ICT) state involving transfer of a full electron charge from the amino to the nitro group yielding a dipole moment of about 50 D. A low-energy, intense absorption band is assigned as a transition to a different ICT state involving a partial electron charge transfer from sulfur to the nitro group.

  10. O-GlcNAc is required for the survival of primed pluripotent stem cells and their reversion to the naïve state.

    PubMed

    Miura, Taichi; Nishihara, Shoko

    2016-10-27

    "Naïve" mouse embryonic stem cells (ESCs) are derived from pre-implantation embryos and possess pluripotency, the ability to differentiate into any cell type of the body. "Primed" mouse epiblast stem cells (EpiSCs) are also pluripotent but are derived from post-implantation embryos. ESC-derived EpiSCs (ESD-EpiSCs) are "primed" pluripotent stem cells and can revert to naïve reverted ESCs (rESCs). O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) is a posttranslational modification in the cytoplasm and nucleus. O-GlcNAc is transferred to serine and threonine residues of proteins by O-GlcNAc transferase (Ogt) and removed from them by O-GlcNAcase (Oga). In naïve ESCs, O-GlcNAc contributes to maintain the undifferentiated state. In the transition from naïve state to primed state, Ogt maintains cell survival, whereas Oga has no function. However, the function of O-GlcNAc in primed ESD-EpiSCs and during the reversion from the primed state to naïve rESCs remains unclear. Here, we show that Ogt is required for the survival of primed ESD-EpiSCs. The expression of cytosolic Oga was significantly increased during induction from naïve ESCs to primed ESD-EpiSCs. Furthermore, both Ogt and Oga were required for the reversion from primed ESD-EpiSCs to naïve rESCs. These findings indicate that O-GlcNAcylation plays an important role in the survival of primed ESD-EpiSCs and in their reversion to naïve rESCs.

  11. The transition from naïve to primed nociceptive state: A novel wind-up protocol in mice.

    PubMed

    Ziv, Nadav Y; Tal, Michael; Shavit, Yehuda

    2016-01-01

    Wind-up (WU) is a progressive, frequency-dependent facilitation of spinal cord neurons in response to repetitive nociceptive stimulation of constant intensity. We identified a new WU-associated phenomenon in naïve mice (not exposed to noxious stimulation immediately prior to WU stimulation), which were subjected to a novel experimental protocol composed of three consecutive trains of WU stimulation. The 1st train produced a typical linear 'wind-up' curve as expected following a repeating series of stimuli; in addition, this 1st train sensitized ('primed') the nociceptive system so that the responses to two subsequent trains (inter-train interval of 10 min) were significantly amplified compared with the response to the 1st train. We named this augmented response potentiation-of-windup, or "PoW". The PoW phenomenon appears to be centrally mediated, as the augmented response was suppressed by administration of an NMDA receptor antagonist (MK-801) and by cutting the spinal cord. Furthermore, the PoW protocol is accompanied by enhanced pain behavior. The 'priming' effect of the 1st train could be mimicked by exposure to natural noxious stimuli prior to the PoW protocol. Presumably, the PoW phenomenon has not been previously reported due to a procedural reason: typically, WU protocols have been executed in 'primed' rather than naïve animals, i.e., animals exposed to nociceptive stimulation prior to the actual WU recording. Our findings indicate that the PoW paradigm can distinguish between 'naïve' and 'primed' states, suggesting its use as a tool for the assessment of central sensitization.

  12. Photoexcited states of biruthenium(II) compounds bridged by 2,2 prime -bis(2-pyridyl)bibenzimidazole or 1,2-bis(2-(2-pyridyl)benzimidazolyl)ethane

    SciTech Connect

    Ohno, Takeshi; Nozaki, Koichi ); Haga, Masaaki )

    1992-02-19

    Charge-transfer (CT) excited states of RuL{sub 2}(L{prime}-L{prime}){sup 2+} and RuL{sub 2}(L{prime}-L{prime}){sup 4+} have been studied by means of emission and transient absorption (TA) spectroscopy at 77-300 K. The bridging ligand (L{prime}-L{prime}) is either 2,2{prime}-bis(2-pyridyl)benzimidazole (bpbimH{sub 2}) or 1,2-bis(2-(2-pyridyl)benzimidazolyl)ethane (dpbime) and L is 2,2{prime}-bipyridine (bpy), 4,4{prime}-dimethyl-2,2{prime}-bipyridine (dmbpy), or 1,10-phenanthroline (phen). Transient absorption (TA) spectra of the ruthenium(II) compounds subjected to laser excitation, whose molar extinction coefficients were determined, are deconvoluted to {pi}-{pi}* bands of L and L{prime}-L{prime} coordinating to Ru(III), L (or L{prime}-L{prime})-to-Ru(III) CT bands, and a {pi}-{pi}* band of (L{prime}-L{prime}){sup {center dot}-} (or L{sup {center dot}-}) by comparison with the absorption spectra of the oxidized compounds (RuL{sub 2}(L{prime}-L{prime}){sup 3+}). The degree of electron population on the ligand decreases in the order bpbimH{sub 2} > bpy {approximately} phen > dpbime > dmbpy in the excited CT states, while there is no discernible difference in the reduction potential between bpbimH{sub 2} (or dpbime) and bpy coordinating to Ru(II). The excitation efficiency of the metal sites in (Ru(bpy){sub 2}){sub 2}(dpbime){sup 4+} is lower than 50% when the laser power was large enough to excite more than 80% of the mononuclear compounds. The low excitation efficiency of the former is ascribed to rapid intramolecular annihilation of the excited states.

  13. Modeling of decomposition activity and priming effect in soil using the versatile index of microbial physiological state

    NASA Astrophysics Data System (ADS)

    Blagodatskiy, Sergey

    2015-04-01

    The implementation of microbial biomass in soil organic matter (SOM) models is still unresolved issue. The approaches using explicit description of microbial biomass (decomposer) interaction with SOM usually cannot be easily verified by means of experimental estimating of total microbial biomass dynamics. Standard experimental methods, such as fumigation extraction or direct microscopic count, does not represent microbial activity (Blagodatskaya and Kuzyakov, 2013), which is essential for the control of decomposition rate. More advanced approaches, explicitly simulating intracellular metabolic activity (Resat et al., 2012) and e.g. production and turnover of extracellular enzymes (Lawrence et al., 2009) are prohibitively complex for the field and larger scales, which are most often under demand for SOM modelling. One possible parsimonious solution is an application of index of microbial physiological state (r), which describes the adaptive variation of the cell composition and metabolic activity by one variable (Panikov, 1995). This variable (r) can reflect the microbial response to the availability of carbon and nitrogen and shift of microbial biomass between active and dormant state (Blagodatsky and Richter, 1998), but also can be used for the description of the effect of external factors, such as temperature and moisture, on microbial activity. This approach is extremely useful for the description of priming effect (Blagodatsky et al., 2010) and the influence of substrate availability and external factors on the size and dynamics of priming. Distinguishing of these two types of driving forces for priming is crucial for modelling of SOM dynamics and steady-state stocks of different SOM pools. I will present the analysis of model response on combination of limiting factors presented as functions controlling the change of microbial physiological state and size of priming effect. Alternatively, the direct effect of the same factors on decomposition rate and priming

  14. Poking Vesicles

    NASA Astrophysics Data System (ADS)

    Powers, Thomas R.; Huber, Greg; Goldstein, Raymond E.

    2000-03-01

    Lipid vesicles can exhibit a variety of interesting shapes when subject to point forcing, as has been demonstrated with laser-tweezed beads and with growing microtubules. Using numerical and analytic techniques, we study the force vs. extension in two regimes. At low forces, the resistance to deformation is primarily entropic, with a tension given by the fluctuating bending modes. At high forces, the thermal wrinkles are smoothed out, and there is bending and stretching which balance in a thin tether, which we treat using boundary-layer techniques. This work was supported in part by the Harvard MRSEC and Army Research Office Grant DAA655-97-1-014 (TRP) and NSF DMR9812526 (GH & REG).

  15. Medium-resolution studies of extreme ultraviolet emission from N2 by electron impact - Vibrational perturbations and cross sections of the c4-prime 1Sigma(+)u and b-prime 1Sigma(+)u states

    NASA Technical Reports Server (NTRS)

    Ajello, Joseph M.; James, Geoffrey K.; Franklin, Brian O.; Shemansky, Donald E.

    1989-01-01

    In a crossed-beam experiment the electron-impact-induced fluorescence spectrum of N2 in the extreme ultraviolet is studied at a spectral resolution of up to 0.03 nm. The optically thin experiment obtained the highest-resolution electron-impact emission spectrum of the Rydberg and valence states of N2. The spectral measurements provide the emission cross sections of each of the vibrational transitions of the Carroll-Yoshino and the Birge-Hopfield-II band systems. Laboratory vibrational-excitation cross sections arising from the mutual perturbation of the c4-prime 1Sigma(+)u and b-prime 1Sigma(+)u states by homogeneous configuration interactions are measured from 10 to 400 eV, and a modified Born approximation analytic model is given for them. The analysis leads to accurate band-system oscillator strengths. The relative emission and excitation cross sections each of the vibrational levels are compared. In addition, low-resolution measurements of the cross section of the atomic dissociation fragments (NI, NII, NIII) from 40 to 102 nm are made, and medium-resolution measurements are made of the emission cross section of the c4 1Pi(u), c5-prime 1Sigma(+)u, c5 1Pi(u), and c6-prime 1Sigma(+)u to X 1Sigma(+)g (0,0) transitions.

  16. Kinetic barriers to SNAREpin assembly in the regulation of membrane docking/priming and fusion

    PubMed Central

    Li, Feng; Tiwari, Neeraj; Rothman, James E.; Pincet, Frederic

    2016-01-01

    Neurotransmission is achieved by soluble NSF attachment protein receptor (SNARE)-driven fusion of readily releasable vesicles that are docked and primed at the presynaptic plasma membrane. After neurotransmission, the readily releasable pool of vesicles must be refilled in less than 100 ms for subsequent release. Here we show that the initial association of SNARE complexes, SNAREpins, is far too slow to support this rapid refilling owing to an inherently high activation energy barrier. Our data suggest that acceleration of this process, i.e., lowering of the barrier, is physiologically necessary and can be achieved by molecular factors. Furthermore, under zero force, a low second energy barrier transiently traps SNAREpins in a half-zippered state similar to the partial assembly that engages calcium-sensitive regulatory machinery. This result suggests that the barrier must be actively raised in vivo to generate a sufficient pause in the zippering process for the regulators to set in place. We show that the heights of the activation energy barriers can be selectively changed by molecular factors. Thus, it is possible to modify, both in vitro and in vivo, the lifespan of each metastable state. This controllability provides a simple model in which vesicle docking/priming, an intrinsically slow process, can be substantially accelerated. It also explains how the machinery that regulates vesicle fusion can be set in place while SNAREpins are trapped in a half-zippered state. PMID:27601655

  17. Synaptic Vesicle Proteins and Active Zone Plasticity

    PubMed Central

    Kittel, Robert J.; Heckmann, Manfred

    2016-01-01

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

  18. Analysis of the Source Physics Experiment SPE4 Prime Using State-Of Parallel Numerical Tools.

    NASA Astrophysics Data System (ADS)

    Vorobiev, O.; Ezzedine, S. M.; Antoun, T.; Glenn, L.

    2015-12-01

    This work describes a methodology used for large scale modeling of wave propagation from underground chemical explosions conducted at the Nevada National Security Site (NNSS) fractured granitic rock. We show that the discrete natures of rock masses as well as the spatial variability of the fabric of rock properties are very important to understand ground motions induced by underground explosions. In order to build a credible conceptual model of the subsurface we integrated the geological, geomechanical and geophysical characterizations conducted during recent test at the NNSS as well as historical data from the characterization during the underground nuclear test conducted at the NNSS. Because detailed site characterization is limited, expensive and, in some instances, impossible we have numerically investigated the effects of the characterization gaps on the overall response of the system. We performed several computational studies to identify the key important geologic features specific to fractured media mainly the joints characterized at the NNSS. We have also explored common key features to both geological environments such as saturation and topography and assess which characteristics affect the most the ground motion in the near-field and in the far-field. Stochastic representation of these features based on the field characterizations has been implemented into LLNL's Geodyn-L hydrocode. Simulations were used to guide site characterization efforts in order to provide the essential data to the modeling community. We validate our computational results by comparing the measured and computed ground motion at various ranges for the recently executed SPE4 prime experiment. We have also conducted a comparative study between SPE4 prime and previous experiments SPE1 and SPE3 to assess similarities and differences and draw conclusions on designing SPE5.

  19. Open Syntaxin Docks Synaptic Vesicles

    PubMed Central

    Olsen, Shawn; Jorgensen, Erik M

    2007-01-01

    Synaptic vesicles dock to the plasma membrane at synapses to facilitate rapid exocytosis. Docking was originally proposed to require the soluble N-ethylmaleimide–sensitive fusion attachment protein receptor (SNARE) proteins; however, perturbation studies suggested that docking was independent of the SNARE proteins. We now find that the SNARE protein syntaxin is required for docking of all vesicles at synapses in the nematode Caenorhabditis elegans. The active zone protein UNC-13, which interacts with syntaxin, is also required for docking in the active zone. The docking defects in unc-13 mutants can be fully rescued by overexpressing a constitutively open form of syntaxin, but not by wild-type syntaxin. These experiments support a model for docking in which UNC-13 converts syntaxin from the closed to the open state, and open syntaxin acts directly in docking vesicles to the plasma membrane. These data provide a molecular basis for synaptic vesicle docking. PMID:17645391

  20. N-cadherin expression level distinguishes reserved versus primed states of hematopoietic stem cells.

    PubMed

    Haug, Jeffrey S; He, Xi C; Grindley, Justin C; Wunderlich, Joshua P; Gaudenz, Karin; Ross, Jason T; Paulson, Ariel; Wagner, Kathryn P; Xie, Yucai; Zhu, Ruihong; Yin, Tong; Perry, John M; Hembree, Mark J; Redenbaugh, Erin P; Radice, Glenn L; Seidel, Christopher; Li, Linheng

    2008-04-10

    Osteoblasts expressing the homophilic adhesion molecule N-cadherin form a hematopoietic stem cell (HSC) niche. Therefore, we examined how N-cadherin expression in HSCs relates to their function. We found that bone marrow (BM) cells highly expressing N-cadherin (N-cadherin(hi)) are not stem cells, being largely devoid of a Lineage(-)Sca1(+)cKit(+) population and unable to reconstitute hematopoietic lineages in irradiated recipient mice. Instead, long-term HSCs form distinct populations expressing N-cadherin at intermediate (N-cadherin(int)) or low (N-cadherin(lo)) levels. The minority N-cadherin(lo) population can robustly reconstitute the hematopoietic system, express genes that may prime them to mobilize, and predominate among HSCs mobilized from BM to spleen. The larger N-cadherin(int) population performs poorly in reconstitution assays when freshly isolated but improves in response to overnight in vitro culture. Their expression profile and lower cell-cycle entry rate suggest N-cadherin(int) cells are being held in reserve. Thus, differential N-cadherin expression reflects functional distinctions between two HSC subpopulations.

  1. Photoionization of alkylphenothiazines in vesicles: Effects of the alkyl chain length and the vesicle surface charge

    SciTech Connect

    Sakaguchi, Masato; Hu, Ming; Kevan, L. )

    1990-01-25

    The photoionization of alkylphenothiazine (AP = alkylphenothiazine) in vesicles were observed by electron spin resonance (ESR) and electron spin echo modulation (ESEM) methods. Alkylphenothiazine derivatives including sodium 10-methylphenothiazinesulfonate (C{sub 1}PSO{sub 3}Na), sodium 10-dodecylphenothiazinesulfonate (C{sub 12}PSO{sub 3}Na), sodium 3-(10{prime}-phenothiazinyl)propane-1-sulfonate (PC{sub 3}SO{sub 3}Na), sodium 6-(10{prime}-phenothiazinyl)hexane-1-sulfonate (PC{sub 6}SO{sub 3}Na), and sodium 12-(10{prime}-phenothiazinyl)dodecane-1-sulfonate (PC{sub 12}SO{sub 3} Na) were synthesized and used to study the effects of the alkyl chain length, the position of the sulfonate group, and the vesicle surface charge on the photoionization. A single ESR spectrum due to the alkylphenothiazine cation radicals (AP{sup +}) was observed from rapidly frozen AP in dioctadecyldimethylammonium chloride (DODAC) or dihexadecyl phosphate (DHP) vesicles photoirradiated for 10 min with {lambda} > 300 nm. In DODAC vesicles with a positive surface charge, the photoionization yield of PC{sub 12}SO{sub 3}Na with a sulfonate group at the dodecyl chain end is higher than that of C{sub 12}PSO{sub 3}Na with a sulfonate group on the phenothiazine ring. The photoionization yields of AP having the sulfonate group at the alkyl chain end in DODAC vesicles increase with decreasing alkyl chain length. The highest photoionization yield was obtained from PC{sub 3}SO{sub 3}Na, which has the shortest alkyl chain in this study and has the sulfonate group at the end of the propyl chain. The photoionization yield of AP in DHP vesicles with a negative surface charge was not changed by added alkyl chains or the position of the sulfonate group in AP. The results are discussed in terms of the alkyl chain length, the position of the sulfonate group, and the vesicle surface charge.

  2. Heparan sulfation-dependent fibroblast growth factor signaling maintains embryonic stem cells primed for differentiation in a heterogeneous state.

    PubMed

    Lanner, Fredrik; Lee, Kian Leong; Sohl, Marcus; Holmborn, Katarina; Yang, Henry; Wilbertz, Johannes; Poellinger, Lorenz; Rossant, Janet; Farnebo, Filip

    2010-02-01

    Embryonic stem (ES) cells continuously decide whether to maintain pluripotency or differentiate. While exogenous leukemia inhibitory factor and BMP4 perpetuate a pluripotent state, less is known about the factors initiating differentiation. We show that heparan sulfate (HS) proteoglycans are critical coreceptors for signals inducing ES cell differentiation. Genetic targeting of NDST1 and NDST2, two enzymes required for N-sulfation of proteoglycans, blocked differentiation. This phenotype was rescued by HS presented in trans or by soluble heparin. NaClO(3) (-), which reduces sulfation of proteoglycans, potently blocked differentiation of wild-type cells. Mechanistically, N-sulfation was identified to be critical for functional autocrine fibroblast growth factor 4 (FGF4) signaling. Microarray analysis identified the pluripotency maintaining transcription factors Nanog, KLF2/4/8, Tbx3, and Tcf3 to be negatively regulated, whereas markers of differentiation such as Gbx2, Dnmt3b, FGF5, and Brachyury were induced by sulfation-dependent FGF receptor (FGFR) signaling. We show that several of these genes are heterogeneously expressed in ES cells, and that targeting of heparan sulfation or FGFR-signaling facilitated a homogenous Nanog/KLF4/Tbx3 positive ES cell state. This finding suggests that the recently discovered heterogeneous state of ES cells is regulated by HS-dependent FGFR signaling. Similarly, culturing blastocysts with NaClO(3) (-) eliminated GATA6-positive primitive endoderm progenitors generating a homogenous Nanog-positive inner cell mass. Functionally, reduction of sulfation robustly improved de novo ES cell derivation efficiency. We conclude that N-sulfated HS is required for FGF4 signaling to maintain ES cells primed for differentiation in a heterogeneous state. Inhibiting this pathway facilitates a more naïve ground state.

  3. Granulosa cell-oocyte interactions: the phosphorylation of specific proteins in mouse oocytes at the germinal vesicle stage is dependent upon the differentiative state of companion somatic cells

    SciTech Connect

    Cecconi, S.; Tatone, C.; Buccione, R.; Mangia, F.; Colonna, R. )

    1991-05-01

    The role of granulosa cells in the regulation of mouse ovarian oocyte metabolism was investigated. Fully grown antral oocytes, isolated from surrounding cumulus cells, were cultured on monolayers of preantral granulosa cells in the presence of dbcAMP to prevent the resumption of meiosis. Under these conditions metabolic cooperativity was established between the two cell types as early as 1 hr after seeding. Moreover, cocultured oocytes phosphorylated two polypeptides of 74 and 21 kDa which are normally phosphorylated in follicle-enclosed growing oocytes but not in cumulus cell-enclosed fully grown oocytes at the germinal vesicle stage. When cocultured oocytes were allowed to resume meiosis, the 74 and 21 kDa proteins were synthesized but no longer phosphorylated even though intercellular coupling between the two cell types was maintained during radiolabeling. It appears therefore: (a) that the different protein kinase activity of growing and fully grown germinal vesicle-stage mouse oocytes is related to the differentiative state of granulosa cells, and (b) that the regulation of oocyte protein phosphorylation activity by granulosa cells is dependent on the meiotic stage of the oocyte.

  4. Theoretical description of the 2A/double prime/ and 2A/prime/ states of the peroxyformyl radical. [for air pollution and Mars atmospheric studies

    NASA Technical Reports Server (NTRS)

    Winter, N. W.; Goddard, W. A., III; Bender, C. F.

    1975-01-01

    Simple orbital ideas are used to describe the lowest two states of the peroxyformyl radical, and ab initio Hartree-Fock calculations in these states are reported. It is found that both states may be formed exothermically by association of O2 and HCO in their ground states; however, the excited state may decompose readily to OH and CO2. The possible role of such processes in oxidation of aldelydes is discussed.

  5. Morphological docking of secretory vesicles

    PubMed Central

    2010-01-01

    Calcium-dependent secretion of neurotransmitters and hormones is essential for brain function and neuroendocrine-signaling. Prior to exocytosis, neurotransmitter-containing vesicles dock to the target membrane. In electron micrographs of neurons and neuroendocrine cells, like chromaffin cells many synaptic vesicles (SVs) and large dense-core vesicles (LDCVs) are docked. For many years the molecular identity of the morphologically docked state was unknown. Recently, we resolved the minimal docking machinery in adrenal medullary chromaffin cells using embryonic mouse model systems together with electron-microscopic analyses and also found that docking is controlled by the sub-membrane filamentous (F-)actin. Currently it is unclear if the same docking machinery operates in synapses. Here, I will review our docking assay that led to the identification of the LDCV docking machinery in chromaffin cells and also discuss whether identical docking proteins are required for SV docking in synapses. PMID:20577884

  6. An ab initio calculation of the zero-field splitting parameters of the 3A-double prime state of formaldehyde

    NASA Technical Reports Server (NTRS)

    Davidson, E. R.; Ellenbogen, J. C.; Langhoff, S. R.

    1980-01-01

    The spin dipole-dipole and spin-orbit contributions to the zero-field splitting of the 3A-double prime state of formaldehyde have been evaluated at the excited state experimental geometry. Ab initio CI wave functions were generated from a Dunning double zeta plus polarization bases set using 3A-double prime rhf orbitals. Twelve states of each symmetry were used to evaluate the second-order spin-orbit effect. The resulting values of D and E were 0.19 and 0.03 kayser with the principal magnetic axes rotated 36 deg from the CO bond. The values of alpha and beta relative to the inertial axes were calculated to be 0.03 and 0.01 kayser compared to the experimental values of 0.05 plus or minus 0.01 and 0.02 plus or minus 0.02 kayser.

  7. The Affective Regulation of Cognitive Priming

    PubMed Central

    Storbeck, Justin; Clore, Gerald L.

    2008-01-01

    Semantic and affective priming are classic effects observed in cognitive and social psychology, respectively. We discovered that affect regulates such priming effects. In Experiment 1, positive and negative moods were induced prior to one of three priming tasks; evaluation, categorization, or lexical decision. As predicted, positive affect led to both affective priming (evaluation task) and semantic priming (category and lexical decision tasks). However, negative affect inhibited such effects. In Experiment 2, participants in their natural affective state completed the same priming tasks as in Experiment 1. As expected, affective priming (evaluation task) and category priming (categorization and lexical decision tasks) were observed in such resting affective states. Hence, we conclude that negative affect inhibits semantic and affective priming. These results support recent theoretical models, which suggest that positive affect promotes associations among strong and weak concepts, and that negative affect impairs such associations (Kuhl, 2000; Clore & Storbeck, 2006). PMID:18410195

  8. Use of BABA and INA As Activators of a Primed State in the Common Bean (Phaseolus vulgaris L.)

    PubMed Central

    Martínez-Aguilar, Keren; Ramírez-Carrasco, Gabriela; Hernández-Chávez, José Luis; Barraza, Aarón; Alvarez-Venegas, Raúl

    2016-01-01

    To survive in adverse conditions, plants have evolved complex mechanisms that “prime” their defense system to respond and adapt to stresses. Their competence to respond to such stresses fundamentally depends on its capacity to modulate the transcriptome rapidly and specifically. Thus, chromatin dynamics is a mechanism linked to transcriptional regulation and enhanced defense in plants. For example, in Arabidopsis, priming of the SA-dependent defense pathway is linked to histone lysine methylation. Such modifications could create a memory of the primary infection that is associated with an amplified gene response upon exposure to a second stress-stimulus. In addition, the priming status of a plant for induced resistance can be inherited to its offspring. However, analyses on the molecular mechanisms of generational and transgenerational priming in the common bean (Phaseolus vulagris L.), an economically important crop, are absent. Here, we provide evidence that resistance to P. syringae pv. phaseolicola infection was induced in the common bean with the synthetic priming activators BABA and INA. Resistance was assessed by evaluating symptom appearance, pathogen accumulation, changes in gene expression of defense genes, as well as changes in the H3K4me3 and H3K36me3 marks at the promoter-exon regions of defense-associated genes. We conclude that defense priming in the common bean occurred in response to BABA and INA and that these synthetic activators primed distinct genes for enhanced disease resistance. We hope that an understanding of the molecular changes leading to defense priming and pathogen resistance will provide valuable knowledge for producing disease-resistant crop varieties by exposing parental plants to priming activators, as well as to the development of novel plant protection chemicals that stimulate the plant's inherent disease resistance mechanisms. PMID:27242854

  9. High salt primes a specific activation state of macrophages, M(Na).

    PubMed

    Zhang, Wu-Chang; Zheng, Xiao-Jun; Du, Lin-Juan; Sun, Jian-Yong; Shen, Zhu-Xia; Shi, Chaoji; Sun, Shuyang; Zhang, Zhiyuan; Chen, Xiao-Qing; Qin, Mu; Liu, Xu; Tao, Jun; Jia, Lijun; Fan, Heng-Yu; Zhou, Bin; Yu, Ying; Ying, Hao; Hui, Lijian; Liu, Xiaolong; Yi, Xianghua; Liu, Xiaojing; Zhang, Lanjing; Duan, Sheng-Zhong

    2015-08-01

    High salt is positively associated with the risk of many diseases. However, little is known about the mechanisms. Here we showed that high salt increased proinflammatory molecules, while decreased anti-inflammatory and proendocytic molecules in both human and mouse macrophages. High salt also potentiated lipopolysaccharide-induced macrophage activation and suppressed interleukin 4-induced macrophage activation. High salt induced the proinflammatory aspects by activating p38/cFos and/or Erk1/2/cFos pathways, while inhibited the anti-inflammatory and proendocytic aspects by Erk1/2/signal transducer and activator of transcription 6 pathway. Consistent with the in vitro results, high-salt diet increased proinflammatory gene expression of mouse alveolar macrophages. In mouse models of acute lung injury, high-salt diet aggravated lipopolysaccharide-induced pulmonary macrophage activation and inflammation in lungs. These results identify a novel macrophage activation state, M(Na), and high salt as a potential environmental risk factor for lung inflammation through the induction of M(Na).

  10. Dollar Summary of Prime Contract Awards by Contractor, State or Country, and Place, FY83, Part 3 (Planning Design & Research Engr-Zytron Corp.)

    DTIC Science & Technology

    1983-01-01

    PROCESSING INC BELGIUM 126 126 128 126 PRECISION COMPONENTS DANBURY CONN 30 36 30 30 PRI"E-CONTRACTS BY CONTRACTOR, STATE. AND CITY - (OCT 82 - SEP 83...INC CINCINNATI OHIO 309 309 309 309 PROCEDYNE ASSOCIATES INC NEW BRUNSWICK NEW JERSEY 38 38 38 38 PROCESS CONSTRUCTORS INC PHILADELPHIA PA 67 67 67 67... PROCESS CONTROL TECHNOLOGY INC KELLY AFB TEXAS 206 206 206 206 PROCESS ENSINEERINQ INC NEWINGTON NH 205 205 205 205 PRIME-CONTRACTS BY CONTRACTOR

  11. Adhesion of Polymer Vesicles

    NASA Astrophysics Data System (ADS)

    Lin, John J.; Bates, Frank S.; Hammer, Daniel A.; Silas, James A.

    2005-07-01

    The adhesion and bending modulus of polybutadiene-poly(ethylene oxide) block copolymer vesicles made from a bidisperse mixture of polymers is measured using micropipette aspiration. The adhesion energy between biotinylated vesicles and avidin beads is modeled by incorporating the extension of the adhesive ligands above the surface brush of the vesicle according to the blob model of bidisperse polymer mixtures of Komura and Safran assuming the polymer brush at the surface of the vesicle is compact. The same model accurately reproduces the scaling of the bending modulus with polymer composition.

  12. Differential Regulation of Synaptic Vesicle Tethering and Docking by UNC-18 and TOM-1

    PubMed Central

    Gracheva, Elena O.; Maryon, Ed B.; Berthelot-Grosjean, Martine; Richmond, Janet E.

    2010-01-01

    The assembly of SNARE complexes between syntaxin, SNAP-25 and synaptobrevin is required to prime synaptic vesicles for fusion. Since Munc18 and tomosyn compete for syntaxin interactions, the interplay between these proteins is predicted to be important in regulating synaptic transmission. We explored this possibility, by examining genetic interactions between C. elegans unc-18(Munc18), unc-64(syntaxin) and tom-1(tomosyn). We have previously demonstrated that unc-18 mutants have reduced synaptic transmission, whereas tom-1 mutants exhibit enhanced release. Here we show that the unc-18 mutant release defect is associated with loss of two morphologically distinct vesicle pools; those tethered within 25 nm of the plasma membrane and those docked with the plasma membrane. In contrast, priming defective unc-13 mutants accumulate tethered vesicles, while docked vesicles are greatly reduced, indicating tethering is UNC-18-dependent and occurs in the absence of priming. C. elegans unc-64 mutants phenocopy unc-18 mutants, losing both tethered and docked vesicles, whereas overexpression of open syntaxin preferentially increases vesicle docking, suggesting UNC-18/closed syntaxin interactions are responsible for vesicle tethering. Given the competition between vertebrate tomosyn and Munc18, for syntaxin binding, we hypothesized that C. elegans TOM-1 may inhibit both UNC-18-dependent vesicle targeting steps. Consistent with this hypothesis, tom-1 mutants exhibit enhanced UNC-18 plasma membrane localization and a concomitant increase in both tethered and docked synaptic vesicles. Furthermore, in tom-1;unc-18 double mutants the docked, primed vesicle pool is preferentially rescued relative to unc-18 single mutants. Together these data provide evidence for the differential regulation of two vesicle targeting steps by UNC-18 and TOM-1 through competitive interactions with syntaxin. PMID:21423527

  13. CAPS and Munc13: CATCHRs that SNARE Vesicles

    PubMed Central

    James, Declan J.; Martin, Thomas F. J.

    2013-01-01

    CAPS (Calcium-dependent Activator Protein for Secretion, aka CADPS) and Munc13 (Mammalian Unc-13) proteins function to prime vesicles for Ca2+-triggered exocytosis in neurons and neuroendocrine cells. CAPS and Munc13 proteins contain conserved C-terminal domains that promote the assembly of SNARE complexes for vesicle priming. Similarities of the C-terminal domains of CAPS/Munc13 proteins with Complex Associated with Tethering Containing Helical Rods domains in multi-subunit tethering complexes (MTCs) have been reported. MTCs coordinate multiple interactions for SNARE complex assembly at constitutive membrane fusion steps. We review aspects of these diverse tethering and priming factors to identify common operating principles. PMID:24363652

  14. On the Computing Potential of Intracellular Vesicles

    PubMed Central

    Mayne, Richard; Adamatzky, Andrew

    2015-01-01

    Collision-based computing (CBC) is a form of unconventional computing in which travelling localisations represent data and conditional routing of signals determines the output state; collisions between localisations represent logical operations. We investigated patterns of Ca2+-containing vesicle distribution within a live organism, slime mould Physarum polycephalum, with confocal microscopy and observed them colliding regularly. Vesicles travel down cytoskeletal ‘circuitry’ and their collisions may result in reflection, fusion or annihilation. We demonstrate through experimental observations that naturally-occurring vesicle dynamics may be characterised as a computationally-universal set of Boolean logical operations and present a ‘vesicle modification’ of the archetypal CBC ‘billiard ball model’ of computation. We proceed to discuss the viability of intracellular vesicles as an unconventional computing substrate in which we delineate practical considerations for reliable vesicle ‘programming’ in both in vivo and in vitro vesicle computing architectures and present optimised designs for both single logical gates and combinatorial logic circuits based on cytoskeletal network conformations. The results presented here demonstrate the first characterisation of intracelluar phenomena as collision-based computing and hence the viability of biological substrates for computing. PMID:26431435

  15. On the Computing Potential of Intracellular Vesicles.

    PubMed

    Mayne, Richard; Adamatzky, Andrew

    2015-01-01

    Collision-based computing (CBC) is a form of unconventional computing in which travelling localisations represent data and conditional routing of signals determines the output state; collisions between localisations represent logical operations. We investigated patterns of Ca2+-containing vesicle distribution within a live organism, slime mould Physarum polycephalum, with confocal microscopy and observed them colliding regularly. Vesicles travel down cytoskeletal 'circuitry' and their collisions may result in reflection, fusion or annihilation. We demonstrate through experimental observations that naturally-occurring vesicle dynamics may be characterised as a computationally-universal set of Boolean logical operations and present a 'vesicle modification' of the archetypal CBC 'billiard ball model' of computation. We proceed to discuss the viability of intracellular vesicles as an unconventional computing substrate in which we delineate practical considerations for reliable vesicle 'programming' in both in vivo and in vitro vesicle computing architectures and present optimised designs for both single logical gates and combinatorial logic circuits based on cytoskeletal network conformations. The results presented here demonstrate the first characterisation of intracelluar phenomena as collision-based computing and hence the viability of biological substrates for computing.

  16. The pharmacology of nociceptor priming.

    PubMed

    Kandasamy, Ram; Price, Theodore J

    2015-01-01

    Nociceptors and neurons in the central nervous system (CNS) that receive nociceptive input show remarkable plasticity in response to injury. This plasticity is thought to underlie the development of chronic pain states. Hence, further understanding of the molecular mechanisms driving and maintaining this plasticity has the potential to lead to novel therapeutic approaches for the treatment of chronic pain states. An important concept in pain plasticity is the presence and persistence of "hyperalgesic priming." This priming arises from an initial injury and results in a remarkable susceptibility to normally subthreshold noxious inputs causing a prolonged pain state in primed animals. Here we describe our current understanding of how this priming is manifested through changes in signaling in the primary nociceptor as well as through memory like alterations at CNS synapses. Moreover, we discuss how commonly utilized analgesics, such as opioids, enhance priming therefore potentially contributing to the development of persistent pain states. Finally we highlight where these priming models draw parallels to common human chronic pain conditions. Collectively, these advances in our understanding of pain plasticity reveal a variety of targets for therapeutic intervention with the potential to reverse rather than palliate chronic pain states.

  17. The Pharmacology of Nociceptor Priming

    PubMed Central

    Kandasamy, Ram

    2015-01-01

    Nociceptors and neurons in the central nervous system (CNS) that receive nociceptive input show remarkable plasticity in response to injury. This plasticity is thought to underlie the development of chronic pain states. Hence, further understanding of the molecular mechanisms driving and maintaining this plasticity has the potential to lead to novel therapeutic approaches for the treatment of chronic pain states. An important concept in pain plasticity is the presence and persistence of “hyperalgesic priming.” This priming arises from an initial injury and results in a remarkable susceptibility to normally subthreshold noxious inputs causing a prolonged pain state in primed animals. Here we describe our current understanding of how this priming is manifested through changes in signaling in the primary nociceptor as well as through memory like alterations at CNS synapses. Moreover, we discuss how commonly utilized analgesics, such as opioids, enhance priming therefore potentially contributing to the development of persistent pain states. Finally we highlight where these priming models draw parallels to common human chronic pain conditions. Collectively, these advances in our understanding of pain plasticity reveal a variety of targets for therapeutic intervention with the potential to reverse rather than palliate chronic pain states. PMID:25846612

  18. Vesicle trafficking and cell surface membrane patchiness.

    PubMed Central

    Tang, Q; Edidin, M

    2001-01-01

    Membrane proteins and lipids often appear to be distributed in patches on the cell surface. These patches are often assumed to be membrane domains, arising from specific molecular associations. However, a computer simulation (Gheber and Edidin, 1999) shows that membrane patchiness may result from a combination of vesicle trafficking and dynamic barriers to lateral mobility. The simulation predicts that the steady-state patches of proteins and lipids seen on the cell surface will decay if vesicle trafficking is inhibited. To test this prediction, we compared the apparent sizes and intensities of patches of class I HLA molecules, integral membrane proteins, before and after inhibiting endocytic vesicle traffic from the cell surface, either by incubation in hypertonic medium or by expression of a dominant-negative mutant dynamin. As predicted by the simulation, the apparent sizes of HLA patches increased, whereas their intensities decreased after endocytosis and vesicle trafficking were inhibited. PMID:11423406

  19. Dollar Summary of Prime Contract Awards by Contractor, State or Country, and Place, FY83, Part 1 (A&A Chemical Toilet Rentals-Gillespie Construction Co.)

    DTIC Science & Technology

    1983-01-01

    1L 11w1. MICROCOPY RESOLUTION TESI CHARI NATION. (TUH AO’I NP PRIME-CONTRACTS BY CONTRACTOR, STATE, AND CITY - (OCT 82 - SEP 83) Itir$ ",’? (0(4I.L ARS...INC ORANGE CALIFORNIA 44 14 44 44 FORMA RAPID CONSTRUCTION CORP PONCE PUERTO RCO 224 224 7OPMA RAP10 CONSTRUCTION C00Sp RnOSEVELT ROAD3 PUERTO RCO 60...o R C CONTRACTING INC JACKSONVILLE FLORIDA 58 56 58 58 S R 0 CO WARMINSTER PA 1,084 1,084 1 084 1,084 o R G ENGINEERING INC FORT BUCHANAN PUERTO RCO

  20. The transition of mouse pluripotent stem cells from the naïve to the primed state requires Fas signaling through 3-O sulfated heparan sulfate structures recognized by the HS4C3 antibody

    SciTech Connect

    Hirano, Kazumi; Van Kuppevelt, Toin H.; Nishihara, Shoko

    2013-01-18

    Highlights: ► Fas transcript increases during the transition from the naïve to the primed state. ► 3OST-5 transcript, the HS4C3 epitope synthesis gene, increases during the transition. ► Fas signaling regulates the transition from the naïve to the primed state. ► HS4C3-binding epitope regulates the transition from the naïve to the primed state. ► Fas signaling is regulated by the HS4C3 epitope during the transition. -- Abstract: The characteristics of pluripotent embryonic stem cells of human and mouse are different. The properties of human embryonic stem cells (hESCs) are similar to those of mouse epiblast stem cells (mEpiSCs), which are in a later developmental pluripotency state, the so-called “primed state” compared to mouse embryonic stem cells (mESCs) which are in a naïve state. As a result of the properties of the primed state, hESCs proliferate slowly, cannot survive as single cells, and can only be transfected with genes at low efficiency. Generating hESCs in the naïve state is necessary to overcome these problems and allow their application in regenerative medicine. Therefore, clarifying the mechanism of the transition between the naïve and primed states in pluripotent stem cells is important for the establishment of stable methods of generating naïve state hESCs. However, the signaling pathways which contribute to the transition between the naïve and primed states are still unclear. In this study, we carried out induction from mESCs to mEpiSC-like cells (mEpiSCLCs), and observed an increase in the activation of Fas signaling during the induction. The expression of Fgf5, an epiblast marker, was diminished by inhibition of Fas signaling using the caspase-8 and -3 blocking peptides, IETD and DEVD, respectively. Furthermore, during the induction, we observed increased expression of 3-O sulfated heparan sulfate (HS) structures synthesized by HS 3-O-sulfotransferase (3OST), which are recognized by the HS4C3 antibody (HS4C3-binding epitope

  1. Vesicle extrusion in nanopores

    NASA Astrophysics Data System (ADS)

    Joóos, B. Éla; Bertrand, Martin; Ouellet, S. Ébastien

    2010-03-01

    Monodisperse vesicles of nearly circular shape or liposomes are used as drug delivery systems. Their fabrication involves repeated passage of large vesicles through small pores. At each passage the vesicle ruptures and the fragments reform into smaller vesicles. We report on the last stages of the process where small liposomes are pushed by pressure differences into nano-sized pores, and we study the stress distribution along the lipid bilayer to determine the rupture lines. This is done by performing coarse grained Molecular Dynamics simulations. We have developed a technique to measure the stress in the membrane based on a tessellation of the surface which allows us to monitor the local area per lipid fluctuations. The results show subtle and complex flow phenomena. We can predict the final size distribution after many passages. Comparisons will be made with existing experimental data.

  2. Learning about Primes

    ERIC Educational Resources Information Center

    McEachran, Alec

    2008-01-01

    In this article, the author relates his unhappy experience in learning about prime numbers at secondary school. To introduce primes, a teacher first told students a definition of a prime number, then students were taught how to find prime numbers. Students defined and listed them and at some later point were tested on their memory of both the…

  3. Vesicles in Poiseuille flow.

    PubMed

    Danker, Gerrit; Vlahovska, Petia M; Misbah, Chaouqi

    2009-04-10

    Blood microcirculation critically depends on the migration of red cells towards the flow centerline. We identify theoretically the ratio of the inner over the outer fluid viscosities lambda as a key parameter. At low lambda, the vesicle deforms into a tank-treading ellipsoid shape far away from the flow centerline. The migration is always towards the flow centerline, unlike drops. Above a critical lambda, the vesicle tumbles or breaths and migration is suppressed. A surprising coexistence of two types of shapes at the centerline, a bulletlike and a parachutelike shape, is predicted.

  4. Critical dynamics of vesicle stretching transition in elongational flow.

    PubMed

    Kantsler, Vasiliy; Segre, Enrico; Steinberg, Victor

    2008-07-25

    We present results on the stretching of single tubular vesicles in an elongation flow toward dumbbell shapes, and on their relaxation. A critical strain rate epsilonc exists; for strain rates epsilonvesicle remains tubular but fluctuates, though its steady state extension increases with the strain rate epsilon. Above epsilonc, first a shape transition to dumbbell occurs, and then high order shape modes become unstable, leading to a pearling state. We have quantitatively characterized the transition and found a scaling of epsilonc with the system parameters. A remarkable feature of vesicle tube behavior around the critical point is a slowdown of the vesicle relaxation to the final extended state in the vesicle stretching. Such feature is similar to that found in continuous phase transitions and to the critical effects recently observed for polymer molecules near the coil-stretch transition in elongation flow.

  5. Complexity of vesicle microcirculation

    NASA Astrophysics Data System (ADS)

    Kaoui, B.; Tahiri, N.; Biben, T.; Ez-Zahraouy, H.; Benyoussef, A.; Biros, G.; Misbah, C.

    2011-10-01

    This study focuses numerically on dynamics in two dimensions of vesicles in microcirculation. The method used is based on boundary integral formulation. This study is inspired by the behavior of red blood cells (RBCs) in the microvasculature. Red RBCs carry oxygen from the lungs and deliver it through the microvasculature. The shape adopted by RBCs can affect blood flow and influence oxygen delivery. Our simulation using vesicles (a simple model for RBC) reveals unexpected complexity as compared to the case where a purely unbounded Poiseuille flow is considered [Kaoui, Biros, and Misbah, Phys. Rev. Lett.10.1103/PhysRevLett.103.188101 103, 188101 (2009)]. In sufficiently large channels (in the range of 100μm; the vesicle size and its reduced volume are taken in the range of those of a human RBC), such as arterioles, a slipperlike (asymmetric) shape prevails. A parachutelike (symmetric) shape is adopted in smaller channels (in the range of 20μm, as in venules), but this shape loses stability and again changes to a pronounced slipperlike morphology in channels having a size typical of capillaries (5-10 μm). Stiff membranes, mimicking malaria infection, for example, adopt a centered or off-centered snakelike locomotion instead (the denomination snaking is used for this regime). A general scenario of how and why vesicles adopt their morphologies and dynamics among several distinct possibilities is provided. This finding potentially points to nontrivial RBCs dynamics in the microvasculature.

  6. Prime Contract Awards Alphabetically by Contractor, by State or Country, and Place. Part 16 (Panhandle Food Sales-Ray Gene Electric Co Inc)

    DTIC Science & Technology

    1990-01-01

    AD-A236 466 1990 ANNUAL PRIME CONTRACTS AWARDS ALPHABETICALLY BY CONTRACTOR, BY STATE OR COUNTRY, AND PLACE. PT-16 (PANHANDLE FOOD SALES-RAY GENE ...4 I-l e~ I 0 MM 0)1 47470:14710) 0) 0) 0) I" gene , I LA 4 00 0) co -4 -FF- F- C (D <r- 0CD I~ gene , U0 -4 0. C4 0> 0D-4-4-40 C> C- N0 -4 (n L-4 cn...LW ,U ’L a) I CIme 𔃺" 0)0)100)0)10)0) (00 0 Ei(Cn" cc cc c c c c c c c c c 410mc’ " ::U. C 001. C00 r-0C0C r00 C w izI0’ M" ~ 000000 , . > .0 000 s-s

  7. Descendants of primed Arabidopsis plants exhibit resistance to biotic stress.

    PubMed

    Slaughter, Ana; Daniel, Xavier; Flors, Victor; Luna, Estrella; Hohn, Barbara; Mauch-Mani, Brigitte

    2012-02-01

    An attack of plants by pathogens or treatment with certain resistance-inducing compounds can lead to the establishment of a unique primed state of defense. Primed plants show enhanced defense reactions upon further challenge with biotic or abiotic stress. Here, we report that the primed state in Arabidopsis (Arabidopsis thaliana) is still functional in the next generation without additional treatment. We compared the reactions of Arabidopsis plants that had been either primed with β-amino-butyric acid (BABA) or with an avirulent isolate of the bacteria Pseudomonas syringae pv tomato (PstavrRpt2). The descendants of primed plants showed a faster and higher accumulation of transcripts of defense-related genes in the salicylic acid signaling pathway and enhanced disease resistance upon challenge inoculation with a virulent isolate of P. syringae. In addition, the progeny of primed plants was also more resistant against the oomycete pathogen Hyaloperonospora arabidopsidis. When transgenerationally primed plants were subjected to an additional priming treatment, their descendants displayed an even stronger primed phenotype, suggesting that plants can inherit a sensitization for the priming phenomenon. Interestingly, this primed to be primed phenotype was much reduced in the Arabidopsis β-amino-butyric acid priming mutant ibs1 (induced BABA sterility1). Our results demonstrate that the primed state of plants is transferred to their progeny and confers improved protection from pathogen attack as compared to the descendants of unprimed plants.

  8. Motor Priming in Neurorehabilitation

    PubMed Central

    Stoykov, Mary Ellen; Madhavan, Sangeetha

    2014-01-01

    Priming is a type of implicit learning wherein a stimulus prompts a change in behavior. Priming has been long studied in the field of psychology. More recently, rehabilitation researchers have studied motor priming as a possible way to facilitate motor learning. For example, priming of the motor cortex is associated with changes in neuroplasticity that are associated with improvements in motor performance. Of the numerous motor priming paradigms under investigation, only a few are practical for the current clinical environment, and the optimal priming modalities for specific clinical presentations are not known. Accordingly, developing an understanding of the various types of motor priming paradigms and their underlying neural mechanisms is an important step for therapists in neurorehabilitation. Most importantly, an understanding of the methods and their underlying mechanisms is essential for optimizing rehabilitation outcomes. The future of neurorehabilitation is likely to include these priming methods, which are delivered prior to or in conjunction with primary neurorehabilitation therapies. In this Special Interest article we discuss those priming paradigms that are supported by the greatest amount of evidence including: (i) stimulation-based priming, (ii) motor imagery and action observation, (iii) sensory priming, (iv) movement-based priming, and (v) pharmacological priming. PMID:25415551

  9. Equilibrium of nematic vesicles

    NASA Astrophysics Data System (ADS)

    Napoli, Gaetano; Vergori, Luigi

    2010-11-01

    A variational scheme is proposed which allows the derivation of a concise and elegant formulation of the equilibrium equations for closed fluid membranes, endowed with a nematic microstructure. The nematic order is described by an in-plane nematic director and a degree of orientation, as customary in the theory of uniaxial nematics. The only constitutive ingredient in this scheme is a free-energy density which depends on the vesicle geometry and order parameters. The stress and the couple stress tensors related to this free-energy density are provided. As an application of the proposed scheme, a certain number of special theories are deduced: soap bubbles, lipid vesicles, chiral and achiral nematic membranes, and nematics on curved substrates.

  10. Limited intermixing of synaptic vesicle components upon vesicle recycling.

    PubMed

    Opazo, Felipe; Punge, Annedore; Bückers, Johanna; Hoopmann, Peer; Kastrup, Lars; Hell, Stefan W; Rizzoli, Silvio O

    2010-06-01

    Synaptic vesicles recycle repeatedly in order to maintain synaptic transmission. We have previously proposed that upon exocytosis the vesicle components persist as clusters, which would be endocytosed as whole units. It has also been proposed that the vesicle components diffuse into the plasma membrane and are then randomly gathered into new vesicles. We found here that while strong stimulation (releasing the entire recycling pool) causes the diffusion of the vesicle marker synaptotagmin out of synaptic boutons, moderate stimulation (releasing approximately 19% of all vesicles) is followed by no measurable diffusion. In agreement with this observation, synaptotagmin molecules labeled with different fluorescently tagged antibodies did not appear to mix upon vesicle recycling, when investigated by subdiffraction resolution stimulated emission depletion (STED) microscopy. Finally, as protein diffusion from vesicles has been mainly observed using molecules tagged with pH-sensitive green fluorescent protein (pHluorin), we have also investigated the membrane patterning of several native and pHluorin-tagged proteins. While the native proteins had a clustered distribution, the GFP-tagged ones were diffused in the plasma membrane. We conclude that synaptic vesicle components intermix little, at least under moderate stimulation, possibly because of the formation of clusters in the plasma membrane. We suggest that several pHluorin-tagged vesicle proteins are less well integrated in clusters.

  11. A Molecular Dynamics Simulation of Vesicle Deformation and Rupture in Confined Poiseuille Flow

    NASA Astrophysics Data System (ADS)

    Harman, Alison

    Vesicles are simple structures, but display complex, non-linear dynamics in fluid flow. I investigate the deformation of nanometer-sized vesicles, both fully-inflated and those with excess area, as they travel in tightly confined capillaries. By varying both channel size and flow strength, I simulate vesicles as they transition from steady-state to unstable shapes, and then rupture in strong flow fields. By employing a molecular dynamics model of the vesicle, fluid, and capillary system one is able to rupture the lipid bilayer of these vesicles. This is unique in that most other numerical methods for modelling vesicles are unable to show rupture. The rupture of fully-inflated vesicles is applicable to drug delivery in which the release of the encapsulated medicine needs to be controlled. The deformation and rupture of vesicles with excess area could be applicable to red blood cells which have similar rheological properties.

  12. A Well-Defined Readily Releasable Pool with Fixed Capacity for Storing Vesicles at Calyx of Held

    PubMed Central

    Mahfooz, Kashif; Singh, Mahendra; Renden, Robert; Wesseling, John F.

    2016-01-01

    The readily releasable pool (RRP) of vesicles is a core concept in studies of presynaptic function. However, operating principles lack consensus definition and the utility for quantitative analysis has been questioned. Here we confirm that RRPs at calyces of Held from 14 to 21 day old mice have a fixed capacity for storing vesicles that is not modulated by Ca2+. Discrepancies with previous studies are explained by a dynamic flow-through pool, established during heavy use, containing vesicles that are released with low probability despite being immediately releasable. Quantitative analysis ruled out a posteriori explanations for the vesicles with low release probability, such as Ca2+-channel inactivation, and established unexpected boundary conditions for remaining alternatives. Vesicles in the flow-through pool could be incompletely primed, in which case the full sequence of priming steps downstream of recruitment to the RRP would have an average unitary rate of at least 9/s during heavy use. Alternatively, vesicles with low and high release probability could be recruited to distinct types of release sites; in this case the timing of recruitment would be similar at the two types, and the downstream transition from recruited to fully primed would be much faster. In either case, further analysis showed that activity accelerates the upstream step where vesicles are initially recruited to the RRP. Overall, our results show that the RRP can be well defined in the mathematical sense, and support the concept that the defining mechanism is a stable group of autonomous release sites. PMID:27035349

  13. Shapes of Mixed Phospholipid Vesicles

    PubMed Central

    Aranda-Espinoza, Helim; Maldonado, Amir

    2006-01-01

    We studied the shape of phospholipid vesicles prepared by hydration of a mixture of phosphatidylcholine (SOPC) and phosphatidylserine (SOPS) in different proportions. The aim of the work is to obtain some insight into the influence of the chemical composition of a biomembrane on its shape. The optical microscopy results show that the shape of the vesicles depend on the SOPC:SOPS composition. For low SOPS contents, coiled cylindrical vesicles are observed. The results suggest that specific compositions of the SOPC:SOPS vesicles produce some spontaneous curvature on the membrane and then a coiling instability. PMID:19669461

  14. Dihydrolipoyl dioleoylglycerol antioxidant capacity in phospholipid vesicles.

    PubMed

    Laszlo, Joseph A; Evans, Kervin O; Compton, David L; Appell, Michael

    2012-02-01

    Antioxidants have critical roles in maintaining cellular homeostasis and disease-state prevention. The multi-functional agent α-lipoic acid offers numerous beneficial effects to oxidatively stressed tissues. α-Lipoic acid was enzymatically incorporated into a triglyceride in conjunction with oleic acid, creating lipoyl dioleoylglycerol, and chemically reduced to form dihydrolipoyl dioleoylglycerol. The triglyceride forms of lipoic acid stabilized dioleoylphosphatidylcholine unilamellar liposomal vesicles, as judged by calcein-cobalt leakage. Stabilization resulted from increased packing density of phospholipid acyl chains. Scavenging activity against the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radical was monitored by oxidation of 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C(11)-Bodipy). Dihydrolipoyl dioleoylglycerol in vesicles demonstrated strong antioxidant capacity in comparison to the conventional Trolox standard. Fluorescence quenching measurements indicated the lipoyl moiety of dihydrolipoyl dioleoylglycerol is positioned near the vesicle aqueous/lipid boundary. Treatment of intact vesicles with a nonpenetrating sulfhydryl reagent indicated that 80% of the dihydrolipoyl dioleoylglycerol was available for reaction. Molecular modeling of lipoyl dioleoylglycerol and dihydrolipoyl dioleoylglycerol in a phospholipid layer confirmed the existence of an extended configuration for the molecules that accounts for the interfacial location of the lipoyl moiety, which may allow the antioxidant to readily react with radical species approaching membranes from the aqueous phase.

  15. 2l-nl{prime} x-ray transitions from neonlike charge states of the row 5 metals with 39 {le} Z {le} 46

    SciTech Connect

    Rice, J.E.; Terry, J.L.; Marmar, E.S.; Fournier, K.B.; Goldstein, W.H.; Finkenthal, M.; Safronova, U.I.

    1996-03-18

    X-ray spectra of 2l-2l{prime} transitions with 3 {le} n {le} 12 in the row five transition metals zirconium (Z = 40), niobium (Z = 41), molybdenum (Z = 42) and palladium (Z = 46) from charge states around neonlike have been observed from Alcator C-Mod plasmas. Accurate wavelengths ({+-} .2 m{angstrom}) have been determined by comparison with neighboring argon, chlorine and sulfur lines with well known wavelengths. Line identifications have been made by comparison to ab initio atomic structure calculations, using a fully relativistic, parametric potential code. For neonlike ions, calculated wavelengths and oscillator strengths are tabulated for 2p-nd transitions in Y (Z = 39), Tc (Z = 43), Ru (Z = 44) and Rh (Z = 45) with n = 6 and 7. The magnitude of the configuration interaction between the (2p{sup 5}){sub 1/2}6d{sub 3/2} J = 1 level and the (2p{sup 5}){sub 3/2}7D{sub 5/2} J = 1 levels is demonstrated as a function of atomic number for successive neonlike ions. Measured spectra of selected transitions in the aluminum-, magnesium-, sodium- and fluorine like isosequences are also shown.

  16. Characterization of extracellular vesicles in whole blood: Influence of pre-analytical parameters and visualization of vesicle-cell interactions using imaging flow cytometry.

    PubMed

    Fendl, Birgit; Weiss, René; Fischer, Michael B; Spittler, Andreas; Weber, Viktoria

    2016-09-09

    Extracellular vesicles are central players in intercellular communication and are released from the plasma membrane under tightly regulated conditions, depending on the physiological and pathophysiological state of the producing cell. Their heterogeneity requires a spectrum of methods for isolation and characterization, where pre-analytical parameters have profound impact on vesicle analysis, particularly in blood, since sampling, addition of anticoagulants, as well as post-sampling vesicle generation may influence the outcome. Here, we characterized microvesicles directly in whole blood using a combination of flow cytometry and imaging flow cytometry. We assessed the influence of sample agitation, anticoagulation, and temperature on post-sampling vesicle generation, and show that vesicle counts remained stable over time in samples stored without agitation. Storage with gentle rolling mimicking agitation, in contrast, resulted in strong release of platelet-derived vesicles in blood anticoagulated with citrate or heparin, whereas vesicle counts remained stable upon anticoagulation with EDTA. Using imaging flow cytometry, we could visualize microvesicles adhering to blood cells and revealed an anticoagulant-dependent increase in vesicle-cell aggregates over time. We demonstrate that vesicles adhere preferentially to monocytes and granulocytes in whole blood, while no microvesicles could be visualized on lymphocytes. Our data underscore the relevance of pre-analytical parameters in vesicle analysis and demonstrate that imaging flow cytometry is a suitable tool to study the interaction of extracellular vesicles with their target cells.

  17. Unconventional molecular regulation of synaptic vesicle replenishment in cochlear inner hair cells.

    PubMed

    Vogl, Christian; Cooper, Benjamin H; Neef, Jakob; Wojcik, Sonja M; Reim, Kerstin; Reisinger, Ellen; Brose, Nils; Rhee, Jeong-Seop; Moser, Tobias; Wichmann, Carolin

    2015-02-15

    Ribbon synapses of cochlear inner hair cells (IHCs) employ efficient vesicle replenishment to indefatigably encode sound. In neurons, neuroendocrine and immune cells, vesicle replenishment depends on proteins of the mammalian uncoordinated 13 (Munc13, also known as Unc13) and Ca(2+)-dependent activator proteins for secretion (CAPS) families, which prime vesicles for exocytosis. Here, we tested whether Munc13 and CAPS proteins also regulate exocytosis in mouse IHCs by combining immunohistochemistry with auditory systems physiology and IHC patch-clamp recordings of exocytosis in mice lacking Munc13 and CAPS isoforms. Surprisingly, we did not detect Munc13 or CAPS proteins at IHC presynaptic active zones and found normal IHC exocytosis as well as auditory brainstem responses (ABRs) in Munc13 and CAPS deletion mutants. Instead, we show that otoferlin, a C2-domain protein that is crucial for vesicular fusion and replenishment in IHCs, clusters at the plasma membrane of the presynaptic active zone. Electron tomography of otoferlin-deficient IHC synapses revealed a reduction of short tethers holding vesicles at the active zone, which might be a structural correlate of impaired vesicle priming in otoferlin-deficient IHCs. We conclude that IHCs use an unconventional priming machinery that involves otoferlin.

  18. All Prime Contract Awards by State or Country, Place, and Contractor, FY 85. Part 14 (Aberdeen, North Carolina - Zanesville, Ohio).

    DTIC Science & Technology

    1985-01-01

    j _j 0 0 0 0C/ I/>C uONN.NN.,C,.C,.CC .(1.C,.r’ 0n UO’t c ON c,4~ ON 0 0 4- U1- 1 nO 0 4 o . -- 0. 4 C4N 0 4 (n In i-i &n Gmo V4 Co n C.4 m W ) 0 m) C...0000 ix 𔃺M0-4 1 1. 000 coc c nv)o nm n nm(no non en r 00 M 10on(m0n1C0-m40a)01 M ) -4 ECO -40V04 Co I M10-4 1 I 1r - NN " N4 NN 4N 44 . . NC j A4 1...8217r:mr Co !i±rnet Awc-rd. by State or Country, Place, and Contractor, FY 85, r, 14 (Aberdeen. North Carolina - Zanesville, Ohio). PERSONAL AUTHOR{S

  19. Vesicles with a double bilayer.

    PubMed

    Zawada, Zygmunt H

    2004-01-01

    A modified reverse phase evaporation method was used to prepare intermediate unilamellar vesicles coated with an additional membrane, or large vesicles in which several vesicles were coated with a common membrane. In both kinds of vesicle, the outer and inner membranes are usually of different phospholipid composition. The preparation involves the formation of a double emulsion: vesicles in a buffer are emerged in a low-boiling point organic solution of phospholipids. Then the organic solvent is evaporated during the heating and mixing process. As result large unilamellar vesicles (LUVs), about 100 nm in diameter, were coated with an additional membrane from egg lecithin or dipalmitoylphosphatidylcholine and cholesterol. The highest yield of the coating was about 50%. When DPPC was used for coating above the phase transition temperature Tm, the data suggested the formation of vesicles that were slightly larger than the starting LUVs. It might be concluded that many of these had a double bilayer. If the coating was done below Tm, the micrographs suggested the formation of structures resembling multi-vesicular vesicles. They looked like LUV clusters coated with a common membrane.

  20. Electrical Properties of Phospholipid Vesicles

    PubMed Central

    Schwan, H. P.; Takashima, S.; Miyamoto, V. K.; Stoeckenius, W.

    1970-01-01

    The capacitance of the membrane of phospholipid vesicles and the electrical properties of the vesicle interior have been determined. To this end the electrical properties of phospholipid vesicles have been investigated over a frequency range extending from 1 kHz to 100 MHz. The dielectric behavior is characterized by two dispersions, one placed between 1 kHz and 1 MHz and the other between 1 and 100 MHz. The relaxational behavior at low frequencies is explained by counterion movement tangential to the vesicle surface and a reasonable value for the fixed charge of the vesicles is calculated from the dispersion magnitude. The relaxation at high frequencies is of the Maxwell-Wagner type and appears caused by the phospholipid bilayer bounding the interior phase of the vesicles. It is consistent with the existence of a closed bilayer with a capacitance of about 2 μF/cm2 and an internal phase similar to the vesicle suspending medium. There is no indication of other than normally structured water inside the small vesicles. PMID:5471701

  1. An ERP investigation of orthographic priming with superset primes.

    PubMed

    Ktori, Maria; Midgley, Katherine; Holcomb, Phillip J; Grainger, Jonathan

    2015-01-12

    Prime stimuli formed by inserting unrelated letters in a given target word (called "superset" primes) provide a means to modify the relative positions of the letters shared by prime and target. Here we examined the time-course of superset priming effects in an ERP study using the sandwich-priming paradigm. We compared the effects of superset primes formed by the insertion of unrelated letters (e.g., maurkdet-MARKET), or by the insertion of hyphens (e.g., ma-rk-et-MARKET), with identity priming (e.g., market-MARKET), all measured relative to unrelated control primes. Behavioral data revealed significantly greater priming in the hyphen-insert condition compared with the letter-insert condition. In the ERP signal, letter-insert priming emerged later than hyphen-insert priming and produced a reversed priming effect in the N400 time-window compared with the more typical N400 priming effects seen for both hyphen-insert priming and identity priming. The different pattern of priming effects seen for letter-insert primes and hyphen-insert primes suggests that compared with identity priming, letter superset priming reflects the joint influence of: (1) a disruption in letter position information, and (2) an inhibitory influence of mismatching letters.

  2. Identification of a Munc13-sensitive step in chromaffin cell large dense-core vesicle exocytosis

    PubMed Central

    Man, Kwun Nok M; Imig, Cordelia; Walter, Alexander M; Pinheiro, Paulo S; Stevens, David R; Rettig, Jens; Sørensen, Jakob B; Cooper, Benjamin H; Brose, Nils; Wojcik, Sonja M

    2015-01-01

    It is currently unknown whether the molecular steps of large dense-core vesicle (LDCV) docking and priming are identical to the corresponding reactions in synaptic vesicle (SV) exocytosis. Munc13s are essential for SV docking and priming, and we systematically analyzed their role in LDCV exocytosis using chromaffin cells lacking individual isoforms. We show that particularly Munc13-2 plays a fundamental role in LDCV exocytosis, but in contrast to synapses lacking Munc13s, the corresponding chromaffin cells do not exhibit a vesicle docking defect. We further demonstrate that ubMunc13-2 and Munc13-1 confer Ca2+-dependent LDCV priming with similar affinities, but distinct kinetics. Using a mathematical model, we identify an early LDCV priming step that is strongly dependent upon Munc13s. Our data demonstrate that the molecular steps of SV and LDCV priming are very similar while SV and LDCV docking mechanisms are distinct. DOI: http://dx.doi.org/10.7554/eLife.10635.001 PMID:26575293

  3. The dynamics of adhesion of a pair of vesicles

    NASA Astrophysics Data System (ADS)

    Walter, Johann; Leal, L. Gary

    2012-11-01

    Adhesive interactions within a suspension of vesicles, such as many personal care products, vectors for drug delivery or artificial blood, can lead to aggregation of the vesicles and dramatic changes to the properties of the suspension. We study the adhesion of a pair of unilamellar, charged vesicles under flow, in the presence of a non-adsorbing polymer or micelle creating a depletion attraction force between the vesicles. Simulations are conducted using a numerical model coupling the boundary integral method for the motion of the fluids and a finite element method for the membrane mechanics (resistance to bending and area increase are both taken into account). The dynamics of the drainage process are studied. At steady state, the adhesion energy is found to depend greatly on the ability of the vesicles to increase their surface area. Finally, when the vesicles are separated in an elongational flow, different behaviors are observed depending on the deformability of the vesicles: an increase of the film thickness with a constant contact area, or peeling-off phenomenon where the contact area decreases at constant film thickness.

  4. Reversible Recruitment of a Homeostatic Reserve Pool of Synaptic Vesicles Underlies Rapid Homeostatic Plasticity of Quantal Content

    PubMed Central

    Pinter, Martin J.; Rich, Mark M.

    2016-01-01

    priming/docking of a small homeostatic reserve pool of vesicles that normally have slow-release kinetics. Following priming, the reserve pool of vesicles is released synchronously with the normal readily releasable pool of synaptic vesicles. This is the first description of this unique pool of synaptic vesicles. PMID:26791213

  5. Priming Gestures with Sounds

    PubMed Central

    Lemaitre, Guillaume; Heller, Laurie M.; Navolio, Nicole; Zúñiga-Peñaranda, Nicolas

    2015-01-01

    We report a series of experiments about a little-studied type of compatibility effect between a stimulus and a response: the priming of manual gestures via sounds associated with these gestures. The goal was to investigate the plasticity of the gesture-sound associations mediating this type of priming. Five experiments used a primed choice-reaction task. Participants were cued by a stimulus to perform response gestures that produced response sounds; those sounds were also used as primes before the response cues. We compared arbitrary associations between gestures and sounds (key lifts and pure tones) created during the experiment (i.e. no pre-existing knowledge) with ecological associations corresponding to the structure of the world (tapping gestures and sounds, scraping gestures and sounds) learned through the entire life of the participant (thus existing prior to the experiment). Two results were found. First, the priming effect exists for ecological as well as arbitrary associations between gestures and sounds. Second, the priming effect is greatly reduced for ecologically existing associations and is eliminated for arbitrary associations when the response gesture stops producing the associated sounds. These results provide evidence that auditory-motor priming is mainly created by rapid learning of the association between sounds and the gestures that produce them. Auditory-motor priming is therefore mediated by short-term associations between gestures and sounds that can be readily reconfigured regardless of prior knowledge. PMID:26544884

  6. Recognizing Plant Defense Priming.

    PubMed

    Martinez-Medina, Ainhoa; Flors, Victor; Heil, Martin; Mauch-Mani, Brigitte; Pieterse, Corné M J; Pozo, Maria J; Ton, Jurriaan; van Dam, Nicole M; Conrath, Uwe

    2016-10-01

    Defense priming conditions diverse plant species for the superinduction of defense, often resulting in enhanced pest and disease resistance and abiotic stress tolerance. Here, we propose a guideline that might assist the plant research community in a consistent assessment of defense priming in plants.

  7. Priming Ability Emotional Intelligence

    ERIC Educational Resources Information Center

    Schutte, Nicola S.; Malouff, John M.

    2012-01-01

    Two studies examined whether priming self-schemas relating to successful emotional competency results in better emotional intelligence performance. In the first study participants were randomly assigned to a successful emotional competency self-schema prime condition or a control condition and then completed an ability measure of emotional…

  8. Discovery: Prime Numbers

    ERIC Educational Resources Information Center

    de Mestre, Neville

    2008-01-01

    Prime numbers are important as the building blocks for the set of all natural numbers, because prime factorisation is an important and useful property of all natural numbers. Students can discover them by using the method known as the Sieve of Eratosthenes, named after the Greek geographer and astronomer who lived from c. 276-194 BC. Eratosthenes…

  9. PRIME Lab Radiocarbon Measurements

    NASA Astrophysics Data System (ADS)

    Hillegonds, D. J.; Mueller, K. A.; Ma, X.; Lipschutz, M. E.

    1996-03-01

    The Purdue Rare Isotope Measurement Laboratory (PRIME Lab) is one of three NSF national facilities for accelerator mass spectrometry (AMS), and is the only one capable of determining six cosmogenic radionuclides: 10Be, 14C, 26Al, 36Cl, 41Ca, and 129I. This abstract describes the current status of the radiocarbon analysis program at PRIME Lab.

  10. Building Numbers from Primes

    ERIC Educational Resources Information Center

    Burkhart, Jerry

    2009-01-01

    Prime numbers are often described as the "building blocks" of natural numbers. This article shows how the author and his students took this idea literally by using prime factorizations to build numbers with blocks. In this activity, students explore many concepts of number theory, including the relationship between greatest common factors and…

  11. Membrane Transport in Isolated Vesicles from Sugarbeet Taproot 1

    PubMed Central

    Briskin, Donald P.; Thornley, W. Robert; Wyse, Roger E.

    1985-01-01

    Sealed membrane vesicles were isolated from homogenates of sugarbeet (Beta vulgaris L.) taproot by a combination of differential centrifugation, extraction with KI, and dextran gradient centrifugation. Relative to the KI-extracted microsomes, the content of plasma membranes, mitochondrial membranes, and Golgi membranes was much reduced in the final vesicle fraction. A component of ATPase activity that was inhibited by nitrate co-enriched with the capacity of the vesicles to form a steady state pH gradient during the purification procedure. This suggests that the nitrate-sensitive ATPase may be involved in driving H+-transport, and this is consistent with the observation that H+-transport, in the final vesicle fraction was inhibited by nitrate. Proton transport in the sugarbeet vesicles was substrate specific for ATP, insensitive to sodium vanadate and oligomycin but was inhibited by diethylstilbestrol and N,N′-dicyclohexylcarbodiimide. The formation of a pH gradient in the vesicles was enhanced by halide ions in the sequence I− > Br− > Cl− while F− was inhibitory. These stimulatory effects occur from both a direct stimulation of the ATPase by anions and a reduction in the vesicle membrane potential. In the presence of Cl−, alkali cations reduce the pH gradient relative to that observed with bis-tris-propane, possibly by H+/alkali cation exchange. Based upon the properties of the H+-transporting vesicles, it is proposed that they are most likely derived from the tonoplast so that this vesicle preparation would represent a convenient system for studying the mechanism of transport at this membrane boundary. PMID:16664342

  12. Activation of metabotropic GABA receptors increases the energy barrier for vesicle fusion.

    PubMed

    Rost, Benjamin R; Nicholson, Patrick; Ahnert-Hilger, Gudrun; Rummel, Andreas; Rosenmund, Christian; Breustedt, Joerg; Schmitz, Dietmar

    2011-09-15

    Neurotransmitter release from presynaptic terminals is under the tight control of various metabotropic receptors. We report here that in addition to the regulation of Ca(2+) channel activity, metabotropic GABA(B) receptors (GABA(B)Rs) at murine hippocampal glutamatergic synapses utilize an inhibitory pathway that directly targets the synaptic vesicle release machinery. Acute application of the GABA(B)R agonist baclofen rapidly and reversibly inhibits vesicle fusion, which occurs independently of the SNAP-25 C-terminus. Using applications of hypertonic sucrose solutions, we find that the size of the readily releasable pool remains unchanged by GABA(B)R activation, but the sensitivity of primed vesicles to hypertonic stimuli appears lowered as the response amplitudes at intermediate sucrose concentrations are smaller and release kinetics are slowed. These data show that presynaptic GABA(B)Rs can inhibit neurotransmitter release directly by increasing the energy barrier for vesicle fusion.

  13. Microfluidic isolation of cancer-cell-derived microvesicles from hetergeneous extracellular shed vesicle populations

    PubMed Central

    Santana, Steven M.; Antonyak, Marc A.; Cerione, Richard A.

    2015-01-01

    Extracellular shed vesicles, including exosomes and microvesicles, are disseminated throughout the body and represent an important conduit of cell communication. Cancer-cell-derived microvesicles have potential as a cancer biomarker as they help shape the tumor microenvironment to promote the growth of the primary tumor and prime the metastatic niche. It is likely that, in cancer cell cultures, the two constituent extracellular shed vesicle subpopulations, observed in dynamic light scattering, represent an exosome population and a cancer-cell-specific microvesicle population and that extracellular shed vesicle size provides information about provenance and cargo. We have designed and implemented a novel microfluidic technology that separates microvesicles, as a function of diameter, from heterogeneous populations of cancer-cell-derived extracellular shed vesicles. We measured cargo carried by the microvesicle subpopulation processed through this microfluidic platform. Such analyses could enable future investigations to more accurately and reliably determine provenance, functional activity, and mechanisms of transformation in cancer. PMID:25342569

  14. Synaptic Vesicle Exocytosis

    PubMed Central

    Südhof, Thomas C.; Rizo, Josep

    2011-01-01

    Presynaptic nerve terminals release neurotransmitters by synaptic vesicle exocytosis. Membrane fusion mediating synaptic exocytosis and other intracellular membrane traffic is affected by a universal machinery that includes SNARE (for “soluble NSF-attachment protein receptor”) and SM (for “Sec1/Munc18-like”) proteins. During fusion, vesicular and target SNARE proteins assemble into an α-helical trans-SNARE complex that forces the two membranes tightly together, and SM proteins likely wrap around assembling trans-SNARE complexes to catalyze membrane fusion. After fusion, SNARE complexes are dissociated by the ATPase NSF (for “N-ethylmaleimide sensitive factor”). Fusion-competent conformations of SNARE proteins are maintained by chaperone complexes composed of CSPα, Hsc70, and SGT, and by nonenzymatically acting synuclein chaperones; dysfunction of these chaperones results in neurodegeneration. The synaptic membrane-fusion machinery is controlled by synaptotagmin, and additionally regulated by a presynaptic protein matrix (the “active zone”) that includes Munc13 and RIM proteins as central components. PMID:22026965

  15. Prime Time in the First Grade at the North Gibson School Corporation: The First Four Years. A Longitudinal Evaluation of Indiana's State-Supported Reduced Class Size Program.

    ERIC Educational Resources Information Center

    Gilman, David A.

    The purpose of this study was to investigate the impact of reducing the size of first grade classes on students' achievement. Four groups of first grade students who had participated in Indiana's Project PRIME TIME (average class size of l8 or fewer) for 1 year were compared to one class of first grade students who had received first grade…

  16. Spectroscopic and photophysical properties of complexes of 4{prime}-ferrocenyl-2,2{prime}:6{prime},2{double_prime}-terpyridine and related ligands

    SciTech Connect

    Hutchinson, K.; Morris, J.C.; Nile, T.A.; Walsh, J.L.; Thompson, D.W.; Petersen, J.D.; Schoonover, J.R.

    1999-05-17

    4{prime}-(Ferrocenyl)-2,2{prime}:6{prime}2{double_prime}-terpyridine (Fctpy) and 4{prime}-(4-pyridyl)-2,2{prime}:6{prime},2{double_prime}-terpyridine (pytpy) were prepared from the corresponding ferrocene- and pyridinecarboxaldehyle and 2-acetylpyridine using the Krohnke synthetic methodology. Metal complexes, [M(Fctpy){sub 2}](PF{sub 6}){sub 2} (M = Ru, Fe, Zn), [Ru(tpy)(Fctpy)](PF{sub 6}){sub 2} (tpy = 2,2{prime}:6{prime},6{double_prime}-terpyridine), and [Ru(pytpy){sub 2}](PF{sub 6}){sub 2} were prepared and characterized. Cyclic voltammetric analysis indicated Ru(III/II) and ferrocenium/ferrocene redox couples near expected potentials (Ru{sup III/II} {approximately}1.3 V and ferrocenium/ferrocene {approximately}0.6 V vs Ag/AgCl). In addition to dominant {pi}{sub tpy} {r_arrow} {pi}{sub tpy}{sup *} UV absorptions near 240 and 280 nm and d{sub {pi}}{sup Ru} {r_arrow} {pi}{sub tpy}{sup *} MLCT absorptions around 480 nm, the complexes [Ru(Fctpy){sub 2}](PF{sub 6}){sub 2} and [Ru(tpy)(Fctpy)](PF{sub 6}){sub 2} exhibit an unusual absorption band around 530 nm. Resonance Raman measurements indicate that this band is due to a {sup 1}[(d({pi}){sub Fc}){sup 6}] {r_arrow} {sup 1}[(d({pi}){sub Fc}){sup 5}({pi}{sup *}{sub tpy}{sup Ru}){sup 1}] transition. For [Ru(Fctpy){sub 2}](PF{sub 6}){sub 2} and [Ru(tpy)(Fctpy)](PF{sub 6}){sub 2}, excited-state emission and lifetime measurements indicated an upper-limit emission quantum yield of 0.003 and an upper-limit emission lifetime of 0.025 {micro}s. The influence of the ferrocenyl site on excited-state decay is discussed, and an excited-state energy level diagram is proposed.

  17. Largest known twin primes and Sophie Germain primes

    NASA Astrophysics Data System (ADS)

    Indlekofer, Karl-Heinz; Járai, Antal

    The numbers 242206083* 2^38880+-1 are twin primes. The number p=2375063906985* 2^19380-1 is a Sophie Germain prime, i.e. p and 2p+1 are both primes. For p=4610194180515* 2^ 5056-1, the numbers p, p+2 and 2p+1 are all primes.

  18. Masked priming effect reflects evidence accumulated by the prime.

    PubMed

    Kinoshita, Sachiko; Norris, Dennis

    2010-01-01

    In the same-different match task, masked priming is observed with the same responses but not different responses. Norris and Kinoshita's (2008) Bayesian reader account of masked priming explains this pattern based on the same principle as that explaining the absence of priming for nonwords in the lexical decision task. The pattern of priming follows from the way the model makes optimal decisions in the two tasks; priming does not depend on first activating the prime and then the target. An alternative explanation is in terms of a bias towards responding "same" that exactly counters the facilitatory effect of lexical access. The present study tested these two views by varying both the degree to which the prime predicts the response and the visibility of the prime. Unmasked primes produced effects expected from the view that priming is influenced by the degree to which the prime predicts the response. In contrast, with masked primes, the size of priming for the same response was completely unaffected by predictability. These results rule out response bias as an explanation of the absence of masked priming for different responses and, in turn, indicate that masked priming is not a consequence of automatic lexical access of the prime.

  19. How important is a prime's gestalt for subliminal priming?

    PubMed

    Jaśkowski, Piotr; Slósarek, Maciej

    2007-06-01

    Masked stimuli (primes) can affect the preparation of a motor response to subsequently presented target stimuli. Under some conditions, reactions to the main stimulus can be facilitated (straight priming) or inhibited (inverse priming) when preceded by a compatible prime (calling for the same response). In the majority of studies in which inverse priming was demonstrated arrows pointing left or right were used as prime and targets. There is, however, evidence that arrows are special overlearned stimuli which are processed in a favorable way. Here we report three experiments designated to test whether the "arrowness" of primes/targets is a sufficient condition for inverse priming. The results clearly show that although inverse priming appeared when non-arrow shapes were used, the magnitude of the priming effect was larger with arrows. The possible reasons for this effect are discussed.

  20. Phosphorylation of synaptotagmin-1 controls a post-priming step in PKC-dependent presynaptic plasticity

    PubMed Central

    de Jong, Arthur P. H.; Meijer, Marieke; Saarloos, Ingrid; Cornelisse, Lennart Niels; Toonen, Ruud F. G.; Sørensen, Jakob B.; Verhage, Matthijs

    2016-01-01

    Presynaptic activation of the diacylglycerol (DAG)/protein kinase C (PKC) pathway is a central event in short-term synaptic plasticity. Two substrates, Munc13-1 and Munc18-1, are essential for DAG-induced potentiation of vesicle priming, but the role of most presynaptic PKC substrates is not understood. Here, we show that a mutation in synaptotagmin-1 (Syt1T112A), which prevents its PKC-dependent phosphorylation, abolishes DAG-induced potentiation of synaptic transmission in hippocampal neurons. This mutant also reduces potentiation of spontaneous release, but only if alternative Ca2+ sensors, Doc2A/B proteins, are absent. However, unlike mutations in Munc13-1 or Munc18-1 that prevent DAG-induced potentiation, the synaptotagmin-1 mutation does not affect paired-pulse facilitation. Furthermore, experiments to probe vesicle priming (recovery after train stimulation and dual application of hypertonic solutions) also reveal no abnormalities. Expression of synaptotagmin-2, which lacks a seven amino acid sequence that contains the phosphorylation site in synaptotagmin-1, or a synaptotagmin-1 variant with these seven residues removed (Syt1Δ109–116), supports normal DAG-induced potentiation. These data suggest that this seven residue sequence in synaptotagmin-1 situated in the linker between the transmembrane and C2A domains is inhibitory in the unphosphorylated state and becomes permissive of potentiation upon phosphorylation. We conclude that synaptotagmin-1 phosphorylation is an essential step in PKC-dependent potentiation of synaptic transmission, acting downstream of the two other essential DAG/PKC substrates, Munc13-1 and Munc18-1. PMID:27091977

  1. Phosphorylation of synaptotagmin-1 controls a post-priming step in PKC-dependent presynaptic plasticity.

    PubMed

    de Jong, Arthur P H; Meijer, Marieke; Saarloos, Ingrid; Cornelisse, Lennart Niels; Toonen, Ruud F G; Sørensen, Jakob B; Verhage, Matthijs

    2016-05-03

    Presynaptic activation of the diacylglycerol (DAG)/protein kinase C (PKC) pathway is a central event in short-term synaptic plasticity. Two substrates, Munc13-1 and Munc18-1, are essential for DAG-induced potentiation of vesicle priming, but the role of most presynaptic PKC substrates is not understood. Here, we show that a mutation in synaptotagmin-1 (Syt1(T112A)), which prevents its PKC-dependent phosphorylation, abolishes DAG-induced potentiation of synaptic transmission in hippocampal neurons. This mutant also reduces potentiation of spontaneous release, but only if alternative Ca(2+) sensors, Doc2A/B proteins, are absent. However, unlike mutations in Munc13-1 or Munc18-1 that prevent DAG-induced potentiation, the synaptotagmin-1 mutation does not affect paired-pulse facilitation. Furthermore, experiments to probe vesicle priming (recovery after train stimulation and dual application of hypertonic solutions) also reveal no abnormalities. Expression of synaptotagmin-2, which lacks a seven amino acid sequence that contains the phosphorylation site in synaptotagmin-1, or a synaptotagmin-1 variant with these seven residues removed (Syt1(Δ109-116)), supports normal DAG-induced potentiation. These data suggest that this seven residue sequence in synaptotagmin-1 situated in the linker between the transmembrane and C2A domains is inhibitory in the unphosphorylated state and becomes permissive of potentiation upon phosphorylation. We conclude that synaptotagmin-1 phosphorylation is an essential step in PKC-dependent potentiation of synaptic transmission, acting downstream of the two other essential DAG/PKC substrates, Munc13-1 and Munc18-1.

  2. Overall energy conversion efficiency of a photosynthetic vesicle.

    PubMed

    Sener, Melih; Strumpfer, Johan; Singharoy, Abhishek; Hunter, C Neil; Schulten, Klaus

    2016-08-26

    The chromatophore of purple bacteria is an intracellular spherical vesicle that exists in numerous copies in the cell and that efficiently converts sunlight into ATP synthesis, operating typically under low light conditions. Building on an atomic-level structural model of a low-light-adapted chromatophore vesicle from Rhodobacter sphaeroides, we investigate the cooperation between more than a hundred protein complexes in the vesicle. The steady-state ATP production rate as a function of incident light intensity is determined after identifying quinol turnover at the cytochrome bc1 complex (cytb⁢c1) as rate limiting and assuming that the quinone/quinol pool of about 900 molecules acts in a quasi-stationary state. For an illumination condition equivalent to 1% of full sunlight, the vesicle exhibits an ATP production rate of 82 ATP molecules/s. The energy conversion efficiency of ATP synthesis at illuminations corresponding to 1%-5% of full sunlight is calculated to be 0.12-0.04, respectively. The vesicle stoichiometry, evolutionarily adapted to the low light intensities in the habitat of purple bacteria, is suboptimal for steady-state ATP turnover for the benefit of protection against over-illumination.

  3. Overall energy conversion efficiency of a photosynthetic vesicle

    PubMed Central

    Sener, Melih; Strumpfer, Johan; Singharoy, Abhishek; Hunter, C Neil; Schulten, Klaus

    2016-01-01

    The chromatophore of purple bacteria is an intracellular spherical vesicle that exists in numerous copies in the cell and that efficiently converts sunlight into ATP synthesis, operating typically under low light conditions. Building on an atomic-level structural model of a low-light-adapted chromatophore vesicle from Rhodobacter sphaeroides, we investigate the cooperation between more than a hundred protein complexes in the vesicle. The steady-state ATP production rate as a function of incident light intensity is determined after identifying quinol turnover at the cytochrome bc1 complex (cytb⁢c1) as rate limiting and assuming that the quinone/quinol pool of about 900 molecules acts in a quasi-stationary state. For an illumination condition equivalent to 1% of full sunlight, the vesicle exhibits an ATP production rate of 82 ATP molecules/s. The energy conversion efficiency of ATP synthesis at illuminations corresponding to 1%–5% of full sunlight is calculated to be 0.12–0.04, respectively. The vesicle stoichiometry, evolutionarily adapted to the low light intensities in the habitat of purple bacteria, is suboptimal for steady-state ATP turnover for the benefit of protection against over-illumination. DOI: http://dx.doi.org/10.7554/eLife.09541.001 PMID:27564854

  4. Ellipsoidal Relaxation of Deformed Vesicles

    NASA Astrophysics Data System (ADS)

    Yu, Miao; Lira, Rafael B.; Riske, Karin A.; Dimova, Rumiana; Lin, Hao

    2015-09-01

    Theoretical analysis and experimental quantification on the ellipsoidal relaxation of vesicles are presented. The current work reveals the simplicity and universal aspects of this process. The Helfrich formula is shown to apply to the dynamic relaxation of moderate-to-high tension membranes, and a closed-form solution is derived which predicts the vesicle aspect ratio as a function of time. Scattered data are unified by a time scale, which leads to a similarity behavior, governed by a distinctive solution for each vesicle type. Two separate regimes in the relaxation are identified, namely, the "entropic" and the "constant-tension" regimes. The bending rigidity and the initial membrane tension can be simultaneously extracted from the data analysis, posing the current approach as an effective means for the mechanical analysis of biomembranes.

  5. Temperature-Dependent Differences between Readily Releasable and Reserve Pool Vesicles in Chromaffin Cells

    PubMed Central

    Haynes, Christy L.; Siff, Lauren N.; Wightman, R. Mark

    2007-01-01

    Summary Statistical differences between amperometric traces recorded from chromaffin cells using K+ and Ba2+ secretagogues support the assertion that readily releasable pool (RRP) and reserve pool (RP) vesicles can be probed with pool-specific secretagogues. Release from the RRP was evoked by K+ while release from the RP was evoked by Ba2+. Similar temperature-dependent changes in spike area and half-width for both pools suggest that the content of RRP and RP vesicles is similar and packaged in the same way. Differences between the vesicle pools were revealed in the temperature dependence of spike frequency. While the burst spike frequency of the RRP, which is comprised of pre-docked and primed vesicles, increased 2.8% per °C, the RP spike frequency increased 12% per °C. This difference is attributed to a temperature dependent mobilization of the RP. Furthermore, the RP exhibited more foot events at room temperature than the RRP but this difference was not apparent at 37°C. This trend suggests that RP vesicle membranes have a compromised surface tension compared to RRP vesicles. Collectively, the changes of release characteristics with temperature reveal distinctions between the RRP and the RP. PMID:17467077

  6. Synaptic vesicle distribution by conveyor belt.

    PubMed

    Moughamian, Armen J; Holzbaur, Erika L F

    2012-03-02

    The equal distribution of synaptic vesicles among synapses along the axon is critical for robust neurotransmission. Wong et al. show that the continuous circulation of synaptic vesicles throughout the axon driven by molecular motors ultimately yields this even distribution.

  7. Mechanisms of COPI vesicle formation

    PubMed Central

    Hsu, Victor W.; Yang, Jia-Shu

    2009-01-01

    Coat Protein I (COPI) is one of the most intensely investigated coat complexes. Numerous studies have contributed to a general understanding of how coat proteins act to initiate intracellular vesicular transport. This review highlights key recent findings that have shaped our current understanding of how COPI vesicles are formed. PMID:19854177

  8. Priming with NO controls redox state and prevents cadmium-induced general up-regulation of methionine sulfoxide reductase gene family in Arabidopsis.

    PubMed

    Méndez, Andrea A E; Pena, Liliana B; Benavides, María P; Gallego, Susana M

    2016-12-01

    In the present study we evaluated the pre-treatment (priming) of Arabidopsis thaliana plants with sodium nitroprusside (SNP), a NO-donor, as an interesting approach for improving plant tolerance to cadmium stress. We focused on the cell redox balance and on the methionine sulfoxide reductases (MSR) family as a key component of such response. MSR catalyse the reversible oxidation of MetSO residues back to Met. Five MSRA genes and nine MSRB genes have been identified in A. thaliana, coding for proteins with different subcellular locations. After treating 20 days-old A. thaliana (Col 0) plants with 100 μM CdCl2, increased protein carbonylation in leaf tissue, lower chlorophyll content and higher levels of reactive oxygen species (ROS) in chloroplasts were detected, together with increased accumulation of all MSR transcripts evaluated. Further analysis showed reduction in guaiacol peroxidase activity (GPX) and increased catalase (CAT) activity, with no effect on ascorbate peroxidase (APX) activity. Pre-exposition of plants to 100 μM SNP before cadmium treatment restored redox balance; this seems to be linked to a better performance of antioxidant defenses. Our results indicate that NO priming may be acting as a modulator of plant antioxidant system by interfering in oxidative responses and by preventing up-regulation of MSR genes caused by metal exposure.

  9. Morphology of nematic and smectic vesicles

    PubMed Central

    Xing, Xiangjun; Shin, Homin; Bowick, Mark J.; Yao, Zhenwei; Jia, Lin; Li, Min-Hui

    2012-01-01

    Recent experiments on vesicles formed from block copolymers with liquid-crystalline side chains reveal a rich variety of vesicle morphologies. The additional internal order (“structure”) developed by these self-assembled block copolymer vesicles can lead to significantly deformed vesicles as a result of the delicate interplay between two-dimensional ordering and vesicle shape. The inevitable topological defects in structured vesicles of spherical topology also play an essential role in controlling the final vesicle morphology. Here we develop a minimal theoretical model for the morphology of the membrane structure with internal nematic/smectic order. Using both analytic and numerical approaches, we show that the possible low free energy morphologies include nano-size cylindrical micelles (nano-fibers), faceted tetrahedral vesicles, and ellipsoidal vesicles, as well as cylindrical vesicles. The tetrahedral vesicle is a particularly fascinating example of a faceted liquid-crystalline membrane. Faceted liquid vesicles may lead to the design of supramolecular structures with tetrahedral symmetry and new classes of nano-carriers. PMID:22431595

  10. Prime Retrieval of Motor Responses in Negative Priming

    ERIC Educational Resources Information Center

    Mayr, Susanne; Buchner, Axel; Dentale, Sandra

    2009-01-01

    Three auditory identification experiments were designed to specify the prime-response retrieval model of negative priming (S. Mayr & A. Buchner, 2006), which assumes that the prime response is retrieved in ignored repetition trials and interferes with probe responding. In Experiment 1, shortly before (in Experiment 1A) or after (in Experiment 1B)…

  11. Roles of BLOC-1 and Adaptor Protein-3 Complexes in Cargo Sorting to Synaptic Vesicles

    PubMed Central

    Newell-Litwa, Karen; Salazar, Gloria; Smith, Yoland

    2009-01-01

    Neuronal lysosomes and their biogenesis mechanisms are primarily thought to clear metabolites and proteins whose abnormal accumulation leads to neurodegenerative disease pathology. However, it remains unknown whether lysosomal sorting mechanisms regulate the levels of membrane proteins within synaptic vesicles. Using high-resolution deconvolution microscopy, we identified early endosomal compartments where both selected synaptic vesicle and lysosomal membrane proteins coexist with the adaptor protein complex 3 (AP-3) in neuronal cells. From these early endosomes, both synaptic vesicle membrane proteins and characteristic AP-3 lysosomal cargoes can be similarly sorted to brain synaptic vesicles and PC12 synaptic-like microvesicles. Mouse knockouts for two Hermansky–Pudlak complexes involved in lysosomal biogenesis from early endosomes, the ubiquitous isoform of AP-3 (Ap3b1−/−) and muted, defective in the biogenesis of lysosome-related organelles complex 1 (BLOC-1), increased the content of characteristic synaptic vesicle proteins and known AP-3 lysosomal proteins in isolated synaptic vesicle fractions. These phenotypes contrast with those of the mouse knockout for the neuronal AP-3 isoform involved in synaptic vesicle biogenesis (Ap3b2−/−), in which the content of select proteins was reduced in synaptic vesicles. Our results demonstrate that lysosomal and lysosome-related organelle biogenesis mechanisms regulate steady-state synaptic vesicle protein composition from shared early endosomes. PMID:19144828

  12. Rotavirus interaction with isolated membrane vesicles.

    PubMed

    Ruiz, M C; Alonso-Torre, S R; Charpilienne, A; Vasseur, M; Michelangeli, F; Cohen, J; Alvarado, F

    1994-06-01

    To gain information about the mechanism of epithelial cell infection by rotavirus, we studied the interaction of bovine rotavirus, RF strain, with isolated membrane vesicles from apical membrane of pig enterocytes. Vesicles were charged with high (quenching) concentrations of either carboxyfluorescein or calcein, and the rate of fluorophore release (dequenching) was monitored as a function of time after mixing with purified virus particles. Purified single-shelled particles and untrypsinized double-shelled ones had no effect. Trypsinized double-shelled virions induced carboxyfluorescein release according to sigmoid curves whose lag period and amplitude were a function of virus concentration and depended on both temperature and pH. The presence of 100 mM salts (Tris Cl, NaCl, or KCl) was required, since there was no reaction in isoosmotic salt-free sorbitol media. Other membrane vesicle preparations such as apical membranes of piglet enterocyte and rat placenta syncytiotrophoblasts, basolateral membranes of pig enterocytes, and the undifferentiated plasma membrane of cultured MA104 cells all gave qualitatively similar responses. Inhibition by a specific monoclonal antibody suggests that the active species causing carboxyfluorescein release is VP5*. Ca2+ (1 mM), but not Mg2+, inhibited the reaction. In situ solubilization of the outer capsid of trypsinized double-shelled particles changed release kinetics from sigmoidal to hyperbolic and was not inhibited by Ca2+. Our results indicate that membrane destabilization caused by trypsinized outer capsid proteins of rotavirus leads to fluorophore release. From the data presented here, a hypothetical model of the interaction of the various states of the viral particles with the membrane lipid phase is proposed. Membrane permeabilization induced by rotavirus may be related to the mechanism of entry of the virus into the host cell.

  13. "Fell" Primes "Fall", but Does "Bell" Prime "Ball"? Masked Priming with Irregularly-Inflected Primes

    ERIC Educational Resources Information Center

    Crepaldi, Davide; Rastle, Kathleen; Coltheart, Max; Nickels, Lyndsey

    2010-01-01

    Recent masked priming experiments have brought to light a morphological level of analysis that is exclusively based on the orthographic appearance of words, so that it breaks down corner into corn- and -er, as well as dealer into deal- and -er (Rastle, Davis, & New, 2004). Being insensitive to semantic factors, this morpho-orthographic…

  14. Prime-Time Nutrition.

    ERIC Educational Resources Information Center

    Kaufman, Lois

    1980-01-01

    Presents a study identifying, analyzing, and describing messages on prime-time network television related to food, eating behavior, and ideal body image. Program content and commercials studied present conflicting messages: (1) that we eat in ways almost guaranteed to make us fat, and (2) that we strive to remain too slim. (JMF)

  15. Preparation and Properties of Asymmetric Large Unilamellar Vesicles: Interleaflet Coupling in Asymmetric Vesicles Is Dependent on Temperature but Not Curvature

    PubMed Central

    Cheng, Hui-Ting; London, Erwin

    2011-01-01

    Asymmetry of inner and outer leaflet lipid composition is an important characteristic of eukaryotic plasma membranes. We previously described a technique in which methyl-β-cyclodextrin-induced lipid exchange is used to prepare biological membrane-like asymmetric small unilamellar vesicles (SUVs). Here, to mimic plasma membranes more closely, we used a lipid-exchange-based method to prepare asymmetric large unilamellar vesicles (LUVs), which have less membrane curvature than SUVs. Asymmetric LUVs in which sphingomyelin (SM) or SM + 1-palmitoyl-2-oleoyl-phosphatidylcholine was exchanged into the outer leaflet of vesicles composed of 1,2-dioleoyl-phosphatidylethanolamine (DOPE) and 1-palmitoyl-2-oleoyl-phosphatidylserine (POPS) were prepared with or without cholesterol. Approximately 80–100% replacement of outer leaflet DOPE and POPS was achieved. At room temperature, SM exchange into the outer leaflet increased the inner leaflet lipid order, suggesting significant interleaflet interaction. However, the SM-rich outer leaflet formed an ordered state, melting with a midpoint at ∼37°C. This was about the same value observed in pure SM vesicles, and was significantly higher than that observed in symmetric vesicles with the same SM content, which melted at ∼20°C. In other words, ordered state formation by outer-leaflet SM in asymmetric vesicles was not destabilized by an inner leaflet composed of DOPE and POPS. These properties suggest that the coupling between the physical states of the outer and inner leaflets in these asymmetric LUVs becomes very weak as the temperature approaches 37°C. Overall, the properties of asymmetric LUVs were very similar to those previously observed in asymmetric SUVs, indicating that they do not arise from the high membrane curvature of asymmetric SUVs. PMID:21641312

  16. Masked Repetition Priming Using Magnetoencephalography

    ERIC Educational Resources Information Center

    Monahan, Philip J.; Fiorentino, Robert; Poeppel, David

    2008-01-01

    Masked priming is used in psycholinguistic studies to assess questions about lexical access and representation. We present two masked priming experiments using MEG. If the MEG signal elicited by words reflects specific aspects of lexical retrieval, then one expects to identify specific neural correlates of retrieval that are sensitive to priming.…

  17. Rhizosphere priming: a nutrient perspective

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhizosphere priming is the change in decomposition of soil organic matter (SOM) caused by root activity. Rhizosphere priming plays a crucial role in soil carbon (C) dynamics and their response to global climate change. Rhizosphere priming may be affected by soil nutrient availability, but rhizospher...

  18. Priming Ditransitive Structures in Comprehension

    ERIC Educational Resources Information Center

    Arai, Manabu; van Gompel, Roger P. G.; Scheepers, Cristoph

    2007-01-01

    Many studies have shown evidence for syntactic priming during language production (e.g., Bock, 1986). It is often assumed that comprehension and production share similar mechanisms and that priming also occurs during comprehension (e.g., Pickering & Garrod, 2004). Research investigating priming during comprehension (e.g., Branigan et al., 2005 and…

  19. Rheological properties of a vesicle suspension

    NASA Astrophysics Data System (ADS)

    Guedda, M.; Benlahsen, M.; Misbah, C.

    2014-11-01

    The rheological behavior of a dilute suspension of vesicles in linear shear flow at a finite concentration is analytically examined. In the quasispherical limit, two coupled nonlinear equations that describe the vesicle orientation in the flow and its shape evolution were derived [Phys. Rev. Lett. 96, 028104 (2006), 10.1103/PhysRevLett.96.028104] and serve here as a starting point. Of special interest is to provide, for the first time, an exact analytical prediction of the time-dependent effective viscosity ηeff and normal stress differences N1 and N2. Our results shed light on the effect of the viscosity ratio λ (defined as the inner over the outer fluid viscosities) as the main controlling parameter. It is shown that ηeff,N1 , and N2 either tend to a steady state or describe a periodic time-dependent rheological response, previously reported numerically and experimentally. In particular, the shear viscosity minimum and the cusp singularities of ηeff,N1 , and N2 at the tumbling threshold are brought to light. We also report on rheology properties for an arbitrary linear flow. We were able to obtain a constitutive law in a closed form relating the stress tensor to the strain rate tensor. It is found that the resulting constitutive markedly contrasts with classical laws known for other complex fluids, such as emulsions, capsule suspensions, and dilute polymer solutions (Oldroyd B model). We highlight the main differences between our law and classical laws.

  20. Hemifusion in Synaptic Vesicle Cycle

    PubMed Central

    Kweon, Dae-Hyuk; Kong, Byoungjae; Shin, Yeon-Kyun

    2017-01-01

    In the neuron, early neurotransmitters are released through the fusion pore prior to the complete vesicle fusion. It has been thought that the fusion pore is a gap junction-like structure made of transmembrane domains (TMDs) of soluble N-ethylmaleimide-sensitive-factor attachment protein receptor (SNARE) proteins. However, evidence has accumulated that lipid mixing occurs prior to the neurotransmitter release through the fusion pore lined predominantly with lipids. To explain these observations, the hemifusion, a membrane structure in which two bilayers are partially merged, has emerged as a key step preceding the formation of the fusion pore. Furthermore, the hemifusion appears to be the bona fide intermediate step not only for the synaptic vesicle cycle, but for a wide range of membrane remodeling processes, such as viral membrane fusion and endocytotic membrane fission. PMID:28360835

  1. Phospholipid Vesicles in Materials Science

    SciTech Connect

    Granick, Steve

    2016-05-11

    The objective of this research was to develop the science basis needed to deploy phospholipid vesicles as functional materials in energy contexts. Specifically, we sought to: (1) Develop an integrated molecular-level understanding of what determines their dynamical shape, spatial organization, and responsiveness to complex, time-varying environments; and (2) Develop understanding of their active transportation in crowded environments, which our preliminary measurements in cells suggest may hold design principles for targeting improved energy efficiency in new materials systems. The methods to do this largely involved fluorescence imaging and other spectroscopy involving single particles, vesicles, particles, DNA, and endosomes. An unexpected importance outcome was a new method to image light-emitting diodes during actual operation using super-resolution spectroscopy.

  2. Vesicle Motion during Sustained Exocytosis in Chromaffin Cells: Numerical Model Based on Amperometric Measurements

    PubMed Central

    Jarukanont, Daungruthai; Bonifas Arredondo, Imelda; Femat, Ricardo; Garcia, Martin E.

    2015-01-01

    Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. We developed a numerical model based on Langevin simulations of vesicle motion towards the cell membrane and on the statistical analysis of vesicle arrival times. We also performed amperometric experiments in bovine-adrenal Chromaffin cells under Ba2+ stimulation to capture neurotransmitter releases during sustained exocytosis. In the sustained phase, each amperometric peak can be related to a single release from a new vesicle arriving at the active site. The amperometric signal can then be mapped into a spike-series of release events. We normalized the spike-series resulting from the current peaks using a time-rescaling transformation, thus making signals coming from different cells comparable. We discuss why the obtained spike-series may contain information about the motion of all vesicles leading to release of catecholamines. We show that the release statistics in our experiments considerably deviate from Poisson processes. Moreover, the interspike-time probability is reasonably well described by two-parameter gamma distributions. In order to interpret this result we computed the vesicles’ arrival statistics from our Langevin simulations. As expected, assuming purely diffusive vesicle motion we obtain Poisson statistics. However, if we assume that all vesicles are guided toward the membrane by an attractive harmonic potential, simulations also lead to gamma distributions of the interspike-time probability, in remarkably good agreement with experiment. We also show that

  3. Semantic priming of familiar songs.

    PubMed

    Johnson, Sarah K; Halpern, Andrea R

    2012-05-01

    We explored the functional organization of semantic memory for music by comparing priming across familiar songs both within modalities (Experiment 1, tune to tune; Experiment 3, category label to lyrics) and across modalities (Experiment 2, category label to tune; Experiment 4, tune to lyrics). Participants judged whether or not the target tune or lyrics were real (akin to lexical decision tasks). We found significant priming, analogous to linguistic associative-priming effects, in reaction times for related primes as compared to unrelated primes, but primarily for within-modality comparisons. Reaction times to tunes (e.g., "Silent Night") were faster following related tunes ("Deck the Hall") than following unrelated tunes ("God Bless America"). However, a category label (e.g., Christmas) did not prime tunes from within that category. Lyrics were primed by a related category label, but not by a related tune. These results support the conceptual organization of music in semantic memory, but with potentially weaker associations across modalities.

  4. Primes in Fibonacci n-step and Lucas n-step Sequences

    NASA Astrophysics Data System (ADS)

    Noe, Tony D.; Vos Post, Jonathan

    2005-09-01

    We search for primes in the Fibonacci n-step and Lucas n-step sequences, which are the natural generalizations of the Fibonacci and Lucas numbers. While the Fibonacci n-step sequences are nearly devoid of primes, the Lucas n-step sequences are prime-rich. We tabulate the occurrence of primes in the first 10000 terms for n <= 100. We also state two conjectures about Diophantine equations based on these sequences.

  5. Two primes priming: does feature integration occur before response activation?

    PubMed

    Grainger, Julianne E; Scharnowski, Frank; Schmidt, Thomas; Herzog, Michael H

    2013-07-17

    Responses to a target can be sped up or slowed down by a congruent or incongruent prime, respectively. Even though presentations are rapid, the prime and the target are thought to activate motor responses in strict sequence, with prime activation preceding target activation. In feature fusion, the opposite seems to be the case. For example, a vernier offset to the left is immediately followed by a vernier offset to the right at the same location. The two verniers are not perceived as two elements in sequence but as a single, aligned vernier. Here, we ask the question as to how features are integrated: before or after motor activation? We presented two vernier primes with opposite offset directions preceding a single vernier target. No priming effect occurred when the vernier primes were presented at the same location, indicating that verniers integrate before motor activation. There was also no priming effect when the primes were presented simultaneously at different locations, indicating that there is an integration stage different from the perceptual fusion stage. When the second prime is delayed, it determines priming, even for very long delays. To explain these long integration times, we argue that there is a buffer preceding motor activation.

  6. Deciphering dead-end docking of large dense core vesicles in bovine chromaffin cells.

    PubMed

    Hugo, Sandra; Dembla, Ekta; Halimani, Mahantappa; Matti, Ulf; Rettig, Jens; Becherer, Ute

    2013-10-23

    Large dense core vesicle (LDCV) exocytosis in chromaffin cells follows a well characterized process consisting of docking, priming, and fusion. Total internal reflection fluorescence microscopy (TIRFM) studies suggest that some LDCVs, although being able to dock, are resistant to calcium-triggered release. This phenomenon termed dead-end docking has not been investigated until now. We characterized dead-end vesicles using a combination of membrane capacitance measurement and visualization of LDCVs with TIRFM. Stimulation of bovine chromaffin cells for 5 min with 6 μm free intracellular Ca2+ induced strong secretion and a large reduction of the LDCV density at the plasma membrane. Approximately 15% of the LDCVs were visible at the plasma membrane throughout experiments, indicating they were permanently docked dead-end vesicles. Overexpression of Munc18-2 or SNAP-25 reduced the fraction of dead-end vesicles. Conversely, expressing open-syntaxin increased the fraction of dead-end vesicles. These results indicate the existence of the unproductive target soluble N-ethylmaleimide-sensitive factor attachment protein receptor acceptor complex composed of 2:1 syntaxin-SNAP-25 in vivo. More importantly, they define a novel function for this acceptor complex in mediating dead-end docking.

  7. Hydrothermal syntheses, crystal structures and luminescence properties of zinc(II) and cadmium(II) coordination polymers based on bifunctional 3,2 Prime :6 Prime ,3 Prime Prime -terpyridine-4 Prime -carboxylic acid

    SciTech Connect

    Li, Na; Guo, Hui-Lin; Hu, Huai-Ming; Song, Juan; Xu, Bing; Yang, Meng-Lin; Dong, Fa-Xin; Xue, Gang-Lin

    2013-02-15

    Five new coordination polymers, [Zn{sub 2}(ctpy){sub 2}Cl{sub 2}]{sub n} (1), [Zn{sub 2}(ctpy){sub 2}(ox)(H{sub 2}O){sub 2}]{sub n} (2), [Zn{sub 2}(ctpy)(3-btc)(H{sub 2}O)]{sub n}{center_dot}0.5nH{sub 2}O (3), [Cd(ctpy){sub 2}(H{sub 2}O)]{sub n} (4), [Cd{sub 4}(ctpy){sub 2}(2-btc){sub 2}(H{sub 2}O){sub 2}]{sub n}{center_dot}2nH{sub 2}O (5), (Hctpy=3,2 Prime :6 Prime ,3 Prime Prime -terpyridine-4 Prime -carboxylic acid, H{sub 2}ox=oxalic acid, H{sub 3}(3-btc)=1,3,5-benzenetricarboxylic acid, H{sub 3}(2-btc)=1,2,4-benzenetricarboxylic acid) have been synthesized under hydrothermal conditions and characterized by elemental analysis, IR spectroscopy, and single-crystal X-ray diffraction. Compounds 1-2 are a one-dimensional chain with weak interactions to form 3D supramolecular structures. Compound 3 is a 4-nodal 3D topology framework comprised of binuclear zinc units and (ctpy){sup -} anions. Compound 4 shows two dimensional net. Compound 5 is a (4,5,6)-connected framework with {l_brace}4{sup 4}{center_dot}6{sup 2}{r_brace}{l_brace}4{sup 6}{center_dot}6{sup 4}{r_brace}{sub 2}{l_brace}4{sup 9}{center_dot}6{sup 6}{r_brace} topology. In addition, the thermal stabilities and photoluminescence properties of 1-5 were also studied in the solid state. - Graphical abstract: Five new Zn/Cd compounds with 3,2 Prime :6 Prime ,3 Prime Prime -terpyridine-4 Prime -carboxylic acid were prepared. The photoluminescence and thermal stabilities properties of 1-5 were investigated in the solid state. Highlights: Black-Right-Pointing-Pointer Five new zinc/cadmium metal-organic frameworks have been hydrothermal synthesized. Black-Right-Pointing-Pointer The structural variation is attributed to the diverse metal ions and auxiliary ligand. Black-Right-Pointing-Pointer Compounds 1-5 exhibit 1D ring chain, 2D layer and 3D open-framework, respectively. Black-Right-Pointing-Pointer These compounds exhibit strong solid state luminescence emission at room temperature.

  8. Synaptotagmin-1 and -7 Are Redundantly Essential for Maintaining the Capacity of the Readily-Releasable Pool of Synaptic Vesicles

    PubMed Central

    Burré, Jacqueline; Malenka, Robert C.; Liu, Xinran; Südhof, Thomas C.

    2015-01-01

    In forebrain neurons, Ca2+ triggers exocytosis of readily releasable vesicles by binding to synaptotagmin-1 and -7, thereby inducing fast and slow vesicle exocytosis, respectively. Loss-of-function of synaptotagmin-1 or -7 selectively impairs the fast and slow phase of release, respectively, but does not change the size of the readily-releasable pool (RRP) of vesicles as measured by stimulation of release with hypertonic sucrose, or alter the rate of vesicle priming into the RRP. Here we show, however, that simultaneous loss-of-function of both synaptotagmin-1 and -7 dramatically decreased the capacity of the RRP, again without altering the rate of vesicle priming into the RRP. Either synaptotagmin-1 or -7 was sufficient to rescue the RRP size in neurons lacking both synaptotagmin-1 and -7. Although maintenance of RRP size was Ca2+-independent, mutations in Ca2+-binding sequences of synaptotagmin-1 or synaptotagmin-7—which are contained in flexible top-loop sequences of their C2 domains—blocked the ability of these synaptotagmins to maintain the RRP size. Both synaptotagmins bound to SNARE complexes; SNARE complex binding was reduced by the top-loop mutations that impaired RRP maintenance. Thus, synaptotagmin-1 and -7 perform redundant functions in maintaining the capacity of the RRP in addition to nonredundant functions in the Ca2+ triggering of different phases of release. PMID:26437117

  9. Tcf15 Primes Pluripotent Cells for Differentiation

    PubMed Central

    Davies, Owen R.; Lin, Chia-Yi; Radzisheuskaya, Aliaksandra; Zhou, Xinzhi; Taube, Jessica; Blin, Guillaume; Waterhouse, Anna; Smith, Andrew J.H.; Lowell, Sally

    2013-01-01

    Summary The events that prime pluripotent cells for differentiation are not well understood. Inhibitor of DNA binding/differentiation (Id) proteins, which are inhibitors of basic helix-loop-helix (bHLH) transcription factor activity, contribute to pluripotency by blocking sequential transitions toward differentiation. Using yeast-two-hybrid screens, we have identified Id-regulated transcription factors that are expressed in embryonic stem cells (ESCs). One of these, Tcf15, is also expressed in the embryonic day 4.5 embryo and is specifically associated with a novel subpopulation of primed ESCs. An Id-resistant form of Tcf15 rapidly downregulates Nanog and accelerates somatic lineage commitment. We propose that because Tcf15 can be held in an inactive state through Id activity, it may prime pluripotent cells for entry to somatic lineages upon downregulation of Id. We also find that Tcf15 expression is dependent on fibroblast growth factor (FGF) signaling, providing an explanation for how FGF can prime for differentiation without driving cells out of the pluripotent state. PMID:23395635

  10. Gas Vesicle Nanoparticles for Antigen Display

    PubMed Central

    DasSarma, Shiladitya; DasSarma, Priya

    2015-01-01

    Microorganisms like the halophilic archaeon Halobacterium sp. NRC-1 produce gas-filled buoyant organelles, which are easily purified as protein nanoparticles (called gas vesicles or GVNPs). GVNPs are non-toxic, exceptionally stable, bioengineerable, and self-adjuvanting. A large gene cluster encoding more than a dozen proteins has been implicated in their biogenesis. One protein, GvpC, found on the exterior surface of the nanoparticles, can accommodate insertions near the C-terminal region and results in GVNPs displaying the inserted sequences on the surface of the nanoparticles. Here, we review the current state of knowledge on GVNP structure and biogenesis as well as available studies on immunogenicity of pathogenic viral, bacterial, and eukaryotic proteins and peptides displayed on the nanoparticles. Recent improvements in genetic tools for bioengineering of GVNPs are discussed, along with future opportunities and challenges for development of vaccines and other applications. PMID:26350601

  11. Extrasynaptic vesicle recycling in mature hippocampal neurons.

    PubMed

    Ratnayaka, Arjuna; Marra, Vincenzo; Branco, Tiago; Staras, Kevin

    2011-11-08

    Fast neuronal signalling relies on highly regulated vesicle fusion and recycling at specialized presynaptic terminals. Recently, examples of non-classical neurotransmission have also been reported, where fusion of vesicles can occur at sites remote from conventional synapses. This has potentially broad biological implications, but the underlying mechanisms are not well established. Here we show that a complete vesicle recycling pathway can occur at discrete axonal sites in mature hippocampal neurons and that extrasynaptic fusion is a robust feature of native tissue. We demonstrate that laterally mobile vesicle clusters trafficking between synaptic terminals become transiently stabilized by evoked action potentials and undergo complete but delayed Ca(2+)-dependent fusion along axons. This fusion is associated with dynamic actin accumulation and, subsequently, vesicles can be locally recycled, re-acidified and re-used. Immunofluorescence and ultrastructural work demonstrates that extrasynaptic fusion sites can have apposed postsynaptic specializations, suggesting that mobile vesicle recycling may underlie highly dynamic neuron-neuron communication.

  12. Ultrasound-responsive ultrathin multiblock copolyamide vesicles

    NASA Astrophysics Data System (ADS)

    Huang, Lei; Yu, Chunyang; Huang, Tong; Xu, Shuting; Bai, Yongping; Zhou, Yongfeng

    2016-02-01

    This study reports the self-assembly of novel polymer vesicles from an amphiphilic multiblock copolyamide, and the vesicles show a special structure with an ultrathin wall thickness of about 4.5 nm and a combined bilayer and monolayer packing model. Most interestingly, the vesicles are ultrasound-responsive and can release the encapsulated model drugs in response to ultrasonic irradiation.This study reports the self-assembly of novel polymer vesicles from an amphiphilic multiblock copolyamide, and the vesicles show a special structure with an ultrathin wall thickness of about 4.5 nm and a combined bilayer and monolayer packing model. Most interestingly, the vesicles are ultrasound-responsive and can release the encapsulated model drugs in response to ultrasonic irradiation. Electronic supplementary information (ESI) available: Details of experiments and characterization, and FT-IR, TEM, DPD, FL and micro-DSC results. See DOI: 10.1039/c5nr08596a

  13. Optogenetic Acidification of Synaptic Vesicles and Lysosomes

    PubMed Central

    Grauel, M. Katharina; Wozny, Christian; Bentz, Claudia; Blessing, Anja; Rosenmund, Tanja; Jentsch, Thomas J.; Schmitz, Dietmar; Hegemann, Peter; Rosenmund, Christian

    2016-01-01

    Acidification is required for the function of many intracellular organelles, but methods to acutely manipulate their intraluminal pH have not been available. Here we present a targeting strategy to selectively express the light-driven proton pump Arch3 on synaptic vesicles. Our new tool, pHoenix, can functionally replace endogenous proton pumps, enabling optogenetic control of vesicular acidification and neurotransmitter accumulation. Under physiological conditions, glutamatergic vesicles are nearly full, as additional vesicle acidification with pHoenix only slightly increased the quantal size. By contrast, we found that incompletely filled vesicles exhibited a lower release probability than full vesicles, suggesting preferential exocytosis of vesicles with high transmitter content. Our subcellular targeting approach can be transferred to other organelles, as demonstrated for a pHoenix variant that allows light-activated acidification of lysosomes. PMID:26551543

  14. Reversed Priming Effects May Be Driven by Misperception Rather than Subliminal Processing

    PubMed Central

    Sand, Anders

    2016-01-01

    A new paradigm for investigating whether a cognitive process is independent of perception was recently suggested. In the paradigm, primes are shown at an intermediate signal strength that leads to trial-to-trial and inter-individual variability in prime perception. Here, I used this paradigm and an objective measure of perception to assess the influence of prime identification responses on Stroop priming. I found that sensory states producing correct and incorrect prime identification responses were also associated with qualitatively different priming effects. Incorrect prime identification responses were associated with reversed priming effects but in contrast to previous studies, I interpret this to result from the (mis-)perception of primes rather than from a subliminal process. Furthermore, the intermediate signal strength also produced inter-individual variability in prime perception that strongly influenced priming effects: only participants who on average perceived the primes were Stroop primed. I discuss how this new paradigm, with a wide range of d′ values, is more appropriate when regression analysis on inter-individual identification performance is used to investigate perception-dependent processing. The results of this study, in line with previous results, suggest that drawing conclusions about subliminal processes based on data averaged over individuals may be unwarranted. PMID:26925016

  15. Priming a new identity: self-monitoring moderates the effects of nonself primes on self-judgments and behavior.

    PubMed

    DeMarree, Kenneth G; Wheeler, S Christian; Petty, Richard E

    2005-11-01

    When a construct is primed, people often act in construct-consistent ways. Several accounts for this effect have been offered, including ideomotor theory and a social functional perspective. The authors tested an additional perspective, the Active-Self account, whereby primes can temporarily alter self-perceptions. In Study 1, non-African American participants reported feeling more aggressive on an implicit measure following an African American prime. In Study 2, participants reported feeling luckier on an implicit measure following a number 7 (vs. 13) prime. In both studies, these effects were obtained only for low self-monitors, who are more likely to change self-conceptions in response to diagnostic self-information and to use their internal states in guiding behavior. Study 3 showed that low self-monitors also show larger behavioral effects of primes.

  16. Vesicles

    MedlinePlus

    ... Contact dermatitis (may be caused by poison ivy) Herpes simplex (cold sores, genital herpes ) Herpes zoster (shingles) Impetigo ... Hand, foot, and mouth disease on the soles Herpes simplex - close-up Herpes zoster (shingles) - close-up of ...

  17. Apollo 1 Prime Crew

    NASA Technical Reports Server (NTRS)

    1966-01-01

    Portrait of the Apollo 1 prime crew for first manned Apollo space flight. From left to right are: Edward H. White II, Virgil I. 'Gus' Grissom, and Roger B. Chaffee. On January 27, 1967 at 5:31 p.m. CST (6:31 local time) during a routine simulated launch test onboard the Apollo Saturn V Moon rocket, an electrical short circuit inside the Apollo Command Module ignited the pure oxygen environment and within a matter of seconds all three Apollo 1 crewmembers perished.

  18. Alignment of Synaptic Vesicle Macromolecules with the Macromolecules in Active Zone Material that Direct Vesicle Docking

    PubMed Central

    Xu, Jing; Jung, Jae Hoon; Marshall, Robert M.; McMahan, Uel J.

    2013-01-01

    Synaptic vesicles dock at active zones on the presynaptic plasma membrane of a neuron’s axon terminals as a precondition for fusing with the membrane and releasing their neurotransmitter to mediate synaptic impulse transmission. Typically, docked vesicles are next to aggregates of plasma membrane-bound macromolecules called active zone material (AZM). Electron tomography on tissue sections from fixed and stained axon terminals of active and resting frog neuromuscular junctions has led to the conclusion that undocked vesicles are directed to and held at the docking sites by the successive formation of stable connections between vesicle membrane proteins and proteins in different classes of AZM macromolecules. Using the same nanometer scale 3D imaging technology on appropriately stained frog neuromuscular junctions, we found that ∼10% of a vesicle’s luminal volume is occupied by a radial assembly of elongate macromolecules attached by narrow projections, nubs, to the vesicle membrane at ∼25 sites. The assembly’s chiral, bilateral shape is nearly the same vesicle to vesicle, and nubs, at their sites of connection to the vesicle membrane, are linked to macromolecules that span the membrane. For docked vesicles, the orientation of the assembly’s shape relative to the AZM and the presynaptic membrane is the same vesicle to vesicle, whereas for undocked vesicles it is not. The connection sites of most nubs on the membrane of docked vesicles are paired with the connection sites of the different classes of AZM macromolecules that regulate docking, and the membrane spanning macromolecules linked to these nubs are also attached to the AZM macromolecules. We conclude that the luminal assembly of macromolecules anchors in a particular arrangement vesicle membrane macromolecules, which contain the proteins that connect the vesicles to AZM macromolecules during docking. Undocked vesicles must move in a way that aligns this arrangement with the AZM macromolecules for

  19. All Prime Contract Awards by State or Country, Place, and Contractor, FY 88. Part 12. (Amherst, New Hampshire-White Sands Missile Range, New Mexico).

    DTIC Science & Technology

    1988-01-01

    6w1 M.~hKCUMT ~hSD STATE OR COUNTRY minC meD In COUTTERG FY a. (U) MESNINTON HLUUAATE*S SERVICES (DOD) DC DIRECTOATE FOR INF. 19Wt DIORtSTII-N-PT-12...ORGANIZATION WHSj, Directorate for Informaticn (If applicable) Operations and Reports 6 ,’ADDRESS (City, State , and ZIPCode) 7b. ADDRESS (City, State , and...applicable) 8c. ADDRESS (City, State , and ZIP Code) 10. SOURCE OF FUNDING NUMBERS PROGRAM PROJECT TASK WORK UNIT ELEMENT NO. NO. NO. ACCESSION NO. 11

  20. Transposed-Letter Priming Effects with Masked Subset Primes: A Re-Examination of the "Relative Position Priming Constraint"

    ERIC Educational Resources Information Center

    Stinchcombe, Eric J.; Lupker, Stephen J.; Davis, Colin J.

    2012-01-01

    Three experiments are reported investigating the role of letter order in orthographic subset priming (e.g., "grdn"-GARDEN) using both the conventional masked priming technique as well as the sandwich priming technique in a lexical decision task. In all three experiments, subset primes produced priming with the effect being considerably…

  1. Urinary extracellular vesicles and the kidney: biomarkers and beyond.

    PubMed

    Salih, Mahdi; Zietse, Robert; Hoorn, Ewout J

    2014-06-01

    Extracellular vesicles have been isolated in various body fluids, including urine. The cargo of urinary extracellular vesicles (uEVs) is composed of proteins and nucleic acids reflecting the physiological and possibly pathophysiological state of cells lining the nephron. Because urine is a noninvasive and readily available biofluid, the discovery of uEVs has opened a new field of biomarker research. Their potential use as diagnostic, prognostic, or therapeutic biomarkers for various kidney diseases, including glomerulonephritis, acute kidney injury, tubular disorders, and polycystic kidney disease, is currently being explored. Some challenges, however, remain. These challenges include the need to standardize isolation methods, normalization between samples, and validation of candidate biomarkers. Also, the development of a high-throughput platform to isolate and analyze uEVs, for example, an enzyme-linked immunosorbent assay, is desirable. Here, we review recent studies on uEVs dealing with kidney physiology and pathophysiology. Furthermore, we discuss new and exciting developments regarding vesicles, including their role in cell-to-cell communication and the possibility of using vesicles as a therapy for kidney disorders.

  2. Neuroimaging correlates of negative priming.

    PubMed

    Steel, C; Haworth, E J; Peters, E; Hemsley, D R; Sharma, T; Gray, J A; Pickering, A; Gregory, L; Simmons, A; Bullmore, E T; Williams, S C

    2001-11-16

    Many theoretical accounts of selective attention and memory retrieval include reference to active inhibitory processes, such as those argued to underlie the negative priming effect. fMRI was used in order to investigate the areas of cortical activation associated with Stroop interference, Stroop facilitation and Stroop negative priming tasks. The most significant activation within the negative priming task was within the inferior parietal lobule, left temporal lobe and frontal lobes. Areas of cortical activation are discussed with reference to theoretical accounts of the negative priming effect.

  3. Construction of macroscopic cytomimetic vesicle aggregates based on click chemistry: controllable vesicle fusion and phase separation.

    PubMed

    Jin, Haibao; Huang, Wei; Zheng, Yongli; Zhou, Yongfeng; Yan, Deyue

    2012-07-09

    Vesicle-vesicle aggregation to mimic cell-cell aggregation has attracted much attention. Here, hyperbranched polymer vesicles (branched-polymersomes, BPs) with a cell-like size were selected as model membranes, and the vesicle aggregation process, triggered by click chemistry of the copper-catalysed azide-alkyne cycloaddition reaction, was systematically studied. For this purpose, azide and alkynyl groups were loaded on the membranes of BPs through the co-assembly method to obtain N(3)-BPs and Alk-BPs, respectively. Subsequently, macroscopic vesicle aggregates were obtained when these two kinds of functional BPs were mixed together with the ratio of azide to alkynyl groups of about 1:1. Both the vesicle fusion events and lateral phase separation on the vesicle membrane occurred during such a vesicle aggregation process, and the fusion rate and phase-separation degree could be controlled by adjusting the clickable group content. The vesicle aggregation process with N(3) -micelles as desmosome mimics to connect with Alk-BPs through click-chemistry reaction was also studied, and large-scale vesicle aggregates without vesicle fusion were obtained in this process. The present work has extended the controllable cytomimetic vesicle aggregation process with the use of covalent bonds, instead of noncovalent bonds, as the driving force.

  4. Prime mover progress

    SciTech Connect

    Hennagir, T.

    1995-05-01

    Manufacturers continue to upgrade fluidized bed and steam technologies to meet more stringent emission, efficiency and service challenges from global power market customers. Offshore, suppliers are being pushed to provide quality, low-cost production and significant local competitive presence in nearly all international markets. The ability to meet customer and regulatory demands for emissions across different global market segments remains a priority for prime mover suppliers as they develop environmental technical improvements. Products initially developed for the more stringent European regulations are now finding a market in the US. In developing countries, demand continues for utility-size boiler products, but pollution control equipment costs remain a parallel consideration. Industry improvements to steam cycle equipment, namely turbines, continue to reap benefits in terms of efficiency and operational flexibility. Boiler and steam manufacturers` response to global markets remains right on target as product development strategies are constantly adjusted and analyzed to reach an optimum marketing mix. Innovation and advances in prime mover power equipment technology remain a mainstay of suppliers` ability to meet the needs of a changing, competitive clientele.

  5. Vesicles protect activated acetic acid.

    PubMed

    Todd, Zoe R; House, Christopher H

    2014-10-01

    Abstract Methyl thioacetate, or activated acetic acid, has been proposed to be central to the origin of life and an important energy currency molecule in early cellular evolution. We have investigated the hydrolysis of methyl thioacetate under various conditions. Its uncatalyzed rate of hydrolysis is about 3 orders of magnitude faster (K=0.00663 s(-1); 100°C, pH 7.5, concentration=0.33 mM) than published rates for its catalyzed production, making it unlikely to accumulate under prebiotic conditions. However, our experiments showed that methyl thioacetate was protected from hydrolysis when inside its own hydrophobic droplets. Further, we found that methyl thioacetate protection from hydrolysis was also possible in droplets of hexane and in the membranes of nonanoic acid vesicles. Thus, the hydrophobic regions of prebiotic vesicles and early cell membranes could have offered a refuge for this energetic molecule, increasing its lifetime in close proximity to the reactions for which it would be needed. This model of early energy storage evokes an additional critical function for the earliest cell membranes.

  6. Dielectrophoresis of Functional Phospholipid Vesicles

    NASA Astrophysics Data System (ADS)

    Froude, Victoria; Zhu, Yingxi Elaine

    2008-03-01

    Recently, there has been an emerging interest in using AC-dielectrophoresis (DEP) to transport and assemble phospholipid vesicles (liposomes) and nanoparticles to form functional bio-assemblies where the underlying charge polarization mechanism of colloids in AC fields strongly depends on nano-scaled surface charge. In this work, we study liposomes segregation and aggregation in the presence of nanocolloids and salts in which the biological functionality of liposomes is augmented by the physical functionality of inorganic coating and particles. Liposomes, synthesized by sonication with 1,2-Dioleoyl-sn-Glycero-3-Phosphate (DOPA), are manipulated at varied AC-field frequencies across fabricated micro-electrodes in a quadrapole configuration on glass. We observe the co-assembly of liposome and opposite-charged nanocolloids by confocal microscopy and SEM, where the smaller nanocolloids are captured in between liposome junctions to form stabilized composite vesicles at several distinct frequencies. We observe a strong dependence of the liposome DEP mobility on the number of nanoparticles present in suspension and propose a new mechanism based on charge segregation and charged nanocolloid entrainment in the double layer.

  7. Proteomics of extracellular vesicles: Exosomes and ectosomes.

    PubMed

    Choi, Dong-Sic; Kim, Dae-Kyum; Kim, Yoon-Keun; Gho, Yong Song

    2015-01-01

    Almost all bacteria, archaea, and eukaryotic cells shed extracellular vesicles either constitutively or in a regulated manner. These nanosized membrane vesicles are spherical, bilayered proteolipids that harbor specific subsets of proteins, DNAs, RNAs, and lipids. Recent research has facilitated conceptual advancements in this emerging field that indicate that extracellular vesicles act as intercellular communicasomes by transferring signals to their target cell via surface ligands and delivering receptors and functional molecules. Recent progress in mass spectrometry-based proteomic analyses of mammalian extracellular vesicles derived from diverse cell types and body fluids has resulted in the identification of several thousand vesicular proteins that provide us with essential clues to the molecular mechanisms involved in vesicle cargo sorting and biogenesis. Furthermore, cell-type- or disease-specific vesicular proteins help us to understand the pathophysiological functions of extracellular vesicles and contribute to the discovery of diagnostic and therapeutic target proteins. This review focuses on the high-throughput mass spectrometry-based proteomic analyses of mammalian extracellular vesicles (i.e., exosomes and ectosomes), EVpedia (a free web-based integrated database of high-throughput data for systematic analyses of extracellular vesicles; http://evpedia.info), and the intravesicular protein-protein interaction network analyses of mammalian extracellular vesicles. The goal of this article is to encourage further studies to construct a comprehensive proteome database for extracellular vesicles that will help us to not only decode the biogenesis and cargo-sorting mechanisms during vesicle formation but also elucidate the pathophysiological roles of these complex extracellular organelles.

  8. The role of vesicles in the transport of ferritin through frog endothelium.

    PubMed Central

    Clough, G; Michel, C C

    1981-01-01

    1. The transport of ferritin molecules by endothelial cell vesicles has been quantitatively investigated by electron microscopy. Single mesenteric capillaries of pithed frogs were perfused with solutions containing 6.7 g ferritin 100 ml.-1 for known periods before fixation in situ with osmium tetroxide. 2. Two series of experiments were carried out: in the first series the perfusate contained bovine serum albumin (1.0 g 100 ml.-1); in the second series the perfusate contained no protein other than the ferritin. To assess the molecular radius of ferritin in solution, the free diffusion coefficient of ferritin was measured in the presence and absence of albumin. 3. The free diffusion coefficient of ferritin in saline solution (110 m-mole 1.-1) was found to be 0.35 X 10(-6) cm2 sec-1 at 21 degrees C and was not affected by the presence of bovine serum albumin. This indicates that there is no significant binding of albumin to ferritin in solution and yields a value for the Stokes-Einstein radius of ferritin of 6.1 nm. 4. In all perfusion experiments the percentage of luminal vesicles containing ferritin exceeded the percentage of labelled cytoplasmic vesicles, which in turn exceeded the percentage of labelled abluminal vesicles. 5. Labelling of all vesicle populations was seen after perfusions lasting less than 1 sec. At this time luminal vesicles were more heavily labelled in the absence of albumin. 6. The labelling of luminal vesicles increased with lengthening perfusion times up to 30-40 sec, after which steady levels of labelling were achieved. The rate of rise in luminal labelling and the steady-state levels reached were both greater in the absence of albumin. By contrast cytoplasmic labelling increased above its initial value only after perfusions of longer than 10 sec. 7. In the steady state, labelled cytoplasmic vesicles contained, on average, fewer ferritin molecules than labelled luminal vesicles. This finding is inconsistent with translocation of labelled

  9. 5[prime] to 3[prime] nucleic acid synthesis using 3[prime]-photoremovable protecting group

    DOEpatents

    Pirrung, M.C.; Shuey, S.W.; Bradley, J.C.

    1999-06-01

    The present invention relates, in general, to a method of synthesizing a nucleic acid, and, in particular, to a method of effecting 5[prime] to 3[prime] nucleic acid synthesis. The method can be used to prepare arrays of oligomers bound to a support via their 5[prime] end. The invention also relates to a method of effecting mutation analysis using such arrays. The invention further relates to compounds and compositions suitable for use in such methods.

  10. Investigating Home Primes and Their Families

    ERIC Educational Resources Information Center

    Herman, Marlena; Schiffman, Jay

    2014-01-01

    The process of prime factor splicing to generate home primes raises opportunity for conjecture and exploration. The notion of "home primes" is relatively new in the chronicle of mathematics. Heleen (1996-97) first described a procedure called "prime factor splicing" (PFS). The exploration of home primes is interesting and…

  11. Attention to primes modulates affective priming of pronunciation responses.

    PubMed

    De Houwer, Jan; Randell, Tom

    2002-01-01

    In studies on affective priming of pronunciation responses, two words are presented on each trial and participants are asked to read the second word out loud. Whereas some studies revealed shorter reaction times when the two words had the same valence than when they had a different valence, other studies either found no effect of affective congruence or revealed a reversed effect. In the present experiments, a significant effect of affective congruence only emerged when filler trials were presented in which the prime and target were identical and participants were instructed to attend to the primes (Experiment 2). No effects were found when participants were merely instructed to attend to or ignore the primes (Experiment 1), or when affectively incongruent filler trials were presented and participants were instructed to ignore the primes (Experiment 2).

  12. Phase transition in a stochastic prime-number generator.

    PubMed

    Luque, Bartolo; Lacasa, Lucas; Miramontes, Octavio

    2007-07-01

    We introduce a stochastic algorithm that acts as a prime-number generator. The dynamics of this algorithm gives rise to a continuous phase transition, which separates a phase where the algorithm is able to reduce a whole set of integers into primes and a phase where the system reaches a frozen state with low prime density. We present both numerical simulations and an analytical approach in terms of an annealed approximation, by means of which the data are collapsed. A critical slowing-down phenomenon is also outlined.

  13. Dynamical simulations of vesicle growth and division

    NASA Astrophysics Data System (ADS)

    Ruiz-Herrero, Teresa; Mahadevan, L.

    2015-03-01

    Prebiotic cells constitute a beautiful and intriguing example of self-replicating vesicles. How these cells managed to grow and divide without sophisticated machinery is still an open question. The properties of these primitive vesicles can shed light on the ways modern cells have evolved by exploiting those characteristics to develop their replication mechanisms. The equilibrium configurations of elastic shells are well understood, however the dynamical behavior during growth still lacks of a deep theoretical understanding. To study vesicle growth from a general perspective, we have developed a minimal generic model where vesicles are represented by a 2D spring network and characterized by a minimum set of magnitudes: growth rate, permeability, bending stiffness, viscosity and temperature. We have performed hybrid molecuar dynamic simulations as a function of a reduced set of dimensionless parameters. Three main outcomes were observed: vesicles that grow without division, vesicles that divide symmetrically, and vesicles that act as generators of daughter vesicles. The type of outcome depends on the system parameters and specifically on its dynamics via two timescales. Furthermore, we found sets of parameters where the system shows size homeostasis. TRH was supported by Ramon Areces Foundation.

  14. Functional Advantages of Porphyromonas gingivalis Vesicles

    PubMed Central

    Ho, Meng-Hsuan; Chen, Chin-Ho; Goodwin, J. Shawn; Wang, Bing-Yan; Xie, Hua

    2015-01-01

    Porphyromonas gingivalis is a keystone pathogen of periodontitis. Outer membrane vesicles (OMVs) have been considered as both offense and defense components of this bacterium. Previous studies indicated that like their originating cells, P. gingivalis vesicles, are able to invade oral epithelial cells and gingival fibroblasts, in order to promote aggregation of some specific oral bacteria and to induce host immune responses. In the present study, we investigated the invasive efficiency of P. gingivalis OMVs and compared results with that of the originating cells. Results revealed that 70–90% of human primary oral epithelial cells, gingival fibroblasts, and human umbilical vein endothelial cells carried vesicles from P. gingivalis 33277 after being exposed to the vesicles for 1 h, while 20–50% of the host cells had internalized P. gingivalis cells. We also detected vesicle-associated DNA and RNA and a vesicle-mediated horizontal gene transfer in P. gingivalis strains, which represents a novel mechanism for gene transfer between P. gingivalis strains. Moreover, purified vesicles of P. gingivalis appear to have a negative impact on biofilm formation and the maintenance of Streptococcus gordonii. Our results suggest that vesicles are likely the best offence weapon of P. gingivalis for bacterial survival in the oral cavity and for induction of periodontitis. PMID:25897780

  15. Amyloglucosidase enzymatic reactivity inside lipid vesicles

    PubMed Central

    Li, Mian; Hanford, Michael J; Kim, Jin-Woo; Peeples, Tonya L

    2007-01-01

    Efficient functioning of enzymes inside liposomes would open new avenues for applications in biocatalysis and bioanalytical tools. In this study, the entrapment of amyloglucosidase (AMG) (EC 3.2.1.3) from Aspergillus niger into dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles (MLVs) and large unilamellar vesicles (LUVs) was investigated. Negative-stain, freeze-fracture, and cryo-transmission electron microscopy images verified vesicle formation in the presence of AMG. Vesicles with entrapped AMG were isolated from the solution by centrifugation, and vesicle lamellarity was identified using fluorescence laser confocal microscopy. The kinetics of starch hydrolysis by AMG was modeled for two different systems, free enzyme in aqueous solution and entrapped enzyme within vesicles in aqueous suspension. For the free enzyme system, intrinsic kinetics were described by a Michaelis-Menten kinetic model with product inhibition. The kinetic constants, Vmax and Km, were determined by initial velocity measurements, and Ki was obtained by fitting the model to experimental data of glucose concentration-time curves. Predicted concentration-time curves using these kinetic constants were in good agreement with experimental measurements. In the case of the vesicles, the time-dependence of product (glucose) formation was experimentally determined and simulated by considering the kinetic behavior of the enzyme and the permeation of substrate into the vesicle. Experimental results demonstrated that entrapped enzymes were much more stable than free enyzme. The entrapped enzyme could be recycled with retention of 60% activity after 3 cycles. These methodologies can be useful in evaluating other liposomal catalysis operations. PMID:18271982

  16. Extracellular vesicles in coronary artery disease.

    PubMed

    Boulanger, Chantal M; Loyer, Xavier; Rautou, Pierre-Emmanuel; Amabile, Nicolas

    2017-02-02

    Membrane vesicles released in the extracellular space are composed of a lipid bilayer enclosing soluble cytosolic material and nuclear components. Extracellular vesicles include apoptotic bodies, exosomes, and microvesicles (also known previously as microparticles). Originating from different subcellular compartments, the role of extracellular vesicles as regulators of transfer of biological information, acting locally and remotely, is now acknowledged. Circulating vesicles released from platelets, erythrocytes, leukocytes, and endothelial cells contain potential valuable biological information for biomarker discovery in primary and secondary prevention of coronary artery disease. Extracellular vesicles also accumulate in human atherosclerotic plaques, where they affect major biological pathways, including inflammation, proliferation, thrombosis, calcification, and vasoactive responses. Extracellular vesicles also recapitulate the beneficial effect of stem cells to treat cardiac consequences of acute myocardial infarction, and now emerge as an attractive alternative to cell therapy, opening new avenues to vectorize biological information to target tissues. Although interest in microvesicles in the cardiovascular field emerged about 2 decades ago, that for extracellular vesicles, in particular exosomes, started to unfold a decade ago, opening new research and therapeutic avenues. This Review summarizes current knowledge on the role of extracellular vesicles in coronary artery disease, and their emerging potential as biomarkers and therapeutic agents.

  17. Self-assembled carbohydrate-based vesicles for lectin targeting.

    PubMed

    Dos Santos, Marinalva Cardoso; Micheletto, Yasmine Miguel Serafini; da Silveira, Nadya Pesce; da Silva Pinto, Luciano; Giacomelli, Fernando Carlos; de Lima, Vânia Rodrigues; Frizon, Tiago Elias Allievi; Dal-Bó, Alexandre Gonçalves

    2016-12-01

    This study examined the physicochemical interactions between vesicles formed by phosphatidylcholine (PC) and glycosylated polymeric amphiphile N-acetyl-β-d-glucosaminyl-PEG900-docosanate (C22PEG900GlcNAc) conjugated with Bauhinia variegata lectin (BVL). Lectins are proteins or glycoproteins capable of binding glycosylated membrane components. Accordingly, the surface functionalization by such entities is considered a potential strategy for targeted drug delivery. We observed increased hydrodynamic radii (RH) of PC+C22PEG900GlcNAc vesicles in the presence of lectins, suggesting that this aggregation was due to the interaction between lectins and the vesicular glycosylated surfaces. Furthermore, changes in the zeta potential of the vesicles with increasing lectin concentrations implied that the vesicular glycosylated surfaces were recognized by the investigated lectin. The presence of carbohydrate residues on vesicle surfaces and the ability of the vesicles to establish specific interactions with BVL were further explored using atomic force microscopy (AFM) and small-angle X-ray scattering (SAXS) analysis. The results indicated that the thickness of the hydrophilic layer was to some extent influenced by the presence of lectins. The presence of lectins required a higher degree of polydispersity as indicated by the width parameter of the log-normal distribution of size, which also suggested more irregular structures. Reflectance Fourier transform infrared (HATR-FTIR), differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis.) analyses revealed that the studied lectin preferentially interacted with the choline and carbonyl groups of the lipid, thereby changing the choline orientation and intermolecular interactions. The protein also discretely reduced the intermolecular communication of the hydrophobic acyl chains, resulting in a disordered state.

  18. Small Angle Neutron-Scattering Studies of the Core Structure of Intact Neurosecretory Vesicles.

    NASA Astrophysics Data System (ADS)

    Krueger, Susan Takacs

    Small angle neutron scattering (SANS) was used to study the state of the dense cores within intact neurosecretory vesicles. These vesicles transport the neurophysin proteins, along with their associated hormones, oxytocin or vasopressin, from the posterior pituitary gland to the bloodstream, where the entire vesicle contents are released. Knowledge of the vesicle core structure is important in developing an understanding of this release mechanism. Since the core constituents exist in a dense state at concentrations which cannot be reproduced (in solution) in the laboratory, a new method was developed to determine the core structure from SANS experiments performed on intact neurosecretory vesicles. These studies were complemented by biochemical assays performed to determine the role, if any, played by phospholipids in the interactions between the core constituents. H_2O/D_2 O ratio in the solvent can be adjusted, using the method of contrast variation, such that the scattering due to the vesicle membranes is minimized, thus emphasizing the scattering originating from the cores. The applicability of this method for examining the interior of biological vesicles was tested by performing an initial study on human red blood cells, which are similar in structure to other biological vesicles. Changes in intermolecular hemoglobin interactions, occurring when the ionic strength of the solvent was varied or when the cells were deoxygenated, were examined. The results agreed with those expected for dense protein solutions, indicating that the method developed was suitable for the study of hemoglobin within the cells. Similar SANS studies were then performed on intact neurosecretory vesicles. The experimental results were inconsistent with model calculations which assumed that the cores consisted of small, densely-packed particles or large, globular aggregates. Although a unique model could not be determined, the data suggest that the core constituents form long aggregates of

  19. Representing Numbers: Prime and Irrational

    ERIC Educational Resources Information Center

    Zazkis, Rina

    2005-01-01

    This article draws an analogy between prime and irrational numbers with respect to how these numbers are defined and how they are perceived by learners. Excerpts are presented from two research studies: a study on understanding prime numbers by pre-service elementary school teachers and a study on understanding irrational numbers by pre-service…

  20. Priming Macho Attitudes and Emotions.

    ERIC Educational Resources Information Center

    Beaver, Erik D.; And Others

    1992-01-01

    Investigated the effects of reading one of four priming stimuli stories (control, consenting sex, rape, or family) on males' evaluations of, and emotional reactions to, two videotaped date-rape scenarios. Results supported the concepts of a macho personality and revealed interactive effects for both the rape and family prime. (RJM)

  1. Exodus: Prime Mover

    NASA Technical Reports Server (NTRS)

    Bauer, Nikkol; Conwell, Pete; Johnson, Matt; Shields, Wendy; Thornton, Tim; Tokarz, Rob; Mcmanus, Rich

    1992-01-01

    The Exodus Prime Mover is an overnight package delivery aircraft designed to serve the Northern Hemisphere of Aeroworld. The preliminary design goals originated from the desire to produce a large profit. The two main driving forces throughout the design process were first to reduce the construction man-hours by simplifying the aircraft design, thereby decreasing the total production cost of the aircraft. The second influential factor affecting the design was minimizing the fuel cost during cruise. The lowest fuel consumption occurs at a cruise velocity of 30 ft/s. Overall, it was necessary to balance the economic benefits with the performance characteristics in order to create a profitable product that meets all specified requirements and objectives.

  2. Visualization of SNARE-Mediated Hemifusion between Giant Unilamellar Vesicles Arrested by Myricetin

    PubMed Central

    Heo, Paul; Park, Joon-Bum; Shin, Yeon-Kyun; Kweon, Dae-Hyuk

    2017-01-01

    Neurotransmitters are released within a millisecond after Ca2+ arrives at an active zone. However, the vesicle fusion pathway underlying this synchronous release is yet to be understood. At the center of controversy is whether hemifusion, in which outer leaflets are merged while inner leaflets are still separated, is an on-pathway or off-pathway product of Ca2+-triggered exocytosis. Using the single vesicle fusion assay, we recently demonstrated that hemifusion is an on-pathway intermediate that immediately proceeds to full fusion upon Ca2+ triggering. It has been shown that the flavonoid myricetin arrests soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE)-mediated vesicle fusion at hemifusion, but that the hemifused vesicles spontaneously convert to full fusion when the myricetin clamp is removed by the enzyme laccase. In the present study, we visualized SNARE-mediated hemifusion between two SNARE-reconstituted giant unilamellar vesicles (GUVs) arrested by myricetin. The large size of the GUVs enabled us to directly image the hemifusion between them. When two merging GUVs were labeled with different fluorescent dyes, GUV pairs showed asymmetric fluorescence intensities depending on the position on the GUV pair consistent with what is expected for hemifusion. The flow of lipids from one vesicle to the other was revealed with fluorescence recovery after photobleaching (FRAP), indicating that the two membranes had hemifused. These results support the hypothesis that hemifusion may be the molecular status that primes Ca2+-triggered millisecond exocytosis. This study represents the first imaging of SNARE-driven hemifusion between GUVs.

  3. Thermodynamics and kinetics of vesicles formation processes.

    PubMed

    Guida, Vincenzo

    2010-12-15

    Vesicles are hollow aggregates, composed of bilayers of amphiphilic molecules, dispersed into and filled with a liquid solvent. These aggregates can be formed either as equilibrium or as out of equilibrium meta-stable structures and they exhibit a rich variety of different morphologies. The surprising richness of structures, the vast range of industrial applications and the presence of vesicles in a number of biological systems have attracted the interest of numerous researchers and scientists. In this article, we review both the thermodynamics and the kinetics aspects of the phenomena of formation of vesicles. We start presenting the thermodynamics of bilayer membranes formation and deformation, with the aim of deriving the conditions for the existence of equilibrium vesicles. Specifically, we use the results from continuum thermodynamics to discuss the possibility of formation of stable equilibrium vesicles, from both mixed amphiphiles and single component systems. We also link the bilayer membrane properties to the molecular structure of the starting amphiphiles. In the second part of this article, we focus on the dynamics and kinetics of vesiculation. We review the process of vesicles formation both from planar lamellar phase under shear and from isotropic micelles. In order to clarify the physical mechanisms of vesicles formation, we continuously draw a parallel between emulsification and vesiculation processes. Specifically, we compare the experimental results, the driving forces and the relative scaling laws identified for the two processes. Describing the dynamics of vesicles formation, we also discuss why non equilibrium vesicles can be formed by kinetics control and why they are meta-stable. Understanding how to control the properties, the stability and the formation process of vesicles is of fundamental importance for a vast number of industrial applications.

  4. Analysis of Yeast Extracellular Vesicles.

    PubMed

    Rodrigues, Marcio L; Oliveira, Debora L; Vargas, Gabriele; Girard-Dias, Wendell; Franzen, Anderson J; Frasés, Susana; Miranda, Kildare; Nimrichter, Leonardo

    2016-01-01

    Extracellular vesicles (EV) are important carriers of biologically active components in a number of organisms, including fungal cells. Experimental characterization of fungal EVs suggested that these membranous compartments are likely involved in the regulation of several biological events. In fungal pathogens, these events include mechanisms of disease progression and/or control, suggesting potential targets for therapeutic intervention or disease prophylaxis. In this manuscript we describe methods that have been used in the last 10 years for the characterization of EVs produced by yeast forms of several fungal species. Experimental approaches detailed in this chapter include ultracentrifugation methods for EV fractionation, chromatographic approaches for analysis of EV lipids, microscopy techniques for analysis of both intracellular and extracellular vesicular compartments, interaction of EVs with host cells, and physical chemical analysis of EVs by dynamic light scattering.

  5. Priming the Physician Pipeline: A Regional AHEC's Use of in-state Medical School Data to Guide Its Health Careers Programming.

    PubMed

    Gross, David A; Mattox, Lainey C; Winkleman, Nicole

    2016-01-01

    Amid evidence of its rural, Appalachian students' low application and matriculation rates to in-state medical schools, the Northeast Kentucky Area Health Education Center developed two physician pipeline programs. This report describes the programs' early successes and provides a basis for heightened regional responsibility in the recruitment of medical school applicants.

  6. An Extracellular Subtilase Switch for Immune Priming in Arabidopsis

    PubMed Central

    Mauch-Mani, Brigitte; Gil, Ma José; Vera, Pablo

    2013-01-01

    In higher eukaryotes, induced resistance associates with acquisition of a priming state of the cells for a more effective activation of innate immunity; however, the nature of the components for mounting this type of immunological memory is not well known. We identified an extracellular subtilase from Arabidopsis, SBT3.3, the overexpression of which enhances innate immune responses while the loss of function compromises them. SBT3.3 expression initiates a durable autoinduction mechanism that promotes chromatin remodeling and activates a salicylic acid(SA)-dependent mechanism of priming of defense genes for amplified response. Moreover, SBT3.3 expression-sensitized plants for enhanced expression of the OXI1 kinase gene and activation of MAP kinases following pathogen attack, providing additional clues for the regulation of immune priming by SBT3.3. Conversely, in sbt3.3 mutant plants pathogen-mediated induction of SA-related defense gene expression is drastically reduced and activation of MAP kinases inhibited. Moreover, chromatin remodeling of defense-related genes normally associated with activation of an immune priming response appear inhibited in sbt3.3 plants, further indicating the importance of the extracellular SBT3.3 subtilase in the establishment of immune priming. Our results also point to an epigenetic control in the regulation of plant immunity, since SBT3.3 is up-regulated and priming activated when epigenetic control is impeded. SBT3.3 represents a new regulator of primed immunity. PMID:23818851

  7. The effects of divided attention on auditory priming.

    PubMed

    Mulligan, Neil W; Duke, Marquinn; Cooper, Angela W

    2007-09-01

    Traditional theorizing stresses the importance of attentional state during encoding for later memory, based primarily on research with explicit memory. Recent research has begun to investigate the role of attention in implicit memory but has focused almost exclusively on priming in the visual modality. The present experiments examined the effect of divided attention on auditory implicit memory, using auditory perceptual identification, word-stem completion and word-fragment completion. Participants heard study words under full attention conditions or while simultaneously carrying out a distractor task (the divided attention condition). In Experiment 1, a distractor task with low response frequency failed to disrupt later auditory priming (but diminished explicit memory as assessed with auditory recognition). In Experiment 2, a distractor task with greater response frequency disrupted priming on all three of the auditory priming tasks as well as the explicit test. These results imply that although auditory priming is less reliant on attention than explicit memory, it is still greatly affected by at least some divided-attention manipulations. These results are consistent with research using visual priming tasks and have relevance for hypotheses regarding attention and auditory priming.

  8. Complexin facilitates exocytosis and synchronizes vesicle release in two secretory model systems

    PubMed Central

    Lin, Ming-Yi; Rohan, Joyce G; Cai, Haijiang; Reim, Kerstin; Ko, Chien-Ping; Chow, Robert H

    2013-01-01

    Complexins (Cplxs) are small, SNARE-associated proteins believed to regulate fast, calcium-triggered exocytosis. However, studies have pointed to either an inhibitory and/or facilitatory role in exocytosis, and the role of Cplxs in synchronizing exocytosis is relatively unexplored. Here, we compare the function of two types of complexin, Cplx 1 and 2, in two model systems of calcium-dependent exocytosis. In mouse neuromuscular junctions (NMJs), we find that lack of Cplx 1 significantly reduces and desynchronizes calcium-triggered synaptic transmission; furthermore, high-frequency stimulation elicits synaptic facilitation, instead of normal synaptic depression, and the degree of facilitation is highly sensitive to the amount of cytoplasmic calcium buffering. In Cplx 2-null adrenal chromaffin cells, we also find decreased and desynchronized evoked release, and identify a significant reduction in the vesicle pool close to the calcium channels (immediately releasable pool, IRP). Viral transduction with either Cplx 1 or 2 rescues both the size of the evoked response and the synchronicity of release, and it restores the IRP size. Our findings in two model systems are mutually compatible and indicate a role of Cplx 1 and 2 in facilitating vesicle priming, and also lead to the new hypothesis that Cplxs may synchronize vesicle release by promoting coupling between secretory vesicles and calcium channels. PMID:23401610

  9. All Prime Contract Awards by State or Country, Place, and Contractor, FY 85. Part 5 (Arapahoe, Colorado - Washington, D.C.).

    DTIC Science & Technology

    1985-01-01

    VKS -, :±-rectcrate for Informati n (if applicable) C.prationrs & FPecrts r’( Ar)Z)RESS (City, State, and ZIP Code) lb. ADDRESS (City, State, and ZIP...I --- - -. F- - -4 t c-.. in - IL Vk 0 -41 0 0 0) -1- .- 4 44- 4 - 4- 1- 4- --4 - -4 4- 4-4. 4 0-40 -4 0 4 - 0a-00I 04 4 .1 0~ a - aI 4CJ C M0 4C 4MC...ca oom LLCJ 4 000))1 0 0 000-- 0 -4-O Lon 44CO 00 00 00 00 ca44-0 .- 0-40-4 at) I V C’) I CN N N CJNN .N .N N N’ .N .N .N ENNNNN n < lww IC CM COM CC

  10. Dollar Summary of Prime Contract Awards by State, Place, and Contractor, FY84, Part 1 (Adamsville, Alabama-Santa Clara, California).

    DTIC Science & Technology

    1984-01-01

    INC 167 167 C P I MACHINING CO INC 2,761 2,761 CALCOMP CORPORATION 55 55 CALIFORNIA CMPTR PRDTS INC 201 49 97 30 25 CARTWRIGHT ENGINEERING INC 6,000...UNITED MEDIA INC 28 28 UNITED STATES DEPARTMENT DEF 43 43 WALTER DORWIN TEAGUE INC 254 254 YEAGER MANUFACTURING CORF 36 36 TOTAL - ANAHEIM 1,020,504...POMONA T & T INDUSTRIES 57 57 WALTER DORWIN TEAGUE INC 2,013 2,013

  11. Development and characterization of nanopore system for nano-vesicle analysis

    NASA Astrophysics Data System (ADS)

    Goyal, Gaurav

    Nano-vesicles have recently attracted a lot of attention in research and medical communities and are very promising next-generation drug delivery vehicles. This is due to their biocompatibility, biodegradability and their ability to protect drug cargo and deliver it to site-specific locations, while maintaining the desired pharmacokinetic profile. The interaction of these drug loaded vesicles with the recipient cells via adsorption, endocytosis or receptor mediated internalization involve significant bending and deformation and is governed by mechanical properties of the nano-vesicles. Currently, the mechanical characteristics of nano-vesicles are left unexplored because of the difficulties associated with vesicle analysis at sub-100 nm length scale. The need for a complete understanding of nano-vesicle interaction with each other and the recipient cells warrants development of an analytical tool capable of mechanical investigation of individual vesicles at sub-100 nm scale. This dissertation presents investigation of nano-vesicle deformability using resistive pulse sensing and solid-state nanopore devices. The dissertation is divided into four chapters. Chapter 1 discusses the motivation, specific aims and presents an overview of nanoparticle characterization techniques, resistive pulse sensing background and principles, techniques for fabricating solid-state nanopores, as well the deformation behavior of giant vesicles when placed in electric field. Chapter 2 is dedicated to understanding of the scientific principles governing transport of sub-100 nm particles in dilute solutions. We investigated the translocation of rigid nanoparticles through nanopores at salt concentrations < 50 mM. When using low electrolyte strength, surface effects become predominant and resulted in unconventional current signatures in our experiments. It prompted us to explore the effects of different experimental parameters using Multiphysics simulations, in order to optimize our system

  12. Priming healthy eating. You can't prime all the people all of the time☆

    PubMed Central

    Forwood, Suzanna E.; Ahern, Amy L.; Hollands, Gareth J.; Ng, Yin-Lam; Marteau, Theresa M.

    2015-01-01

    Objective In the context of a food purchasing environment filled with advertising and promotions, and an increased desire from policy makers to guide individuals toward choosing healthier foods, this study tests whether priming methods that use healthy food adverts to increase preference for healthier food generalize to a representative population. MethodsIn two studies (Study 1 n = 143; Study 2 n = 764), participants were randomly allocated to a prime condition, where they viewed fruit and vegetable advertisements, or a control condition, with no advertisements. A subsequent forced choice task assessed preference between fruits and other sweet snacks. Additional measures included current hunger and thirst, dietary restraint, age, gender, education and self-reported weight and height. ResultsIn Study 1, hunger reduced preferences for fruits (OR (95% CI) = 0.38 (0.26–0.56), p < 0.0001), an effect countered by the prime (OR (95% CI) = 2.29 (1.33–3.96), p = 0.003). In Study 2, the effect of the prime did not generalize to a representative population. More educated participants, as used in Study 1, chose more fruit when hungry and primed (OR (95% CI) = 1.42 (1.13–1.79), p = 0.003), while less educated participants' fruit choice was unaffected by hunger or the prime. ConclusionThis study provides preliminary evidence that the effects of adverts on healthy eating choices depend on key individual traits (education level) and states (hunger), do not generalize to a broader population and have the potential to increase health inequalities arising from food choice. PMID:25636234

  13. Polymeric Microgels as Potential Drug Delivery Vesicles

    NASA Astrophysics Data System (ADS)

    McDonough, Ryan; Streletzky, Kiril; Bayachou, Mekki; Peiris, Pubudu

    2010-03-01

    The temperature dependent volume phase change of cross-linked amphiphilic molecules (microgels) suggests their use as drug delivery vesicles. Drug particles aggregate in the slightly hydrophobic microgel interior. They are stored in equilibrium until the critical temperature (Tv) is reached where the volume phase change limits available space, thus expelling the drugs. This loading property of hydroxypropylcellulose (HPC) microgels was tested using amperometric analytical techniques. Small molecules inside microgels do not approach the electrode surface, which decreases current signal. A room temperature (Troom) flow amperometric measurement comparing microgel/paracetamol solution with control paracetamol samples yielded about 20 percent concentration reduction in the microgel sample. Results from the steady-state electrochemical experiment confirm the 20 percent concentration drop in the microgel sample compared to the control sample at Troom. Using the steady-state experiment with a cyclic temperature ramp from Troom to beyond Tv showed that the paracetamol concentration change between the temperature extremes was greater for the microgels than for the controls. An evolving aspect of the study is the characterization of microgel shrinkage from in situ, temperature controlled liquid AFM images as compared to previously completed DLS characterization of the same microgel sample.

  14. Characterization of Alternaria infectoria extracellular vesicles

    PubMed Central

    Silva, Branca M.A.; Prados-Rosales, Rafael; Espadas-Moreno, Javier; Wolf, Julie M.; Luque-Garcia, Jose L.; Gonçalves, Teresa; Casadevall, Arturo

    2015-01-01

    Many fungi use membrane vesicles to transport complex molecules across their cell walls. Like mammalian exosomes, fungal vesicles contain lipids, proteins, and polysaccharides, many of which are associated with virulence. Here we identify and characterize extracellular vesicles (EVs) in Alternaria infectoria, a ubiquitous, environmental filamentous fungus that is also an opportunistic human pathogen. Examination of the A. infectoria EVs revealed a morphology similar to that of vesicles described in other fungal species. Of note, proteomic analysis detected a reduced number of vesicle-associated proteins. There were two prevalent categories among the 20 identified proteins, including the polysaccharide metabolism group, probably related to plant host invasion or biosynthesis/degradation of cell wall components, and the nuclear proteins, especially DNA repair enzymes. We also found enzymes related to pigment synthesis, adhesion to the host cell, and trafficking of vesicles/organelles/molecules. This is the first time EV secretions have been identified in a filamentous fungus. We believe that these vesicles might have a role in virulence. PMID:24576997

  15. Priming against environmental challenges and proteomics in plants: Update and agricultural perspectives

    PubMed Central

    Tanou, Georgia; Fotopoulos, Vasileios; Molassiotis, Athanassios

    2012-01-01

    Priming is the cellular state in which the harmful effects of abiotic stress factors in plants are hindered by pre-exposure to a stimulus, thus resulting in greater survival. It is becoming increasingly evident that priming techniques (e.g., external application of natural or synthetic compounds in plants) can enhance the tolerance of crops to environmental stresses. Innovative systems biology approaches such as proteomics are currently recognized as essential tools to understand the molecular mechanisms underlying plant responses to environmental stimuli and priming phenomena. The few published proteomic studies on priming in the context of environmental stress identify key protein targets and signaling pathways which are being involved in the alleviation of negative effects of stress factors. Since priming is a very promising strategy in modern crop production management, further research is needed in order to establish the global picture of priming phenomena against environmental challenges as well as to characterize specific priming-related protein indicators in plants. PMID:22973291

  16. Prime Contract Awards Alphabetically by Contractor, by State or Country, and Place, FY 85. Part 18 (Seneca Food Corp - Staplex Co Inc).

    DTIC Science & Technology

    1985-01-01

    1965 UNCLASSIFIED DIOR/ST18-BS-PT- 18 F/O 15/5 ML momhmhhhmhmmum mEmhEEohhohmhI mhhmhhEEmhhmhI mhohhhEmhmhhE 2.2: 36 1 .1I 25 1.4 11111.6 MICROCOPY...ontractor, by State cr Countryj, and Place, FY 8r, 18 (Seneca Food Corp - Staplex Co Inc). ~ ~ YO~Q~ E~OT ~ TIM COERE 14DAT OF REPORT (Year, Month, Day) 5...PAGE COUNT - - .FROM _ ___TO ___ 98 A 16 iP.OLiIEMEN’TARY NOTATION COSATi CODES 18 . SUBJECT TERMS (Continue on reverse if necessary and identify by

  17. Vesicle membrane fluctuations at nm resolution

    NASA Astrophysics Data System (ADS)

    Chen, Kejia; Bae, Sung Chul; Min, Chang-Ki; Granick, Steve; Granick Group Team

    2011-03-01

    We measure membrane thermal fluctuations with nanometer spatial resolution and microsecond time resolution, extending a scattering technique used at the Curie Institute to study red blood cell dynamics (Timo Betz et al., Proc. Nat. Acad. Sci. USA 106, 15320, 2009). A laser beam is focused at the leading edge of a phospholipid vesicle membrane and the forward scattered light is detected by a quadrant photodiode. The measurements over 4 orders of magnitude of frequency allow quantification of more complete fluctuation spectra than competing methods, and therefore fuller understanding of the vesicle membrane mechanics. As a proof of concept, we quantify how adsorbed nanoparticles stiffen giant unilamellar vesicles (GUVs).

  18. Extracellular vesicles: Exosomes, microvesicles, and friends

    PubMed Central

    Stoorvogel, Willem

    2013-01-01

    Cells release into the extracellular environment diverse types of membrane vesicles of endosomal and plasma membrane origin called exosomes and microvesicles, respectively. These extracellular vesicles (EVs) represent an important mode of intercellular communication by serving as vehicles for transfer between cells of membrane and cytosolic proteins, lipids, and RNA. Deficiencies in our knowledge of the molecular mechanisms for EV formation and lack of methods to interfere with the packaging of cargo or with vesicle release, however, still hamper identification of their physiological relevance in vivo. In this review, we focus on the characterization of EVs and on currently proposed mechanisms for their formation, targeting, and function. PMID:23420871

  19. Time-dependent quantum wave packet study of the Ar+H{sub 2}{sup +}{yields}ArH{sup +}+H reaction on a new ab initio potential energy surface for the ground electronic state (1{sup 2}A Prime )

    SciTech Connect

    Hu Mei; Liu Xinguo; Tan Ruishan; Li Hongzheng; Xu Wenwu

    2013-05-07

    A new global potential energy surface for the ground electronic state (1{sup 2}A Prime ) of the Ar+H{sub 2}{sup +}{yields}ArH{sup +}+H reaction has been constructed by multi-reference configuration interaction method with Davidson correction and a basis set of aug-cc-pVQZ. Using 6080 ab initio single-point energies of all the regions for the dynamics, a many-body expansion function form has been used to fit these points. The quantum reactive scattering dynamics calculations taking into account the Coriolis coupling (CC) were carried out on the new potential energy surface over a range of collision energies (0.03-1.0 eV). The reaction probabilities and integral cross sections for the title reaction were calculated. The significance of including the CC quantum scattering calculation has been revealed by the comparison between the CC and the centrifugal sudden approximation calculation. The calculated cross section is in agreement with the experimental result at collision energy 1.0 eV.

  20. Space Place Prime

    NASA Technical Reports Server (NTRS)

    Fitzpatrick, Austin J.; Novati, Alexander; Fisher, Diane K.; Leon, Nancy J.; Netting, Ruth

    2013-01-01

    Space Place Prime is public engagement and education software for use on iPad. It targets a multi-generational audience with news, images, videos, and educational articles from the Space Place Web site and other NASA sources. New content is downloaded daily (or whenever the user accesses the app) via the wireless connection. In addition to the Space Place Web site, several NASA RSS feeds are tapped to provide new content. Content is retained for the previous several days, or some number of editions of each feed. All content is controlled on the server side, so features about the latest news, or changes to any content, can be made without updating the app in the Apple Store. It gathers many popular NASA features into one app. The interface is a boundless, slidable- in-any-direction grid of images, unique for each feature, and iconized as image, video, or article. A tap opens the feature. An alternate list mode presents menus of images, videos, and articles separately. Favorites can be tagged for permanent archive. Face - book, Twitter, and e-mail connections make any feature shareable.

  1. Direct Measurement of Pore Dynamics and Leakage Induced by a Model Antimicrobial Peptide in Single Vesicles and Cells.

    PubMed

    Burton, Matthew G; Huang, Qi M; Hossain, Mohammed A; Wade, John D; Palombo, Enzo A; Gee, Michelle L; Clayton, Andrew H A

    2016-06-28

    Antimicrobial peptides are promising therapeutic alternatives to counter growing antimicrobial resistance. Their precise mechanism of action remains elusive, however, particularly with respect to live bacterial cells. We investigated the interaction of a fluorescent melittin analogue with single giant unilamellar vesicles, giant multilamellar vesicles, and bilamellar Gram-negative Escherichia coli (E. coli) bacteria. Time-lapse fluorescence lifetime imaging microscopy was employed to determine the population distribution of the fluorescent melittin analogue between pore state and membrane surface state, and simultaneously measure the leakage of entrapped fluorescent species from the vesicle (or bacterium) interior. In giant unilamellar vesicles, leakage from vesicle interior was correlated with an increase in level of pore states, consistent with a stable pore formation mechanism. In giant multilamellar vesicles, vesicle leakage occurred more gradually and did not appear to correlate with increased pore states. Instead pore levels remained at a low steady-state level, which is more in line with coupled equilibria. Finally, in single bacterial cells, significant increases in pore levels were observed over time, which were correlated with only partial loss of cytosolic contents. These observations suggested that pore formation, as opposed to complete dissolution of membrane, was responsible for the leakage of contents in these systems, and that the bacterial membrane has an adaptive capacity that resists peptide attack. We interpret the three distinct pore dynamics regimes in the context of the increasing physical and biological complexity of the membranes.

  2. Extracellular vesicles in parasitic diseases

    PubMed Central

    Marcilla, Antonio; Martin-Jaular, Lorena; Trelis, Maria; de Menezes-Neto, Armando; Osuna, Antonio; Bernal, Dolores; Fernandez-Becerra, Carmen; Almeida, Igor C.; del Portillo, Hernando A.

    2014-01-01

    Parasitic diseases affect billions of people and are considered a major public health issue. Close to 400 species are estimated to parasitize humans, of which around 90 are responsible for great clinical burden and mortality rates. Unfortunately, they are largely neglected as they are mainly endemic to poor regions. Of relevance to this review, there is accumulating evidence of the release of extracellular vesicles (EVs) in parasitic diseases, acting both in parasite–parasite inter-communication as well as in parasite–host interactions. EVs participate in the dissemination of the pathogen and play a role in the regulation of the host immune systems. Production of EVs from parasites or parasitized cells has been described for a number of parasitic infections. In this review, we provide the most relevant findings of the involvement of EVs in intercellular communication, modulation of immune responses, involvement in pathology, and their potential as new diagnostic tools and therapeutic agents in some of the major human parasitic pathogens. PMID:25536932

  3. The PRIME Lab biomedical program

    NASA Astrophysics Data System (ADS)

    Jackson, George S.; Elmore, David; Rickey, Frank A.; Musameh, Sharif M.; Sharma, Pankaj; Hillegonds, Darren; Coury, Louis; Kissinger, Peter

    2000-10-01

    The biomedical accelerator mass spectrometry (AMS) initiative at PRIME Lab including the status of equipment and sample preparation is described. Several biomedical projects are underway involving one or more of the nuclides: 14C, 26Al and 41Ca. Routine production of CaF 2 and graphite is taking place. Finally, the future direction and plans for improvement of the biomedical program at PRIME Lab are discussed.

  4. MIL-PRIME Program System

    NASA Astrophysics Data System (ADS)

    Rall, F. T., Jr.

    1982-11-01

    The MIL-PRIME Program was developed to provide the Aeronautical Systems Division with a specifications and standards program for the development of new weapon systems. This program was started in 1976 and is still in the process of being implemented. This presentation provides the background and insight into the MIL PRIME System, the use of these new documents in the acquisition process, and the current status of the program.

  5. Psychotherapy Augmentation through Preconscious Priming

    PubMed Central

    Borgeat, François; O’Connor, Kieron; Amado, Danielle; St-Pierre-Delorme, Marie-Ève

    2013-01-01

    Objective: To test the hypothesis that repeated preconscious (masked) priming of personalized positive cognitions could augment cognitive change and facilitate achievement of patients’ goals following a therapy. Methods: Twenty social phobic patients (13 women) completed a 36-weeks study beginning by 12 weeks of group behavioral therapy. After the therapy, they received 6 weeks of preconscious priming and 6 weeks of a control procedure in a randomized cross-over design. The Priming condition involved listening twice daily with a passive attitude to a recording of individualized formulations of appropriate cognitions and attitudes masked by music. The Control condition involved listening to an indistinguishable recording where the formulations had been replaced by random numbers. Changes in social cognitions were measured by the Social Interaction Self Statements Test (SISST). Results: Patients improved following therapy. The Priming procedure was associated with increased positive cognitions and decreased negative cognitions on the SISST while the Control procedure was not. The Priming procedure induced more cognitive change when applied immediately after the group therapy. Conclusion: An effect of priming was observed on social phobia related cognitions in the expected direction. This self administered addition to a therapy could be seen as an augmentation strategy. PMID:23508724

  6. Ceramidase Regulates Synaptic Vesicle Exocytosis and Trafficking

    PubMed Central

    Rohrbough, Jeffrey; Rushton, Emma; Palanker, Laura; Woodruff, Elvin; Matthies, Heinrich J. G.; Acharya, Usha; Acharya, Jairaj K.; Broadie, Kendal

    2009-01-01

    A screen for Drosophila synaptic dysfunction mutants identified slug-a-bed (slab). The slab gene encodes ceramidase, a central enzyme in sphingolipid metabolism and regulation. Sphingolipids are major constituents of lipid rafts, membrane domains with roles in vesicle trafficking, and signaling pathways. Null slab mutants arrest as fully developed embryos with severely reduced movement. The SLAB protein is widely expressed in different tissues but enriched in neurons at all stages of development. Targeted neuronal expression of slab rescues mutant lethality, demonstrating the essential neuronal function of the protein. C5-ceramide applied to living preparations is rapidly accumulated at neuromuscular junction (NMJ) synapses dependent on the SLAB expression level, indicating that synaptic sphingolipid trafficking and distribution is regulated by SLAB function. Evoked synaptic currents at slab mutant NMJs are reduced by 50–70%, whereas postsynaptic glutamate-gated currents are normal, demonstrating a specific presynaptic impairment. Hypertonic saline-evoked synaptic vesicle fusion is similarly impaired by 50–70%, demonstrating a loss of readily releasable vesicles. In addition, FM1-43 dye uptake is reduced in slab mutant presynaptic terminals, indicating a smaller cycling vesicle pool. Ultrastructural analyses of mutants reveal a normal vesicle distribution clustered and docked at active zones, but fewer vesicles in reserve regions, and a twofold to threefold increased incidence of vesicles linked together and tethered at the plasma membrane. These results indicate that SLAB ceramidase function controls presynaptic terminal sphingolipid composition to regulate vesicle fusion and trafficking, and thus the strength and reliability of synaptic transmission. PMID:15356190

  7. Concentration-Independent Spontaneously Forming Biomimetric Vesicles

    NASA Astrophysics Data System (ADS)

    Nieh, M.-P.; Harroun, T. A.; Raghunathan, V. A.; Glinka, C. J.; Katsaras, J.

    2003-10-01

    In this Letter we present small-angle neutron scattering data from a biomimetic system composed of the phospholipids dimyristoyl and dihexanoyl phosphorylcholine (DMPC and DHPC, respectively). Doping DMPC-DHPC multilamellar vesicles with either the negatively charged lipid dimyristoyl phosphorylglycerol (DMPG, net charge -1) or the divalent cation, calcium (Ca2+), leads to the spontaneous formation of energetically stabilized monodisperse unilamellar vesicles whose radii are concentration independent and in contrast with previous experimental observations.

  8. Test Sequence Priming in Recognition Memory

    ERIC Educational Resources Information Center

    Johns, Elizabeth E.; Mewhort, D. J. K.

    2009-01-01

    The authors examined priming within the test sequence in 3 recognition memory experiments. A probe primed its successor whenever both probes shared a feature with the same studied item ("interjacent priming"), indicating that the study item like the probe is central to the decision. Interjacent priming occurred even when the 2 probes did…

  9. The PrimeTime Expose: Reactions and Recommendations.

    ERIC Educational Resources Information Center

    Child Care Information Exchange, 1991

    1991-01-01

    Describes "Day Care Nightmares," a two-part expose on ABC's PrimeTime Live, broadcast in June 1991, in which negative claims were made about the state of day care in the United States. A variety of advocates, policymakers, and child care providers present their reactions to the shows and their recommendations on what action can be taken.…

  10. Immune priming in arthropods: an update focusing on the red flour beetle.

    PubMed

    Milutinović, Barbara; Peuß, Robert; Ferro, Kevin; Kurtz, Joachim

    2016-08-01

    Immune priming has now been demonstrated in a wide range of invertebrate species. Studies testing this phenomenon largely differ in terms of experimental design, host-parasite combinations, agents used for priming, and in particular the degree of demonstrated specificity of the primed response. This review provides an overview of known and putative mechanisms underlying broad-spectrum and specific immune priming in arthropods. We focus on insects and particularly the red flour beetle Tribolium castaneum, where priming has been demonstrated within and across generations. We will also draw attention to the relevance of routes of priming and infection, which can occur septically and orally, with largely differing physiology. For oral priming, an involvement of gut microbiota was demonstrated in mosquitoes and flour beetles. Generally, a primed state could result from long-lasting immune activation or a form of memory that does not entail lingering immune components. Moreover, the primed state could also be of a qualitatively different kind than the challenge response. Finally, we will consider that there should be natural variation in priming capability, and therefore a possibility to study this trait with experimental evolution approaches.

  11. Oxidative and other posttranslational modifications in extracellular vesicle biology.

    PubMed

    Szabó-Taylor, Katalin; Ryan, Brent; Osteikoetxea, Xabier; Szabó, Tamás G; Sódar, Barbara; Holub, Marcsilla; Németh, Andrea; Pálóczi, Krisztina; Pállinger, Éva; Winyard, Paul; Buzás, Edit I

    2015-04-01

    Extracellular vesicles including exosomes, microvesicles and apoptotic vesicles, are phospholipid bilayer surrounded structures secreted by cells universally, in an evolutionarily conserved fashion. Posttranslational modifications such as oxidation, citrullination, phosphorylation and glycosylation play diverse roles in extracellular vesicle biology. Posttranslational modifications orchestrate the biogenesis of extracellular vesicles. The signals extracellular vesicles transmit between cells also often function via modulating posttranslational modifications of target molecules, given that extracellular vesicles are carriers of several active enzymes catalysing posttranslational modifications. Posttranslational modifications of extracellular vesicles can also contribute to disease pathology by e.g. amplifying inflammation, generating neoepitopes or carrying neoepitopes themselves.

  12. Locus of single-prime negative priming: the role of perceptual form.

    PubMed

    Chao, Hsuan-Fu

    2013-07-01

    Single-prime negative priming refers to the phenomenon in which after a single prime word is briefly presented, repeating it as the probe target results in a delay in responding to the target. The present study investigated the locus of this negative priming effect. Experiment 1 showed that repeating the identity of the prime produced a negative priming effect but merely repeating the response of the prime did not. Experiment 2 showed that the negative priming effect transformed into positive priming when the probe distractor was absent. Experiments 3 and 4 further revealed that single-prime negative priming was observed when the perceptual form was repeated. Taken together, these results suggest that single-prime negative priming involves a perceptual locus.

  13. Differential detergent sensitivity of extracellular vesicle subpopulations.

    PubMed

    Osteikoetxea, Xabier; Sódar, Barbara; Németh, Andrea; Szabó-Taylor, Katalin; Pálóczi, Krisztina; Vukman, Krisztina V; Tamási, Viola; Balogh, Andrea; Kittel, Ágnes; Pállinger, Éva; Buzás, Edit Irén

    2015-10-14

    Extracellular vesicles (including exosomes, microvesicles and apoptotic bodies) are currently attracting rapidly increasing attention from various fields of biology due to their ability to carry complex information and act as autocrine, paracrine and even endocrine intercellular messengers. In the present study we investigated the sensitivity of size-based subpopulations of extracellular vesicles to different concentrations of detergents including sodium dodecyl sulphate, Triton X-100, Tween 20 and deoxycholate. We determined the required detergent concentration that lysed each of the vesicle subpopulations secreted by Jurkat, THP-1, MiaPaCa and U937 human cell lines. We characterized the vesicles by tunable resistive pulse sensing, flow cytometry and transmission electron microscopy. Microvesicles and apoptotic bodies were found to be more sensitive to detergent lysis than exosomes. Furthermore, we found evidence that sodium dodecyl sulphate and Triton X-100 were more effective in vesicle lysis at low concentrations than deoxycholate or Tween 20. Taken together, our data suggest that a combination of differential detergent lysis with tunable resistive pulse sensing or flow cytometry may prove useful for simple and fast differentiation between exosomes and other extracellular vesicle subpopulations as well as between vesicular and non-vesicular structures.

  14. Horizontal Transmission of Cytosolic Sup35 Prions by Extracellular Vesicles

    PubMed Central

    Liu, Shu; Hossinger, André; Hofmann, Julia P.; Denner, Philip

    2016-01-01

    ABSTRACT Prions are infectious protein particles that replicate by templating their aggregated state onto soluble protein of the same type. Originally identified as the causative agent of transmissible spongiform encephalopathies, prions in yeast (Saccharomyces cerevisiae) are epigenetic elements of inheritance that induce phenotypic changes of their host cells. The prototype yeast prion is the translation termination factor Sup35. Prions composed of Sup35 or its modular prion domain NM are heritable and are transmitted vertically to progeny or horizontally during mating. Interestingly, in mammalian cells, protein aggregates derived from yeast Sup35 NM behave as true infectious entities that employ dissemination strategies similar to those of mammalian prions. While transmission is most efficient when cells are in direct contact, we demonstrate here that cytosolic Sup35 NM prions are also released into the extracellular space in association with nanometer-sized membrane vesicles. Importantly, extracellular vesicles are biologically active and are taken up by recipient cells, where they induce self-sustained Sup35 NM protein aggregation. Thus, in mammalian cells, extracellular vesicles can serve as dissemination vehicles for protein-based epigenetic information transfer. PMID:27406566

  15. On the Control of Single-Prime Negative Priming: The Effects of Practice and Time Course

    ERIC Educational Resources Information Center

    Chao, Hsuan-Fu

    2009-01-01

    Single-prime negative priming refers to the phenomenon wherein repetition of a prime as the probe target results in delayed response. Sometimes this effect has been found to be contingent on participants' unawareness of the primes, and sometimes it has not. Further, sometimes this effect has been found to be eliminated when the prime could predict…

  16. The Intervenor Effect in Masked Priming: How Does Masked Priming Survive across an Intervening Word?

    ERIC Educational Resources Information Center

    Forster, Kenneth I.

    2009-01-01

    Four masked priming experiments are reported investigating the effect of inserting an unrelated word between the masked prime and the target. When the intervening word is visible, identity priming is reduced to the level of one-letter-different form priming, but form priming is largely unaffected. However, when the intervening word is itself…

  17. Transfer of oleic acid between albumin and phospholipid vesicles

    SciTech Connect

    Hamilton, J.A.; Cistola, D.P.

    1986-01-01

    The net transfer of oleic acid between egg phosphatidylcholine unilamellar vesicles and bovine serum albumin has been monitored by TC NMR spectroscopy and 90% isotopically substituted (1- TC)oleic acid. The carboxyl chemical shifts of oleic acid bound to albumin were different from those for oleic acid in phospholipid vesicles. Therefore, in mixtures of donor particles, the equilibrium distribution of oleic acid was determined from chemical shift and peak intensity data without separation of donor and acceptor particles. In a system containing equal masses of albumin and phospholipid and a stoichiometry of 4-5 mol of oleic acid per mol of albumin, the oleic acid distribution was pH dependent, with greater than or equal to80% of the oleic acid associated with albumin at pH 7.4; association was greater than or equal to90% at pH 8.0. Decreasing the pH below 7.4 markedly decreased the proportion of fatty acid bound to albumin. The distribution was reversible with pH and was independent of whether vesicles or albumin acted as a donor. These data suggest that pH may strongly influence the partitioning of fatty acid between cellular membranes and albumin. The TC NMR method is also advantageous because it provides information about the structural environments of oleic acid bound to albumin or phospholipid, the ionization state of oleic acid in each environment, and the structural integrity of the vesicles. In addition, minimum and maximum limits for the exchange rates of oleic acid among different environments were obtained from the NMR data.

  18. Structure formation of lipid membranes: Membrane self-assembly and vesicle opening-up to octopus-like micelles

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2013-02-01

    We briefly review our recent studies on self-assembly and vesicle rupture of lipid membranes using coarse-grained molecular simulations. For single component membranes, lipid molecules self-assemble from random gas states to vesicles via disk-shaped clusters. Clusters aggregate into larger clusters, and subsequently the large disks close into vesicles. The size of vesicles are determined by kinetics than by thermodynamics. When a vesicle composed of lipid and detergent types of molecules is ruptured, a disk-shaped micelle called bicelle can be formed. When both surfactants have negligibly low critical micelle concentration, it is found that bicelles connected with worm-like micelles are also formed depending on the surfactant ratio and spontaneous curvature of the membrane monolayer.

  19. Repetition priming and cortical arousal in healthy aging and Alzheimer's disease.

    PubMed

    Kane, Amy E; Festa, Elena K; Salmon, David P; Heindel, William C

    2015-04-01

    Repetition priming refers to a form of implicit memory in which prior exposure to a stimulus facilitates the subsequent processing of the same or a related stimulus. One frequently used repetition priming task is word-stem completion priming. In this task, participants complete a series of beginning word stems with the first word that comes to mind after having viewed, in an unrelated context, words that can complete some of the stems. Patients with Alzheimer's disease (AD) exhibit a significant deficit in word-stem completion priming, but the neural mechanisms underlying this deficit have yet to be identified. The present study examined the possibility that the word-stem completion priming deficit in AD is due to disruption of ascending neuromodulatory systems that mediate cortical arousal by comparing word-stem completion priming and behavioral measures of spatial orienting and phasic alerting. Results showed that in healthy elderly controls higher levels of phasic alerting were associated with a sharpening of the temporal dynamics of priming across two delay intervals: those with higher levels of alerting showed more immediate priming but less delayed priming than those with lesser levels of alerting. In patients with AD, priming was impaired despite intact levels of phasic alerting and spatial orienting, and group status rather than individual levels of alerting or orienting predicted the magnitude of their stem-completion priming. Furthermore, the change in priming across delays they displayed was not related to level of alerting or orienting. These findings support the role of the noradrenergic projection system in modulating the level of steady-state cortical activation (or "cortical tonus") underlying both phasic alerting and the temporal dynamics of repetition priming. However, impaired priming in patients with AD does not appear to be due to disruption of this neuromodulatory system.

  20. Transmembrane flux and receptor desensitization measured with membrane vesicles. Homogeneity of vesicles investigated by computer simulation.

    PubMed Central

    Cash, D J; Langer, R M; Subbarao, K; Bradbury, J R

    1988-01-01

    The use of membrane vesicles to make quantitative studies of transmembrane transport and exchange processes involves an assumption of homogeneity of the membrane vesicles. In studies of 86Rb+ exchange mediated by acetylcholine receptor from the electric organ of Electrophorus electricus and of 36Cl- exchange mediated by GABA receptor from rat brain, measurements of ion exchange and receptor desensitization precisely followed first order kinetics in support of this assumption. In other measurements a biphasic decay of receptor activity was seen. To elucidate the molecular properties of receptors from such measurements it is important to appreciate what the requirements of vesicle monodispersity are for meaningful results and what the effect of vesicle heterogeneity would be. The experiments were simulated with single vesicle populations with variable defined size distributions as well as with mixtures of different populations of vesicles. The properties of the receptors and their density in the membrane could be varied. Different receptors could be present on the same or different membrane vesicles. The simulated measurements were not very sensitive to size dispersity. A very broad size distribution of a single vesicle population was necessary to give rise to detectable deviations from first order kinetics or errors in the determined kinetic constants. Errors could become significant with mixtures of different vesicle populations, where the dispersity in initial ion exchange rate constant, proportional to the receptor concentration per internal volume, became large. In this case the apparent rate of receptor desensitization would diverge in opposite directions from the input value when measured by two different methods, suggesting an experimental test for such kinetic heterogeneity. A biphasic decrease of receptor activity could not be attributed to vesicle heterogeneity and must be due to desensitization processes with different rates. Significant errors would not

  1. Procoagulant and immunogenic properties of melanoma exosomes, microvesicles and apoptotic vesicles

    PubMed Central

    Muhsin-Sharafaldine, Morad-Rémy; Saunderson, Sarah C.; Dunn, Amy C.; Faed, James M.; Kleffmann, Torsten; McLellan, Alexander D.

    2016-01-01

    Extracellular vesicles (EV) are lipid particles released from eukaryotic cells into the extracellular fluid. Depending on the cell type or mechanism of release, vesicles vary in form and function and exert distinct functions in coagulation and immunity. Tumor cells may constitutively shed vesicles known as exosomes or microvesicles (MV). Alternatively, apoptosis induces the release of apoptotic blebs or vesicles (ApoV) from the plasma membrane. EV have been implicated in thrombotic events (the second highest cause of death in cancer patients) and tumor vesicles contribute to the anti-cancer immune response. In this study, we utilized the well characterized B16 melanoma model to determine the molecular composition and procoagulant and immunogenic potential of exosomes, MV and ApoV. Distinct patterns of surface and cytoplasmic molecules (tetraspanins, integrins, heat shock proteins and histones) were expressed between the vesicle types. Moreover, in vitro coagulation assays revealed that membrane-derived vesicles, namely MV and ApoV, were more procoagulant than exosomes–with tissue factor and phosphatidylserine critical for procoagulant activity. Mice immunized with antigen-pulsed ApoV and challenged with B16 tumors were protected out to 60 days, while lower protection rates were afforded by MV and exosomes. Together the results demonstrate distinct phenotypic and functional differences between vesicle types, with important procoagulant and immunogenic functions emerging for membrane-derived MV and ApoV versus endosome-derived exosomes. This study highlights the potential of EV to contribute to the prothrombotic state, as well as to anti-cancer immunity. PMID:27462921

  2. Interleukin 12 acts directly on CD4+ T cells to enhance priming for interferon gamma production and diminishes interleukin 4 inhibition of such priming.

    PubMed Central

    Seder, R A; Gazzinelli, R; Sher, A; Paul, W E

    1993-01-01

    Naive CD4+ T cells produce interleukin 2 (IL-2) but little IL-4 or interferon gamma (IFN-gamma). In vitro, they develop into IL-4 or IFN-gamma producers depending on the conditions of the priming culture. Using T-cell receptor transgenic CD4+ T cells, the role of IL-12 and IL-4 in antigen-specific priming was examined. IL-12 substantially enhanced the ability of naive CD4+ T cells to develop into cells that produced IFN-gamma upon restimulation. However, it was not essential since anti-IL-12 antibodies failed to block the priming for IFN-gamma observed in the absence of exogenous IL-12. When both IL-12 and IL-4 were present in the priming culture, IL-12 did not inhibit priming for IL-4 production. In contrast, IL-4 diminished but did not abolish priming for IFN-gamma production. In an accessory cell-independent priming system, IL-12 strikingly augmented priming for IFN-gamma production, indicating that it acts directly on T cells. IFN-gamma itself did not enhance priming for IFN-gamma production in either accessory cell-dependent or independent systems. In an accessory cell-dependent system, the IL-12-mediated enhancement was not blocked by adding neutralizing anti-IFN-gamma monoclonal antibody. However, in an accessory cell-independent system, anti-IFN-gamma antibody did inhibit priming for IFN-gamma production leaving open a role for IFN-gamma in the priming process. These data indicate that IL-12 has a major effect on the inductive phase of T-cell priming by enhancing commitment to IFN-gamma production and thus can profoundly influence the state of immunity that develops. Images Fig. 4 PMID:7901851

  3. How did bacterial ancestors reproduce? Lessons from L-form cells and giant lipid vesicles: multiplication similarities between lipid vesicles and L-form bacteria.

    PubMed

    Briers, Yves; Walde, Peter; Schuppler, Markus; Loessner, Martin J

    2012-12-01

    In possible scenarios on the origin of life, protocells represent the precursors of the first living cells. To study such hypothetical protocells, giant vesicles are being widely used as a simple model. Lipid vesicles can undergo complex morphological changes enabling self-reproduction such as growth, fission, and extra- and intravesicular budding. These properties of vesicular systems may in some way reflect the mechanism of reproduction used by protocells. Moreover, remarkable similarities exist between the morphological changes observed in giant vesicles and bacterial L-form cells, which represent bacteria that have lost their rigid cell wall, but retain the ability to reproduce. L-forms feature a dismantled cellular structure and are unable to carry out classical binary fission. We propose that the striking similarities in morphological transitions of L-forms and giant lipid vesicles may provide insights into primitive reproductive mechanisms and contribute to a better understanding of the origin and evolution of mechanisms of cell reproduction. Editor's suggested further reading in BioEssays Synthesizing artificial cells from giant unilamellar vesicles: State-of-the art in the development of microfluidic technology Abstract.

  4. A two phase field model for tracking vesicle-vesicle adhesion.

    PubMed

    Gu, Rui; Wang, Xiaoqiang; Gunzburger, Max

    2016-11-01

    A multi-phase-field model for simulating the adhesion between two vesicles is constructed. Two phase field functions are introduced to simulate each of the two vesicles. An energy model is defined which accounts for the elastic bending energy of each vesicle and the contact potential energy between the two vesicles; the vesicle volume and surface area constraints are imposed using a penalty method. Numerical results are provided to verify the efficacy of our model and to provide visual illustrations of the different types of contact. The method can be adjusted to solve endocytosis problems by modifying the bending rigidity coefficients of the two elastic bending energies. The method can also be extended to simulate multi-cell adhesions, one example of which is erythrocyte rouleaux. A comparison with laboratory observations demonstrates the effectiveness of the multi-phase field approach.

  5. Coarse-grained simulation of lipid vesicles with ``n-atic'' orientational order

    NASA Astrophysics Data System (ADS)

    Geng, Jun; Selinger, Jonathan; Selinger, Robin

    2012-02-01

    We perform coarse-grained simulation studies of fluid lipid vesicles with in-plane ``n-atic'' orientational order associated with the shape of lipid head group, to test the theoretical predictions of Park, Lubensky and MacKintosh [1] for resulting vesicle shape and defect structures. Our simulation model uses a single layer coarse-grained implicit-solvent approach proposed by Yuan et al [2], with addition of an extra vector degree of freedom representing in-plane orientational order. We carry out simulation studies for n=1 to 6, examining in each case the spatial distribution of defects and resulting deformation of the vesicle. An initially spherical vesicle (genus zero) with n-atic order has a ground state with 2n vortices of strength 1/n, as expected, but the observed equilibrium shapes are sometimes quite different from those predicted theoretically. For the n=1 case, we find that the vesicle may become trapped in a disordered, long-lived metastable state with extra +/- defects whose pair-annihilation is inhibited by local changes in membrane curvature, and thus may never reach its predicted ground state. [4pt] [1] J. Park, T. C. Lubensky, and F. C. MacKintosh, Europhys. Lett. 20, 279 (1992)[0pt] [2] H. Yuan, C. Huang, Ju Li, G. Lykotrafitis, and S. Zhang, Phys. Rev. E 82, 011905 (2010)

  6. Priming and Habituation for Faces: Individual Differences and Inversion Effects

    ERIC Educational Resources Information Center

    Rieth, Cory A.; Huber, David E.

    2010-01-01

    Immediate repetition priming for faces was examined across a range of prime durations in a threshold identification task. Similar to word repetition priming results, short duration face primes produced positive priming whereas long duration face primes eliminated or reversed this effect. A habituation model of such priming effects predicted that…

  7. Oligomerizations of deoxyadenosine bis-phosphates and of their 3-prime-5-prime, 3-prime-3-prime, and 5-prime-5-prime dimers - Effects of a pyrophosphate-linked, poly(T) analog

    NASA Technical Reports Server (NTRS)

    Visscher, J.; Bakker, C. G.; Schwartz, Alan W.

    1990-01-01

    The effect of a 3-prime-5-prime pyrophosphate-linked oligomer of pTp on oligomerizations of pdAp and of its 3-prime-5-prime, 3-prime-3-prime, and 5-prime-5-prime dimers was investigated, using HPLC to separate the reaction mixtures; peak detection was by absorbance monitoring at 254 nm. It was expected that the dimers would form stable complexes with the template, with the degree of stability depending upon the internal linkage of each dimer. It was found that, although the isomers differ substantially in their oligomerization behavior in the absence of template, the analog-template catalyzes the oligomerization to about the same extent in all three cases.

  8. Tube extrusion from permeabilized giant vesicles

    NASA Astrophysics Data System (ADS)

    Borghi, N.; Kremer, S.; Askovic, V.; Brochard-Wyart, F.

    2006-08-01

    This letter reports the permeabilization effects of chemical additives on mechanical properties of Giant Unilamellar Vesicles (GUVs). We use a surfactant, Tween 20, inducing transient pores and a protein, Streptolysin O, inducing permanent pores in the membrane. Lipid tubes are extracted from GUVs anchored onto the tip of a micro-needle and submitted to hydrodynamic flows. On bare vesicles, tube extrusion is governed by the entropic elasticity of the membrane. The vesicle tension increases until it balances the flow velocity U and the tube reaches a stationary length. In permeabilized vesicles, the membrane tension is maintained at a constant value σp by the permeation of inner solution through nanometric pores. This allows extrusion of "infinite" tubes at constant velocity that never reach a stationary length. Tween-20 preliminary results suggest that σp strongly depends on surfactant concentration. For Streptolysin O, we have measured σp vs. U and found two regimes: a "high-porosity" regime for U > Up0 and a "low-porosity" regime for U < Up0, where Up0 is related to the number of pores on the vesicle surface.

  9. Getting to know the extracellular vesicle glycome.

    PubMed

    Gerlach, Jared Q; Griffin, Matthew D

    2016-04-01

    Extracellular vesicles (EVs) are a diverse population of complex biological particles with diameters ranging from approximately 20 to 1000 nm. Tremendous interest in EVs has been generated following a number of recent, high-profile reports describing their potential utility in diagnostic, prognostic, drug delivery, and therapeutic roles. Subpopulations, such as exosomes, are now known to directly participate in cell-cell communication and direct material transfer. Glycomics, the 'omic' portion of the glycobiology field, has only begun to catalog the surface oligosaccharide and polysaccharide structures and also the carbohydrate-binding proteins found on and inside EVs. The EV glycome undoubtedly contains vital clues essential to better understanding the function, biogenesis, release and transfer of vesicles, however getting at this information is technically challenging and made even more so because of the small physical size of the vesicles and the typically minute yield from physiological-scale biological samples. Vesicle micro-heterogeneity which may be related to specific vesicle origins and functions presents a further challenge. A number of primary studies carried out over the past decade have turned up specific and valuable clues regarding the composition and roles of glycan structures and also glycan binding proteins involved EV biogenesis and transfer. This review explores some of the major EV glycobiological research carried out to date and discusses the potential implications of these findings across the life sciences.

  10. Synaptic vesicle recycling: steps and principles

    PubMed Central

    Rizzoli, Silvio O

    2014-01-01

    Synaptic vesicle recycling is one of the best-studied cellular pathways. Many of the proteins involved are known, and their interactions are becoming increasingly clear. However, as for many other pathways, it is still difficult to understand synaptic vesicle recycling as a whole. While it is generally possible to point out how synaptic reactions take place, it is not always easy to understand what triggers or controls them. Also, it is often difficult to understand how the availability of the reaction partners is controlled: how the reaction partners manage to find each other in the right place, at the right time. I present here an overview of synaptic vesicle recycling, discussing the mechanisms that trigger different reactions, and those that ensure the availability of reaction partners. A central argument is that synaptic vesicles bind soluble cofactor proteins, with low affinity, and thus control their availability in the synapse, forming a buffer for cofactor proteins. The availability of cofactor proteins, in turn, regulates the different synaptic reactions. Similar mechanisms, in which one of the reaction partners buffers another, may apply to many other processes, from the biogenesis to the degradation of the synaptic vesicle. PMID:24596248

  11. Lysozyme binding onto cat-anionic vesicles.

    PubMed

    Bonincontro, A; Spigone, E; Ruiz Peña, M; Letizia, C; La Mesa, C

    2006-12-15

    Mixing aqueous sodium dodecylsulfate with cetyltrimethylammonium bromide solutions in mole ratios close to (1.7/1.0) allows the formation of cat-anionic vesicles with an excess of negative charges on the outer surface. The vesicular dispersions are mixed with lysozyme, and interact electrostatically with the positive charges on the protein, forming lipo-plexes. Dielectric relaxation, zeta-potential, and light scattering indicate the occurrence of interactions between vesicles and the protein. According to CD, the vesicle-adsorbed protein retains its native conformation. Binding and surface saturation, inferred by dielectric relaxation and zeta-potential, fulfil a charge neutralisation stoichiometry. Adsorbed lysozyme promotes the vesicle clustering and is concomitant with the lipo-plexes flocculation. Above the charge neutralisation threshold, lysozyme in excess remains dispersed in molecular form. Attempts were made to determine in what conditions protein release from the vesicles occurs. Accordingly, the full neutralisation of sodium dodecylsulfate in excess by cetyltrimethylammonium bromide ensures the lipo-plexes break-up, the precipitation of the mixed surfactants and the protein release in native form.

  12. Activation of calcineurin by phosphotidylserine containing vesicles

    SciTech Connect

    Politino, M.; King, M.M.

    1986-05-01

    Calcineurin (CaN) is a Ca/sup 2 +/- and calmodulin-regulated phosphatase. Recent findings suggested an association of CaN with biological membranes and prompted the present investigation into the interactions of the phosphatase with phospholipids in vitro. In the absence of calmodulin, sonicated preparations of phosphatidylserine (PS) provided a five-fold activation of the Ni- and Mn-supported activities of CaN towards (/sup 32/P) histone Hl; activation in the presence of calmodulin was much less pronounced. Half-maximal activation in the absence of calmodulin required approximately 0.1 mg/ml of PS. Activation of CaN was also observed with mixed vesicles of phosphatidylcholine (PC) containing 20% PS but not with PC alone, or with phosphatidylethanolamine (PE). Molecular sieve chromatography on Ultrogel AcA 34 provided further evidence that CaN associates with phospholipid vesicles composed of PS, or PC containing 20% PS, but not with vesicles of PC or PE. Complete association with medium sized vesicles of PS and PC/PS required Ca/sup 2 +/ ions; in the absence of the metal ion at least 60% of the enzyme failed to interact with the lipids while the remainder preferentially migrated with larger vesicles. These results suggest a role for Ca/sup 2 +/ in regulating CaN's interaction with phospholipids.

  13. Structural Priming: A Critical Review

    PubMed Central

    Pickering, Martin J.; Ferreira, Victor S.

    2009-01-01

    Repetition is a central phenomenon of behavior, and researchers make extensive use of it to illuminate psychological functioning. In the language sciences, a ubiquitous form of such repetition is structural priming, a tendency to repeat or better process a current sentence because of its structural similarity to a previously experienced (“prime”) sentence (Bock, 1986). The recent explosion of research in structural priming has made it the dominant means of investigating the processes involved in the production (and increasingly, comprehension) of complex expressions such as sentences. This review considers its implications for the representation of syntax and the mechanisms of production, comprehension, and their relationship. It then addresses the potential functions of structural priming, before turning to its implications for first language acquisition, bilingualism, and aphasia We close with theoretical and empirical recommendations for future investigations. PMID:18444704

  14. Priming in Systemic Plant Immunity

    SciTech Connect

    Jung, Ho Won; Tschaplinski, Timothy J; Wang, Lin; Glazebrook, Jane; Greenberg, Jean T.

    2009-01-01

    Upon local infection, plants possess inducible systemic defense responses against their natural enemies. Bacterial infection results in the accumulation to high levels of the mobile metabolite C9-dicarboxylic acid azelaic acid in the vascular sap of Arabidopsis. Azelaic acid confers local and systemic resistance against Pseudomonas syringae. The compound primes plants to strongly accumulate salicylic acid (SA), a known defense signal, upon infection. Mutation of a gene induced by azelaic acid (AZI1) results in the specific loss in plants of systemic immunity triggered by pathogen or azelaic acid and of the priming of SA induction. AZI1, a predicted secreted protein, is also important for generating vascular sap that confers disease resistance. Thus, azelaic acid and AZI1 comprise novel components of plant systemic immunity involved in priming defenses.

  15. Symmetry breaking of vesicle shapes in Poiseuille flow.

    PubMed

    Farutin, Alexander; Misbah, Chaouqi

    2011-07-01

    Vesicle behavior under unbounded axial Poiseuille flow is studied analytically. Our study reveals subtle features of the dynamics. It is established that there exists a stable off-centerline steady-state solution for low enough flow strength. This solution appears as a symmetry-breaking bifurcation upon lowering the flow strength and includes slipper shapes, which are characteristic of red blood cells in the microvasculature. A stable axisymmetric solution exists for any flow strength provided the excess area is small enough. It is shown that the mechanism of the symmetry breaking depends on the geometry of the flow: The bifurcation is subcritical in axial Poiseuille flow and supercritical in planar flow.

  16. Two synaptobrevin molecules are sufficient for vesicle fusion in central nervous system synapses.

    PubMed

    Sinha, Raunak; Ahmed, Saheeb; Jahn, Reinhard; Klingauf, Jurgen

    2011-08-23

    Exocytosis of synaptic vesicles (SVs) during fast synaptic transmission is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex assembly formed by the coil-coiling of three members of this protein family: vesicle SNARE protein, synaptobrevin 2 (syb2), and the presynaptic membrane SNAREs syntaxin-1A and SNAP-25. However, it is controversially debated how many SNARE complexes are minimally needed for SV priming and fusion. To quantify this effective number, we measured the fluorescence responses from single fusing vesicles expressing pHluorin (pHl), a pH-sensitive variant of GFP, fused to the luminal domain of the vesicular SNARE syb2 (spH) in cultured hippocampal neurons lacking endogenous syb2. Fluorescence responses were quantal, with the unitary signals precisely corresponding to single pHluorin molecules. Using this approach we found that two copies of spH per SV fully rescued evoked fusion whereas SVs expressing only one spH were unable to rapidly fuse upon stimulation. Thus, two syb2 molecules and likely two SNARE complexes are necessary and sufficient for SV fusion during fast synaptic transmission.

  17. Carbon Nanotubes Mediate Fusion of Lipid Vesicles.

    PubMed

    Bhaskara, Ramachandra M; Linker, Stephanie M; Vögele, Martin; Köfinger, Jürgen; Hummer, Gerhard

    2017-02-28

    The fusion of lipid membranes is opposed by high energetic barriers. In living organisms, complex protein machineries carry out this biologically essential process. Here we show that membrane-spanning carbon nanotubes (CNTs) can trigger spontaneous fusion of small lipid vesicles. In coarse-grained molecular dynamics simulations, we find that a CNT bridging between two vesicles locally perturbs their lipid structure. Their outer leaflets merge as the CNT pulls lipids out of the membranes, creating an hourglass-shaped fusion intermediate with still intact inner leaflets. As the CNT moves away from the symmetry axis connecting the vesicle centers, the inner leaflets merge, forming a pore that completes fusion. The distinct mechanism of CNT-mediated membrane fusion may be transferable, providing guidance in the development of fusion agents, e.g., for the targeted delivery of drugs or nucleic acids.

  18. Directed vesicle transport by diffusio-osmosis

    NASA Astrophysics Data System (ADS)

    Michler, D.; Shahidzadeh, N.; Sprik, R.; Bonn, D.

    2015-04-01

    We present a study on surfactant vesicles that spontaneously move towards an oil droplet that is deposited on a glass substrate. Tracer particles in the surfactant solution show that the motion is not self-propelled: the vesicles are entrained by a macroscopic hydrodynamic flow. Measurements of the flow velocity suggest that the flow is of diffusio-osmotic nature. The surfactant is observed to move into the oil phase which creates a gradient in ion concentration in the vicinity of the droplet. As the diffusion coefficients of the surfactant's co- and counter-ions differ, a charge separation takes place and an electric field arises. This electric field then generates a hydrodynamic flow along the charged glass substrate in which the vesicles are entrained.

  19. Dynamics of fibers growing inside soft vesicles

    NASA Astrophysics Data System (ADS)

    Marenduzzo, D.; Orlandini, E.

    2007-11-01

    We present 3D stochastic dynamic simulations of the growth of a semiflexible polymer inside a soft vesicle. We find that very stiff fibers stall soon and lock the membrane into a strongly deformed prolate shape. Fibers of intermediate stiffness buckle and form a toroidal configuration which distorts the membrane into an oblate shape. Finally, more flexible polymers form massive spool-like condensates with ordered domains, while the vesicle inflates isotropically. We discuss our results with respect to observations on cell shape in sickle red blood cells, developing erythrocytes, and genome packing inside bacteriophages. We quantify how the force felt by the fiber tip, and the vesicle aspect ratio, change during growth, and we discuss possible "synthetic biology" experiments to validate our results.

  20. Ciliary Extracellular Vesicles: Txt Msg Organelles.

    PubMed

    Wang, Juan; Barr, Maureen M

    2016-04-01

    Cilia are sensory organelles that protrude from cell surfaces to monitor the surrounding environment. In addition to its role as sensory receiver, the cilium also releases extracellular vesicles (EVs). The release of sub-micron sized EVs is a conserved form of intercellular communication used by all three kingdoms of life. These extracellular organelles play important roles in both short and long range signaling between donor and target cells and may coordinate systemic responses within an organism in normal and diseased states. EV shedding from ciliated cells and EV-cilia interactions are evolutionarily conserved phenomena, yet remarkably little is known about the relationship between the cilia and EVs and the fundamental biology of EVs. Studies in the model organisms Chlamydomonas and Caenorhabditis elegans have begun to shed light on ciliary EVs. Chlamydomonas EVs are shed from tips of flagella and are bioactive. Caenorhabditis elegans EVs are shed and released by ciliated sensory neurons in an intraflagellar transport-dependent manner. Caenorhabditis elegans EVs play a role in modulating animal-to-animal communication, and this EV bioactivity is dependent on EV cargo content. Some ciliary pathologies, or ciliopathies, are associated with abnormal EV shedding or with abnormal cilia-EV interactions. Until the 21st century, both cilia and EVs were ignored as vestigial or cellular junk. As research interest in these two organelles continues to gain momentum, we envision a new field of cell biology emerging. Here, we propose that the cilium is a dedicated organelle for EV biogenesis and EV reception. We will also discuss possible mechanisms by which EVs exert bioactivity and explain how what is learned in model organisms regarding EV biogenesis and function may provide insight to human ciliopathies.

  1. Ciliary extracellular vesicles: Txt msg orgnlls

    PubMed Central

    Wang, Juan; Barr, Maureen M.

    2016-01-01

    Cilia are sensory organelles that protrude from cell surfaces to monitor the surrounding environment. In addition to its role as sensory receiver, the cilium also releases extracellular vesicles (EVs). The release of sub-micron sized EVs is a conserved form of intercellular communication used by all three kingdoms of life. These extracellular organelles play important roles in both short and long range signaling between donor and target cells and may coordinate systemic responses within an organism in normal and diseased states. EV shedding from ciliated cells and EV-cilia interactions are evolutionarily conserved phenomena, yet remarkably little is known about the relationship between the cilia and EVs and the fundamental biology of EVs. Studies in the model organisms Chlamydomonas and C. elegans have begun to shed light on ciliary EVs. Chlamydomonas EVs are shed from tips of flagella and are bioactive. C. elegans EVs are shed and released by ciliated sensory neurons in an intraflagellar transport (IFT)-dependent manner. C. elegans EVs play a role in modulating animal-to-animal communication, and this EV bioactivity is dependent on EV cargo content. Some ciliary pathologies, or ciliopathies, are associated with abnormal EV shedding or with abnormal cilia-EV interactions, suggest the cilium may be an important organelle as an EV donor or as an EV target. Until the past few decades, both cilia and EVs were ignored as vestigial or cellular junk. As research interest in these two organelles continues to gain momentum, we envision a new field of cell biology emerging. Here, we propose that the cilium is a dedicated organelle for EV biogenesis and EV reception. We will also discuss possible mechanisms by which EVs exert bioactivity and explain how what is learned in model organisms regarding EV biogenesis and function may provide insight to human ciliopathies. PMID:26983828

  2. Elastic energy of polyhedral bilayer vesicles

    NASA Astrophysics Data System (ADS)

    Haselwandter, Christoph A.; Phillips, Rob

    2011-06-01

    In recent experiments [M. Dubois, B. Demé, T. Gulik-Krzywicki, J.-C. Dedieu, C. Vautrin, S. Désert, E. Perez, and T. Zemb, Nature (London)NATUAS0028-083610.1038/35079541 411, 672 (2001)] the spontaneous formation of hollow bilayer vesicles with polyhedral symmetry has been observed. On the basis of the experimental phenomenology it was suggested [M. Dubois, V. Lizunov, A. Meister, T. Gulik-Krzywicki, J. M. Verbavatz, E. Perez, J. Zimmerberg, and T. Zemb, Proc. Natl. Acad. Sci. USAPNASA60027-842410.1073/pnas.0400837101 101, 15082 (2004)] that the mechanism for the formation of bilayer polyhedra is minimization of elastic bending energy. Motivated by these experiments, we study the elastic bending energy of polyhedral bilayer vesicles. In agreement with experiments, and provided that excess amphiphiles exhibiting spontaneous curvature are present in sufficient quantity, we find that polyhedral bilayer vesicles can indeed be energetically favorable compared to spherical bilayer vesicles. Consistent with experimental observations we also find that the bending energy associated with the vertices of bilayer polyhedra can be locally reduced through the formation of pores. However, the stabilization of polyhedral bilayer vesicles over spherical bilayer vesicles relies crucially on molecular segregation of excess amphiphiles along the ridges rather than the vertices of bilayer polyhedra. Furthermore, our analysis implies that, contrary to what has been suggested on the basis of experiments, the icosahedron does not minimize elastic bending energy among arbitrary polyhedral shapes and sizes. Instead, we find that, for large polyhedron sizes, the snub dodecahedron and the snub cube both have lower total bending energies than the icosahedron.

  3. Structure of a micropipette-aspirated vesicle determined from the bending-energy model.

    PubMed

    Chen, Jeff Z Y

    2012-10-01

    The structure of the system consisting of an aspirating pipette and an aspirated vesicle is investigated with fixed total vesicle volume, total vesicle surface area, and aspirated volume fraction, based on the bending-energy model. Through an energetic consideration, the usage of an aspirated volume fraction can be converted to the aspirating pressure for the determination of a phase diagram; the procedure identifies a first-order transition, between a weakly aspirated state and the strongly aspirated state, as the pressure increases. The physical properties of the system are obtained from minimization of the bending energy by an implementation of the simulated annealing Monte Carlo procedure, which searches for a minimum in a multivariable space. An analysis of the hysteresis effects indicates that the experimentally observed aspirating and releasing critical pressures are related to the location of the spinodal points.

  4. On the control of single-prime negative priming: the effects of practice and time course.

    PubMed

    Chao, Hsuan-Fu

    2009-09-01

    Single-prime negative priming refers to the phenomenon wherein repetition of a prime as the probe target results in delayed response. Sometimes this effect has been found to be contingent on participants' unawareness of the primes, and sometimes it has not. Further, sometimes this effect has been found to be eliminated when the prime could predict the following probe target, and sometimes it has not. An integrative account is postulated to account for these findings. Three experiments supported this account by demonstrating that (a) regardless of the proportion of prime repetition, negative priming was the default effect; (b) the control mechanism was triggered to activate the prime after there was enough practice for the detection of the contingency between the prime and probe; and (c) it took time for the control mechanism to overcome the negative-priming effect and produce a positive-priming effect.

  5. Toroidal membrane vesicles in spherical confinement.

    PubMed

    Bouzar, Lila; Menas, Ferhat; Müller, Martin Michael

    2015-09-01

    We investigate the morphology of a toroidal fluid membrane vesicle confined inside a spherical container. The equilibrium shapes are assembled in a geometrical phase diagram as a function of scaled area and reduced volume of the membrane. For small area the vesicle can adopt its free form. When increasing the area, the membrane cannot avoid contact and touches the confining sphere along a circular contact line, which extends to a zone of contact for higher area. The elastic energies of the equilibrium shapes are compared to those of their confined counterparts of spherical topology to predict under which conditions a topology change is favored energetically.

  6. Toroidal membrane vesicles in spherical confinement

    NASA Astrophysics Data System (ADS)

    Bouzar, Lila; Menas, Ferhat; Müller, Martin Michael

    2015-09-01

    We investigate the morphology of a toroidal fluid membrane vesicle confined inside a spherical container. The equilibrium shapes are assembled in a geometrical phase diagram as a function of scaled area and reduced volume of the membrane. For small area the vesicle can adopt its free form. When increasing the area, the membrane cannot avoid contact and touches the confining sphere along a circular contact line, which extends to a zone of contact for higher area. The elastic energies of the equilibrium shapes are compared to those of their confined counterparts of spherical topology to predict under which conditions a topology change is favored energetically.

  7. Short-term synaptic depression and stochastic vesicle dynamics reduce and shape neuronal correlations.

    PubMed

    Rosenbaum, Robert; Rubin, Jonathan E; Doiron, Brent

    2013-01-01

    Correlated neuronal activity is an important feature in many neural codes, a neural correlate of a variety of cognitive states, as well as a signature of several disease states in the nervous system. The cellular and circuit mechanics of neural correlations is a vibrant area of research. Synapses throughout the cortex exhibit a form of short-term depression where increased presynaptic firing rates deplete neurotransmitter vesicles, which transiently reduces synaptic efficacy. The release and recovery of these vesicles are inherently stochastic, and this stochasticity introduces variability into the conductance elicited by depressing synapses. The impact of spiking and subthreshold membrane dynamics on the transfer of neuronal correlations has been studied intensively, but an investigation of the impact of short-term synaptic depression and stochastic vesicle dynamics on correlation transfer is lacking. We find that short-term synaptic depression and stochastic vesicle dynamics can substantially reduce correlations, shape the timescale over which these correlations occur, and alter the dependence of spiking correlations on firing rate. Our results show that short-term depression and stochastic vesicle dynamics need to be taken into account when modeling correlations in neuronal populations.

  8. A small pool of vesicles maintains synaptic activity in vivo

    PubMed Central

    Denker, Annette; Bethani, Ioanna; Kröhnert, Katharina; Körber, Christoph; Horstmann, Heinz; Wilhelm, Benjamin G.; Barysch, Sina V.; Kuner, Thomas; Neher, Erwin; Rizzoli, Silvio O.

    2011-01-01

    Chemical synapses contain substantial numbers of neurotransmitter-filled synaptic vesicles, ranging from approximately 100 to many thousands. The vesicles fuse with the plasma membrane to release neurotransmitter and are subsequently reformed and recycled. Stimulation of synapses in vitro generally causes the majority of the synaptic vesicles to release neurotransmitter, leading to the assumption that synapses contain numerous vesicles to sustain transmission during high activity. We tested this assumption by an approach we termed cellular ethology, monitoring vesicle function in behaving animals (10 animal models, nematodes to mammals). Using FM dye photooxidation, pHluorin imaging, and HRP uptake we found that only approximately 1–5% of the vesicles recycled over several hours, in both CNS synapses and neuromuscular junctions. These vesicles recycle repeatedly, intermixing slowly (over hours) with the reserve vesicles. The latter can eventually release when recycling is inhibited in vivo but do not seem to participate under normal activity. Vesicle recycling increased only to ≈5% in animals subjected to an extreme stress situation (frog predation on locusts). Synapsin, a molecule binding both vesicles and the cytoskeleton, may be a marker for the reserve vesicles: the proportion of vesicles recycling in vivo increased to 30% in synapsin-null Drosophila. We conclude that synapses do not require numerous reserve vesicles to sustain neurotransmitter release and thus may use them for other purposes, examined in the accompanying paper. PMID:21903928

  9. Prime Suspect, Second Row Center

    ERIC Educational Resources Information Center

    Laird, Ellen A.

    2011-01-01

    His father had been hacked to death in his own bed with an ax the previous November. His mother was similarly brutalized and left for dead with her husband but survived. On the last Monday of that August, after several months and many investigative twists, turns, and fumbles, there sat the son--the prime suspect--in Ellen Laird's literature class,…

  10. The Search for Prime Numbers.

    ERIC Educational Resources Information Center

    Pomerance, Carl

    1982-01-01

    Until recently the testing of a 100-digit number to determine whether it is prime or composite could have taken a century. However, in the past two years a method has been developed enabling a computer to determine the primality of an arbitrary number in about 40 seconds of running time. (Author/JN)

  11. Evaluative Priming in the Pronunciation Task.

    PubMed

    Klauer, Karl Christoph; Becker, Manuel; Spruyt, Adriaan

    2016-01-01

    We replicated and extended a study by Spruyt and Hermans (2008) in which picture primes engendered an evaluative-priming effect on the pronunciation of target words. As preliminary steps, we assessed data reproducibility of the original study, conducted Pilot Study I to identify highly semantically related prime-target pairs, reanalyzed the original data excluding such pairs, conducted Pilot Study II to demonstrate that we can replicate traditional associative priming effects in the pronunciation task, and conducted Pilot Study III to generate relatively unrelated sets of prime pictures and target words. The main study comprised three between-participants conditions: (1) a close replication of the original study, (2) the same condition excluding highly related prime-target pairs, and (3) a condition based on the relatively unrelated sets of prime pictures and target words developed in Pilot Study III. There was little evidence for an evaluative priming effect independent of semantic relatedness.

  12. The Impact of Emotional Priming on MMPI-2 Scale Scores

    ERIC Educational Resources Information Center

    Lee, Tayla T. C.; Forbey, Johnathan D.; Ritchey, Kristin A.

    2011-01-01

    The current study investigated potential emotional priming effects on Minnesota Multiphasic Personality Inventory-2 (MMPI-2) scale scores. Participants included 98 college students who completed a personal narrative intended to induce temporary mood states, the MMPI-2, and a mood rating inventory. Results of the mood manipulation indicated that…

  13. Prime Minister Pierre Trudeau and Cruise Missile Testing.

    ERIC Educational Resources Information Center

    Bennett, Paul W.

    1986-01-01

    Based on the 1983 controversy over cruise missile testing by the United States over Canadian air space, this article provides the text of an open letter to the people and an interview by Prime Minister Trudeau. Parenthetical comments inserted by the author point out contradiction contained in the two documents. (JDH)

  14. Bioaccumulation of dietary 2,2{prime},4,4{prime},5,5{prime}-hexachlorobiphenyl and induction of hepatic arylhydrocarbon hydroxylase in rainbow trout (Oncorhynchus mykiss)

    SciTech Connect

    Costa, E.G. da; Curtis, L.R.

    1995-10-01

    Juvenile rainbow trout (Oncorhynchus mykiss) were fed either 5 or 20 {micro}g, 2,2{prime},4,4{prime},5,5{prime}-hexachlorobiphenyl (245-HxCB)/g diet (wet wt.) for 4,8, or 12 weeks. Hepatic xenobiotic-metabolizing enzyme activities and dietary 245-HxCB accumulation in liver, muscle, and remaining carcass were determined. Liver-to-body weight ratios were not altered by either of the two 245-HxCB concentrations. Relative growth rate increased with time but was not altered by 245-HxCB concentration. Bioaccumulation of 245-HxCB was dose and time dependent in all tissues without reaching apparent stayed state. Hepatic arylhydrocarbon hydroxylase (AHH) activities increased with 245-HxCB dose and with time. Ethoxyresorufin-O-deethylase (EROD) activities also increased in fish fed 20 {micro}g 245-HxCB/g diet. No 245-HxCB-induced changes in uridine diphosphoglucuronosyl transferase (UDP-DT) or NADPH-dytochrome-c reductase (NCCR) activities were determined. High-resolution GC-MS analysis of the 245-HxCB standard revealed trace (0.4--0.5%) contamination by two mono-ortho pentachlorobiphenyls (PnCBs): 2,3,3{prime},4,4{prime}-PnCB and 2,3,4,4{prime},5-PnCB. Total liver accumulation of these contaminants was inversely related with corresponding EROD and AHH activities and estimated to contribute minimally to their induction. Results from this study suggested that long-term dietary 245-HxCB exposure induced cytochrome P4501A activities in rainbow trout liver.

  15. Calcium uptake by intestinal brush border membrane vesicles. Comparison with in vivo calcium transport.

    PubMed Central

    Schedl, H P; Wilson, H D

    1985-01-01

    In prior studies, we examined kinetics of steady state in vivo transepithelial calcium transport in rat and hamster. The present studies related calcium uptake by the brush border to in vivo transport. We measured calcium uptake by brush border membrane vesicles from the two species. In the rat, our prior in vivo studies had shown that (a) calcium transport was mediated, (b) no nonmediated component was detectable, and (c) Vmax was 2.5 times greater in proximal than distal small intestine. In brush border membrane vesicles from the rat, Vmax for the saturable component of calcium uptake was again 2.5 times greater in proximal than distal intestine. Contrasting with in vivo studies, a major nonsaturable component was present in vesicles from proximal and distal small intestine. In the hamster, our previous in vivo studies had shown (1) both mediated and nonmediated components of calcium transport, (2) greater nonmediated transport in proximal than distal small intestines, and (3) Vmax for calcium transport twice as great in distal as in proximal small intestine. In the present study with brush border membrane vesicles from hamster, Vmax for saturable calcium transport was again twice as great in distal as in proximal small intestine. However, nonsaturable calcium transport rates relative to saturable rates were much greater with vesicles than in in vivo studies, and were greater in vesicles from distal than proximal small intestine. Since rates of saturable calcium uptake by brush border membrane vesicles parallel corresponding in vivo mediated transport rates, we conclude that the segmental rates of calcium transport in rat and hamster could be determined by brush border function. PMID:2997294

  16. Competition Effects in Phonological Priming: The Role of Mismatch Position between Primes and Targets

    ERIC Educational Resources Information Center

    Dufour, Sophie; Peereman, Ronald

    2009-01-01

    In three experiments, we examined lexical competition effects using the phonological priming paradigm in a shadowing task. Experiments 1A and 1B showed that an inhibitory priming effect occurred when the primes mismatched the targets on the last phoneme (/bagar/-/bagaj/). In contrast, a facilitatory priming effect was observed when the primes…

  17. Evidence for Episodic Retrieval of Inadequate Prime Responses in Auditory Negative Priming

    ERIC Educational Resources Information Center

    Mayr, Susanne; Buchner, Axel

    2006-01-01

    Four experiments are reported in which the mechanisms underlying auditory negative priming were investigated. In Experiments 1A and 1B, preprime-prime intervals and prime-probe intervals were manipulated.The ratio between the 2 intervals determined the size of the negative priming effect. Results are compatible with the episodic retrieval account,…

  18. Affective Priming with Associatively Acquired Valence

    ERIC Educational Resources Information Center

    Aguado, Luis; Pierna, Manuel; Saugar, Cristina

    2005-01-01

    Three experiments explored the effect of affectively congruent or incongruent primes on evaluation responses to positive or negative valenced targets (the "affective priming" effect). Experiment 1 replicated the basic affective priming effect with Spanish nouns: reaction time for evaluative responses (pleasant/unpleasant) were slower on…

  19. Priming Lexical Stress in Reading Italian Aloud

    ERIC Educational Resources Information Center

    Sulpizio, Simone; Job, Remo; Burani, Cristina

    2012-01-01

    Two experiments using a lexical priming paradigm investigated how stress information is processed in reading Italian words. In both experiments, prime and target words either shared the stress pattern or they had different stress patterns. We expected that lexical activation of the prime would favour the assignment of congruent stress to the…

  20. Structural Priming and Second Language Learning

    ERIC Educational Resources Information Center

    Shin, Jeong-Ah; Christianson, Kiel

    2012-01-01

    Structural priming (or syntactic priming) is a speaker's tendency to reuse the same structural pattern as one that was previously encountered (Bock, 1986). This study investigated (a) whether the implicit learning processes involved in long-lag structural priming lead to differential second language (L2) improvement in producing two structural…

  1. Spatial priming in ecologically relevant reference frames.

    PubMed

    Tower-Richardi, Sarah M; Leber, Andrew B; Golomb, Julie D

    2016-01-01

    In recent years, researchers have observed many phenomena demonstrating how the visual system exploits spatial regularities in the environment in order to benefit behavior. In this paper, we question whether spatial priming can be considered one such phenomenon. Spatial priming is defined as a response time facilitation to a visual search target when its spatial position has been repeated in recent trials (Maljkovic & Nakayama, 1996, Perception & Psychophysics, 58, 977-991). Does this priming serve a behaviorally adaptive role or is it merely a byproduct of ongoing visual processing? Critically, an adaptive priming mechanism must actively transform visual inputs from native retinotopic (eye-centered) coordinates into ecologically relevant coordinates, e.g., spatiotopic (world-centered) and/or object-centered. In Experiment 1, we tested this hypothesis by having participants move their eyes between trials, which dissociated retinotopic and spatiotopic frames of reference. Results showed only weak retinotopic priming, but robust spatiotopic priming. The second experiment again had participants move their eyes between trials but also manipulated the placement of a grouped array of display objects from trial to trial. This allowed us to measure not just retinotopic and spatiotopic priming, but object-centered priming as well. Results from this experiment did not yield retinotopic priming but showed robust spatiotopic and object-centered priming. These findings demonstrate that spatial priming operates within ecologically relevant coordinate systems, and the findings support the notion that spatial priming serves an adaptive role in human behavior.

  2. Affective Priming with Auditory Speech Stimuli

    ERIC Educational Resources Information Center

    Degner, Juliane

    2011-01-01

    Four experiments explored the applicability of auditory stimulus presentation in affective priming tasks. In Experiment 1, it was found that standard affective priming effects occur when prime and target words are presented simultaneously via headphones similar to a dichotic listening procedure. In Experiment 2, stimulus onset asynchrony (SOA) was…

  3. Semantic priming with product verification but not production.

    PubMed

    Campbell, Jamie I D

    2011-12-01

    Campbell and Reynvoet (2009) found that time to name a single-digit target was about 8 ms faster if preceded by a near prime (±1) compared to a far prime (at least ±3) when prime-digit pairs were interleaved with number comparisons (9↑3; name larger) and not when they were interleaved with multiplication problems (9×3; state product). This is consistent with the claim by previous researchers that magnitude comparison can enable a semantic pathway for digit naming whereas number-fact retrieval can inhibit it. To pursue this, the current study compared priming in the context of multiplication production (9×3=?) versus multiplication verification (e.g., 9×3=24, true or false). Multiplication production, but not verification, may inhibit semantic digit naming to reduce naming-related interference with verbal number production. Indeed, semantic priming of digit naming occurred only in verification and not production blocks. This supports the conclusion that multiplication production can inhibit semantic mediation of digit naming, which is enabled in other number processing tasks (e.g., comparison, verification) that do not compete with naming for verbal number production processes.

  4. Preparation of vesicles entrapped lycopene extract.

    PubMed

    Luxsuwong, Dhitaree; Indranupakorn, Ratana; Wongtrakul, Paveena

    2014-01-01

    Lycopene, a lipophilic carotenoid, has been known as an effective antioxidant in supporting the cutaneous defensive system. However, it is unstable when exposed to light and water. In this study, lycopene was isolated from tomatoes and a vesicular delivery system was developed to entrap and stabilize the lycopene in the aqueous system. A simple process, maceration in ethyl acetate, was used to extract lycopene from the tomatoes. The extract was then chromatographed on the Sephadex LH20 column using acetone as a solvent system to yield 995 μg of lycopene per gram of dried tomato weight. The vesicular delivery system was prepared from a combination of ascorbic acid-6-palmitate (AP), cholesterol and dicetyl phosphate using a thin film hydration method. The formulation was composed of AP, cholesterol and dicetyl phosphate at a 44:44:12 molar ratio and with 2.12 μmol/ml of the isolated lycopene. Both blank vesicles and lycopene loaded vesicles were kept for a period of 3 months at 4±2°C and at the room temperature (28±2°C) to evaluate the effect of the encapsulation on the characteristic of the vesicles and on the antioxidant activity of the encapsulated lycopene. The result implied that lycopene could be stabilized in the vesicles and its scavenging activity against DPPH free radicals was superior to that of the free lycopene solution.

  5. Matrix vesicles: Are they anchored exosomes?

    PubMed

    Shapiro, Irving M; Landis, William J; Risbud, Makarand V

    2015-10-01

    Numerous studies have documented that matrix vesicles are unique extracellular membrane-bound microparticles that serve as initial sites for mineral formation in the growth plate and most other vertebrate mineralizing tissues. Microparticle generation is not confined to hard tissues, as cells in soft tissues generate similar structures; numerous studies have shown that a common type of extracellular particle, termed an exosome, a product of the endosomal pathway, shares many characteristics of matrix vesicles. Indeed, analyses of size, morphology and lipid and protein content indicate that matrix vesicles and exosomes are homologous structures. Such a possibility impacts our understanding of the biogenesis, processing and function of matrix vesicles (exosomes) in vertebrate hard tissues and explains in part how cells control the earliest stages of mineral deposition. Moreover, since exosomes influence a spectrum of functions, including cell-cell communication, it is suggested that this type of microparticle may provide a mechanism for the transfer of signaling molecules between cells within the growth plate and thereby regulate endochondral bone development and formation.

  6. The Bretherton Problem for a Vesicle

    NASA Astrophysics Data System (ADS)

    Barakat, Joseph; Spann, Andrew; Shaqfeh, Eric

    2016-11-01

    The motion of a lipid bilayer vesicle through a circular tube is investigated by singular perturbation theory in the limit of vanishing clearance. The vesicle is treated as a sac of fluid enclosed by a thin, elastic sheet that admits a bending stiffness. It is assumed that the vesicle is axisymmetric and swollen to a near-critical volume such that the clearance "e" between the membrane and the tube wall is very small. In this limit, bending resistance is of negligible importance compared to the isotropic tension, allowing the vesicle to be treated as a "no-slip bubble." The effective membrane tension is found to scale inversely with "e" raised to the 3/2 power with a comparatively weak Marangoni gradient. The extra pressure drop is found to have a leading contribution due to the cylindrical midsection, which scales inversely with "e," as well as a correction due to the end caps, which scales inversely with the square root of "e." The apparent viscosity is predicted as a unique function of the geometry. The theory exhibits excellent agreement with a simplified, "quasi-parallel" theory and with direct numerical simulations using the boundary element method. The results of this work are compared to those for bubbles, rigid particles, and red blood cells in confined flows.

  7. Vesicle dynamics in shear and capillary flows

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi; Gompper, Gerhard

    2005-11-01

    The deformation of vesicles in flow is studied by a mesoscopic simulation technique, which combines multi-particle collision dynamics for the solvent with a dynamically triangulated surface model for the membrane. Shape transitions are investigated both in simple shear flows and in cylindrical capillary flows. We focus on reduced volumes, where the discocyte shape of fluid vesicles is stable, and the prolate shape is metastable. In simple shear flow at low membrane viscosity, the shear induces a transformation from discocyte to prolate with increasing shear rate, while at high membrane viscosity, the shear induces a transformation from prolate to discocyte, or tumbling motion accompanied by oscillations between these two morphologies. In capillary flow, at small flow velocities the symmetry axis of the discocyte is found not to be oriented perpendicular to the cylinder axis. With increasing flow velocity, a transition to a prolate shape occurs for fluid vesicles, while vesicles with shear-elastic membranes (like red blood cells) transform into a coaxial parachute-like shape.

  8. Compartmentalization and Transport in Synthetic Vesicles

    PubMed Central

    Schmitt, Christine; Lippert, Anna H.; Bonakdar, Navid; Sandoghdar, Vahid; Voll, Lars M.

    2016-01-01

    Nanoscale vesicles have become a popular tool in life sciences. Besides liposomes that are generated from phospholipids of natural origin, polymersomes fabricated of synthetic block copolymers enjoy increasing popularity, as they represent more versatile membrane building blocks that can be selected based on their specific physicochemical properties, such as permeability, stability, or chemical reactivity. In this review, we focus on the application of simple and nested artificial vesicles in synthetic biology. First, we provide an introduction into the utilization of multicompartmented vesosomes as compartmentalized nanoscale bioreactors. In the bottom-up development of protocells from vesicular nanoreactors, the specific exchange of pathway intermediates across compartment boundaries represents a bottleneck for future studies. To date, most compartmented bioreactors rely on unspecific exchange of substrates and products. This is either based on changes in permeability of the coblock polymer shell by physicochemical triggers or by the incorporation of unspecific porin proteins into the vesicle membrane. Since the incorporation of membrane transport proteins into simple and nested artificial vesicles offers the potential for specific exchange of substances between subcompartments, it opens new vistas in the design of protocells. Therefore, we devote the main part of the review to summarize the technical advances in the use of phospholipids and block copolymers for the reconstitution of membrane proteins. PMID:26973834

  9. Vesicle deformation by microtubules: A phase diagram

    NASA Astrophysics Data System (ADS)

    Emsellem, Virginie; Cardoso, Olivier; Tabeling, Patrick

    1998-10-01

    The experimental investigation of vesicles deformed by the growth of encapsulated microtubules shows that the axisymmetric morphologies can be classified into ovals, lemons, φ, cherries, dumbbells, and pearls. A geometrical phase diagram is established. Numerical minimization of the elastic energy of the membrane reproduces satisfactorily well the observed morphologies and the corresponding phase diagram.

  10. Amino Acid Neurotransmitters; Mechanisms of Their Uptake into Synaptic Vesicles

    DTIC Science & Technology

    1991-08-01

    same neuron, at least from the cerebellar Golgi cell terminals. It should be kept in mind that the uptake of noradrenaline and dopamine in synaptic...vesicles prepared from rat brain is relatively non-specific. Noradrenaline containing vesicles can take up noradrenaline, dopamine and serotonin. In...also shown that the vesicles isolated from corpus striatum exhibited the same ratio of uptake of dopamine /noradrenaline as did vesicles from cerebral

  11. Automatic non-associative semantic priming: episodic affective priming of naming responses.

    PubMed

    Spruyt, Adriaan; Hermans, Dirk; De Houwer, Jan; Eelen, Paul

    2004-05-01

    Affective priming for associatively unrelated prime-target pairs was investigated using (a) the naming task, (b) a short stimulus onset asynchrony (250 ms), and (c) primes that had acquired their affective connotation during a differential evaluative conditioning procedure. Despite the fact that the primes and the targets were related on the affective dimension only, significant priming emerged. This finding indicates that mere affective overlap is sufficient to produce automatic priming. As such, our results are in line with theoretical accounts of automatic priming that are based on semantic relatedness.

  12. Semantic priming in the prime task effect: evidence of automatic semantic processing of distractors.

    PubMed

    Marí-Beffa, P; Fuentes, L J; Catena, A; Houghton, G

    2000-06-01

    The automaticity of the semantic processing of words has been questioned because of the reduction of semantic priming when the prime word is processed nonsemantically--for example, in letter search (the prime task effect). In two experiments, prime distractor words produced semantic priming in a subsequent lexical decision task, but with the direction of priming (positive or negative) depending on the prime task. Lexico-semantic tasks produced negative semantic priming, whereas letter search produced positive semantic priming. These results are discussed in terms of task-based inhibition. We argue that, given the results from the distractors, the absence of semantic priming does not indicate an absence of semantic activation but reflects the action of control processes on prepotent responses when less practiced responses are needed.

  13. Calcium regulates vesicle replenishment at the cone ribbon synapse.

    PubMed

    Babai, Norbert; Bartoletti, Theodore M; Thoreson, Wallace B

    2010-11-24

    Cones release glutamate-filled vesicles continuously in darkness, and changing illumination modulates this release. Because sustained release in darkness is governed by vesicle replenishment rates, we analyzed how cone membrane potential regulates replenishment. Synaptic release from cones was measured by recording postsynaptic currents in Ambystoma tigrinum horizontal or OFF bipolar cells evoked by depolarization of simultaneously voltage-clamped cones. We measured replenishment after attaining a steady state between vesicle release and replenishment using trains of test pulses. Increasing Ca(2+) currents (I(Ca)) by changing the test step from -30 to -10 mV increased replenishment. Lengthening -30 mV test pulses to match the Ca(2+) influx during 25 ms test pulses to -10 mV produced similar replenishment rates. Reducing Ca(2+) driving force by using test steps to +30 mV slowed replenishment. Using UV flashes to reverse inhibition of I(Ca) by nifedipine accelerated replenishment. Increasing [Ca(2+)](i) by flash photolysis of caged Ca(2+) also accelerated replenishment. Replenishment, but not the initial burst of release, was enhanced by using an intracellular Ca(2+) buffer of 0.5 mm EGTA rather than 5 mm EGTA, and diminished by 1 mm BAPTA. This suggests that although release and replenishment exhibited similar Ca(2+) dependencies, release sites are <200 nm from Ca(2+) channels but replenishment sites are >200 nm away. Membrane potential thus regulates replenishment by controlling Ca(2+) influx, principally by effects on replenishment mechanisms but also by altering releasable pool size. This in turn provides a mechanism for converting changes in light intensity into changes in sustained release at the cone ribbon synapse.

  14. In vitro fusion between Saccharomyces cerevisiae secretory vesicles and cytoplasmic-side-out plasma membrane vesicles.

    PubMed Central

    Arrastua, Lorena; San Sebastian, Eider; Quincoces, Ana F; Antony, Claude; Ugalde, Unai

    2003-01-01

    The final step in the secretory pathway, which is the fusion event between secretory vesicles and the plasma membrane, was reconstructed using highly purified secretory vesicles and cytoplasmic-side-out plasma membrane vesicles from the yeast Saccharomyces cerevisiae. Both organelle preparations were obtained from a sec 6-4 temperature-sensitive mutant. Fusion was monitored by means of a fluorescence assay based on the dequenching of the lipophilic fluorescent probe octadecylrhodamine B-chloride (R18). The probe was incorporated into the membrane of secretory vesicles, and it diluted in unlabelled cytoplasmic-side-out plasma membrane vesicles as the fusion process took place. The obtained experimental dequenching curves were found by mathematical analysis to consist of two independent but simultaneous processes. Whereas one of them reflected the fusion process between both vesicle populations as confirmed by its dependence on the assay conditions, the other represented a non-specific transfer of the probe. The fusion process may now be examined in detail using the preparation, validation and analytical methods developed in this study. PMID:12435271

  15. Formation of polyhedral vesicles and polygonal membrane tubes induced by banana-shaped proteins

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2015-12-01

    The shape transformations of fluid membranes induced by curved protein rods are studied using meshless membrane simulations. The rod assembly at low rod density induces a flat membrane tube and oblate vesicle. It is found that the polyhedral shapes are stabilized at high rod densities. The discrete shape transition between triangular and buckled discoidal tubes is obtained and their curvature energies are analyzed by a simple geometric model. For vesicles, triangular hosohedron and elliptic-disk shapes are formed in equilibrium, whereas tetrahedral and triangular prism shapes are obtained as metastable states.

  16. Particle filtering for tracking of GLUT4 vesicles in TIRF microscpy

    NASA Astrophysics Data System (ADS)

    Wu, Xiangping; Liu, Xiaofang; Xu, Wenglong; Yan, Dandan; Chen, Yongli

    2009-10-01

    GLUT4 is responsible for insulin-stimulated glucose uptake into fat cells and description of the dynamic behavior of it can give insight in some working mechanisms and structures of these cells. Quantitative analysis of the dynamical process requires tracking of hundreds of GLUT4 vesicles characterized as bright spots in noisy image sequences. In this paper, a 3D tracking algorithm built in Bayesian probabilistic framework is put forward, combined with the unique features of the TIRF microscopy. A brightness-correction procedure is firstly applied to ensure that the intensity of a vesicle is constant along time and is only affected by spatial factors. Then, tracking is formalized as a state estimation problem and a developed particle filter integrated by a sub-optimizer that steers the particles towards a region with high likelihood is used. Once each tracked vesicle is located in image plane, the depth information of a granule can be indirectly inferred according to the exponential relationship between its intensity and its vertical position. The experimental results indicate that the vesicles are tracked well under different motion styles. More, the algorithm provides the depth information of the tracked vesicle.

  17. The effect of spontaneous curvature on a two-phase vesicle

    PubMed Central

    Cox, Geoffrey; Lowengrub, John

    2015-01-01

    Vesicles are membrane-bound structures commonly known for their roles in cellular transport and the shape of a vesicle is determined by its surrounding membrane (lipid bilayer). When the membrane is composed of different lipids, it is natural for the lipids of similar molecular structure to migrate towards one another (via spinodal decomposition), creating a multi-phase vesicle. In this article, we consider a two-phase vesicle model which is driven by nature’s propensity to maintain a minimal state of elastic energy. The model assumes a continuum limit, thereby treating the membrane as a closed three-dimensional surface. The main purpose of this study is to reveal the complexity of the Helfrich two-phase vesicle model with non-zero spontaneous curvature and provide further evidence to support the relevance of spontaneous curvature as a modelling parameter. In this paper, we illustrate the complexity of the Helfrich two-phase model by providing multiple examples of undocumented solutions and energy hysteresis. We also investigate the influence of spontaneous curvature on morphological effects and membrane phenomena such as budding and fusion. PMID:26097287

  18. Dissecting docking and tethering of secretory vesicles at the target membrane

    PubMed Central

    Toonen, Ruud F; Kochubey, Olexiy; de Wit, Heidi; Gulyas-Kovacs, Attila; Konijnenburg, Bas; Sørensen, Jakob B; Klingauf, Jurgen; Verhage, Matthijs

    2006-01-01

    Secretory vesicles dock at their target in preparation for fusion. Using single-vesicle total internal reflection fluorescence microscopy in chromaffin cells, we show that most approaching vesicles dock only transiently, but that some are captured by at least two different tethering modes, weak and strong. Both vesicle delivery and tethering depend on Munc18-1, a known docking factor. By decreasing the amount of cortical actin by Latrunculin A application, morphological docking can be restored artificially in docking-deficient munc18-1 null cells, but neither strong tethering nor fusion, demonstrating that morphological docking is not sufficient for secretion. Deletion of the t-SNARE and Munc18-1 binding partner syntaxin, but not the v-SNARE synaptobrevin/VAMP, also reduces strong tethering and fusion. We conclude that docking vesicles either undock immediately or are captured by minimal tethering machinery and converted in a munc18-1/syntaxin-dependent, strongly tethered, fusion-competent state. PMID:16902411

  19. Material Studies of Lipid Vesicles in the Lα and Lα-Gel Coexistence Regimes

    PubMed Central

    Shoemaker, Scott D.; Vanderlick, T. Kyle

    2003-01-01

    In this work, we utilize micropipette aspiration and fluorescence imaging to examine the material properties of lipid vesicles made from mixtures of palmitoyloleoylphosphocholine (POPC) and dipalmitoylphosphatidylcholine (DPPC). At elevated temperatures/low DPPC fractions, these lipids are in a miscible liquid crystalline (Lα) state, whereas at lower temperatures/higher DPPC fractions they phase-separate into Lα and gel phases. We show that the elastic modulus, K, and critical tension, τc, of Lα vesicles are independent of DPPC fraction. However, as the sample temperature is increased from 15°C to 45°C, we measure decreases in both K and τc of 20% and 50%, respectively. The elasticity change is likely driven by a change in interfacial tension. We describe the reduction in critical tension using a simple model of thermally activated membrane pores. Vesicles with two-phase coexistence exhibit material properties that differ from Lα vesicles including critical tensions that are 20–40% lower. Fluorescence imaging of phase coexistent POPC/DPPC vesicles shows that the DPPC-rich domains exist in an extended network structure that exhibits characteristics of a solid. This gel network explains many of the unusual material properties of two-phase membranes. PMID:12547781

  20. Comparison of Prime Movers Suitable for USMC Expeditionary Power Sources

    SciTech Connect

    Theiss, T.J.

    2000-04-18

    This report documents the results of the ORNL investigation into prime movers that would be desirable for the construction of a power system suitable for the United States Marine Corps (USMC) expeditionary forces under Operational Maneuvers From The Sea (OMFTS) doctrine. Discrete power levels of {approx}1, 5, 15, and 30 kW are considered. The only requirement is that the prime mover consumes diesel fuel. A brief description is given for the prime movers to describe their basic scientific foundations and relative advantages and disadvantages. A list of key attributes developed by ORNL has been weighted by the USMC to indicate the level of importance. A total of 14 different prime movers were scored by ORNL personnel in four size ranges (1,5, 15, & 30 kW) for their relative strength in each attribute area. The resulting weighted analysis was used to indicate which prime movers are likely to be suitable for USMC needs. No single engine or prime mover emerged as the clear-cut favorite but several engines scored as well or better than the diesel engine. At the higher load levels (15 & 30 kW), the results indicate that the open Brayton (gas turbine) is a relatively mature technology and likely a suitable choice to meet USMC needs. At the lower power levels, the situation is more difficult and the market alone is not likely to provide an optimum solution in the time frame desired (2010). Several prime movers should be considered for future developments and may be satisfactory; specifically, the Atkinson cycle, the open Brayton cycle (gas turbine), the 2-stroke diesel. The rotary diesel and the solid oxide fuel cell should be backup candidates. Of all these prime movers, the Atkinson cycle may well be the most suitable for this application but is an immature technology. Additional demonstrations of this engine will be conducted at ORNL. If this analysis is positive, then the performance of a generator set using this engine, the open Brayton and the 2-stroke diesel should

  1. Microfluidic approaches for isolation, detection, and characterization of extracellular vesicles: Current status and future directions.

    PubMed

    Gholizadeh, Shima; Shehata Draz, Mohamed; Zarghooni, Maryam; Sanati-Nezhad, Amir; Ghavami, Saeid; Shafiee, Hadi; Akbari, Mohsen

    2017-05-15

    Extracellular vesicles (EVs) are cell-derived vesicles present in body fluids that play an essential role in various cellular processes, such as intercellular communication, inflammation, cellular homeostasis, survival, transport, and regeneration. Their isolation and analysis from body fluids have a great clinical potential to provide information on a variety of disease states such as cancer, cardiovascular complications and inflammatory disorders. Despite increasing scientific and clinical interest in this field, there are still no standardized procedures available for the purification, detection, and characterization of EVs. Advances in microfluidics allow for chemical sampling with increasingly high spatial resolution and under precise manipulation down to single molecule level. In this review, our objective is to give a brief overview on the working principle and examples of the isolation and detection methods with the potential to be used for extracellular vesicles. This review will also highlight the integrated on-chip systems for isolation and characterization of EVs.

  2. Interfacial Recognition of Acetylcholine by an Amphiphilic p-Sulfonatocalix[8]arene Derivative Incorporated into Dimyristoyl Phosphatidylcholine Vesicles

    PubMed Central

    Jin, Takashi; Fujii, Fumihiko; Ooi, Yasuhiro

    2008-01-01

    Dodecyl ether derivatives 1-3 of p-sulfonatocalix[n]arene were incorporated into dimyristoyl phosphatidylcholine (DMPC) vesicles, and their binding abilities for acetylcholine (ACh) were examined by using steady-state fluorescence/fluorescence anisotropy and fluorescence correlation spectroscopy (FCS). For the detection of ACh binding to the DMPC vesicles containing 5 mol % of 1-3, competitive fluorophore displacement experiments were performed, where rhodamine 6G (Rh6G) was used as a fluorescent guest. The addition of Rh6G to the DMPC vesicles containing 3 resulted in a decrease in the fluorescence intensity of Rh6G with an increase of its fluorescence anisotropy, indicating that Rh6G binds to the DMPC-3 vesicles. In the case of DMPC-1 and DMPC-2 vesicles, significant changes in the fluorescence spectra of Rh6G were not observed. When ACh was added to the DMPC-3 vesicles in the presence of Rh6G ([3]/[Rh6G]=100), the fluorescence intensity of Rh6G increased with a decrease in its fluorescence anisotropy. From the analysis of fluorescence titration data, the association constants were determined to be 7.1×105 M-1 for Rh6G-3 complex and 1.1×102 M-1 for ACh-3 complex at the DMPC-3 vesicles. To get a direct evidence for the binding of Rh6G and its displacement by ACh at the DMPC-3 vesicles, diffusion times of the Rh6G were measured by using FCS. Binding selectivity of the DMPC-3 vesicles for ACh, choline, GABA, l-aspartic acid,l-glutamic acid, l-arginine, l-lysine, l-histamine and ammonium chloride was also evaluated using FCS. PMID:27873899

  3. Polyurethane Self-Priming Topcoats

    DTIC Science & Technology

    1992-06-23

    Approved for public release; distribution is unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT A corrosion -resistant coating which can be applied directly...57] ABSTRACT A corrosion -resistant coating which can be applied directly to a surface as a self-priming topcoat compris- ing from about 10 to 90... corrosion -inhibiting pigments consisting The invention described herein mav be manufactured 5 «»eniially of ar alkaline earth metal or zinc phosphate

  4. Maglev ready for prime time.

    SciTech Connect

    Rote, D. M.; Johnson, L. R.; Energy Systems

    2003-01-01

    Putting Maglev on Track' (Issues, Spring 1990) observed that growing airline traffic and associated delays were already significant and predicted that they would worsen. The article argued that a 300-mile-per-hour (mph) magnetic levitation (maglev) system integrated into airport and airline operations could be a part of the solution. Maglev was not ready for prime time in 1990, but it is now.

  5. KAPAO Prime: Design and Simulation

    NASA Astrophysics Data System (ADS)

    McGonigle, Lorcan; Choi, P. I.; Severson, S. A.; Spjut, E.

    2013-01-01

    KAPAO (KAPAO A Pomona Adaptive Optics instrument) is a dual-band natural guide star adaptive optics system designed to measure and remove atmospheric aberration over UV-NIR wavelengths from Pomona College’s telescope atop Table Mountain. We present here, the final optical system, KAPAO Prime, designed in Zemax Optical Design Software that uses custom off-axis paraboloid mirrors (OAPs) to manipulate light appropriately for a Shack-Hartman wavefront sensor, deformable mirror, and science cameras. KAPAO Prime is characterized by diffraction limited imaging over the full 81” field of view of our optical camera at f/33 as well as over the smaller field of view of our NIR camera at f/50. In Zemax, tolerances of 1% on OAP focal length and off-axis distance were shown to contribute an additional 4 nm of wavefront error (98% confidence) over the field of view of our optical camera; the contribution from surface irregularity was determined analytically to be 40nm for OAPs specified to λ/10 surface irregularity (632.8nm). Modeling of the temperature deformation of the breadboard in SolidWorks revealed 70 micron contractions along the edges of the board for a decrease of 75°F when applied to OAP positions such displacements from the optimal layout are predicted to contribute an additional 20 nanometers of wavefront error. Flexure modeling of the breadboard due to gravity is on-going. We hope to begin alignment and testing of KAPAO Prime in Q1 2013.

  6. KAPAO Prime: Design and Simulation

    NASA Astrophysics Data System (ADS)

    McGonigle, Lorcan

    2012-11-01

    KAPAO (KAPAO A Pomona Adaptive Optics instrument) is a dual-band natural guide star adaptive optics system designed to measure and remove atmospheric aberration from Pomona College's telescope atop Table Mountain. We present here, the final optical system, referred to as Prime, designed in Zemax Optical Design Software. Prime is characterized by diffraction limited imaging over the full 73'' field of view of our Andor Camera at f/33 as well as for our NIR Xenics camera at f/50. In Zemax, tolerances of 1% on OAP focal length and off-axis distance were shown to contribute an additional 4 nm of wavefront error (98% confidence) over the field of view of the Andor camera; the contribution from surface irregularity was determined analytically to be 40nm for OAPs specified to l/10 surface irregularity. Modeling of the temperature deformation of the breadboard in SolidWorks revealed 70 micron contractions along the edges of the board for a decrease of 75 F; when applied to OAP positions such displacements from the optimal layout are predicted to contribute an additional 20 nanometers of wavefront error. Flexure modeling of the breadboard due to gravity is on-going. We hope to begin alignment and testing of ``Prime'' in Q1 2013.

  7. What determines the direction of subliminal priming

    PubMed Central

    Jaśkowski, Piotr; Verleger, Rolf

    2008-01-01

    Masked stimuli (primes) can affect the preparation of a motor response to subsequently presented target stimuli. Reactions to the target can be facilitated (straight priming) or inhibited (inverse priming) when preceded by a compatible prime (calling for the same response) and also when preceded by an incompatible prime. Several hypotheses are currently under debate. These are the self-inhibition (SI) hypothesis, the object-updating (OU) hypothesis, and mask-triggered inhibition (MTI) hypothesis. All assume that the initial activation of the motor response is elicited by the prime according to its identity. This activation inevitably leads to straight priming in some cases and the mechanisms involved are undisputed. The hypotheses differ, however, as to why inverse priming occurs. The self-inhibition (SI) hypothesis assumes that the motor activation elicited by a prime is automatically followed by an inhibition phase, leading to inverse priming if three conditions are fulfilled: perceptual evidence for the prime has to be sufficiently strong, it has to be immediately removed by the mask, and the delay between the prime and target has to be long enough for inhibition to become effective. The object-updating (OU) hypothesis assumes that inverse priming is triggered by the mask, provided that it contains features calling for the alternative response (i.e. the one contrasting with the response induced by the prime). The MTI hypothesis assumes that the inhibitory phase is triggered by each successive stimulus which does not support the perceptual hypothesis provided by the prime. Based mostly on our own experiments, we argue that (1) attempts to manipulate the three factors required by the SI hypothesis imply changes of other variables and that (2) indeed, other variables seem to affect priming: prime-mask perceptual interaction and temporal position of the mask. These observations are in favor of the MTI hypothesis. A limiting factor for all three hypotheses is that

  8. Physicochemical characterization of drug-loaded rigid and elastic vesicles.

    PubMed

    Uchino, Tomonobu; Lefeber, Fons; Gooris, Gert; Bouwstra, Joke

    2011-06-30

    Ketorolac loaded rigid and elastic vesicles were prepared by sonication and the physicochemical properties of the drug loaded-vesicle formulations were examined. Rigid and elastic vesicles were prepared from the double chain surfactant sucrose-ester laurate (L-595) and the single chain surfactant octaoxyethylene-laurate ester (PEG-8-L). Sulfosuccinate (TR-70) was used as a negative charge inducer. Evaluation of the prepared vesicle was performed by dynamic light scattering, extrusion and by (1)H NMR (T(2) relaxation studies). The vesicles mean size varied between 90 and 150 nm. The elasticity of the vesicles was enhanced with increasing PEG-8-L/L-595 ratio, while an increase in loading of ketorolac resulted in a reduction in vesicle elasticity. (1)H NMR measurements showed that the molecular mobility of ketorolac was restricted, which indicates that ketorolac molecules were entrapped within the vesicle bilayers. The T(2) values of the aromatic protons of ketorolac increased gradually at higher PEG-8-L levels, indicating that ketorolac mobility increased in the vesicle bilayer. The chemical stability of ketorolac was dramatically improved in the vesicle formulation compared to a buffer solution. The strong interactions of ketorolac with the bilayers of the vesicles might be the explanation for this increased stability of ketorolac.

  9. Fewer active motors per vesicle may explain slowed vesicle transport in chick motoneurons after three days in vitro.

    PubMed

    Macosko, Jed C; Newbern, Jason M; Rockford, Jean; Chisena, Ernest N; Brown, Charlotte M; Holzwarth, George M; Milligan, Carol E

    2008-05-23

    Vesicle transport in cultured chick motoneurons was studied over a period of 3 days using motion-enhanced differential interference contrast (MEDIC) microscopy, an improved version of video-enhanced DIC. After 3 days in vitro (DIV), the average vesicle velocity was about 30% less than after 1 DIV. In observations at 1, 2 and 3 DIV, larger vesicles moved more slowly than small vesicles, and retrograde vesicles were larger than anterograde vesicles. The number of retrograde vesicles increased relative to anterograde vesicles after 3 DIV, but this fact alone could not explain the decrease in velocity, since the slowing of vesicle transport in maturing motoneurons was observed independently for both anterograde and retrograde vesicles. In order to better understand the slowing trend, the distance vs. time trajectories of individual vesicles were examined at a frame rate of 8.3/s. Qualitatively, these trajectories consisted of short (1-2 s) segments of constant velocity, and the changes in velocity between segments were abrupt (<0.2 s). The trajectories were therefore fit to a series of connected straight lines. Surprisingly, the slopes of theses lines, i.e. the vesicle velocities, were often found to be multiples of ~0.6 mum/s. The velocity histogram showed multiple peaks, which, when fit with Gaussians using a least squares minimization, yielded an average spacing of 0.57 mum/s (taken as the slope of a fit to peak position vs. peak number, R(2)=0.994). We propose that the abrupt velocity changes occur when 1 or 2 motors suddenly begin or cease actively participating in vesicle transport. Under this hypothesis, the decrease in average vesicle velocity observed for maturing motoneurons is due to a decrease in the average number of active motors per vesicle.

  10. Ultrastructural and functional fate of recycled vesicles in hippocampal synapses

    PubMed Central

    Rey, Stephanie A.; Smith, Catherine A.; Fowler, Milena W.; Crawford, Freya; Burden, Jemima J.; Staras, Kevin

    2015-01-01

    Efficient recycling of synaptic vesicles is thought to be critical for sustained information transfer at central terminals. However, the specific contribution that retrieved vesicles make to future transmission events remains unclear. Here we exploit fluorescence and time-stamped electron microscopy to track the functional and positional fate of vesicles endocytosed after readily releasable pool (RRP) stimulation in rat hippocampal synapses. We show that most vesicles are recovered near the active zone but subsequently take up random positions in the cluster, without preferential bias for future use. These vesicles non-selectively queue, advancing towards the release site with further stimulation in an actin-dependent manner. Nonetheless, the small subset of vesicles retrieved recently in the stimulus train persist nearer the active zone and exhibit more privileged use in the next RRP. Our findings reveal heterogeneity in vesicle fate based on nanoscale position and timing rules, providing new insights into the origins of future pool constitution. PMID:26292808

  11. Reactive block copolymer vesicles with an epoxy wall.

    PubMed

    Zhu, Hui; Liu, Qingchun; Chen, Yongming

    2007-01-16

    Recently, block copolymer vesicles have attracted considerable attention because of their properties in encapsulation and release. To explore their applications in biorelated fields, functionalization of the polymer vesicle is necessary. Herein, a reactive unilamellar vesicle is reported by self-assembly of poly(ethylene oxide)-block-poly(glycidyl methacrylate) copolymer (PEO-b-PGMA) in solution. When water was added into the PEO-b-PGMA solution in THF, unilamellar vesicles were produced. If hydrophobic primary amine additives, such as hexamethylenediamine (HDA) and dodecylamine (DA), were introduced during block copolymer assembling, the vesicular morphology remained unchanged; instead, the amines reacted with the epoxys and the vesicles were fixed by cross-linking. Furthermore, when 3-aminopropyl trimethoxysilane (APS) was applied, the organic/inorganic hybrid vesicles were obtained, which were stable against the solvent change. Therefore, this research not only supplies a new way to fix the vesicular morphology but also a reactive vesicle scaffold for introducing functional species.

  12. Relational integrativity of prime-target pairs moderates congruity effects in evaluative priming.

    PubMed

    Ihmels, Max; Freytag, Peter; Fiedler, Klaus; Alexopoulos, Theodore

    2016-05-01

    In evaluative priming, positive or negative primes facilitate reactions to targets that share the same valence. While this effect is commonly explained as reflecting invariant structures in semantic long-term memory or in the sensorimotor system, the present research highlights the role of integrativity in evaluative priming. Integrativity refers to the ease of integrating two concepts into a new meaningful compound representation. In extended material tests using paired comparisons from two pools of positive and negative words, we show that evaluative congruity is highly correlated with integrativity. Therefore, in most priming studies, congruity and integrativity are strongly confounded. When both aspects are disentangled by manipulating congruity and integrativity orthogonally, three priming experiments show that evaluative-priming effects were confined to integrative prime-target pairs. No facilitation of prime-congruent targets was obtained for non-integrative stimuli. These findings are discussed from a broader perspective on priming conceived as flexible, context-dependent, and serving a generative adaptation function.

  13. Focus on Extracellular Vesicles: Introducing the Next Small Big Thing.

    PubMed

    Kalra, Hina; Drummen, Gregor P C; Mathivanan, Suresh

    2016-02-06

    Intercellular communication was long thought to be regulated exclusively through direct contact between cells or via release of soluble molecules that transmit the signal by binding to a suitable receptor on the target cell, and/or via uptake into that cell. With the discovery of small secreted vesicular structures that contain complex cargo, both in their lumen and the lipid membrane that surrounds them, a new frontier of signal transduction was discovered. These "extracellular vesicles" (EV) were initially thought to be garbage bags through which the cell ejected its waste. Whilst this is a major function of one type of EV, i.e., apoptotic bodies, many EVs have intricate functions in intercellular communication and compound exchange; although their physiological roles are still ill-defined. Additionally, it is now becoming increasingly clear that EVs mediate disease progression and therefore studying EVs has ignited significant interests among researchers from various fields of life sciences. Consequently, the research effort into the pathogenic roles of EVs is significantly higher even though their protective roles are not well established. The "Focus on extracellular vesicles" series of reviews highlights the current state of the art regarding various topics in EV research, whilst this review serves as an introductory overview of EVs, their biogenesis and molecular composition.

  14. Pretransitional effects in dimyristoylphosphatidylcholine vesicle membranes: optical dynamometry study.

    PubMed Central

    Dimova, R; Pouligny, B; Dietrich, C

    2000-01-01

    We used micron-sized latex spheres to probe the phase state and the viscoelastic properties of dimyristoylphosphatidylcholine (DMPC) bilayers as a function of temperature. One or two particles were manipulated and stuck to a DMPC giant vesicle by means of an optical trap. Above the fluid-gel main transition temperature, T(m) congruent with 23.4 degrees C, the particles could move on the surface of the vesicle, spontaneously (Brownian motion) or driven by an external force, either gravity or the laser beam's radiation pressure. From the analysis of the particle motions, we deduced the values of the membrane hydrodynamic shear viscosity, eta(s), and found that it would increase considerably near T(m). Below T(m), the long-distance motion of the particles was blocked. We performed experiments with two particles stuck on the membrane. By optical dynamometry, we measured the elastic resistance of the membrane to a variation in the interparticle distance and found that it would decrease considerably (down to zero) when the temperature was increased to T(m). We propose an interpretation relating the elastic response to the membrane curvature modulus, k(C). In this scheme, the two-bead dynamometry experiments provide a direct measurement of k(C) in the P'(beta) phase of lipid bilayers. PMID:10866960

  15. Docking of Secretory Vesicles Is Syntaxin Dependent

    PubMed Central

    de Wit, Heidi; Cornelisse, L. Niels; Toonen, Ruud F.G.; Verhage, Matthijs

    2006-01-01

    Secretory vesicles dock at the plasma membrane before they undergo fusion. Molecular docking mechanisms are poorly defined but believed to be independent of SNARE proteins. Here, we challenged this hypothesis by acute deletion of the target SNARE, syntaxin, in vertebrate neurons and neuroendocrine cells. Deletion resulted in fusion arrest in both systems. No docking defects were observed in synapses, in line with previous observations. However, a drastic reduction in morphologically docked secretory vesicles was observed in chromaffin cells. Syntaxin-deficient chromaffin cells showed a small reduction in total and plasma membrane staining for the docking factor Munc18-1, which appears insufficient to explain the drastic reduction in docking. The sub-membrane cortical actin network was unaffected by syntaxin deletion. These observations expose a docking role for syntaxin in the neuroendocrine system. Additional layers of regulation may have evolved to make syntaxin redundant for docking in highly specialized systems like synaptic active zones. PMID:17205130

  16. In vitro formation of gap junction vesicles.

    PubMed

    Goodenough, D A

    1976-02-01

    A method is described that uses trypsin digestion combined with collagenase-hyaluronidase which produces a population of gap junction vesicles. The hexagonal lattice of subunits ("connexons") comprising the gapjunctions appears unaltered by various structural criteria and by buoyant density measurements. The gap junction vesciles are closed by either a single or a double profile of nonjunctional "membrane," which presents a smooth, particle-free fracture face. Horseradish peroxidase and cytochrome c studies have revealed that about 20% of the gap junction vesicles are impermeable to proteins 12,000 daltons or larger. The increased purity of the trypsinized junction preparation suggests that one of the disulfide reduction products of the gap-junction principal protein may be a nonjunctional contaminating peptide. The gap junction appears to be composed of a single 18,000-dalton protein, connexin, which may be reduced to a single 9,000-dalton peak. The number of peptides in this reduced peak are still unknown.

  17. Soft vesicles in the synthesis of hard materials.

    PubMed

    Dong, Renhao; Liu, Weimin; Hao, Jingcheng

    2012-04-17

    Vesicles of surfactants in aqueous solution have received considerable attention because of their use as simple model systems for biological membranes and their applications in various fields including colloids, pharmaceuticals, and materials. Because of their architecture, vesicles could prove useful as "soft" templates for the synthesis of "hard materials". The vesicle phase, however, has been challenging and difficult to work with in the construction of hard materials. In the solution-phase synthesis of various inorganic or macromolecular materials, templating methods provide a powerful strategy to control the size, morphology, and composition of the resulting micro- and nanostructures. In comparison with hard templates, soft templates are generally constructed using amphiphilic molecules, especially surfactants and amphiphilic polymers. These types of compounds offer advantages including the wide variety of available templates, simple fabrication processes under mild conditions, and easy removal of the templates with less damage to the final structures. Researchers have used many ordered molecular aggregates such as vesicles, micelles, liquid crystals, emulsion droplets, and lipid nanotubes as templates or structure-directing agents to control the synthesis or assembly hard micro- and nanomaterials composed from inorganic compounds or polymers. In addition to their range of sizes and morphologies, vesicles present unique structures that can simultaneously supply different microenvironments for the growth and assembly of hard materials: the inner chamber of vesicles, the outer surface of the vesicles, and the space between bilayers. Two main approaches for applying vesicles in the field of hard materials have been explored: (i) in situ synthesis of micro- or nanomaterials within a specific microenvironment by vesicle templating and (ii) the assembly or incorporation of guest materials during the formation of vesicles. This Account provides an in-depth look at

  18. Microbial stress priming: a meta-analysis.

    PubMed

    Andrade-Linares, Diana R; Lehmann, Anika; Rillig, Matthias C

    2016-04-01

    Microbes have to cope with complex and dynamic environments, making it likely that anticipatory responses provide fitness benefits. Mild, previous stressors can prepare microbes (stress priming) to further and potentially damaging stressors (triggering). We here quantitatively summarize the findings from over 250 trials of 34 studies including bacteria and fungi, demonstrating that priming to stress has a beneficial impact on microbial survival. In fact, survival of primed microbes was about 10-fold higher compared with that in non-primed microbes. Categorical moderators related to microbial taxonomy and the kind of stress applied as priming or as triggering revealed significant differences of priming effect size among 14 different microbial species, 6 stress categories and stressor combination. We found that priming by osmotic, physiological and temperature stress had the highest positive effect sizes on microbial response. Cross-protection was evident for physiological, temperature and pH stresses. Microbes are better prepared against triggering by oxidative, temperature and osmotic stress. Our finding of an overall positive mean effect of priming regardless of the microbial system and particular stressor provides unprecedentedly strong evidence of the broad ecological significance of microbial stress priming. These results further suggest that stress priming may be an important factor in shaping microbial communities.

  19. Wrath of God: religious primes and punishment.

    PubMed

    McKay, Ryan; Efferson, Charles; Whitehouse, Harvey; Fehr, Ernst

    2011-06-22

    Recent evidence indicates that priming participants with religious concepts promotes prosocial sharing behaviour. In the present study, we investigated whether religious priming also promotes the costly punishment of unfair behaviour. A total of 304 participants played a punishment game. Before the punishment stage began, participants were subliminally primed with religion primes, secular punishment primes or control primes. We found that religious primes strongly increased the costly punishment of unfair behaviours for a subset of our participants--those who had previously donated to a religious organization. We discuss two proximate mechanisms potentially underpinning this effect. The first is a 'supernatural watcher' mechanism, whereby religious participants punish unfair behaviours when primed because they sense that not doing so will enrage or disappoint an observing supernatural agent. The second is a 'behavioural priming' mechanism, whereby religious primes activate cultural norms pertaining to fairness and its enforcement and occasion behaviour consistent with those norms. We conclude that our results are consistent with dual inheritance proposals about religion and cooperation, whereby religions harness the byproducts of genetically inherited cognitive mechanisms in ways that enhance the survival prospects of their adherents.

  20. The temporal dynamics of visual object priming.

    PubMed

    Ko, Philip C; Duda, Bryant; Hussey, Erin P; Mason, Emily J; Ally, Brandon A

    2014-11-01

    Priming reflects an important means of learning that is mediated by implicit memory. Importantly, priming occurs for previously viewed objects (item-specific priming) and their category relatives (category-wide priming). Two distinct neural mechanisms are known to mediate priming, including the sharpening of a neural object representation and the retrieval of stimulus-response mappings. Here, we investigated whether the relationship between these neural mechanisms could help explain why item-specific priming generates faster responses than category-wide priming. Participants studied pictures of everyday objects, and then performed a difficult picture identification task while we recorded event-related potentials (ERP). The identification task gradually revealed random line segments of previously viewed items (Studied), category exemplars of previously viewed items (Exemplar), and items that were not previously viewed (Unstudied). Studied items were identified sooner than Unstudied items, showing evidence of item-specific priming, and importantly Exemplar items were also identified sooner than Unstudied items, showing evidence of category-wide priming. Early activity showed sustained neural suppression of parietal activity for both types of priming. However, these neural suppression effects may have stemmed from distinct processes because while category-wide neural suppression was correlated with priming behavior, item-specific neural suppression was not. Late activity, examined with response-locked ERPs, showed additional processes related to item-specific priming including neural suppression in occipital areas and parietal activity that was correlated with behavior. Together, we conclude that item-specific and category-wide priming are mediated by separate, parallel neural mechanisms in the context of the current paradigm. Temporal differences in behavior are determined by the timecourses of these distinct processes.

  1. ATP: The crucial component of secretory vesicles.

    PubMed

    Estévez-Herrera, Judith; Domínguez, Natalia; Pardo, Marta R; González-Santana, Ayoze; Westhead, Edward W; Borges, Ricardo; Machado, José David

    2016-07-12

    The colligative properties of ATP and catecholamines demonstrated in vitro are thought to be responsible for the extraordinary accumulation of solutes inside chromaffin cell secretory vesicles, although this has yet to be demonstrated in living cells. Because functional cells cannot be deprived of ATP, we have knocked down the expression of the vesicular nucleotide carrier, the VNUT, to show that a reduction in vesicular ATP is accompanied by a drastic fall in the quantal release of catecholamines. This phenomenon is particularly evident in newly synthesized vesicles, which we show are the first to be released. Surprisingly, we find that inhibiting VNUT expression also reduces the frequency of exocytosis, whereas the overexpression of VNUT drastically increases the quantal size of exocytotic events. To our knowledge, our data provide the first demonstration that ATP, in addition to serving as an energy source and purinergic transmitter, is an essential element in the concentration of catecholamines in secretory vesicles. In this way, cells can use ATP to accumulate neurotransmitters and other secreted substances at high concentrations, supporting quantal transmission.

  2. ATP: The crucial component of secretory vesicles

    PubMed Central

    Estévez-Herrera, Judith; Domínguez, Natalia; Pardo, Marta R.; González-Santana, Ayoze; Westhead, Edward W.; Borges, Ricardo; Machado, José David

    2016-01-01

    The colligative properties of ATP and catecholamines demonstrated in vitro are thought to be responsible for the extraordinary accumulation of solutes inside chromaffin cell secretory vesicles, although this has yet to be demonstrated in living cells. Because functional cells cannot be deprived of ATP, we have knocked down the expression of the vesicular nucleotide carrier, the VNUT, to show that a reduction in vesicular ATP is accompanied by a drastic fall in the quantal release of catecholamines. This phenomenon is particularly evident in newly synthesized vesicles, which we show are the first to be released. Surprisingly, we find that inhibiting VNUT expression also reduces the frequency of exocytosis, whereas the overexpression of VNUT drastically increases the quantal size of exocytotic events. To our knowledge, our data provide the first demonstration that ATP, in addition to serving as an energy source and purinergic transmitter, is an essential element in the concentration of catecholamines in secretory vesicles. In this way, cells can use ATP to accumulate neurotransmitters and other secreted substances at high concentrations, supporting quantal transmission. PMID:27342860

  3. Proton pump of clathrin-coated vesicles

    SciTech Connect

    Xie, X.

    1985-01-01

    Clathrin-coated vesicles were prepared from bovine brain catalyze ATP-driven proton translocation and a /sup 32/Pi-ATP exchange reaction. N-ethylmaleimide (NEM) at 1 mM and dicyclohexylcarbodiimide (DCCD) at 0.5 mM inhibit the pump completely, whereas neither vanadate, efrapeptin, NaN/sub 3/, nor mitochondrial ATPase inhibitor has an effect. The coated vesicle proton pump is characterized by ATP specificity. dATP, but no other nucleotide, can replace ATP as a substrate. The pump is electrogenic and the electrogenicity is neutralized by chloride or bromide serving as co-ions. ATP-driven proton translocation can be observed in the absence of chloride, provided that the membrane potential is collapsed by K/sup +/ moving out in the presence of valinomycin. Chloride transport can be observed independent of proton movements in the absence of ATP, provided that an inside positive membrane potential is generated by K/sup +/ influx in the presence of valinomycin. The proton-translocating ATPase of coated vesicles was solubilized with a nonionic detergent polyoxyethylene 9 lauryl ether, and purified about 700 fold to near homogeneity. During purification the enzymatic activity was lost. A purified brain phospholipid fraction restored the activity and was subsequently identified as phosphatidylserine.

  4. The reserve pool of synaptic vesicles acts as a buffer for proteins involved in synaptic vesicle recycling

    PubMed Central

    Denker, Annette; Kröhnert, Katharina; Bückers, Johanna; Neher, Erwin; Rizzoli, Silvio O.

    2011-01-01

    Presynaptic nerve terminals contain between several hundred vesicles (for example in small CNS synapses) and several tens of thousands (as in neuromuscular junctions). Although it has long been assumed that such high numbers of vesicles are required to sustain neurotransmission during conditions of high demand, we found that activity in vivo requires the recycling of only a few percent of the vesicles. However, the maintenance of large amounts of reserve vesicles in many evolutionarily distinct species suggests that they are relevant for synaptic function. We suggest here that these vesicles constitute buffers for soluble accessory proteins involved in vesicle recycling, preventing their loss into the axon. Supporting this hypothesis, we found that vesicle clusters contain a large variety of proteins needed for vesicle recycling, but without an obvious function within the clusters. Disrupting the clusters by application of black widow spider venom resulted in the diffusion of numerous soluble proteins into the axons. Prolonged stimulation and ionomycin application had a similar effect, suggesting that calcium influx causes the unbinding of soluble proteins from vesicles. Confirming this hypothesis, we found that isolated synaptic vesicles in vitro sequestered soluble proteins from the cytosol in a process that was inhibited by calcium addition. We conclude that the reserve vesicles support neurotransmission indirectly, ensuring that soluble recycling proteins are delivered upon demand during synaptic activity. PMID:21903923

  5. Release of canine parvovirus from endocytic vesicles.

    PubMed

    Suikkanen, Sanna; Antila, Mia; Jaatinen, Anne; Vihinen-Ranta, Maija; Vuento, Matti

    2003-11-25

    Canine parvovirus (CPV) is a small nonenveloped virus with a single-stranded DNA genome. CPV enters cells by clathrin-mediated endocytosis and requires an acidic endosomal step for productive infection. Virion contains a potential nuclear localization signal as well as a phospholipase A(2) like domain in N-terminus of VP1. In this study we characterized the role of PLA(2) activity on CPV entry process. PLA(2) activity of CPV capsids was triggered in vitro by heat or acidic pH. PLA(2) inhibitors inhibited the viral proliferation suggesting that PLA(2) activity is needed for productive infection. The N-terminus of VP1 was exposed during the entry, suggesting that PLA(2) activity might have a role during endocytic entry. The presence of drugs modifying endocytosis (amiloride, bafilomycin A(1), brefeldin A, and monensin) caused viral proteins to remain in endosomal/lysosomal vesicles, even though the drugs were not able to inhibit the exposure of VP1 N-terminal end. These results indicate that the exposure of N-terminus of VP1 alone is not sufficient to allow CPV to proliferate. Some other pH-dependent changes are needed for productive infection. In addition to blocking endocytic entry, amiloride was able to block some postendocytic steps. The ability of CPV to permeabilize endosomal membranes was demonstrated by feeding cells with differently sized rhodamine-conjugated dextrans together with the CPV in the presence or in the absence of amiloride, bafilomycin A(1), brefeldin A, or monensin. Dextran with a molecular weight of 3000 was released from vesicles after 8 h of infection, while dextran with a molecular weight of 10,000 was mainly retained in vesicles. The results suggest that CPV infection does not cause disruption of endosomal vesicles. However, the permeability of endosomal membranes apparently changes during CPV infection, probably due to the PLA(2) activity of the virus. These results suggest that parvoviral PLA(2) activity is essential for productive

  6. Controlled deformation of vesicles by flexible structured media

    PubMed Central

    Zhang, Rui; Zhou, Ye; Martínez-González, José A.; Hernández-Ortiz, Juan P.; Abbott, Nicholas L.; de Pablo, Juan J.

    2016-01-01

    Liquid crystalline (LC) materials, such as actin or tubulin networks, are known to be capable of deforming the shape of cells. Here, elements of that behavior are reproduced in a synthetic system, namely, a giant vesicle suspended in a LC, which we view as a first step toward the preparation of active, anisotropic hybrid systems that mimic some of the functionality encountered in biological systems. To that end, we rely on a coupled particle-continuum representation of deformable networks in a nematic LC represented at the level of a Landau–de Gennes free energy functional. Our results indicate that, depending on its elastic properties, the LC is indeed able to deform the vesicle until it reaches an equilibrium, anisotropic shape. The magnitude of the deformation is determined by a balance of elastic and surface forces. For perpendicular anchoring at the vesicle, a Saturn ring defect forms along the equatorial plane, and the vesicle adopts a pancake-like, oblate shape. For degenerate planar anchoring at the vesicle, two boojum defects are formed at the poles of the vesicle, which adopts an elongated, spheroidal shape. During the deformation, the volume of the topological defects in the LC shrinks considerably as the curvature of the vesicle increases. These predictions are confirmed by our experimental observations of spindle-like shapes in experiments with giant unilamellar vesicles with planar anchoring. We find that the tension of the vesicle suppresses vesicle deformation, whereas anchoring strength and large elastic constants promote shape anisotropy. PMID:27532056

  7. Polymerization of hydrogels inside self-assembled block copolymer vesicles.

    PubMed

    Gaspard, Jeffery; Hahn, Mariah S; Silas, James A

    2009-11-17

    Block copolymer vesicles are powerful tools for investigating cell adhesion since they display the fluid, deformable, semipermeable membrane properties of lipid vesicles while having greater chemical and mechanical stability. The aim of the present study was to fabricate block copolymer vesicles containing hydrogel interiors in order to extend achievable vesicle properties and, thereby, their range of cell-like behaviors. Block copolymer vesicles based on poly(butadiene-b-ethylene oxide) were demonstrated to compartmentalize and retain acrylamide solutions through particle dialysis and to allow for subsequent in situ hydrogel polymerization. Small molecule leakage studies of the resulting particles indicated that the cross-link density of the hydrogel interiors had minimal impact on vesicle permeability to small molecules (<430 Da) relative to vesicle membrane properties. In contrast, particle deformation analyses indicated that initial vesicle surface approach and adhesion was dominated by its membrane properties, whereas its ultimate deformation was primarily governed by the hydrogel interior. Thus, the hydrogel-containing vesicles allowed orthogonal control of particle surface and mechanical properties. Analysis of particle behavior in terms of Gibb's free energy minimization indicated that vesicle adhesion energy, membrane tension, and internal osmotic pressure dominated particle adhesion and deformation. Combined, the present work demonstrates the potential for designing compartmentalized, hierarchical polymer-based cell mimics with broadly tunable dynamic-mechanical properties and surface properties.

  8. Extracellular Vesicles in Renal Diseases: More than Novel Biomarkers?

    PubMed Central

    Le, Thu H.

    2016-01-01

    Extracellular vesicles from the urine and circulation have gained significant interest as potential diagnostic biomarkers in renal diseases. Urinary extracellular vesicles contain proteins from all sections of the nephron, whereas most studied circulating extracellular vesicles are derived from platelets, immune cells, and the endothelium. In addition to their diagnostic role as markers of kidney and vascular damage, extracellular vesicles may have functional significance in renal health and disease by facilitating communication between cells and protecting against kidney injury and bacterial infection in the urinary tract. However, the current understanding of extracellular vesicles has derived mostly from studies with very small numbers of patients or in vitro data. Moreover, accurate assessment of these vesicles remains a challenge, in part because of a lack of consensus in the methodologies to measure extracellular vesicles and the inability of most techniques to capture the entire size range of these vesicles. However, newer techniques and standardized protocols to improve the detection of extracellular vesicles are in development. A clearer understanding of the composition and biology of extracellular vesicles will provide insights into their pathophysiologic, diagnostic, and therapeutic roles. PMID:26251351

  9. Focus on Extracellular Vesicles: Therapeutic Potential of Stem Cell-Derived Extracellular Vesicles.

    PubMed

    Zhang, Bin; Yeo, Ronne Wee Yeh; Tan, Kok Hian; Lim, Sai Kiang

    2016-02-06

    The intense research focus on stem and progenitor cells could be attributed to their differentiation potential to generate new cells to replace diseased or lost cells in many highly intractable degenerative diseases, such as Alzheimer disease, multiple sclerosis, and heart diseases. However, experimental and clinical studies have increasingly attributed the therapeutic efficacy of these cells to their secretion. While stem and progenitor cells secreted many therapeutic molecules, none of these molecules singly or in combination could recapitulate the functional effects of stem cell transplantations. Recently, it was reported that extracellular vesicles (EVs) could recapitulate the therapeutic effects of stem cell transplantation. Based on the observations reported thus far, the prevailing hypothesis is that stem cell EVs exert their therapeutic effects by transferring biologically active molecules such as proteins, lipids, mRNA, and microRNA from the stem cells to injured or diseased cells. In this respect, stem cell EVs are similar to EVs from other cell types. They are both primarily vehicles for intercellular communication. Therefore, the differentiating factor is likely due to the composition of their cargo. The cargo of EVs from different cell types are known to include a common set of proteins and also proteins that reflect the cell source of the EVs and the physiological or pathological state of the cell source. Hence, elucidation of the stem cell EV cargo would provide an insight into the multiple physiological or biochemical changes necessary to affect the many reported stem cell-based therapeutic outcomes in a variety of experimental models and clinical trials.

  10. On Anti-Elite Prime Numbers

    NASA Astrophysics Data System (ADS)

    M"Uller, Tom

    2007-09-01

    An odd prime number p is called anti-elite if only finitely many Fermat numbers are quadratic non-residues to p. This concept is the exact opposite to that of elite prime numbers. We study some fundamental properties of anti-elites and show that there are infinitely many of them. A computational search among all the numbers up to 100 billion yielded 84 anti-elite primes.

  11. All Elite Primes Up to 250 Billion

    NASA Astrophysics Data System (ADS)

    Chaumont, Alain; Müller, Tom

    2006-08-01

    A prime number p is called elite if only finitely many Fermat numbers 2^(2^n)+1 are quadratic residues of p. Previously only the interval up to 10^9 was systematically searched for elite primes and 16 such primes were found. We extended this research up to 2.5*10^11 and found five further elites, among which 1,151,139,841 is the smallest and 171,727,482,881 the largest.

  12. Prime Graph Components of Finite Simple Groups

    NASA Astrophysics Data System (ADS)

    Kondrat'ev, A. S.

    1990-02-01

    Let G be a finite group and π(G) the set of prime factors of its order. The prime graph of G is the graph with vertex-set π(G), two vertices p and q being joined by an edge whenever G contains an element of order pq. This article contains an explicit description of the primes in each of the connected components of the prime graphs of the finite simple groups of Lie type of even characteristic. This solves question 9.16 of the Kourovka Notebook. Bibliography: 15 titles.

  13. Focus on Extracellular Vesicles: Development of Extracellular Vesicle-Based Therapeutic Systems

    PubMed Central

    Ohno, Shin-ichiro; Drummen, Gregor P. C.; Kuroda, Masahiko

    2016-01-01

    Many types of cells release phospholipid membrane vesicles thought to play key roles in cell-cell communication, antigen presentation, and the spread of infectious agents. Extracellular vesicles (EVs) carry various proteins, messenger RNAs (mRNAs), and microRNAs (miRNAs), like a “message in a bottle” to cells in remote locations. The encapsulated molecules are protected from multiple types of degradative enzymes in body fluids, making EVs ideal for delivering drugs. This review presents an overview of the potential roles of EVs as natural drugs and novel drug-delivery systems. PMID:26861303

  14. Transformation priming helps to disambiguate sudden changes of sensory inputs.

    PubMed

    Pastukhov, Alexander; Vivian-Griffiths, Solveiga; Braun, Jochen

    2015-11-01

    Retinal input is riddled with abrupt transients due to self-motion, changes in illumination, object-motion, etc. Our visual system must correctly interpret each of these changes to keep visual perception consistent and sensitive. This poses an enormous challenge, as many transients are highly ambiguous in that they are consistent with many alternative physical transformations. Here we investigated inter-trial effects in three situations with sudden and ambiguous transients, each presenting two alternative appearances (rotation-reversing structure-from-motion, polarity-reversing shape-from-shading, and streaming-bouncing object collisions). In every situation, we observed priming of transformations as the outcome perceived in earlier trials tended to repeat in subsequent trials and this repetition was contingent on perceptual experience. The observed priming was specific to transformations and did not originate in priming of perceptual states preceding a transient. Moreover, transformation priming was independent of attention and specific to low level stimulus attributes. In summary, we show how "transformation priors" and experience-driven updating of such priors helps to disambiguate sudden changes of sensory inputs. We discuss how dynamic transformation priors can be instantiated as "transition energies" in an "energy landscape" model of the visual perception.

  15. Priming effects in marine sediments

    NASA Astrophysics Data System (ADS)

    Gontikaki, Evina; Thornton, Barry; Witte, Ursula

    2013-04-01

    Continental margin sediments (<2000 m) cover merely 15 % of the ocean's seafloor but are responsible for more than 70 % of the global benthic mineralization. Understanding when these systems act as a source or sink of carbon (C) is thus of primary importance if we are to produce reliable global C budgets and predict the effects of future perturbations on the global C cycle. The chemical nature of organic matter (OM) is thought to be one of the major controls on the degradation/preservation balance in sediments; labile and refractory OM pools degrade at different rates but not independently. Priming effects (PE), i.e. changes in the decomposition of refractory organic matter following inputs of labile OM, have the potential to alter the C budget in sediments but have been largely ignored by marine scientists. Climate-driven changes in primary production, and land erosion and run-off are likely to change the quantity and composition of organic matter inputs in marine ecosystems and influence the magnitude and direction of PEs in seawater and sediments. Here, we attempt to evaluate the importance of priming effects on C cycling in marine sediments by use of labelled substrates of different quantity and quality in stable isotope tracer experiments and argue that PEs need to be incorporated in global change models.

  16. Adhesive interactions between vesicles in the strong adhesion limit

    PubMed Central

    Ramachandran, Arun; Anderson, Travers H.; Leal, L. Gary; Israelachvili, Jacob N.

    2010-01-01

    We consider the adhesive interaction energy between a pair of vesicles in the strong adhesion limit, in which bending forces play a negligible role in determining vesicle shape compared to forces due to membrane stretching. Although force-distance or energy distance relationships characterizing adhesive interactions between fluid bilayers are routinely measured using the surface forces apparatus, the atomic force microscope and the biomembrane force probe, the interacting bilayers in these methods are supported on surfaces (e.g. mica sheet) and cannot be deformed. However, it is known that in a suspension, vesicles composed of the same bilayer can deform by stretching or bending, and can also undergo changes in volume. Adhesively interacting vesicles can thus form flat regions in the contact zone, which will result in an enhanced interaction energy as compared to rigid vesicles. The focus of this paper is to examine the magnitude of the interaction energy between adhesively interacting, deformed vesicles relative to free, undeformed vesicles as a function of the intervesicle separation. The modification of the intervesicle interaction energy due to vesicle deformability can be calculated knowing the undeformed radius of the vesicles, R0, the bending modulus kb, the area expansion modulus Ka, and the adhesive minimum WP(0) and separation DP(0) in the energy of interaction between two flat bilayers, which can be obtained from the force-distance measurements made using the above supported-bilayer methods. For vesicles with constant volumes, we show that adhesive potentials between non-deforming bilayers such as ∣WP(0)∣∼5×10−4mJ/m2, which are ordinarily considered weak in colloidal physics literature, can result in significantly deep (>10×) energy minima due to increase in vesicle area and flattening in the contact region. If the osmotic expulsion of water across the vesicles driven by the tense, stretched membrane in the presence of an osmotically active

  17. Effect of Ca2+ on Vesicle Fusion on Solid Surface: An In vitro Model of Protein-Accelerated Vesicle Fusion

    NASA Astrophysics Data System (ADS)

    Shinozaki, Youichi; Siitonen, Ari M.; Sumitomo, Koji; Furukawa, Kazuaki; Torimitsu, Keiichi

    2008-07-01

    Lipid vesicle fusion is an important reaction in the cell. Calcium ions (Ca2+) participate in various important biological events including the fusion of vesicles with cell membranes in cells. We studied the effect of Ca2+ on the fusion of egg yolk phosphatidylcholine/brain phosphatidylserine (eggPC/brainPS) lipid vesicles on a mica substrate with fast scanning atomic force microscopy (AFM). When unattached and unfused lipid vesicles on mica were rinsed away, discrete patches of fused vesicles were observed under high Ca2+ concentrations. At 0 mM Ca2+, lipid vesicles were fused on mica and formed continuous supported lipid bilayers (SLBs) covering almost the entire mica surface. The effect of Ca2+ on SLB formation was offset by a Ca2+ chelating agent. When lipid vesicles were added during AFM observation, vesicles fused on mica and covered almost all areas even under high Ca2+ concentrations. These results indicate that force between AFM tip and vesicles overcomes the Ca2+-reduced fusion of lipid vesicles.

  18. Covalent attachment of lipid vesicles to a fluid supported bilayer allows observation of DNA-mediated vesicle interactions

    PubMed Central

    van Lengerich, Bettina; Rawle, Robert J.; Boxer, Steven G.

    2010-01-01

    Specific membrane interactions such as lipid vesicle docking and fusion can be mediated by synthetic DNA-lipid conjugates as a model for the protein-driven processes that are ubiquitous in biological systems. Here we present a method of tethering vesicles to a supported lipid bilayer that allows simultaneous deposition of cognate vesicle partners displaying complementary DNA, resulting in well-mixed populations of tethered vesicles that are laterally mobile. Vesicles are covalently attached to a supporting lipid bilayer using a DNA-templated click reaction; then DNA-mediated interactions between tethered vesicles are triggered by spiking the salt concentration. These interactions, such as docking and fusion, can then be observed for individual vesicles as they collide on the surface. The architecture of this new system also permits control over the number of lipid anchors that tether the vesicle to the supporting bilayer. The diffusion coefficient of tethered vesicles anchored by two lipids is approximately 1.6-fold slower than that of vesicles anchored only with a single lipid, consistent with a simple physical model. PMID:20180548

  19. Large vesicles record pathways of degassing at basalic volcanoes

    SciTech Connect

    Polacci, M.; Baker, D.R.; Bai, L.; Mancini, L.

    2008-10-08

    Volcanic degassing is directly linked to magma dynamics and controls the style of eruptive activity. To better understand how gas is transported within basaltic magma we perform a 3D investigation of vesicles preserved in scoria from the 2005 activity at Stromboli volcano (Italy). We find that clasts are characterized by the ubiquitous occurrence of one to a few large vesicles, exhibiting mostly irregular, tortuous, channel-like textures, orders of magnitude greater in volume than all the other vesicles in the sample. We compare observations on natural samples with results from numerical simulations and experimental investigations of vesicle size distributions and demonstrate that this type of vesicle invariably forms in magmas with vesicularities > 0.30 (and possibly > 0.10). We suggest that large vesicles represent pathways used by gas to flow non-explosively to the surface and that they indicate the development of an efficient system that sustains persistent degassing in basaltic systems.

  20. Gram-negative and Gram-positive bacterial extracellular vesicles.

    PubMed

    Kim, Ji Hyun; Lee, Jaewook; Park, Jaesung; Gho, Yong Song

    2015-04-01

    Like mammalian cells, Gram-negative and Gram-positive bacteria release nano-sized membrane vesicles into the extracellular environment either in a constitutive manner or in a regulated manner. These bacterial extracellular vesicles are spherical bilayered proteolipids enriched with bioactive proteins, lipids, nucleic acids, and virulence factors. Recent progress in this field supports the critical pathophysiological functions of these vesicles in both bacteria-bacteria and bacteria-host interactions. This review provides an overview of the current understanding on Gram-negative and Gram-positive bacterial extracellular vesicles, especially regarding the biogenesis, components, and functions in poly-species communities. We hope that this review will stimulate additional research in this emerging field of bacterial extracellular vesicles and contribute to the development of extracellular vesicle-based diagnostic tools and effective vaccines against pathogenic Gram-negative and Gram-positive bacteria.

  1. Tight coupling between positive and reversed priming in the masked prime paradigm

    PubMed Central

    Boy, Frederic; Sumner, Petroc

    2011-01-01

    When associations between certain visual stimuli and particular actions are learnt, those stimuli become capable of automatically and unconsciously activating their associated action plans. Such sensorimotor priming is assumed to be fundamental for efficient responses, and can be reliably measured in masked prime studies even when the primes are not consciously perceived. However, when the delay between prime and target is increased, reversed priming effects are often found instead (the negative compatibility effect, NCE). The main accounts of the NCE assume that it too is a sensorimotor phenomenon, predicting that it should occur only when the initial positive priming phase also occurs. Alternatively, reversed priming may reflect a perceptual process entirely independent from positive motor priming (which is simply evident at a different temporal delay), in which case no dependency is expected between the NCE and positive priming. We tested these predictions while new sensorimotor associations were learnt, and when learnt associations were suddenly reversed. We found a remarkable symmetry between positive and reversed priming during all such learning phases, supporting the idea that reversed priming represents a sensorimotor process that is contingent on, and automatically follows, the positive priming phase. We discuss also whether the NCE mechanism is subject to a trigger threshold. PMID:20695707

  2. Longer prime presentation decreases picture-word cross-domain priming.

    PubMed

    Miyoshi, Kiyofumi; Kimura, Yusuke; Ashida, Hiroshi

    2015-01-01

    A short prime presentation has been shown to provide a greater priming magnitude, whereas a longer prime presentation results in a lower priming magnitude. In Experiment 1, we attempted to replicate the decrease of priming using word stimuli. Words were presented in both prime and test sessions, and participants judged whether each stimulus was natural or manmade. In Experiment 2, we employed a cross-domain priming paradigm to assess the impact of prime duration on non-perceptual processes. Pictures were presented in prime sessions, and their semantically matched words were presented in test sessions. We did not observe a significant decrease in priming in Experiment 1. However, we found that 2000 ms of prime exposure led to weaker cross-domain priming when compared with 250 ms of the exposure in Experiment 2. The results suggest that the longer presentation of pictures causes a non-perceptual adaptation effect. This effect may occur at conceptual, linguistic, and/or response-related levels.

  3. Unconscious congruency priming from unpracticed words is modulated by prime-target semantic relatedness.

    PubMed

    Ortells, Juan J; Marí-Beffa, Paloma; Plaza-Ayllón, Vanesa

    2013-03-01

    Participants performed a 2-choice categorization task on visible word targets that were preceded by novel (unpracticed) prime words. The prime words were presented for 33 ms and followed either immediately (Experiments 1-3) or after a variable delay (Experiments 1 and 4) by a pattern mask. Both subjective and objective measures of prime visibility were used in all experiments. On 80% of the trials the primes and targets belonged to different categories (incongruent trials), whereas in the remaining 20% (congruent trials) they could be either strong or weak semantically related category members. Positive congruency effects (reaction times faster on congruent than on incongruent trials) were consistently found, but only when the mask immediately followed the primes, and participants reported being unaware of the identity of the primes. Primes followed by a delayed mask (such that participants reported being aware of their identity) produced either nonreliable facilitation or reliable reversed priming (strategic), depending on whether the prime-target stimulus onset asynchrony was either short (200 ms; Experiments 1 and 4) or long (1,000 ms; Experiment 4). Facilitatory priming with immediate mask was found strong (a) even for participants who performed at chance in prime visibility tests; and (b) for high but not for weakly semantically related category coordinates, irrespective of category size (animals, body parts). These findings provide evidence that unconscious congruency priming by unpracticed words from large stimulus sets critically depends on associative strength and/or semantic similarity between category coexemplars.

  4. Comparative Transcriptional Profiling of Primed and Non-primed Rice Seedlings under Submergence Stress

    PubMed Central

    Hussain, Saddam; Yin, Hanqi; Peng, Shaobing; Khan, Faheem A.; Khan, Fahad; Sameeullah, Muhammad; Hussain, Hafiz A.; Huang, Jianliang; Cui, Kehui; Nie, Lixiao

    2016-01-01

    Submergence stress is a limiting factor for direct-seeded rice systems in rainfed lowlands and flood-prone areas of South and Southeast Asia. The present study demonstrated that submergence stress severely hampered the germination and seedling growth of rice, however, seed priming alleviated the detrimental effects of submergence stress. To elucidate the molecular basis of seed priming-induced submergence tolerance, transcriptome analyses were performed using 4-day-old primed (selenium-Se and salicylic acid-SA priming) and non-primed rice seedlings under submergence stress. Genomewide transcriptomic profiling identified 2371 and 2405 transcripts with Se- and SA-priming, respectively, that were differentially expressed in rice compared with non-priming treatment under submergence. Pathway and gene ontology term enrichment analyses revealed that genes involved in regulation of secondary metabolism, development, cell, transport, protein, and metal handling were over-represented after Se- or SA-priming. These coordinated factors might have enhanced the submergence tolerance and maintained the better germination and vigorous seedling growth of primed rice seedlings. It was also found that many genes involved in cellular and metabolic processes such as carbohydrate metabolism, cellular, and metabolic biosynthesis, nitrogen compound metabolic process, transcription, and response to oxidative stress were induced and overlapped in seed priming treatments, a finding which reveals the common mechanism of seed priming-induced submergence tolerance. Taken together, these results may provide new avenues for understanding and advancing priming-induced responses to submergence tolerance in crop plants. PMID:27516766

  5. The Effect of Prime Duration in Masked Orthographic Priming Depends on Neighborhood Distribution

    ERIC Educational Resources Information Center

    Robert, Christelle; Mathey, Stephanie

    2012-01-01

    A lexical decision task was used with a masked priming procedure to investigate whether and to what extent neighborhood distribution influences the effect of prime duration in masked orthographic priming. French word targets had two higher frequency neighbors that were either distributed over two letter positions (e.g., "LOBE/robe-loge")…

  6. Tight Coupling between Positive and Reversed Priming in the Masked Prime Paradigm

    ERIC Educational Resources Information Center

    Boy, Frederic; Sumner, Petroc

    2010-01-01

    When associations between certain visual stimuli and particular actions are learned, those stimuli become capable of automatically and unconsciously activating their associated action plans. Such sensorimotor priming is assumed to be fundamental for efficient responses, and can be reliably measured in masked prime studies even when the primes are…

  7. Unconscious Congruency Priming from Unpracticed Words Is Modulated by Prime-Target Semantic Relatedness

    ERIC Educational Resources Information Center

    Ortells, Juan J.; Mari-Beffa, Paloma; Plaza-Ayllon, Vanesa

    2013-01-01

    Participants performed a 2-choice categorization task on visible word targets that were preceded by novel (unpracticed) prime words. The prime words were presented for 33 ms and followed either immediately (Experiments 1-3) or after a variable delay (Experiments 1 and 4) by a pattern mask. Both subjective and objective measures of prime visibility…

  8. On the robustness of prime response retrieval processes: evidence from auditory negative priming without probe interference.

    PubMed

    Mayr, Susanne; Buchner, Axel

    2014-02-01

    Visual negative priming has been shown to depend on the presence of probe distractors, a finding that has been traditionally seen to support the episodic retrieval model of negative priming; however, facilitated prime-to-probe contingency learning might also underlie this effect. In four sound identification experiments, the role of probe distractor interference in auditory negative priming was investigated. In each experiment, a group of participants was exposed to probe distractor interference while another group ran the task in the absence of probe distractors. Experiments 1A, 1B, and 1C varied in the extent to which fast versus accurate responding was required. Between Experiments 1 and 2, the spatial cueing of the to-be-attended ear was varied. Whereas participants switched ears from prime to probe in Experiment 1, they kept a stable attentional focus throughout Experiment 2. For trials with probe distractors, a negative priming effect was present in all experiments. For trials without probe distractors, the only ubiquitous after-effect of ignoring a prime distractor was an increase of prime response errors in ignored repetition compared to control trials, indicating that prime response retrieval processes took place. Whether negative priming beyond this error increase was found depended on the stability of the attentional focus. The findings suggest that several mechanisms underlie auditory negative priming with the only robust one being prime response retrieval.

  9. Amount of Priming in the Difference of Mental Transformation

    ERIC Educational Resources Information Center

    Kanamori, Nobuhiro; Yagi, Akihiro

    2005-01-01

    We examined in detail effects of priming in 2 mental rotation strategies: spinning (rotating in a picture plane) and flipping (rotating in depth around a horizontal axis) by using a priming paradigm of Kanamori and Yagi (2002). The priming paradigm included prime and probe tasks within 1 trial. In the prime task, 16 participants were asked to…

  10. Inverse Target- and Cue-Priming Effects of Masked Stimuli

    ERIC Educational Resources Information Center

    Mattler, Uwe

    2007-01-01

    The processing of a visual target that follows a briefly presented prime stimulus can be facilitated if prime and target stimuli are similar. In contrast to these positive priming effects, inverse priming effects (or negative compatibility effects) have been found when a mask follows prime stimuli before the target stimulus is presented: Responses…

  11. The development of gamma-gamma-prime lamellar structures in a nickel-base superalloy during elevated temperature mechanical testing

    NASA Technical Reports Server (NTRS)

    Mackay, R. A.; Ebert, L. J.

    1985-01-01

    The kinetics of the formation and subsequent development of the directional coarsening of the gamma-prime precipitate in model Ni-Al-Mo-Ta superalloy single crystals are examined during tensile creep under various stress levels at 982 and 1038 C. Special attention is given to the gamma and gamma-prime relation to creep time and strain in order to trace the changing gamma-gamma-prime morphology. Directional coarsening of gamma-prime is found to begin during primary creep and its rate is shown to increase with an increase in temperature or stress level. The length of gamma-prime thickness increased linearly with time up to a plateau reached after the onset of steady state creep. The raft thickness, equal to the gamma-prime size, remained constant at this initial value up through the onset of the tertiary creep. The interlaminar spacing indicates the stability of directionally coarsened structure.

  12. Tension-induced pore formation and leakage in adhering vesicles

    NASA Astrophysics Data System (ADS)

    Lenz, P.; Johnson, J. M.; Chan, Y.-H. M.; Boxer, S. G.

    2006-08-01

    The influence of inclusion-induced tension on pore formation is studied theoretically and experimentally. It is shown that fluorescently labeled lipids can enhance pore formation and induce leakage of adhering vesicles. These effects are more pronounced for smaller vesicles. The theoretical predictions are confirmed by experimental two-color fluorescent data. Finally, the influence of the pore formation dynamics on rupture processes of vesicles is analyzed yielding a new picture of the transition to bilayer disks.

  13. Soil Primes the Seed: Epigenetic Landscape Drives Tumor Behavior.

    PubMed

    Whitson, Ramon J; Oro, Anthony E

    2017-02-02

    Whether invasive tumor phenotypes like EMT arise from oncogenic drivers or from priming of the pre-tumor cell of origin remains unknown. In this issue of Cell Stem Cell, Latil et al. (2017) show that the pre-tumor niche establishes a chromatin state predisposing squamous cell carcinomas to undergo EMT and metastasis, suggesting that the pre-tumor epigenome has prognostic value.

  14. ICAM-1 on exosomes from mature dendritic cells is critical for efficient naive T-cell priming.

    PubMed

    Segura, Elodie; Nicco, Carole; Lombard, Bérangère; Véron, Philippe; Raposo, Graça; Batteux, Frédéric; Amigorena, Sebastian; Théry, Clotilde

    2005-07-01

    Exosomes are secreted vesicles formed in late endocytic compartments. Immature dendritic cells (DCs) secrete exosomes, which transfer functional major histocompatibility complex (MHC)-peptide complexes to other DCs. Since immature and mature DCs induce different functional T-cell responses (ie, tolerance versus priming), we asked whether DC maturation also influenced the priming abilities of their exosomes. We show that exosomes secreted by lipopolysaccharide (LPS)-treated mature DCs are 50- to 100-fold more potent to induce antigen-specific T-cell activation in vitro than exosomes from immature DCs. In vitro, exosomes from mature DCs transfer to B lymphocytes the ability to prime naive T cells. In vivo, only mature exosomes trigger effector T-cell responses, leading to fast skin graft rejection. Proteomic and biochemical analyses revealed that mature exosomes are enriched in MHC class II, B7.2, intercellular adhesion molecule 1 (ICAM-1), and bear little milk-fat globule-epidermal growth factor-factor VIII (MFG-E8) as compared with immature exosomes. Functional analysis using DC-derived exosomes from knock-out mice showed that MHC class II and ICAM-1 are required for mature exosomes to prime naive T cells, whereas B7.2 and MFG-E8 are dispensable. Therefore, changes in protein composition and priming abilities of exosomes reflect the maturation signals received by DCs.

  15. Sugar uptake by intestinal basolateral membrane vesicles.

    PubMed

    Wright, E M; van Os, C H; Mircheff, A K

    1980-03-27

    A high yield of membrane vesicles was prepared from the basolateral surface of rat intestinal cells using an N2 cavitation bomb and density gradient centrifugation. The membranes were enriched 10-fold and were free of significatn contamination by brush border membranes and mitochondria. The rate of D-E114C]glucose and L-E13H]glucose uptake into the vesicle was measured using a rapid filtration technique. D-Glucose equilibrated within the vesicles with a half-time 1/25th that for L-glucose. The stereospecific uptake exhibited saturation kinetics with a Km of approx. 44 mM and a V of approx. 110 nmol . mg-1 min-1 at 10 degrees C. The activation energy for the process was 14 kcal . mol-1 below 15 degrees C and it approached 3 kcal . mol-1 above 22 degrees C. Carrier-mediated uptake was eliminated in the presence of 1 mM HgCl2 and 0.5 mM phloretin. The rate of transport was unaffected by the absence or presence of sodium concentration gradients. Competition studies demonstrated that all sugars with the D-glucose pyranose ring chair conformation shared the transport system, and that, with the possible exception of the -OH group at carbon No. 1, there were no specific requirements for an equatorial -OH group at any position in the pyranose ring. In the case of alpha-methyl-D-glucoside its inability to share the D-glucose transport system may be due to steric hindrance posed by the -OCH3 group rather than by a specific requirement for a free hydroxyl group at the position in the ring. It is concluded that sugars are transported across the basolateral membrane of the intestinal epithelium by a facilitated diffusion system reminiscent of that in human red blood cells.

  16. A Paradox of Syntactic Priming: Why Response Tendencies Show Priming for Passives, and Response Latencies Show Priming for Actives

    PubMed Central

    Segaert, Katrien; Menenti, Laura; Weber, Kirsten; Hagoort, Peter

    2011-01-01

    Speakers tend to repeat syntactic structures across sentences, a phenomenon called syntactic priming. Although it has been suggested that repeating syntactic structures should result in speeded responses, previous research has focused on effects in response tendencies. We investigated syntactic priming effects simultaneously in response tendencies and response latencies for active and passive transitive sentences in a picture description task. In Experiment 1, there were priming effects in response tendencies for passives and in response latencies for actives. However, when participants' pre-existing preference for actives was altered in Experiment 2, syntactic priming occurred for both actives and passives in response tendencies as well as in response latencies. This is the first investigation of the effects of structure frequency on both response tendencies and latencies in syntactic priming. We discuss the implications of these data for current theories of syntactic processing. PMID:22022352

  17. Noisy Nonlinear Dynamics of Vesicles in Flow

    NASA Astrophysics Data System (ADS)

    Abreu, David; Seifert, Udo

    2013-06-01

    We present a model for the dynamics of fluid vesicles in linear flow which consistently includes thermal fluctuations and nonlinear coupling between different modes. At the transition between tank treading and tumbling, we predict a trembling motion which is at odds with the known deterministic motions and for which thermal noise is strongly amplified. In particular, highly asymmetric shapes are observed even though the deterministic flow only allows for axisymmetric ones. Our results explain quantitatively recent experimental observations [Levant and Steinberg, Phys. Rev. Lett. 109, 268103 (2012)PRLTAO0031-9007].

  18. Immunological properties of seminal vesicle fluid.

    PubMed

    Veselský, L

    1981-08-01

    Protective significance of some seminal plasma components is described. Lactoferrin is characterized as a primary defense factor against microbial invasion. The agglutinating factor in seminal vesicle fluid may prevent premature elimination of the spermatozoa by leukocytes infiltrating into the female genital tract. The protease inhibitors neutralize the activity of the proteases, thereby protecting the tissues and spermatozoa against proteolytic degradation. Antigens absorbed to spermatozoa during the ejaculation may protect the spermatozoa against the immune system of female reproductive tract. Ejaculated spermatozoa contain immunosuppressive substances that inhibit cell-mediated cytotoxicity as well as lymphocyte response to allogenic cells. These substances may constitute the system that prevents immune damage of spermatozoa.

  19. One version of direct response priming requires automatization of the relevant associations but not awareness of the prime.

    PubMed

    Klapp, Stuart T

    2015-07-01

    Priming is the influence of one event on performance during a second event. One type of priming is known as semantic priming because it biases interpretation of the subsequent stimulus. Another type, direct response priming, biases responding directly without semantic mediation. Research reviewed in this article indicates that two versions of the second type, direct response priming, can be distinguished. One version, explicit priming, requires awareness of the prime. The other version, associative response priming, occurs even if the prime is masked and not phenomenally visible. This version, which is attributed to associations relating specific sensory events to movements of particular muscles, is enabled only if the association has previously been automatized by brief practice in which the to-be-primed response is made to the stimulus that subsequently appears as the prime. Associative response priming can be explained by a simple stimulus-response interpretation; other varieties of priming are more theoretically challenging.

  20. Replicating vesicles as models of primitive cell growth and division.

    PubMed

    Hanczyc, Martin M; Szostak, Jack W

    2004-12-01

    Primitive cells, lacking the complex bio-machinery present in modern cells, would have had to rely on the self-organizing properties of their components and on interactions with their environment to achieve basic cellular functions such as growth and division. Many bilayer-membrane vesicles, depending on their composition and environment, can exhibit complex morphological changes such as growth, fusion, fission, budding, internal vesicle assembly and vesicle-surface interactions. The rich dynamic properties of these vesicles provide interesting models of how primitive cellular replication might have occurred in response to purely physical and chemical forces.

  1. Floating Escherichia coli by expressing cyanobacterial gas vesicle genes

    NASA Astrophysics Data System (ADS)

    Wang, Tianhe; Kang, Li; Li, Jiaheng; Wu, Wenjie; Zhang, Peiran; Gong, Minghao; Lai, Weihong; Zhang, Chunyan; Chang, Lei; Peng, Yong; Yang, Zhongzhou; Li, Lian; Bao, Yingying; Xu, Haowen; Zhang, Xiaohua; Sui, Zhenghong; Yang, Guanpin; Wang, Xianghong

    2015-02-01

    Gas vesicles are hollow, air-filled polyprotein structures that provide the buoyancy to cells. They are found in a variety of prokaryotes. In this study, we isolated a partial gas vesicle protein gene cluster containing gvpA and gvpC20Ψ from Planktothrix rubescens, and inserted it into an expression vector and expressed it in E. coli. The gas vesicle was developed in bacterial cells, which made bacterial cells to float on medium surface. We also amplified gvpA and gvpC20Ψ separately and synthesized an artificial operon by fusing these two genes with the standardized gene expression controlling elements of E. coli. The artificial operon was expressed in E. coli, forming gas vesicles and floating bacteria cells. Our findings verified that the whole set of genes and the overall structure of gas vesicle gene cluster are not necessary for developing gas vesicles in bacteria cells. Two genes, gvpA and gvpC20Ψ, of the gas vesicle gene cluster are sufficient for synthesizing an artificial operon that can develop gas vesicles in bacteria cells. Our findings provided a wide range of applications including easing the harvest of cultured microalgae and bacteria, as well as enriching and remediating aquatic pollutants by constructing gas vesicles in their cells.

  2. Dynamics of multicomponent vesicles in a viscous fluid

    SciTech Connect

    Sohn, Jin Sun Tseng, Y-H Li Shuwang Voigt, Axel Lowengrub, John S.

    2010-01-01

    We develop and investigate numerically a thermodynamically consistent model of two-dimensional multicomponent vesicles in an incompressible viscous fluid. The model is derived using an energy variation approach that accounts for different lipid surface phases, the excess energy (line energy) associated with surface phase domain boundaries, bending energy, spontaneous curvature, local inextensibility and fluid flow via the Stokes equations. The equations are high-order (fourth order) nonlinear and nonlocal due to incompressibility of the fluid and the local inextensibility of the vesicle membrane. To solve the equations numerically, we develop a nonstiff, pseudo-spectral boundary integral method that relies on an analysis of the equations at small scales. The algorithm is closely related to that developed very recently by Veerapaneni et al. [81] for homogeneous vesicles although we use a different and more efficient time stepping algorithm and a reformulation of the inextensibility equation. We present simulations of multicomponent vesicles in an initially quiescent fluid and investigate the effect of varying the average surface concentration of an initially unstable mixture of lipid phases. The phases then redistribute and alter the morphology of the vesicle and its dynamics. When an applied shear is introduced, an initially elliptical vesicle tank-treads and attains a steady shape and surface phase distribution. A sufficiently elongated vesicle tumbles and the presence of different surface phases with different bending stiffnesses and spontaneous curvatures yields a complex evolution of the vesicle morphology as the vesicle bends in regions where the bending stiffness and spontaneous curvature are small.

  3. Searching for Contracting Patterns over Time: Do Prime Contractor and Subcontractor Relations Follow Similar Patterns for Professional Services Provision?

    ERIC Educational Resources Information Center

    Ponomariov, Branco; Kingsley, Gordon; Boardman, Craig

    2011-01-01

    This paper compares over a 12-year period (1) patterns of contracting between a state transportation agency and its prime contractors providing engineering design services with (2) patterns between these prime contractors and their subcontractors. We find evidence of different contracting patterns at each level that emerge over time and coexist in…

  4. The International Society for Extracellular Vesicles launches the first massive open online course on extracellular vesicles

    PubMed Central

    Lässer, Cecilia; Théry, Clotilde; Buzás, Edit I.; Mathivanan, Suresh; Zhao, Weian; Gho, Yong Song; Lötvall, Jan

    2016-01-01

    The International Society for Extracellular Vesicles (ISEV) has organised its first educational online course for students and beginners in the field of extracellular vesicles (EVs). This course, “Basics of Extracellular Vesicles,” uses recorded lectures from experts in the field and will be open for an unlimited number of participants. The course is divided into 5 modules and can be accessed at www.coursera.org/learn/extracellular-vesicles. The first module is an introduction to the field covering the nomenclature and history of EVs. Module 2 focuses on the biogenesis and uptake mechanisms of EVs, as well as their RNA, protein and lipid cargo. Module 3 covers the collection and processing of cell culture media and body fluids such as blood, breast milk, cerebrospinal fluid and urine prior to isolation of EVs. Modules 4 and 5 present different isolation methods and characterisation techniques utilised in the EV field. Here, differential ultracentrifugation, size-exclusion chromatography, density gradient centrifugation, kit-based precipitation, electron microscopy, cryo-electron microscopy, flow cytometry, atomic-force microscopy and nanoparticle-tracking analysis are covered. This first massive open online course (MOOC) on EVs was launched on 15 August 2016 at the platform “Coursera” and is free of charge. PMID:27989272

  5. The International Society for Extracellular Vesicles launches the first massive open online course on extracellular vesicles.

    PubMed

    Lässer, Cecilia; Théry, Clotilde; Buzás, Edit I; Mathivanan, Suresh; Zhao, Weian; Gho, Yong Song; Lötvall, Jan

    2016-01-01

    The International Society for Extracellular Vesicles (ISEV) has organised its first educational online course for students and beginners in the field of extracellular vesicles (EVs). This course, "Basics of Extracellular Vesicles," uses recorded lectures from experts in the field and will be open for an unlimited number of participants. The course is divided into 5 modules and can be accessed at www.coursera.org/learn/extracellular-vesicles. The first module is an introduction to the field covering the nomenclature and history of EVs. Module 2 focuses on the biogenesis and uptake mechanisms of EVs, as well as their RNA, protein and lipid cargo. Module 3 covers the collection and processing of cell culture media and body fluids such as blood, breast milk, cerebrospinal fluid and urine prior to isolation of EVs. Modules 4 and 5 present different isolation methods and characterisation techniques utilised in the EV field. Here, differential ultracentrifugation, size-exclusion chromatography, density gradient centrifugation, kit-based precipitation, electron microscopy, cryo-electron microscopy, flow cytometry, atomic-force microscopy and nanoparticle-tracking analysis are covered. This first massive open online course (MOOC) on EVs was launched on 15 August 2016 at the platform "Coursera" and is free of charge.

  6. Phonological Priming in Children's Picture Naming.

    ERIC Educational Resources Information Center

    Brooks, Patricia J.; MacWhinney, Brian

    2000-01-01

    Two experiments examined phonological priming in children and adults using a cross-modal picture-word interference task. Pictures of familiar objects were presented on a computer screen, while interfering words were presented over headphones. Results indicate that priming effects reach a peak during a time when articulatory information is being…

  7. Perceptual learning can reverse subliminal priming effects.

    PubMed

    Przekoracka-Krawczyk, Anna; Jaśkowski, Piotr

    2007-05-01

    Masked primes presented prior to a target can result in inverse priming (i.e., benefits on trials in which the prime and the target are mapped onto opposite responses). In five experiments, time-of-task effects on subliminal priming of motor responses were investigated. First, we replicated Klapp and Hinkley's (2002) finding that the priming effect is initially straight (i.e., it benefits congruent trials, in which the prime and targets are mapped onto the same response) or absent, and only later reverses (i.e., faster responses in incongruent than in congruent trials). We show that the presentation of the mask plays a crucial role in this reversal and that the reversal occurs later if the mask pattern is very complex. We suggest that perceptual learning improves the recognition of task-relevant features. Once recognized, these features can trigger the preparation of the alternative response and/or inhibit the prime-activated response. These findings support an active role of the mask in priming.

  8. Morphological Priming Survives a Language Switch

    ERIC Educational Resources Information Center

    Verdonschot, Rinus G.; Middelburg, Renee; Lensink, Saskia E.; Schiller, Niels O.

    2012-01-01

    In a long-lag morphological priming experiment, Dutch (L1)-English (L2) bilinguals were asked to name pictures and read aloud words. A design using non-switch blocks, consisting solely of Dutch stimuli, and switch-blocks, consisting of Dutch primes and targets with intervening English trials, was administered. Target picture naming was facilitated…

  9. Negative Priming in Free Recall Reconsidered

    ERIC Educational Resources Information Center

    Hanczakowski, Maciej; Beaman, C. Philip; Jones, Dylan M.

    2016-01-01

    Negative priming in free recall is the finding of impaired memory performance when previously ignored auditory distracters become targets of encoding and retrieval. This negative priming has been attributed to an aftereffect of deploying inhibitory mechanisms that serve to suppress auditory distraction and minimize interference with learning and…

  10. Visual Priming of Inverted and Rotated Objects

    ERIC Educational Resources Information Center

    Knowlton, Barbara J.; McAuliffe, Sean P.; Coelho, Chase J.; Hummel, John E.

    2009-01-01

    Object images are identified more efficiently after prior exposure. Here, the authors investigated shape representations supporting object priming. The dependent measure in all experiments was the minimum exposure duration required to correctly identify an object image in a rapid serial visual presentation stream. Priming was defined as the change…

  11. Can False Memories Prime Problem Solutions?

    ERIC Educational Resources Information Center

    Howe, Mark L.; Garner, Sarah R.; Dewhurst, Stephen A.; Ball, Linden J.

    2010-01-01

    Previous research has suggested that false memories can prime performance on related implicit and explicit memory tasks. The present research examined whether false memories can also be used to prime higher order cognitive processes, namely, insight-based problem solving. Participants were asked to solve a number of compound remote associate task…

  12. 7 CFR 29.2290 - Premature primings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Premature primings. 29.2290 Section 29.2290 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... 21) § 29.2290 Premature primings. Ground leaves harvested before reaching complete growth...

  13. Priming by the variability of visual information

    PubMed Central

    Michael, Elizabeth; de Gardelle, Vincent; Summerfield, Christopher

    2014-01-01

    According to recent theories, perception relies on summary representations that encode statistical information about the sensory environment. Here, we used perceptual priming to characterize the representations that mediate categorization of a complex visual array. Observers judged the average shape or color of a target visual array that was preceded by an irrelevant prime array. Manipulating the variability of task-relevant and task-irrelevant feature information in the prime and target orthogonally, we found that observers were faster to respond when the variability of feature information in the prime and target arrays matched. Critically, this effect occurred irrespective of whether the element-by-element features in the prime and target array overlapped or not, and was even present when prime and target features were drawn from opposing categories. This “priming by variance” phenomenon occurred with prime–target intervals as short as 100 ms. Further experiments showed that this effect did not depend on resource allocation, and occurred even when prime and target did not share the same spatial location. These results suggest that human observers adapt to the variability of visual information, and provide evidence for the existence of a low-level mechanism by which the range or dispersion of visual information is rapidly extracted. This information may in turn help to set the gain of neuronal processing during perceptual choice. PMID:24821803

  14. Priming Addition Facts with Semantic Relations

    ERIC Educational Resources Information Center

    Bassok, Miriam; Pedigo, Samuel F.; Oskarsson, An T.

    2008-01-01

    Results from 2 relational-priming experiments suggest the existence of an automatic analogical coordination between semantic and arithmetic relations. Word pairs denoting object sets served as primes in a task that elicits "obligatory" activation of addition facts (5 + 3 activates 8; J. LeFevre, J. Bisanz, & L. Mrkonjic, 1988). Semantic relations…

  15. The Actual Gemini 9 Prime Crew

    NASA Technical Reports Server (NTRS)

    1966-01-01

    The Gemini 9 backup crew members are, Commander, Thomas P. Stafford and pilot Eugene A. Cernan. The back-up crew became the prime crew when on February 28, 1966 the prime crew for the Gemini 9 mission were killed when their twin seat T- 38 trainer jet aircraft crashed into a building during a landing approach in bad weather.

  16. Does Verb Bias Modulate Syntactic Priming?

    ERIC Educational Resources Information Center

    Bernolet, Sarah; Hartsuiker, Robert J.

    2010-01-01

    In a corpus analysis of spontaneous speech Jaeger and Snider (2007) found that the strength of structural priming is correlated with verb alternation bias. This finding is consistent with an implicit learning account of syntactic priming: because the implicit learning model implemented by Chang (2002), Chang, Dell, and Bock (2006), and Chang,…

  17. Syntactic priming in American Sign Language.

    PubMed

    Hall, Matthew L; Ferreira, Victor S; Mayberry, Rachel I

    2015-01-01

    Psycholinguistic studies of sign language processing provide valuable opportunities to assess whether language phenomena, which are primarily studied in spoken language, are fundamentally shaped by peripheral biology. For example, we know that when given a choice between two syntactically permissible ways to express the same proposition, speakers tend to choose structures that were recently used, a phenomenon known as syntactic priming. Here, we report two experiments testing syntactic priming of a noun phrase construction in American Sign Language (ASL). Experiment 1 shows that second language (L2) signers with normal hearing exhibit syntactic priming in ASL and that priming is stronger when the head noun is repeated between prime and target (the lexical boost effect). Experiment 2 shows that syntactic priming is equally strong among deaf native L1 signers, deaf late L1 learners, and hearing L2 signers. Experiment 2 also tested for, but did not find evidence of, phonological or semantic boosts to syntactic priming in ASL. These results show that despite the profound differences between spoken and signed languages in terms of how they are produced and perceived, the psychological representation of sentence structure (as assessed by syntactic priming) operates similarly in sign and speech.

  18. Phasic Affective Modulation of Semantic Priming

    ERIC Educational Resources Information Center

    Topolinski, Sascha; Deutsch, Roland

    2013-01-01

    The present research demonstrates that very brief variations in affect, being around 1 s in length and changing from trial to trial independently from semantic relatedness of primes and targets, modulate the amount of semantic priming. Implementing consonant and dissonant chords (Experiments 1 and 5), naturalistic sounds (Experiment 2), and visual…

  19. Transient outward K+ channels in vesicles derived from frog skeletal muscle plasma membranes.

    PubMed Central

    Camacho, J; Delay, M J; Vazquez, M; Argüello, C; Sánchez, J A

    1996-01-01

    Whole-cell voltage-clamp experiments were performed in vesicles derived from frog skeletal muscle plasma membranes. Capacitance measurements showed that these vesicles lack invaginations. In solutions containing K+, transient outward currents with reversal potentials close to EK were recorded with a maximum potassium conductance of 0.3 mS/cm2. These currents inactivated in a voltage-dependent manner with a time constant of decay that reached a limiting value of 26 ms at large depolarizations. The steady-state inactivation reached half-maximum values at -66 mV. Transient currents were completely blocked with 5 mM 4-aminopyridine. Single-channel recordings made in inside-out excised patches from the vesicles had ensemble averages with characteristics similar to those of the macroscopic currents, although with significantly faster inactivation time constants. The single-channel chord conductance was 21 pS when the pipette and bath solutions contained 2.5 mM and 120 mM KCl, respectively. It is concluded that these vesicles contain potassium channels that are very similar to A channels found in neurons and other cells. Images FIGURE 8 PMID:8804601

  20. Does translation involve structural priming?

    PubMed

    Maier, Robert M; Pickering, Martin J; Hartsuiker, Robert J

    2017-08-01

    When asked to translate utterances, people might merely make sure that their translations have the same meaning as the source, but they might also maintain aspects of sentence form across languages. We report two experiments in which English-German and German-English bilinguals (without specialist translator training) repeated German ditransitive sentences whose meaning was compatible with more than one grammatical form or translated them into English. Participants almost invariably repeated the sentences accurately, thereby retaining the grammatical structure. Importantly, Experiment 1 found that they tended to repeat grammatical form across languages. Experiment 2 included a condition with sentences that had no grammatical equivalent form in English; here participants tended to persist in the order of thematic roles. We argue that cross-linguistic structural priming plays a major role in the act of translation.

  1. The Apollo 11 Prime Crew

    NASA Technical Reports Server (NTRS)

    1969-01-01

    Portrait of the prime crew of the Apollo 11 lunar landing mission. From left to right they are: Commander, Neil A. Armstrong, Command Module Pilot, Michael Collins, and Lunar Module Pilot, Edwin E. Aldrin Jr. On July 20th 1969 at 4:18 PM, EDT the Lunar Module 'Eagle' landed in a region of the Moon called the Mare Tranquillitatis, also known as the Sea of Tranquillity. After securing his spacecraft, Armstrong radioed back to earth: 'Houston, Tranquility Base here, the Eagle has landed'. At 10:56 p.m. that same evening and witnessed by a worldwide television audience, Neil Armstrong stepped off the 'Eagle's landing pad onto the lunar surface and said: 'That's one small step for a man, one giant leap for mankind.' He became the first human to set foot upon the Moon.

  2. Inhibition by Levorphanol and Related Drugs of Amino Acid Transport by Isolated Membrane Vesicles from Escherichia coli

    PubMed Central

    Holland, Mary J. C.; Simon, Eric J.

    1975-01-01

    Levorphanol inhibits the transport of the amino acids proline and lysine by cytoplasmic membrane vesicles derived from Escherichia coli. The degree of inhibition increases with increasing levorphanol concentration and ranges from 26% at 10−6 M levorphanol to 92% at 10−3 M levorphanol. The effect is independent of the energy source, since levorphanol inhibits proline uptake to the same extent in the presence of 20 mM d-lactate or 20 mM succinate and in the absence of an exogenous energy source. Levorphanol does not irreversibly alter the ability of membrane vesicles to transport proline, since incubation of membrane vesicles for 15 min in the presence of 0.25 mM levorphanol, a concentration which inhibits proline transport by more than 75%, has no effect on the rate of proline transport by these vesicles once the drug is removed. Both the maximum velocity and the Km of proline transport are modified by levorphanol, hence, the type of inhibition produced by levorphanol is mixed. The inhibitor constant (Ki) for levorphanol inhibition of proline transport is approximately 3 × 10−4 M. Membrane vesicles incubated in the presence of levorphanol accumulate much less proline at the steady state than do control vesicles. Furthermore, the addition of levorphanol to membrane vesicles preloaded to the steady state with proline produces a marked net efflux of proline. Levorphanol does not block either temperature-induced efflux or exchange of external proline with [14C]proline present in the intravesicular pool. Dextrorphan, the enantiomorph of levorphanol, and levallorphan, the N-allyl analogue of levorphanol, inhibit proline and lysine transport in a similar manner. Possible mechanisms of the effects of these drugs on cell membranes are discussed. PMID:1096802

  3. Object color affects identification and repetition priming.

    PubMed

    Uttl, Bob; Graf, Peter; Santacruz, Pilar

    2006-10-01

    We investigated the influence of color on the identification of both non-studied and studied objects. Participants studied black and white and color photos of common objects and memory was assessed with an identification test. Consistent with our meta-analysis of prior research, we found that objects were easier to identify from color than from black and white photos. We also found substantial priming in all conditions, and study-to-test changes in an object's color reduced the magnitude of priming. Color-specific priming effects were large for color-complex objects, but minimal for color-simple objects. The pattern and magnitude of priming effects was not influenced either by the extent to which an object always appears in the same color (i.e., whether a color is symptomatic of an object) or by the object's origin (natural versus fabricated). We discuss the implications of our findings for theoretical accounts of object perception and repetition priming.

  4. Priming analogical reasoning with false memories.

    PubMed

    Howe, Mark L; Garner, Sarah R; Threadgold, Emma; Ball, Linden J

    2015-08-01

    Like true memories, false memories are capable of priming answers to insight-based problems. Recent research has attempted to extend this paradigm to more advanced problem-solving tasks, including those involving verbal analogical reasoning. However, these experiments are constrained inasmuch as problem solutions could be generated via spreading activation mechanisms (much like false memories themselves) rather than using complex reasoning processes. In three experiments we examined false memory priming of complex analogical reasoning tasks in the absence of simple semantic associations. In Experiment 1, we demonstrated the robustness of false memory priming in analogical reasoning when backward associative strength among the problem terms was eliminated. In Experiments 2a and 2b, we extended these findings by demonstrating priming on newly created homonym analogies that can only be solved by inhibiting semantic associations within the analogy. Overall, the findings of the present experiments provide evidence that the efficacy of false memory priming extends to complex analogical reasoning problems.

  5. J/. psi. and. psi. prime production with 800 GeV protons

    SciTech Connect

    Moss, J.M.

    1990-01-01

    The yields of J/{psi} and {psi}{prime} vector meson states have been measured for 800 GeV protons incident on deuterium, carbon, calcium, iron and tungsten targets. A depletion of the yield per nucleon from heavy nuclei is observed for both J/{psi} and {psi}{prime} production. This depletion exhibits a strong dependence on x{sub F} and p{sub t}. 24 refs., 3 figs.

  6. Individual Differences in Working Memory Capacity Modulates Semantic Negative Priming from Single Prime Words

    PubMed Central

    Ortells, Juan J.; Noguera, Carmen; Álvarez, Dolores; Carmona, Encarna; Houghton, George

    2016-01-01

    The present study investigated whether semantic negative priming from single prime words depends on the availability of cognitive control resources. Participants with high vs. low working memory capacity (as assessed by their performance in complex span and attentional control tasks) were instructed to either attend to or ignore a briefly presented single prime word that was followed by either a semantically related or unrelated target word on which participants made a lexical decision. Individual differences in working memory capacity (WMC) mainly affected the processing of the ignored primes, but not the processing of the attended primes: While the latter produced reliable positive semantic priming for both high- and low-WMC participants, the former gave rise to reliable semantic negative priming only for high WMC participants, with low WMC participants showing the opposite positive priming effect. The present results extend previous findings in demonstrating that (a) single negative priming can reliably generalize to semantic associates of the prime words, and (b) a differential availability of cognitive control resources can reliably modulate the negative priming effect at a semantic level of representation. PMID:27621716

  7. Shifting Evaluation Windows: Predictable Forward Primes with Long SOAs Eliminate the Impact of Backward Primes

    PubMed Central

    Fockenberg, Daniel A.; Koole, Sander L.; Lakens, Daniël; Semin, Gün R.

    2013-01-01

    Recent work suggests that people evaluate target stimuli within short and flexible time periods called evaluation windows. Stimuli that briefly precede a target (forward primes) or briefly succeed a target (backward primes) are often included in the target's evaluation. In this article, the authors propose that predictable forward primes act as “go” signals that prepare target processing, such that earlier forward primes pull the evaluation windows forward in time. Earlier forward primes may thus reduce the impact of backward primes. This shifting evaluation windows hypothesis was tested in two experiments using an evaluative decision task with predictable (vs. unpredictable) forward and backward primes. As expected, a longer time interval between a predictable forward prime and a target eliminated backward priming. In contrast, the time interval between an unpredictable forward primes and a target had no effects on backward priming. These findings suggest that predictable features of dynamic stimuli can shape target extraction by determining which information is included (or excluded) in rapid evaluation processes. PMID:23359809

  8. The effect of prime duration in masked orthographic priming depends on neighborhood distribution.

    PubMed

    Robert, Christelle; Mathey, Stéphanie

    2012-06-01

    A lexical decision task was used with a masked priming procedure to investigate whether and to what extent neighborhood distribution influences the effect of prime duration in masked orthographic priming. French word targets had two higher frequency neighbors that were either distributed over two letter positions (e.g., LOBE/robe-loge) or concentrated on a single letter position (e.g., FARD/tard-lard). Word targets were preceded by their highest frequency neighbor or by a control prime. Four prime durations were compared (27, 39, 53, and 67 ms). Results showed that the inhibitory priming effect found for words with distributed neighbors at a 67-ms stimulus onset asynchrony (SOA) was canceled when prime duration decreased. In contrast, no priming effect was found in any of the four prime durations for words with concentrated neighbors. Simulations run on the word materials revealed that the interactive activation model (McClelland & Rumelhart, 1981) captured the increasing inhibitory priming effect in the distributed neighbor condition but failed to capture the loss of priming in the concentrated neighbor condition.

  9. Correlation between prime duration and semantic priming effect: evidence from N400 effect.

    PubMed

    Liu, B; Wu, G; Meng, X; Dang, J

    2013-05-15

    When the prime word is masked and flashed very quickly, its presence cannot be perceived by the subjects, but it can still accelerate the processing of the subsequent relevant target stimulus. This is known as the masked priming effect. Adopting a similar experimental paradigm, in this study we take the gray scale pictures describing ordinary objects as prime stimuli and the environmental sounds as target stimuli. The subjects are then asked to classify the target sounds to investigate whether incongruent target sounds elicit the N400 semantic priming effect. At the same time, the prime-mask withdraw value is made at these time intervals: 16ms, 33ms, 50ms, 66ms and 83ms, so as to reveal whether there exists a correlation between the prime duration and the priming effect. The result shows that, at each time interval, the processing of sounds that are congruent with prime pictures is sped up and sounds that are incongruent with prime pictures trigger the N400 effect. The amplitude of the N400 effect is enlarged proportionally with the time interval and gradually approaches the point of saturation. Given this, it is believed that the N400 effect reflects automatic semantic processing, that consciousness is not a prerequisite of semantic priming, and that early visual information has an accumulative priming effect on target sounds.

  10. Individual Differences in Working Memory Capacity Modulates Semantic Negative Priming from Single Prime Words.

    PubMed

    Ortells, Juan J; Noguera, Carmen; Álvarez, Dolores; Carmona, Encarna; Houghton, George

    2016-01-01

    The present study investigated whether semantic negative priming from single prime words depends on the availability of cognitive control resources. Participants with high vs. low working memory capacity (as assessed by their performance in complex span and attentional control tasks) were instructed to either attend to or ignore a briefly presented single prime word that was followed by either a semantically related or unrelated target word on which participants made a lexical decision. Individual differences in working memory capacity (WMC) mainly affected the processing of the ignored primes, but not the processing of the attended primes: While the latter produced reliable positive semantic priming for both high- and low-WMC participants, the former gave rise to reliable semantic negative priming only for high WMC participants, with low WMC participants showing the opposite positive priming effect. The present results extend previous findings in demonstrating that (a) single negative priming can reliably generalize to semantic associates of the prime words, and (b) a differential availability of cognitive control resources can reliably modulate the negative priming effect at a semantic level of representation.

  11. Proteomics of Aggregatibacter actinomycetemcomitans Outer Membrane Vesicles.

    PubMed

    Kieselbach, Thomas; Zijnge, Vincent; Granström, Elisabeth; Oscarsson, Jan

    2015-01-01

    Aggregatibacter actinomycetemcomitans is an oral and systemic pathogen associated with aggressive forms of periodontitis and with endocarditis. Outer membrane vesicles (OMVs) released by this species have been demonstrated to deliver effector proteins such as cytolethal distending toxin (CDT) and leukotoxin (LtxA) into human host cells and to act as triggers of innate immunity upon carriage of NOD1- and NOD2-active pathogen-associated molecular patterns (PAMPs). To improve our understanding of the pathogenicity-associated functions that A. actinomycetemcomitans exports via OMVs, we studied the proteome of density gradient-purified OMVs from a rough-colony type clinical isolate, strain 173 (serotype e) using liquid chromatography-tandem mass spectrometry (LC-MS/MS). This analysis yielded the identification of 151 proteins, which were found in at least three out of four independent experiments. Data are available via ProteomeXchange with identifier PXD002509. Through this study, we not only confirmed the vesicle-associated release of LtxA, and the presence of proteins, which are known to act as immunoreactive antigens in the human host, but we also identified numerous additional putative virulence-related proteins in the A. actinomycetemcomitans OMV proteome. The known and putative functions of these proteins include immune evasion, drug targeting, and iron/nutrient acquisition. In summary, our findings are consistent with an OMV-associated proteome that exhibits several offensive and defensive functions, and they provide a comprehensive basis to further disclose roles of A. actinomycetemcomitans OMVs in periodontal and systemic disease.

  12. Nonenzymatic glycation of phosphatidylethanolamine in erythrocyte vesicles

    SciTech Connect

    Patkowska, M.J.; Horowitz, M.I.

    1986-05-01

    Unsealed inside-out and right-side out vesicles were prepared from human red cells. The vesicles were incubated with D-glucose (/sup 14/C(U)) and sodium cyanoborohydride in phosphate buffer, pH 7.4. After incubation, lipids were extracted with 1-butanol and non-lipid contaminants removed by Sephadex G-25 chromatography. Phosphatidylethanolamine-sorbitol was purified by chromatography on columns of silicic acid and phenylboronate agarose gel. Phospholipase C (B. cereus) liberated phosphoethanolamine-sorbitol (I) which comigrated on TLC with synthetic I prepared by reductive condensation of phosphoethanolamine and D-glucose and also with the product of phospholipase C (B. cereus) hydrolysis of reference phosphatidylethanolamine-sorbitol. Exposure of I to alkaline phosphatase (E. coli) gave P/sub i/ and ethanolamine-sorbitol (II) which comigrated on TLC with synthetic II prepared by reductive condensation of ethanolamine and D-glucose or by phospholipase D hydrolysis of reference phosphatidylethanolamine-sorbitol. These studies demonstrate that vesicular phosphatidylethanolamine can be reductively glycated and illustrate the applicability of both phospholipase C and phospholipase D in characterizing glycated phosphoglycerides.

  13. Tetraspanins in extracellular vesicle formation and function.

    PubMed

    Andreu, Zoraida; Yáñez-Mó, María

    2014-01-01

    Extracellular vesicles (EVs) represent a novel mechanism of intercellular communication as vehicles for intercellular transfer of functional membrane and cytosolic proteins, lipids, and RNAs. Microvesicles, ectosomes, shedding vesicles, microparticles, and exosomes are the most common terms to refer to the different kinds of EVs based on their origin, composition, size, and density. Exosomes have an endosomal origin and are released by many different cell types, participating in different physiological and/or pathological processes. Depending on their origin, they can alter the fate of recipient cells according to the information transferred. In the last two decades, EVs have become the focus of many studies because of their putative use as non-invasive biomarkers and their potential in bioengineering and clinical applications. In order to exploit this ability of EVs many aspects of their biology should be deciphered. Here, we review the mechanisms involved in EV biogenesis, assembly, recruitment of selected proteins, and genetic material as well as the uptake mechanisms by target cells in an effort to understand EV functions and their utility in clinical applications. In these contexts, the role of proteins from the tetraspanin superfamily, which are among the most abundant membrane proteins of EVs, will be highlighted.

  14. Bacterial outer membrane vesicles and vaccine applications.

    PubMed

    Acevedo, Reinaldo; Fernández, Sonsire; Zayas, Caridad; Acosta, Armando; Sarmiento, Maria Elena; Ferro, Valerie A; Rosenqvist, Einar; Campa, Concepcion; Cardoso, Daniel; Garcia, Luis; Perez, Jose Luis

    2014-01-01

    Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A Good Manufacturing Practice (GMP) process was developed at the Finlay Institute to produce OMV from N. meningitidis serogroup B (dOMVB) using detergent extraction. Subsequently, OMV from N. meningitidis, serogroup A (dOMVA), serogroup W (dOMVW), and serogroup X (dOMVX) were obtained using this process. More recently, the extraction process has also been applied effectively for obtaining OMV on a research scale from Vibrio cholerae (dOMVC), Bordetella pertussis (dOMVBP), Mycobacterium smegmatis (dOMVSM), and BCG (dOMVBCG). The immunogenicity of the OMV has been evaluated for specific antibody induction, and together with functional bactericidal and challenge assays in mice has shown their protective potential. dOMVB has been evaluated with non-neisserial antigens, including with a herpes virus type 2 glycoprotein, ovalbumin, and allergens. In conclusion, OMV are proving to be more versatile than first conceived and remain an important technology for development of vaccine candidates.

  15. Bacterial Outer Membrane Vesicles and Vaccine Applications

    PubMed Central

    Acevedo, Reinaldo; Fernández, Sonsire; Zayas, Caridad; Acosta, Armando; Sarmiento, Maria Elena; Ferro, Valerie A.; Rosenqvist, Einar; Campa, Concepcion; Cardoso, Daniel; Garcia, Luis; Perez, Jose Luis

    2014-01-01

    Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A Good Manufacturing Practice (GMP) process was developed at the Finlay Institute to produce OMV from N. meningitidis serogroup B (dOMVB) using detergent extraction. Subsequently, OMV from N. meningitidis, serogroup A (dOMVA), serogroup W (dOMVW), and serogroup X (dOMVX) were obtained using this process. More recently, the extraction process has also been applied effectively for obtaining OMV on a research scale from Vibrio cholerae (dOMVC), Bordetella pertussis (dOMVBP), Mycobacterium smegmatis (dOMVSM), and BCG (dOMVBCG). The immunogenicity of the OMV has been evaluated for specific antibody induction, and together with functional bactericidal and challenge assays in mice has shown their protective potential. dOMVB has been evaluated with non-neisserial antigens, including with a herpes virus type 2 glycoprotein, ovalbumin, and allergens. In conclusion, OMV are proving to be more versatile than first conceived and remain an important technology for development of vaccine candidates. PMID:24715891

  16. A kinetic study of stimulus-induced vesicle recycling in electromotor nerve terminals using labile and stable vesicle markers.

    PubMed

    Agoston, D V; Dowe, G H; Fiedler, W; Giompres, P E; Roed, I S; Walker, J H; Whittaker, V P; Yamaguchi, T

    1986-11-01

    The kinetics of recovery, by recycling electromotor synaptic vesicles, of the biophysical parameters of the reserve population has been studied in perfused blocks of electric organ of Torpedo marmorata prestimulated in vivo, followed by density gradient separation of the extracted vesicles in a zonal rotor using labile (acetylcholine and ATP) and stable (proteoglycan) vesicle markers. Stimulation in vivo at 0.15 Hz for 3.3 h depleted tissue acetylcholine much less than stimulation at 1 Hz for 1 h but nevertheless generated a much larger pool of recycled vesicles that recovered more slowly. At the lower rate of stimulation, recovery of the biophysical characteristics of the reserve population by the recycled vesicles, identified by their content of newly synthesized transmitter, was essentially complete by 8 h. The stable proteoglycan marker was immunochemically assayed and was bimodally distributed in the vesicle-containing portion of the density gradient even in experiments with unstimulated or recovered tissue. The second peak corresponded with that of newly synthesized transmitter and was thus identified as containing the recycled vesicles. Its normalized acetylcholine/proteoglycan ratio was lower than that of the first peak, which is consistent with earlier findings that recycled vesicles, before recovery, are only partially loaded with transmitter. However, as expected, the proportion of total vesicular proteoglycan and acetylcholine associated with the recycled vesicle fraction was very much lower in preparations derived from unstimulated or recovered tissue than in those from recently stimulated tissue.

  17. Movement-Based Priming: Clinical Applications and Neural Mechanisms.

    PubMed

    Stoykov, Mary Ellen; Corcos, Daniel Montie; Madhavan, Sangeetha

    2017-01-01

    Priming can be described as behavior change generated by preceding stimuli. Although various types of priming have been long studied in the field of psychology, priming that targets motor cortex is a relatively new topic of research in the fields of motor control and rehabilitation. In reference to a rehabilitation intervention, priming is categorized as a restorative approach. There are a myriad of possible priming approaches including noninvasive brain stimulation, motor imagery, and sensory-based priming, to name a few. The authors report on movement-based priming which, compared to other priming types, is less frequently examined and under reported. Movement-based priming includes, but is not limited to, bilateral motor priming, unilateral priming, and aerobic exercise. Clinical and neural mechanistic aspects of movement-based priming techniques are explored.

  18. Masked priming by misspellings: Word frequency moderates the effects of SOA and prime-target similarity.

    PubMed

    Burt, Jennifer S

    2016-02-01

    University students made lexical decisions to eight- or nine-letter words preceded by masked primes that were the target, an unrelated word, or a typical misspelling of the target. At a stimulus onset asynchrony (SOA) of 47 ms, primes that were misspellings of the target produced a priming benefit for low-, medium-, and high-frequency words, even when the misspelled primes were changed to differ phonologically from their targets. At a longer SOA of 80 ms, misspelled primes facilitated lexical decisions only to medium- and low-frequency targets, and a phonological change attenuated the benefit for medium-frequency targets. The results indicate that orthographic similarity can be preserved over changes in letter position and word length, and that the priming effect of misspelled words at the shorter SOA is orthographically based. Orthographic-priming effects depend on the quality of the orthographic learning of the target word.

  19. Reading a standing wave: figure-ground-alternation masking of primes in evaluative priming.

    PubMed

    Bermeitinger, Christina; Kuhlmann, Michael; Wentura, Dirk

    2012-09-01

    We propose a new masking technique for masking word stimuli. Drawing on the phenomena of metacontrast and paracontrast, we alternately presented two prime displays of the same word with the background color in one display matching the font color in the other display and vice versa. The sequence of twenty alterations (spanning approx. 267 ms) was sandwich-masked by structure masks. Using this masking technique, we conducted evaluative priming experiments with positive and negative target and prime words. Significant priming effects were found - for primes and targets drawn from the same as well as from different word sets. Priming effects were independent of prime discrimination performance in direct tests and they were still significant after the sample was restricted to those participants who showed random responding in the direct test.

  20. Stimulus-driven changes in the direction of neural priming during visual word recognition.

    PubMed

    Pas, Maciej; Nakamura, Kimihiro; Sawamoto, Nobukatsu; Aso, Toshihiko; Fukuyama, Hidenao

    2016-01-15

    Visual object recognition is generally known to be facilitated when targets are preceded by the same or relevant stimuli. For written words, however, the beneficial effect of priming can be reversed when primes and targets share initial syllables (e.g., "boca" and "bono"). Using fMRI, the present study explored neuroanatomical correlates of this negative syllabic priming. In each trial, participants made semantic judgment about a centrally presented target, which was preceded by a masked prime flashed either to the left or right visual field. We observed that the inhibitory priming during reading was associated with a left-lateralized effect of repetition enhancement in the inferior frontal gyrus (IFG), rather than repetition suppression in the ventral visual region previously associated with facilitatory behavioral priming. We further performed a second fMRI experiment using a classical whole-word repetition priming paradigm with the same hemifield procedure and task instruction, and obtained well-known effects of repetition suppression in the left occipito-temporal cortex. These results therefore suggest that the left IFG constitutes a fast word processing system distinct from the posterior visual word-form system and that the directions of repetition effects can change with intrinsic properties of stimuli even when participants' cognitive and attentional states are kept constant.

  1. Phonological and Orthographic Overlap Effects in Fast and Masked Priming

    PubMed Central

    Frisson, Steven; Bélanger, Nathalie N.; Rayner, Keith

    2014-01-01

    We investigated how orthographic and phonological information is activated during reading, using a fast priming task, and during single word recognition, using masked priming. Specifically, different types of overlap between prime and target were contrasted: high orthographic and high phonological overlap (track-crack), high orthographic and low phonological overlap (bear-gear), or low orthographic and high phonological overlap (fruit-chute). In addition, we examined whether (orthographic) beginning overlap (swoop-swoon) yielded the same priming pattern as end (rhyme) overlap (track-crack). Prime durations were 32 and 50ms in the fast priming version, and 50ms in the masked priming version, and mode of presentation (prime and target in lower case) was identical. The fast priming experiment showed facilitatory priming effects when both orthography and phonology overlapped, with no apparent differences between beginning and end overlap pairs. Facilitation was also found when prime and target only overlapped orthographically. In contrast, the masked priming experiment showed inhibition for both types of end overlap pairs (with and without phonological overlap), and no difference for begin overlap items. When prime and target only shared principally phonological information, facilitation was only found with a long prime duration in the fast priming experiment, while no differences were found in the masked priming version. These contrasting results suggest that fast priming and masked priming do not necessarily tap into the same type of processing. PMID:24365065

  2. Epigenetic Control of Defense Signaling and Priming in Plants.

    PubMed

    Espinas, Nino A; Saze, Hidetoshi; Saijo, Yusuke

    2016-01-01

    Immune recognition of pathogen-associated molecular patterns or effectors leads to defense activation at the pathogen challenged sites. This is followed by systemic defense activation at distant non-challenged sites, termed systemic acquired resistance (SAR). These inducible defenses are accompanied by extensive transcriptional reprogramming of defense-related genes. SAR is associated with priming, in which a subset of these genes is kept at a poised state to facilitate subsequent transcriptional regulation. Transgenerational inheritance of defense-related priming in plants indicates the stability of such primed states. Recent studies have revealed the importance and dynamic engagement of epigenetic mechanisms, such as DNA methylation and histone modifications that are closely linked to chromatin reconfiguration, in plant adaptation to different biotic stresses. Herein we review current knowledge regarding the biological significance and underlying mechanisms of epigenetic control for immune responses in plants. We also argue for the importance of host transposable elements as critical regulators of interactions in the evolutionary "arms race" between plants and pathogens.

  3. Epigenetic Control of Defense Signaling and Priming in Plants

    PubMed Central

    Espinas, Nino A.; Saze, Hidetoshi; Saijo, Yusuke

    2016-01-01

    Immune recognition of pathogen-associated molecular patterns or effectors leads to defense activation at the pathogen challenged sites. This is followed by systemic defense activation at distant non-challenged sites, termed systemic acquired resistance (SAR). These inducible defenses are accompanied by extensive transcriptional reprogramming of defense-related genes. SAR is associated with priming, in which a subset of these genes is kept at a poised state to facilitate subsequent transcriptional regulation. Transgenerational inheritance of defense-related priming in plants indicates the stability of such primed states. Recent studies have revealed the importance and dynamic engagement of epigenetic mechanisms, such as DNA methylation and histone modifications that are closely linked to chromatin reconfiguration, in plant adaptation to different biotic stresses. Herein we review current knowledge regarding the biological significance and underlying mechanisms of epigenetic control for immune responses in plants. We also argue for the importance of host transposable elements as critical regulators of interactions in the evolutionary “arms race” between plants and pathogens. PMID:27563304

  4. Vesicle Stability and Dynamics: An Undergraduate Biochemistry Laboratory

    ERIC Educational Resources Information Center

    Del Bianco, Cristina; Torino, Domenica; Mansy, Sheref S.

    2014-01-01

    A laboratory exercise is described that helps students learn about lipid self-assembly by making vesicles under different solution conditions. Concepts covering the chemical properties of different lipids, the dynamics of lipids, and vesicle stability are explored. Further, the described protocol is easy and cheap to implement. One to two…

  5. Schwannoma, a rare tumor of the seminal vesicle

    PubMed Central

    Carrasquinho, Eduardo; Ferreira, Marco; Afonso, Ana; Ferrito, Fernando

    2011-01-01

    We present a rare case of a schwannoma of the seminal vesicle that occurred in a 43-year-old male with symptoms of the lower urinary tract. Ultrasonography and magnetic resonance imaging documented a solid mass in the patient's left seminal vesicle. A transvesical approach with a transtrigonal midline incision was successfully performed. The microscopic aspect was compatible with schwannoma. PMID:24578861

  6. A new role for myosin II in vesicle fission.

    PubMed

    Flores, Juan A; Balseiro-Gomez, Santiago; Cabeza, Jose M; Acosta, Jorge; Ramirez-Ponce, Pilar; Ales, Eva

    2014-01-01

    An endocytic vesicle is formed from a flat plasma membrane patch by a sequential process of invagination, bud formation and fission. The scission step requires the formation of a tubular membrane neck (the fission pore) that connects the endocytic vesicle with the plasma membrane. Progress in vesicle fission can be measured by the formation and closure of the fission pore. Live-cell imaging and sensitive biophysical measurements have provided various glimpses into the structure and behaviour of the fission pore. In the present study, the role of non-muscle myosin II (NM-2) in vesicle fission was tested by analyzing the kinetics of the fission pore with perforated-patch clamp capacitance measurements to detect single vesicle endocytosis with millisecond time resolution in peritoneal mast cells. Blebbistatin, a specific inhibitor of NM-2, dramatically increased the duration of the fission pore and also prevented closure during large endocytic events. Using the fluorescent markers FM1-43 and pHrodo Green dextran, we found that NM-2 inhibition greatly arrested vesicle fission in a late phase of the scission event when the pore reached a final diameter of ∼ 5 nm. Our results indicate that loss of the ATPase activity of myosin II drastically reduces the efficiency of membrane scission by making vesicle closure incomplete and suggest that NM-2 might be especially relevant in vesicle fission during compound endocytosis.

  7. Microscopic evaluation of vesicles shed by erythrocytes at elevated temperatures.

    PubMed

    Moore, Timothy; Sorokulova, Iryna; Pustovyy, Oleg; Globa, Ludmila; Pascoe, David; Rudisill, Mary; Vodyanoy, Vitaly

    2013-11-01

    The images of human erythrocytes and vesicles were analyzed by a light microscopy system with spatial resolution of better than 90 nm. The samples were observed in an aqueous environment and required no freezing, dehydration, staining, shadowing, marking, or any other manipulation. Temperature elevation resulted in significant concentration increase of structurally transformed erythrocytes (echinocytes) and vesicles in the blood. The process of vesicle separation from spiculated erythrocytes was video recorded in real time. At a temperature of 37°C, mean vesicle concentrations and diameters were found to be 1.50 ± 0.35 × 10(6) vesicles per microliter and 0.365 ± 0.065 μm, respectively. The vesicle concentration increased approximately threefold as the temperature increased from 37 to 40°C. It was estimated that 80% of all vesicles found in the blood are smaller than 0.4 μm. Accurate account of vesicle numbers and dimensions suggest that 86% of the lost erythrocyte material is lost not by vesiculation but by another, as yet, unknown mechanism.

  8. Membrane trafficking: decoding vesicle identity with contrasting chemistries.

    PubMed

    Frost, Adam

    2011-10-11

    Proteins involved in membrane traffic must distinguish between different classes of vesicles. New work now shows that α-synuclein and ALPS motifs represent two extreme types of amphipathic helix that are tuned to detect both the curvature of transport vesicles as well as their bulk lipid content.

  9. Slow Sedimentation and Deformability of Charged Lipid Vesicles

    PubMed Central

    Rey Suárez, Iván; Leidy, Chad; Téllez, Gabriel; Gay, Guillaume; Gonzalez-Mancera, Andres

    2013-01-01

    The study of vesicles in suspension is important to understand the complicated dynamics exhibited by cells in in vivo and in vitro. We developed a computer simulation based on the boundary-integral method to model the three dimensional gravity-driven sedimentation of charged vesicles towards a flat surface. The membrane mechanical behavior was modeled using the Helfrich Hamiltonian and near incompressibility of the membrane was enforced via a model which accounts for the thermal fluctuations of the membrane. The simulations were verified and compared to experimental data obtained using suspended vesicles labelled with a fluorescent probe, which allows visualization using fluorescence microscopy and confers the membrane with a negative surface charge. The electrostatic interaction between the vesicle and the surface was modeled using the linear Derjaguin approximation for a low ionic concentration solution. The sedimentation rate as a function of the distance of the vesicle to the surface was determined both experimentally and from the computer simulations. The gap between the vesicle and the surface, as well as the shape of the vesicle at equilibrium were also studied. It was determined that inclusion of the electrostatic interaction is fundamental to accurately predict the sedimentation rate as the vesicle approaches the surface and the size of the gap at equilibrium, we also observed that the presence of charge in the membrane increases its rigidity. PMID:23874582

  10. A New Role for Myosin II in Vesicle Fission

    PubMed Central

    Cabeza, Jose M.; Acosta, Jorge; Ramirez-Ponce, Pilar; Ales, Eva

    2014-01-01

    An endocytic vesicle is formed from a flat plasma membrane patch by a sequential process of invagination, bud formation and fission. The scission step requires the formation of a tubular membrane neck (the fission pore) that connects the endocytic vesicle with the plasma membrane. Progress in vesicle fission can be measured by the formation and closure of the fission pore. Live-cell imaging and sensitive biophysical measurements have provided various glimpses into the structure and behaviour of the fission pore. In the present study, the role of non-muscle myosin II (NM-2) in vesicle fission was tested by analyzing the kinetics of the fission pore with perforated-patch clamp capacitance measurements to detect single vesicle endocytosis with millisecond time resolution in peritoneal mast cells. Blebbistatin, a specific inhibitor of NM-2, dramatically increased the duration of the fission pore and also prevented closure during large endocytic events. Using the fluorescent markers FM1-43 and pHrodo Green dextran, we found that NM-2 inhibition greatly arrested vesicle fission in a late phase of the scission event when the pore reached a final diameter of ∼ 5 nm. Our results indicate that loss of the ATPase activity of myosin II drastically reduces the efficiency of membrane scission by making vesicle closure incomplete and suggest that NM-2 might be especially relevant in vesicle fission during compound endocytosis. PMID:24959909

  11. Investigating how vesicle size influences vesicle adsorption on titanium oxide: a competition between steric packing and shape deformation.

    PubMed

    Ferhan, Abdul Rahim; Jackman, Joshua A; Cho, Nam-Joon

    2017-01-18

    Understanding the adsorption behavior of lipid vesicles at solid-liquid interfaces is important for obtaining fundamental insights into soft matter adsorbates as well as for practical applications such as supported lipid bilayer (SLB) fabrication. While the process of SLB formation has been highly scrutinized, less understood are the details of vesicle adsorption without rupture, especially at high surface coverages. Herein, we tackle this problem by employing simultaneous quartz crystal microbalance-dissipation (QCM-D) and localized surface plasmon resonance (LSPR) measurements in order to investigate the effect of vesicle size (84-211 nm diameter) on vesicle adsorption onto a titanium oxide surface. Owing to fundamental differences in the measurement principles of the two techniques as well as a mismatch in probing volumes, it was possible to determine both the lipid mass adsorbed near the sensor surface as well as the total mass of adsorbed lipid and hydrodynamically coupled solvent in the adsorbed vesicle layer as a whole. With increasing vesicle size, the QCM-D frequency signal exhibited monotonic behavior reaching an asymptotic value, whereas the QCM-D energy dissipation signal continued to increase according to the vesicle size. In marked contrast, the LSPR-tracked lipid mass near the sensor surface followed a parabolic trend, with the greatest corresponding measurement response occurring for intermediate-size vesicles. The findings reveal that the maximum extent of adsorbed vesicles contacting a solid surface occurs at an intermediate vesicle size due to the competing influences of vesicle deformation and steric packing. Looking forward, such information can be applied to control the molecular self-assembly of phospholipid assemblies as well as provide the basis for investigating deformable, soft matter adsorbates.

  12. A Preferable Method for the Formation of Vesicles from Lamellar Liquid Crystals Using Chemical Additives.

    PubMed

    Enomoto, Yasutaka; Imai, Yoko; Tajima, Kazuo

    2017-01-01

    We present a method for vesicle formation from lamellar liquid crystals (LCs) using a cationic amphiphilic substance, namely 2-hydroxyethyl di(alkanol)oxyethyl methylammonium methylsulfate (DEAE). Vesicle formation from the DEAE lamellar dispersion occurred via a two-step chemical addition. This method required neither additional mechanical energy nor the use of special solvents. The transition was solubilized using an organic substance (e.g., limonene) in the lamellar DEAE LC, after which, a small amount of inorganic salt was added to the solubilized lamellar LC dispersion with gentle stirring. The viscosity of the DEAE dispersion following salt addition decreased sharply from 10(5) mPa·s to 10(2) mPa·s, and the DEAE dispersion was converted into a high fluidity liquid. Several organic substances were examined as potential solubilizates to initiate the lamellar-vesicle transition. Inorganic salts were also examined as transition triggers using various types of electrolytes; only neutral salts were effective as trigger additives. Dissociation of inorganic salts yielded anions, which inserted between the DEAE bilayer membranes and induced OH(-) ion exchange. In addition, a number of cations simultaneously formed ion pairs with the DEAE counter ions (CH3SO4(-) ions). However, as the amount of solubilized organic substances in the DEAE bilayer membrane decreased over time, the vesicles were transformed into lamellar LCs once again. The DEAE states in each step were measured by monitoring the zeta potential, pH, viscosity, and by examination of scanning electron microscopy and atomic force microscopy images. A possible molecular mechanism for the lamellar-vesicle transition of DEAE was proposed.

  13. Re-naming D Double Prime

    NASA Technical Reports Server (NTRS)

    Chao, Benjamin F.

    1999-01-01

    "Knowledge about the dynamics of the D double prime region is a key to unlock some fundamental mysteries of the Earth heat engine which governs a wide range of global geophysical processes from tectonics to geodynamo." This benign sentence makes complete sense to many geophysicists. But for many others, it makes sense all except the odd nomenclature "D double prime". One knows about the crust, upper and lower mantle, outer and inner core, but where is the D double prime region? What meaning does it try to convey? Where is D prime region, or D, or A, B, C regions for that matter, and are there higher-order primes? How does such an odd name come about anyway? D double prime, or more "simply" D", is a generic designation given to the thin shell, about 200 km thick, of the lowermost mantle just above the core-mantle boundary inside the Earth. Incidentally, whether D" is "simpler" than "D double prime" depends on whether you are pronouncing it or writing/typing it; and D" can be confusing to readers in distinguishing quotation marks (such as in the above sentences) and second derivatives, and to word processors in spelling check and indexing.

  14. Explicit representation of microbes, enzymes, mineral surfaces, and isotopic tracers helps explain soil organic carbon decomposition and priming

    NASA Astrophysics Data System (ADS)

    Zhu, X.; Tang, J.; Riley, W. J.; Wallenstein, M. D.; Cotrufo, M. F.

    2015-12-01

    Increased plant carbon inputs from CO2 fertilization could accelerate native soil organic carbon (SOC) decomposition through the priming effect. Although this increase in SOC turnover rate due to priming might have important consequences for SOC dynamics, there are large uncertainties in the sign and magnitude of priming, as well as large challenges in identifying underlying mechanisms. Current SOC models, mostly based on first-order decomposition representations, do no represent many important biotic and abiotic processes, including the priming effect. The incorporation of explicit biotic and abiotic interactions in modeling SOC decomposition may improve our ability to accurately predict SOC dynamics. In this study, we (1) develop a microbe-explicit SOC decomposition model to simulate SOC turnovers and priming and (2) test the model with a soil incubation experiment with 14C-labeled glucose addition. We report (1) the evolutions of modeled carbon pools, (2) the fate of 14C labeled glucose addition, (3) the model performance compared to observations, (4) the transient behavior of priming components, and (5) an analysis of the effects of carbon input magnitudes and frequencies on the priming effect. Here are some findings from our model-experiment analyses: (1) the inclusion of an extracellular oxidative metabolism (EXOMET) in addition to intracellular microbial respiration helps improve the model performance; (2) priming is dominated by intracellular microbial respiration at the beginning of incubation (~ first 5 days) but later on dominated by EXOMET, which explains observed long-term sustaining priming; (3) the varying magnitudes of glucose addition do not change the magnitude of priming per unit addition; (4) the varying frequencies of glucose addition change the magnitude of priming per unit addition, but with contrast changing patterns for non-steady-state and steady state simulations; (5) constant annual total glucose addition shift the system to another

  15. Electrophysiology reveals semantic priming at a short SOA irrespective of depth of prime processing.

    PubMed

    Küper, Kristina; Heil, Martin

    2009-04-03

    The otherwise robust behavioral semantic priming effect is reduced to the point of being absent when a letter search has to be performed on the prime word. As a result the automaticity of semantic activation has been called into question. It is unclear, however, in how far automatic processes are even measurable in the letter search priming paradigm as the prime task necessitates a long prime-probe stimulus-onset asynchrony (SOA). In a modified procedure, a short SOA can be realized by delaying the prime task response until after participants have made a lexical decision on the probe. While the absence of lexical decision priming has already been demonstrated in this design it seems premature to draw any definite conclusions from this purely behavioral result since event related potential (ERP) measures have been shown to be a more sensitive index of semantic activation. Using the modified paradigm we thus recorded ERP in addition to lexical decision times. Stimuli were presented at two different SOAs (240 ms vs. 840 ms) and participants performed either a grammatical discrimination (Experiment 1) or a letter search (Experiment 2) on the prime. Irrespective of prime task, the modulation of the N400, the ERP correlate of semantic activation, provided clear-cut evidence of semantic processing at the short SOA. Implications for theories of semantic activation as well as the constraints of the delayed prime task procedure are discussed.

  16. TRPM7 facilitates cholinergic vesicle fusion with the plasma membrane.

    PubMed

    Brauchi, Sebastian; Krapivinsky, Grigory; Krapivinsky, Luba; Clapham, David E

    2008-06-17

    TRPM7, of the transient receptor potential (TRP) family, is both an ion channel and a kinase. Previously, we showed that TRPM7 is located in the membranes of acetylcholine (ACh)-secreting synaptic vesicles of sympathetic neurons, forms a molecular complex with proteins of the vesicular fusion machinery, and is critical for stimulated neurotransmitter release. Here, we targeted pHluorin to small synaptic-like vesicles (SSLV) in PC12 cells and demonstrate that it can serve as a single-vesicle plasma membrane fusion reporter. In PC12 cells, as in sympathetic neurons, TRPM7 is located in ACh-secreting SSLVs. TRPM7 knockdown by siRNA, or abolishing channel activity by expression of a dominant negative TRPM7 pore mutant, decreased the frequency of spontaneous and voltage-stimulated SSLV fusion events without affecting large dense core vesicle secretion. We conclude that the conductance of TRPM7 across the vesicle membrane is important in SSLV fusion.

  17. Placental Extracellular Vesicles and Feto-Maternal Communication

    PubMed Central

    Tong, M.; Chamley, L.W.

    2015-01-01

    The human placenta is an anatomically unique structure that extrudes a variety of extracellular vesicles into the maternal blood (including syncytial nuclear aggregates, microvesicles, and nanovesicles). Large quantities of extracellular vesicles are produced by the placenta in both healthy and diseased pregnancies. Since their first description more than 120 years ago, placental extracellular vesicles are only now being recognized as important carriers for proteins, lipids, and nucleic acids, which may play a crucial role in feto-maternal communication. Here, we summarize the current literature on the cargos of placental extracellular vesicles and the known effects of such vesicles on maternal cells/systems, especially those of the maternal immune and vascular systems. PMID:25635060

  18. Human placental coated vesicles contain receptor-bound transferrin.

    PubMed Central

    Booth, A G; Wilson, M J

    1981-01-01

    Human placental coated vesicles have been purified by a method involving sucrose-density-gradient centrifugation and treatment with wheat-germ agglutinin. These preparations were free of contamination by placental microvillus fragments. Crossed immunoelectrophoresis demonstrated that the coated vesicles contained a single serum protein, which was identified as transferrin. This transferrin was only observed after the vesicles were treated with a non-ionic detergent, and its behaviour during crossed hydrophobic-interaction immunoelectrophoresis suggested that a large proportion of it was receptor-bound. No other serum proteins, including immunoglobulin G, could be detected in these preparations. Receptor-bound transferrin was the only antigen common to placental coated vesicles and microvilli, implying that other plasma-membrane proteins are excluded from the region of membrane involved in coated-vesicle formation. Images PLATE 2 PLATE 1 Fig. 1. Fig. 2. Fig. 3. PMID:6272755

  19. Interaction of a potyviral VPg with anionic phospholipid vesicles

    SciTech Connect

    Rantalainen, Kimmo I.; Christensen, Peter A.; Hafren, Anders; Otzen, Daniel E.; Kalkkinen, Nisse; Maekinen, Kristiina

    2009-12-05

    The viral genome-linked protein (VPg) of Potato virus A (PVA) is a multifunctional protein that belongs to a class of intrinsically disordered proteins. Typically, this type of protein gains a more stable structure upon interactions or posttranslational modifications. In a membrane lipid strip overlay binding assay, PVA VPg was found to bind phosphatidylserine (PS), but not phosphatidylcholine (PC). According to circular dichroism spectroscopy, the secondary structure of PVA VPg was stabilized upon interactions with PS and phosphatidylglycerol (PG), but not with PC vesicles. It is possible that this stabilization favored the formation of alpha-helical structures. Limited tryptic digestion showed that the interaction with anionic vesicles protected certain, otherwise accessible, trypsin cleavage sites. An electron microscopy study revealed that interaction with VPg substantially increased the vesicle diameter and caused the formation of pore or plaque-like electron dense spots on the vesicle surface, which gradually led to disruption of the vesicles.

  20. A light-scattering characterization of membrane vesicles.

    PubMed Central

    Selser, J C; Yeh, Y; Baskin, R J

    1976-01-01

    A technique has been developed in this paper which enables quasi-elastic laser light scattering to be used to accurately and quantitatively measure the average vesicle diffusion coefficient and the relative dispersion in the diffusion coefficient about this average for dilute polydisperse vesicle suspensions. This technique relies on a theoretical analysis of a modified form of the Z-averaged diffusion coefficient. This modified Z-averaged diffusion coefficient explicitly incorporates vesicle size, structure, and polydispersity in a description of the scattered light autocorrelation spectrum. Light-scattering experiments were performed on a dilute, lobster sarcoplasmic reticulum vesicle suspension and the measured average diffusion coefficient and the diffusion coefficient relative dispersion about this average were determined with accuracies of 2 and 10%, respectively. A comparison of vesicle size inferred from light-scattering results was made with size results from electron microscopic analysis of the same sample. Images FIGURE 2 FIGURE 3 FIGURE 4 PMID:1252585

  1. Redox-Reactive Membrane Vesicles produced by Shewanella

    SciTech Connect

    Gorby, Yuri A.; McLean, Jeffrey S.; Korenevsky, Anton A.; Rosso, Kevin M.; El-Naggar, Mohamed Y.; Beveridge, Terrance J.

    2008-06-01

    Dissimilatory iron reducing bacteria produce and release membrane vesicles with diameters ranging from 50 to 250 nm. The vesicles, which arise from the outer membrane of these Gram-negative bacteria, lack DNA but contain proteins that catalyze the reduction of ferric iron and other multivalent heavy metals and radionuclides. This enzymatic process results in the formation of nano-size biogenic mineral assemblages that resemble nanofossils. Under low-shear conditions, membrane vesicles are commonly tethered to intact cells by electrically conductive filaments known as bacterial nanowires. The functional role of membrane vesicles and associated nanowires is not known, but the potential for mineralized vesicles that morphologically resemble nanofossils to serve as paleontological indicators of early life on earth and as biosignatures of like on other planets is recognized.

  2. Recognition and tethering of transport vesicles at the Golgi apparatus.

    PubMed

    Witkos, Tomasz M; Lowe, Martin

    2017-02-23

    The Golgi apparatus occupies a central position within the secretory pathway where it is a hub for vesicle trafficking. Distinct classes of transport vesicles traffic diverse cargoes into and out of this organelle, as well as between the different Golgi subcompartments. A key feature of Golgi trafficking is the specific recognition of transport vesicles at the different regions of the Golgi apparatus, required for the correct cargo delivery. Specificity is ensured by coiled-coil golgins and multi-subunit tethering complexes (MTCs), which act together to capture vesicles and promote their subsequent fusion with the Golgi membrane. In this review we discuss our current understanding of how golgins and MTCs function together to mediate the specific recognition of vesicles at the Golgi apparatus.

  3. Charge-reversal instability in mixed bilayer vesicles

    NASA Astrophysics Data System (ADS)

    Chen, Yi; Nelson, Philip

    2000-08-01

    Bilayer vesicles form readily from mixtures of charged and neutral surfactants. When such a mixed vesicle binds an oppositely charged object, its membrane partially demixes: the adhesion zone recruits more charged surfactants from the rest of the membrane. Given an unlimited supply of adhering objects one might expect the vesicle to remain attractive until it was completely covered. Contrary to this expectation, we show that a vesicle can instead exhibit adhesion saturation, partitioning spontaneously into an attractive zone with definite area fraction, and a repulsive zone. The latter zone rejects additional incoming objects because counterions on the interior of the vesicle migrate there, effectively reversing the membrane's charge. The effect is strongest at high surface charge densities, low ionic strength, and with thin, impermeable membranes. Adhesion saturation in such a situation has recently been observed experimentally [H. Aranda-Espinoza et al., Science 285, 394 (1999)].

  4. Permeability of human erythrocyte membrane vesicles to alkali cations.

    PubMed

    Sze, H; Solomon, A K

    1979-02-02

    The permeability of inside-out and right-side-out vesicles from erythrocyte membranes to inorganic cations was determined quantitatively. Using 86Rb as a K analog, we have measured the rate constant of 86Rb efflux from vesicles under equilibrium exchange conditions, using a dialysis procedure. The permeability coefficients of the vesicles to Rb are only about an order of magnitude greater than that of whole erythrocytes. Furthermore, we have measured many of the specialized transport systems known to exist in erythrocytes and have shown that glucose, sulfate, ATP-dependent Ca and ATP-dependent Na transport activities are retained by the vesicle membranes. These results suggest that inside-out and right-side-out vesicles can be used effectively to study transport properties of erythrocyte membranes.

  5. Interaction between silicon dioxide and dipalmitoylphosphatidylcholine (DPPC) vesicles

    SciTech Connect

    Mohd, Hur Munawar Kabir; Ahmad, Ainee Fatimah; Radiman, Shahidan; Mohamed, Faizal; Rosli, Nur Ratasha Alia Md; Ayob, Muhammad Taqiyuddin Mawardi; Rahman, Irman Abdul

    2014-09-03

    Many of the cellular process depend on the ability of the membrane to separate areas while allowing exchange and tightly regulated transport of material within and across the membrane to occur, which is the driving principle behind cell communication. The complexity of biological membranes has motivated the development of a wide variety of simpler model systems whose size, geometry and composition can be tailored with precision. This study was conducted to investigate the interactions between silica nanoparticles and Dipalmitoylphosphatidylcholine (DPPC) vesicles. The size range of DPPC vesicles formed was from 50 to 150 nm. Concentration of silica added to the vesicles was varied from 0.25 to 1.5 mg/ml. The change in vesicle size distribution, localization and positioning of silica nanoparticles in vesicles was studied via transmission electron microscopy (TEM) and differential scanning calorimetry (DSC)

  6. Vesicle coats: structure, function, and general principles of assembly.

    PubMed

    Faini, Marco; Beck, Rainer; Wieland, Felix T; Briggs, John A G

    2013-06-01

    The transport of proteins and lipids between distinct cellular compartments is conducted by coated vesicles. These vesicles are formed by the self-assembly of coat proteins on a membrane, leading to collection of the vesicle cargo and membrane bending to form a bud. Scission at the bud neck releases the vesicle. X-ray crystallography and electron microscopy (EM) have recently generated models of isolated coat components and assembled coats. Here, we review these data to present a structural overview of the three main coats: clathrin, COPII, and COPI. The three coats have similar function, common ancestry, and structural similarities, but exhibit fundamental differences in structure and assembly. We describe the implications of structural similarities and differences for understanding the function, assembly principles, and evolution of vesicle coats.

  7. Translocation of an Incompressible Vesicle through a Pore

    PubMed Central

    Shojaei, Hamid R.; Muthukumar, Murugappan

    2016-01-01

    We have derived the free energy landscape for the translocation of a single vesicle through a narrow pore by accounting for bending and stretching of the vesicle, and the deformation of the vesicle by the pore. Emergence of a free energy barrier for translocation is a general result, and the magnitude of the barrier is calculated in terms of the various material parameters. The extent of the reduction in the barrier by the presence of an external constant force is calculated. Using the Fokker–Planck formalism, we have calculated the average translocation time corresponding to the various free energy landscapes representing different parameter sets. The dependencies of the average translocation time on the strength of the external force, vesicle size, bending and stretching moduli of the vesicle, and radius and length of the pore are derived, and the computed results are discussed. PMID:27089012

  8. AC-electric field dependent electroformation of giant lipid vesicles.

    PubMed

    Politano, Timothy J; Froude, Victoria E; Jing, Benxin; Zhu, Yingxi

    2010-08-01

    Giant vesicles of larger than 5 microm, which have been of intense interest for their potential as drug delivery vehicles and as a model system for cell membranes, can be rapidly formed from a spin-coated lipid thin film under an electric field. In this work, we explore the AC-field dependent electroformation of giant lipid vesicles in aqueous media over a wide range of AC-frequency from 1 Hz to 1 MHz and peak-to-peak field strength from 0.212 V/mm to 40 V/mm between two parallel conducting electrode surfaces. By using fluorescence microscopy, we perform in-situ microscopic observations of the structural evolution of giant vesicles formed from spin-coated lipid films under varied uniform AC-electric fields. The real-time observation of bilayer bulging from the lipid film, vesicle growth and fusing further examine the critical role of AC-induced electroosmotic flow of surrounding fluids for giant vesicle formation. A rich AC-frequency and field strength phase diagram is obtained experimentally to predict the AC-electroformation of giant unilamellar vesicles (GUVs) of l-alpha-phosphatidylcholine, where a weak dependence of vesicle size on AC-frequency is observed at low AC-field voltages, showing decreased vesicle size with a narrowed size distribution with increased AC-frequency. Formation of vesicles was shown to be constrained by an upper field strength of 10 V/mm and an upper AC-frequency of 10 kHz. Within these parameters, giant lipid vesicles were formed predominantly unilamellar and prevalent across the entire electrode surfaces.

  9. Aggregation of phospholipid vesicles by water-soluble polymers.

    PubMed Central

    Meyuhas, D; Nir, S; Lichtenberg, D

    1996-01-01

    Water-soluble polymers such as dextran and polyethylene glycol are known to induce aggregation and size growth of phospholipid vesicles. The present study addresses the dependence of these processes on vesicle size and concentration, polymer molecular weight, temperature, and compartmentalization of the vesicles and polymers, using static and dynamic light scattering. Increasing the molecular weight of the polymers resulted in a reduction of the concentration of polymer needed for induction of aggregation of small unilamellar vesicles. The aggregation was fully reversible (by dilution), within a few seconds, up to a polymer concentration of at least 20 wt %. At relatively low phosphatidylcholine (PC) concentrations (up to approximately 1 mM), increasing the PC concentration resulted in faster kinetics of aggregation and reduced the threshold concentration of polymer required for rapid aggregation (CA). At higher PC concentrations, CA was only slightly dependent on the concentration of PC and was approximately equal to the overlapping concentration of the polymer (C*). The extent of aggregation was similar at 37 and 4 degrees C. Aggregation of large unilamellar vesicles required a lower polymer concentration, probably because aggregation occurs in a secondary minimum (without surface contact). In contrast to experiments in which the polymers were added directly to the vesicles, dialysis of the vesicles against polymer-containing solutions did not induce aggregation. Based on this result, it appears that exclusion of polymer from the hydration sphere of vesicles and the consequent depletion of polymer molecules from clusters of aggregated vesicles play the central role in the induction of reversible vesicle aggregation. The results of all the other experiments are consistent with this conclusion. PMID:8913598

  10. Microfluidic filtration system to isolate extracellular vesicles from blood.

    PubMed

    Davies, Ryan T; Kim, Junho; Jang, Su Chul; Choi, Eun-Jeong; Gho, Yong Song; Park, Jaesung

    2012-12-21

    Extracellular vesicles are released by various cell types, particularly tumor cells, and may be potential targets for blood-based cancer diagnosis. However, studies performed on blood-borne vesicles to date have been limited by lack of effective, standardized purification strategies. Using in situ prepared nanoporous membranes, we present a simple strategy employing a microfluidic filtration system to isolate vesicles from whole blood samples. This method can be applied to purify nano-sized particles from blood allowing isolation of intact extracellular vesicles, avoiding the need for laborious and potentially damaging centrifugation steps or overly specific antibody-based affinity purification. Porous polymer monoliths were integrated as membranes into poly(methyl methacrylate) microfluidic chips by benchtop UV photopolymerization through a mask, allowing precise positioning of membrane elements while preserving simplicity of device preparation. Pore size could be manipulated by changing the ratio of porogenic solvent to prepolymer solution, and was tuned to a size proper for extraction of vesicles. Using the membrane as a size exclusion filter, we separated vesicles from cells and large debris by injecting whole blood under pressure through the microfluidic device. To enhance isolation purity, DC electrophoresis was employed as an alternative driving force to propel particles across the filter and increase the separation efficiency of vesicles from proteins. From the whole blood of melanoma-grown mice, we isolated extracellular vesicles and performed RT-PCR to verify their contents of RNA. Melan A mRNA derived from melanoma tumor cells were found enriched in filtered samples, confirming the recovery of vesicles via their cargo. This filtration system can be incorporated into other on-chip processes enabling integrated sample preparation for the downstream analysis of blood-based extracellular vesicles.

  11. Vesicle dynamics during the atmospheric entry heating of cosmic spherules

    NASA Astrophysics Data System (ADS)

    Genge, M. J.

    2017-03-01

    Cosmic spherules are unique igneous objects that form by melting due to gas drag heating during atmospheric entry heating. Vesicles are an important component of many cosmic spherules since they suggest their precursors had finite volatile contents. Vesicle abundances in spherules decrease through the series porphyritic, glassy, barred, to cryptocrystalline spherules. Anomalous hollow spherules, with large off-center vesicles occur in both porphyritic and glassy spheres. Numerical simulation of the dynamic behavior of vesicles during atmospheric flight is presented that indicates vesicles rapidly migrate due to deceleration and separate from nonporphyritic particles. Modest rotation rates of tens of radians s-1 are, however, sufficient to impede loss of vesicles and may explain the presence of small solitary vesicles in barred, cryptocrystalline and glassy spherules. Rapid rotation at spin rates of several thousand radians s-1 are required to concentrate vesicles at the rotational axis and leads to rapid growth by coalescence and either separation or retention depending on the orientation of the rotational axis. Complex rapid rotations that concentrate vesicles in the core of particles are proposed as a mechanism for the formation of hollow spherules. High vesicle contents in porphyritic spherules suggest volatile-rich precursors; however, calculation of volatile retention indicates these have lost >99.9% of volatiles to degassing prior to melting. The formation of hollow spherules, by rapid spin, necessarily implies preatmospheric rotations of several thousand radians s-1. These particles are suggested to represent immature dust, recently released from parent bodies, in which rotations have not been slowed by magnetic damping.

  12. Dissociated backward priming effects in lexical decision and pronunciation tasks.

    PubMed

    Kahan, T A; Neely, J H; Forsythe, W J

    1999-03-01

    Backward priming was examined at 150- and 500-msec prime-target stimulus onset asynchronies (SOAs) using visually presented primes and targets in lexical decision and pronunciation tasks. Two kinds of backward relations were used: compound items for which targets and primes formed a word in the backward direction (e.g., prime: HOP; target: bell), and noncompound items for which targets and primes did not form a word but were associatively related in the backward but not the forward direction (e.g., prime: BABY; target: stork). Results showed that backward priming effects were equivalent for compounds and noncompounds. However, for lexical decisions, backward priming occurred at both SOAs, whereas for pronunciation, it occurred only at the 150-msec SOA. We discuss how this SOA-dissociated backward priming effect in lexical decision and pronunciation tasks poses a serious challenge for all theories of semantic priming.

  13. Extracellular Vesicles in Chronic Obstructive Pulmonary Disease

    PubMed Central

    Kadota, Tsukasa; Fujita, Yu; Yoshioka, Yusuke; Araya, Jun; Kuwano, Kazuyoshi; Ochiya, Takahiro

    2016-01-01

    Chronic obstructive pulmonary disease (COPD) is characterized by the progression of irreversible airflow limitation and is a leading cause of morbidity and mortality worldwide. Although several crucial mechanisms of COPD pathogenesis have been studied, the precise mechanism remains unknown. Extracellular vesicles (EVs), including exosomes, microvesicles, and apoptotic bodies, are released from almost all cell types and are recognized as novel cell–cell communication tools. They have been shown to carry and transfer a wide variety of molecules, such as microRNAs, messenger RNAs, and proteins, which are involved in physiological functions and the pathology of various diseases. Recently, EVs have attracted considerable attention in pulmonary research. In this review, we summarize the recent findings of EV-mediated COPD pathogenesis. We also discuss the potential clinical usefulness of EVs as biomarkers and therapeutic agents for the treatment of COPD. PMID:27801806

  14. Biomimetic proteolipid vesicles for targeting inflamed tissues

    NASA Astrophysics Data System (ADS)

    Molinaro, R.; Corbo, C.; Martinez, J. O.; Taraballi, F.; Evangelopoulos, M.; Minardi, S.; Yazdi, I. K.; Zhao, P.; De Rosa, E.; Sherman, M. B.; de Vita, A.; Toledano Furman, N. E.; Wang, X.; Parodi, A.; Tasciotti, E.

    2016-09-01

    A multitude of micro- and nanoparticles have been developed to improve the delivery of systemically administered pharmaceuticals, which are subject to a number of biological barriers that limit their optimal biodistribution. Bioinspired drug-delivery carriers formulated by bottom-up or top-down strategies have emerged as an alternative approach to evade the mononuclear phagocytic system and facilitate transport across the endothelial vessel wall. Here, we describe a method that leverages the advantages of bottom-up and top-down strategies to incorporate proteins derived from the leukocyte plasma membrane into lipid nanoparticles. The resulting proteolipid vesicles--which we refer to as leukosomes--retained the versatility and physicochemical properties typical of liposomal formulations, preferentially targeted inflamed vasculature, enabled the selective and effective delivery of dexamethasone to inflamed tissues, and reduced phlogosis in a localized model of inflammation.

  15. Biomimetic proteolipid vesicles for targeting inflamed tissues

    PubMed Central

    Molinaro, R.; Corbo, C.; Martinez, J. O.; Taraballi, F.; Evangelopoulos, M.; Minardi, S.; Yazdi, I.K.; Zhao, P.; De Rosa, E.; Sherman, M.; De Vita, A.; Furman, N.E. Toledano; Wang, X.; Parodi, A.; Tasciotti, E.

    2016-01-01

    A multitude of micro- and nanoparticles have been developed to improve the delivery of systemically administered pharmaceuticals, which are subject to a number of biological barriers that limit their optimal biodistribution. Bioinspired drug-delivery carriers formulated by bottom-up or top-down strategies have emerged as an alternative approach to evade the mononuclear phagocytic system and facilitate the transport across the endothelial vessel wall. Here, we describe a method that leverages the advantages of bottom-up and top-down strategies to incorporate proteins derived from the leukocyte plasma membrane into lipid nanoparticles. The resulting proteolipid vesicles - which we refer to as leukosomes - retained the versatility and physicochemical properties typical of liposomal formulations, preferentially targeted inflamed vasculature, enabled the selective and effective delivery of dexamethasone to inflamed tissues, and reduced phlogosis in a localized model of inflammation. PMID:27213956

  16. Vesicular Glutamate Transporter 1 Orchestrates Recruitment of Other Synaptic Vesicle Cargo Proteins during Synaptic Vesicle Recycling*

    PubMed Central

    Pan, Ping-Yue; Marrs, Julia; Ryan, Timothy A.

    2015-01-01

    A long standing question in synaptic physiology is how neurotransmitter-filled vesicles are rebuilt after exocytosis. Among the first steps in this process is the endocytic retrieval of the transmembrane proteins that are enriched in synaptic vesicles (SVs). At least six types of transmembrane proteins must be recovered, but the rules for how this multiple cargo selection is accomplished are poorly understood. Among these SV cargos is the vesicular glutamate transporter (vGlut). We show here that vGlut1 has a strong influence on the kinetics of retrieval of half of the known SV cargos and that specifically impairing the endocytosis of vGlut1 in turn slows down other SV cargos, demonstrating that cargo retrieval is a collective cargo-driven process. Finally, we demonstrate that different cargos can be retrieved in the same synapse with different kinetics, suggesting that additional post-endocytic sorting steps likely occur in the nerve terminal. PMID:26224632

  17. Mathematical modeling of vesicle drug delivery systems 2: targeted vesicle interactions with cells, tumors, and the body.

    PubMed

    Ying, Chong T; Wang, Juntian; Lamm, Robert J; Kamei, Daniel T

    2013-02-01

    Vesicles have been studied for several years in their ability to deliver drugs. Mathematical models have much potential in reducing time and resources required to engineer optimal vesicles, and this review article summarizes these models that aid in understanding the ability of targeted vesicles to bind and internalize into cancer cells, diffuse into tumors, and distribute in the body. With regard to binding and internalization, radiolabeling and surface plasmon resonance experiments can be performed to determine optimal vesicle size and the number and type of ligands conjugated. Binding and internalization properties are also inputs into a mathematical model of vesicle diffusion into tumor spheroids, which highlights the importance of the vesicle diffusion coefficient and the binding affinity of the targeting ligand. Biodistribution of vesicles in the body, along with their half-life, can be predicted with compartmental models for pharmacokinetics that include the effect of targeting ligands, and these predictions can be used in conjunction with in vivo models to aid in the design of drug carriers. Mathematical models can prove to be very useful in drug carrier design, and our hope is that this review will encourage more investigators to combine modeling with quantitative experimentation in the field of vesicle-based drug delivery.

  18. Glioblastoma extracellular vesicles: reservoirs of potential biomarkers

    PubMed Central

    Redzic, Jasmina S; Ung, Timothy H; Graner, Michael W

    2014-01-01

    Glioblastoma multiforme (GBM) is the most frequent and most devastating of the primary central nervous system tumors, with few patients living beyond 2 years postdiagnosis. The damage caused by the disease and our treatments for the patients often leave them physically and cognitively debilitated. Generally, GBMs appear after very short clinical histories and are discovered by imaging (using magnetic resonance imaging [MRI]), and the diagnosis is validated by pathology, following surgical resection. The treatment response and diagnosis of tumor recurrence are also tracked by MRI, but there are numerous problems encountered with these monitoring modalities, such as ambiguous interpretation and forms of pseudoprogression. Diagnostic, prognostic, and predictive biomarkers would be an immense boon in following treatment schemes and in determining recurrence, which often requires an invasive intracranial biopsy to verify imaging data. Extracellular vesicles (EVs) are stable, membrane-enclosed, virus-sized particles released from either the cell surface or from endosomal pathways that lead to the systemic release of EVs into accessible biofluids, such as serum/plasma, urine, cerebrospinal fluid, and saliva. EVs carry a wide variety of proteins, nucleic acids, lipids, and other metabolites, with many common features but with enough individuality to be able to identify the cell of origin of the vesicles. These components, if properly interrogated, could allow for the identification of tumor-derived EVs in biofluids, indicating tumor progression, relapse, or treatment failure. That knowledge would allow clinicians to continue with treatment regimens that were actually effective or to change course if the therapies were failing. Here, we review the features of GBM EVs, in terms of EV content and activities that may lead to the use of EVs as serially accessible biomarkers for diagnosis and treatment response in neuro-oncology. PMID:24634586

  19. Synaptic vesicle chips to assay botulinum neurotoxins

    PubMed Central

    2005-01-01

    BoNTs (botulinum neurotoxins), considered to be the most toxic of all biological substances, inhibit neurotransmission through proteolytic cleavage of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins [VAMP (vesicle-associated membrane protein, or synaptobrevin), SNAP-25 (25 kDa synaptosome-associated protein) or syntaxin]. Expansion in the use of BoNTs as therapeutic and cosmetic agents, and the potential threat they constitute as biological weapons, underlines the need for rapid and sensitive in vitro assays. Here, we present new automatized bioassays to detect VAMP cleavage by BoNT/B and F. Western blotting and SPR (surface plasmon resonance) methods revealed that BoNT/B and F totally cleave their substrate on immunoisolated SVs (synaptic vesicles). Real-time monitoring of the immunocapture of native SVs from crude lysates on SPR sensor chips enabled the detection of picogram amounts of different SV proteins. Pre-incubation of a membrane fraction containing SVs with BoNT specifically inhibited capture by anti-VAMP antibodies, and amounts as low as 0.1 pg of BoNT/B were detected. This automated SPR assay is approx. 200 times more sensitive, and 25 times more rapid, than the in vivo BoNT/B test currently used. Moreover, the method can be performed using a few thousand cultured neurons and constitutes a new screening assay for inhibitors. Our data indicate that native VAMP is an optimal substrate for in vitro BoNT assays that can be monitored by SPR. PMID:16011482

  20. Biological reference materials for extracellular vesicle studies.

    PubMed

    Valkonen, S; van der Pol, E; Böing, A; Yuana, Y; Yliperttula, M; Nieuwland, R; Laitinen, S; Siljander, P R M

    2017-02-15

    Extracellular vesicles (EVs) mediate normal physiological homeostasis and pathological processes by facilitating intercellular communication. Research of EVs in basic science and clinical settings requires both methodological standardization and development of reference materials (RM). Here, we show insights and results of biological RM development for EV studies. We used a three-step approach to find and develop a biological RM. First, a literature search was done to find candidates for biological RMs. Second, a questionnaire was sent to EV researchers querying the preferences for RM and their use. Third, a biological RM was selected, developed, characterized, and evaluated. The responses to the survey demonstrated a clear and recognized need for RM optimized for the calibration of EV measurements. Based on the literature, naturally occurring and produced biological RM, such as virus particles and liposomes, were proposed as RM. However, none of these candidate RMs have properties completely matching those of EVs, such as size and refractive index distribution. Therefore, we evaluated the use of nanoerythrosomes (NanoE), vesicles produced from erythrocytes, as a potential biological RM. The strength of NanoE is their resemblance to EVs. Compared to the erythrocyte-derived EVs (eryEVs), NanoE have similar morphology, a similar refractive index (1.37), larger diameter (70% of the NanoE are over 200nm), and increased positive staining for CD235a and lipids (Di-8-ANEPPS) (58% and 67% in NanoE vs. 21% and 45% in eryEVs, respectively). Altogether, our results highlight the general need to develop and validate new RM with similar physical and biochemical properties as EVs to standardize EV measurements between instruments and laboratories.

  1. Structures of yeast vesicle trafficking proteins.

    PubMed Central

    Tishgarten, T.; Yin, F. F.; Faucher, K. M.; Dluhy, R. A.; Grant, T. R.; Fischer von Mollard, G.; Stevens, T. H.; Lipscomb, L. A.

    1999-01-01

    In protein transport between organelles, interactions of v- and t-SNARE proteins are required for fusion of protein-containing vesicles with appropriate target compartments. Mammalian SNARE proteins have been observed to interact with NSF and SNAP, and yeast SNAREs with yeast homologues of NSF and SNAP proteins. This observation led to the hypothesis that, despite low sequence homology, SNARE proteins are structurally similar among eukaryotes. SNARE proteins can be classified into two groups depending on whether they interact with SNARE binding partners via conserved glutamine (Q-SNAREs) or arginine (R-SNAREs). Much of the published structural data available is for SNAREs involved in exocytosis (either in yeast or synaptic vesicles). This paper describes circular dichroism, Fourier transform infrared spectroscopy, and dynamic light scattering data for a set of yeast v- and t-SNARE proteins, Vti1p and Pep12p, that are Q-SNAREs involved in intracellular trafficking. Our results suggest that the secondary structure of Vti1p is highly alpha-helical and that Vti1p forms multimers under a variety of solution conditions. In these respects, Vti1p appears to be distinct from R-SNARE proteins characterized previously. The alpha-helicity of Vti1p is similar to that of Q-SNARE proteins characterized previously. Pep12p, a Q-SNARE, is highly alpha-helical. It is distinct from other Q-SNAREs in that it forms dimers under many of the solution conditions tested in our experiments. The results presented in this paper are among the first to suggest heterogeneity in the functioning of SNARE complexes. PMID:10595551

  2. The PRISM/PRIME Project

    NASA Astrophysics Data System (ADS)

    Barlow, R. J.

    2011-09-01

    Lepton Flavour violation is predicted by many theories beyond the standard model. In the muon sector such a violation entails not only direct μ→eγ decay but also the conversion process μ→e. To measure this to high precision requires a large number of muons of very similar energy, and this is difficult to achieve from a muon target with conventional beam optics. PRISM is an FFAG system designed to accept large numbers of muons ( 10/sec) with a wide range of energies, and render them monochromatic by accelerating the less energetic muons and decelerating the more energetic ones. To preserve Liouville's theorem, this is accompanied by a broadening in the timing of the muons, hence the name 'Phase Rotated Intense Slow Muon source.' The principles of this device have been demonstrated and components prototyped. PRIME is a detector (PRISM Muon Electron Conversion) which has been designed to stop 20 MeV bunches of muons in a thin foil, giving a very clean signal and reaching a background sensitivity of 10, four orders of magnitude better than today's limits and probing the interesting region for BSM theories.

  3. Dynamic control of synaptic vesicle replenishment and short-term plasticity by Ca(2+)-calmodulin-Munc13-1 signaling.

    PubMed

    Lipstein, Noa; Sakaba, Takeshi; Cooper, Benjamin H; Lin, Kun-Han; Strenzke, Nicola; Ashery, Uri; Rhee, Jeong-Seop; Taschenberger, Holger; Neher, Erwin; Brose, Nils

    2013-07-10

    Short-term synaptic plasticity, the dynamic alteration of synaptic strength during high-frequency activity, is a fundamental characteristic of all synapses. At the calyx of Held, repetitive activity eventually results in short-term synaptic depression, which is in part due to the gradual exhaustion of releasable synaptic vesicles. This is counterbalanced by Ca(2+)-dependent vesicle replenishment, but the molecular mechanisms of this replenishment are largely unknown. We studied calyces of Held in knockin mice that express a Ca(2+)-Calmodulin insensitive Munc13-1(W464R) variant of the synaptic vesicle priming protein Munc13-1. Calyces of these mice exhibit a slower rate of synaptic vesicle replenishment, aberrant short-term depression and reduced recovery from synaptic depression after high-frequency stimulation. Our data establish Munc13-1 as a major presynaptic target of Ca(2+)-Calmodulin signaling and show that the Ca(2+)-Calmodulin-Munc13-1 complex is a pivotal component of the molecular machinery that determines short-term synaptic plasticity characteristics.

  4. Mediated priming in the cerebral hemispheres.

    PubMed

    Livesay, Kay; Burgess, Curt

    2003-11-01

    The present experiment investigates hemispheric differences in mediated priming. Theories of lexical representation have argued for an asymmetrical coding between the right and left hemispheres ([Beeman, 1998]), claiming that the right hemisphere is more diffusely represented compared to the left hemisphere. Thus, the right hemisphere activates a larger semantic field compared to the left hemisphere. Mediated (two-step) priming is an ideal task to examine this representational claim, because of the distant nature of the prime-target pairs. Results showed no difference in the magnitude of priming (both mediated and direct) between the right and left hemispheres. These results suggest that the lexical representation of the two hemispheres is equivalent, not asymmetrical as Beeman suggests.

  5. Preserved conceptual priming in Alzheimer's disease.

    PubMed

    Martins, Carla A R; Lloyd-Jones, Toby J

    2006-10-01

    We assessed Alzheimer's disease (AD) and healthy older adult control (HC) group performance on: (1) a conceptual priming task, in which participants had to make a semantic decision as to whether a degraded picture of an object encountered previously belonged to the category of living or non-living things; and (2) a recognition memory task. The AD group showed a dissociation between impaired performance on the recognition task and preserved priming for semantic decisions to degraded pictures. We argue that it is not whether priming is conceptual or perceptual that is important for the observation of priming in AD, rather it is the nature of the response that is required (c.f., Gabrieli et al., 1999).

  6. Negative Priming in Free Recall Reconsidered

    PubMed Central

    2015-01-01

    Negative priming in free recall is the finding of impaired memory performance when previously ignored auditory distracters become targets of encoding and retrieval. This negative priming has been attributed to an aftereffect of deploying inhibitory mechanisms that serve to suppress auditory distraction and minimize interference with learning and retrieval of task-relevant information. In 6 experiments, we tested the inhibitory account of the effect of negative priming in free recall against alternative accounts. We found that ignoring auditory distracters is neither sufficient nor necessary to produce the effect of negative priming in free recall. Instead, the effect is more readily accounted for by a buildup of proactive interference occurring whenever 2 successively presented lists of words are drawn from the same semantic category. PMID:26595066

  7. Russian Prime Minister Calls the Station Crew

    NASA Video Gallery

    Russian Prime Minister Vladimir Putin called the International Space Station from the Russian Mission Control Center in Korolev, Russia, on Jan. 11, 2011. Putin also offered his condolences to ISS ...

  8. NASA and OPTIMUS PRIME Team Up

    NASA Video Gallery

    NASA and OPTIMUS PRIME have teamed up to educate! Kids everywhere created videos showing how NASA technology is truly more than meets the eye, and now you can vote on your favorite! Visit http://ip...

  9. OVATION Prime Model and "Aurorasaurus" Auroral Observations

    NASA Video Gallery

    This video shows the auroral oval, as modeled using OVATION Prime (2013), along with citizen science reports collected by the Aurorasaurus project for the St. Patrick’s Day storm over March 17-19, ...

  10. Degree of handedness and priming: further evidence for a distinction between production and identification priming mechanisms.

    PubMed

    LaVoie, Donna J; Olbinski, Brianna; Palmer, Shayna

    2015-01-01

    The distinction between implicit and explicit forms of memory retrieval is long-standing, and important to the extent it reveals how different neural architecture supports different aspects of memory function. Similarly, distinctions have been made between kinds of repetition priming, a form of implicit memory retrieval. This study focuses on the production-identification (ID) priming distinction, which delineates priming tasks involving verification of stimulus features as compared to priming tasks that require use of a cue to guide response retrieval. Studies investigating this dissociation in dementia or similar patient populations indicate that these forms of priming may differ in their neural bases. The current study looks at degree of handedness as a way of investigating inferred neural architecture supporting these two forms of priming. A growing body of research indicates that degree of handedness (consistent, or CH, versus inconsistent, or ICH) is associated with greater interhemispheric interaction and functional access to right hemisphere processing in ICH, with superior performance seen in ICH on memory tasks reliant on this processing. Arguments about the theoretical mechanisms underlying ID and production forms of perceptual priming tasks suggest that performance on these tasks will differ as a function of degree of handedness. We tested this question in a group of CH and ICH young adults, who were asked to study lists of words prior to performing a production priming task (word stem completion, WSC), a perceptual word ID task, and a word stem cued recall task. While both handedness groups exhibited reliable priming across tasks, WSC priming was greater in ICH than CH participants, with ID priming not differing between groups. This dissociation supports the argument that production and ID forms of priming have different underlying neural bases.

  11. A molecular beacon-based DNA switch for reversible pH sensing in vesicles and live cells.

    PubMed

    Narayanaswamy, Nagarjun; Nair, Raji R; Suseela, Y V; Saini, Deepak Kumar; Govindaraju, T

    2016-07-05

    In this Communication, a molecular beacon-based DNA switch (LMB) is developed as an efficient and reversible pH sensing probe. Remarkably, LMB exhibited reversible structural transition between the closed (molecular beacon) and open (A-motif) states very efficiently in synthetic vesicles and live cells without the need for any transfection agents.

  12. Membrane vesicle production by Chlamydia trachomatis as an adaptive response

    PubMed Central

    Frohlich, Kyla M.; Hua, Ziyu; Quayle, Alison J.; Wang, Jin; Lewis, Maria E.; Chou, Chau-wen; Luo, Miao; Buckner, Lyndsey R.; Shen, Li

    2014-01-01

    Bacteria have evolved specific adaptive responses to cope with changing environments. These adaptations include stress response phenotypes with dynamic modifications of the bacterial cell envelope and generation of membrane vesicles (MVs). The obligate intracellular bacterium, Chlamydia trachomatis, typically has a biphasic lifestyle, but can enter into an altered growth state typified by morphologically aberrant chlamydial forms, termed persistent growth forms, when induced by stress in vitro. How C. trachomatis can adapt to a persistent growth state in host epithelial cells in vivo is not well understood, but is an important question, since it extends the host-bacterial relationship in vitro and has thus been indicated as a survival mechanism in chronic chlamydial infections. Here, we review recent findings on the mechanistic aspects of bacterial adaptation to stress with a focus on how C. trachomatis remodels its envelope, produces MVs, and the potential important consequences of MV production with respect to host-pathogen interactions. Emerging data suggest that the generation of MVs may be an important mechanism for C. trachomatis intracellular survival of stress, and thus may aid in the establishment of a chronic infection in human genital epithelial cells. PMID:24959424

  13. Periodicity In The Intervals Between Primes

    DTIC Science & Technology

    2015-07-02

    statistically strong periodicity is identified in the counting function giving the total number of intervals of a certain size. The nature of the periodic...positive intervals among the first n<=10^6 prime numbers as a probe of the global nature of the sequence of primes. A statistically strong periodicity is...Let x = x1, x2, . . . be an increasing sequence of real numbers which may be either finite or infinitely long. Throughout the following every bold

  14. Integrative Priming of Compositional and Locative Relations

    PubMed Central

    Jones, Lara L.; Wurm, Lee H.; Calcaterra, Ryan D.; Ofen, Noa

    2017-01-01

    Integrative priming refers to the facilitated recognition of a target word (bench) as a real word following a prime (park). Prior integrative priming studies have used a wide variety of integrative relations including temporal (summer rain), topical (travel book), locative (forest river), and compositional (peach pie) relations. Yet differences in the types of integrative relations may yield differences in the underlying explanatory processes of integrative priming. In this study, we compared the magnitude, time course, and three theoretically based correlates of integrative priming for compositional (stone table) and locative (patio table) pairs in a lexical decision task across four stimulus onset asynchronies (SOAs; 50, 300, 800, and 1,600 ms). Based on the Complementary Role Activation theory, integrative ratings (the extent to which the prime and target can be combined into a meaningful phrase) were predicted to facilitate target RTs. Based on the Embodied Conceptual Combination (ECCo) theory, the local co-occurrence of the prime and target, and the ability to perceptually simulate (visually experience) the prime-target pair were tested as predictors. In comparison to unrelated pairs (nose table), target RTs were faster for the compositional and locative pairs, though did not differ between these relations. In support of the Complementary Role Activation theory, integrative ratings predicted target RTs above and beyond our control variables. In support of the ECCo theory, co-occurrence emerged as an early predictor of target RTs, and visual experience ratings was a reliable predictor at the 300 ms SOA, though only for the compositional relations. PMID:28360872

  15. On the relation between conceptual priming, neural priming, and novelty assessment.

    PubMed

    Habib, R

    2001-07-01

    A consistently reported finding in functional neuroimaging studies which compare processing of new information to processing of old information is a reduction in blood flow, and hence neural activity, associated with the old condition. This deactivation has been labeled neural priming. Some investigators have hypothesized that neural priming is the physiological mechanism underlying conceptual priming--a facilitation in the semantic processing of repeated information. Others, however, have hypothesized that neural priming reflects novelty assessment--a mechanism which minimizes the probability that redundant information will be stored in long-term memory. In this paper, the conceptual priming and novelty assessment hypotheses are compared and contrasted in order to ask, and tentatively answer, the question: Are conceptual priming and novelty assessment cognitively and neurophysiologically distinct? Based on a review of the literature, it is suggested that whereas novelty assessment and conceptual priming are distinct cognitive entities, they cannot be presently separated neurophysiologically. That is, some novelty assessment deactivations may in fact reflect priming, and some priming deactivations may in fact reflect novelty assessment.

  16. Non-cognate translation priming in masked priming lexical decision experiments: A meta-analysis.

    PubMed

    Wen, Yun; van Heuven, Walter J B

    2016-09-09

    The masked translation priming paradigm has been widely used in the last 25 years to investigate word processing in bilinguals. Motivated by studies reporting mixed findings, in particular for second language (L2) to first language (L1) translation priming, we conducted, for the first time in the literature, a meta-analysis of 64 masked priming lexical decision experiments across 24 studies to assess the effect sizes of L1-L2 and L2-L1 non-cognate translation priming effects in bilinguals. Our meta-analysis also investigated the influence of potential moderators of translation priming effects. The results provided clear evidence of significant translation priming effects for both directions, with L1-L2 translation priming significantly larger than L2-L1 translation priming (i.e., effect size of 0.86 vs. 0.31). The analyses also revealed that L1-L2 translation effect sizes were moderated by the interval between prime and target (ISI), whereas L2-L1 translation effect sizes were modulated by the number of items per cell. Theoretical and methodological implications of this meta-analysis are discussed and recommendations for future studies are provided.

  17. Bmp4 from the optic vesicle specifies murine retina formation.

    PubMed

    Huang, Jie; Liu, Ying; Oltean, Alina; Beebe, David C

    2015-06-01

    Previous studies of mouse embryos concluded that after the optic vesicle evaginates from the ventral forebrain and contacts the surface ectoderm, signals from the ectoderm specify the distal region of the optic vesicle to become retina and signals from the optic vesicle induce the lens. Germline deletion of Bmp4 resulted in failure of lens formation. We performed conditional deletion of Bmp4 from the optic vesicle to test the function of Bmp4 in murine eye development. The optic vesicle evaginated normally and contacted the surface ectoderm. Lens induction did not occur. The optic cup failed to form and the expression of retina-specific genes decreased markedly in the distal optic vesicle. Instead, cells in the prospective retina expressed genes characteristic of the retinal pigmented epithelium. We conclude that Bmp4 is required for retina specification in mice. In the absence of Bmp4, formation of the retinal pigmented epithelium is the default differentiation pathway of the optic vesicle. Differences in the signaling pathways required for specification of the retina and retinal pigmented epithelium in chicken and mouse embryos suggest major changes in signaling during the evolution of the vertebrate eye.

  18. Epsin1 modulates synaptic vesicle retrieval capacity at CNS synapses

    PubMed Central

    Kyung, Jae Won; Bae, Jae Ryul; Kim, Dae-Hwan; Song, Woo Keun; Kim, Sung Hyun

    2016-01-01

    Synaptic vesicle retrieval is an essential process for continuous maintenance of neural information flow after synaptic transmission. Epsin1, originally identified as an EPS15-interacting protein, is a major component of clathrin-mediated endocytosis. However, the role of Epsin1 in synaptic vesicle endocytosis at CNS synapses remains elusive. Here, we showed significantly altered synaptic vesicle endocytosis in neurons transfected with shRNA targeting Epsin1 during/after neural activity. Endocytosis was effectively restored by introducing shRNA-insensitive Epsin1 into Epsin1-depleted neurons. Domain studies performed on neurons in which domain deletion mutants of Epsin1 were introduced after Epsin1 knockdown revealed that ENTH, CLAP, and NPFs are essential for synaptic vesicle endocytosis, whereas UIMs are not. Strikingly, the efficacy of the rate of synaptic vesicle retrieval (the “endocytic capacity”) was significantly decreased in the absence of Epsin1. Thus, Epsin1 is required for proper synaptic vesicle retrieval and modulates the endocytic capacity of synaptic vesicles. PMID:27557559

  19. Observations of Calcium Dynamics in Cortical Secretory Vesicles

    PubMed Central

    Raveh, Adi; Valitsky, Michael; Shani, Liora; Coorssen, Jens R.; Blank, Paul S.; Zimmerberg, Joshua; Rahamimoff, Rami

    2012-01-01

    SUMMARY Calcium (Ca2+) dynamics were evaluated in fluorescently labeled sea urchin secretory vesicles using confocal microscopy. 71% of the vesicles examined exhibited one or more transient increases in the fluorescence signal that was damped in time. The detection of transient increases in signal was dependent upon the affinity of the fluorescence indicator; the free Ca2+ concentration in the secretory vesicles was estimated to be in the range of ~10 – 100 μM. Non-linear stochastic analysis revealed the presence of extra variance in the Ca2+ dependent fluorescence signal. This noise process increased linearly with the amplitude of the Ca2+ signal. Both the magnitude and spatial properties of this noise process were dependent upon the activity of vesicle p-type (Cav2.1) Ca2+ channels. Blocking the p-type Ca2+ channels with ω-agatoxin decreased signal variance, and altered the spatial noise pattern within the vesicle. These fluorescence signal properties are consistent with vesicle Ca2+ dynamics and not simply due to obvious physical properties such as gross movement artifacts or pH driven changes in Ca2+ indicator fluorescence. The results suggest that the free Ca2+ content of cortical secretory vesicles is dynamic; this property may modulate the exocytotic fusion process. PMID:22831912

  20. Formation and size distribution of self-assembled vesicles

    PubMed Central

    Huang, Changjin; Quinn, David; Suresh, Subra

    2017-01-01

    When detergents and phospholipid membranes are dispersed in aqueous solutions, they tend to self-assemble into vesicles of various shapes and sizes by virtue of their hydrophobic and hydrophilic segments. A clearer understanding of such vesiculation processes holds promise for better elucidation of human physiology and disease, and paves the way to improved diagnostics, drug development, and drug delivery. Here we present a detailed analysis of the energetics and thermodynamics of vesiculation by recourse to nonlinear elasticity, taking into account large deformation that may arise during the vesiculation process. The effects of membrane size, spontaneous curvature, and membrane stiffness on vesiculation and vesicle size distribution were investigated, and the critical size for vesicle formation was determined and found to compare favorably with available experimental evidence. Our analysis also showed that the critical membrane size for spontaneous vesiculation was correlated with membrane thickness, and further illustrated how the combined effects of membrane thickness and physical properties influenced the size, shape, and distribution of vesicles. These findings shed light on the formation of physiological extracellular vesicles, such as exosomes. The findings also suggest pathways for manipulating the size, shape, distribution, and physical properties of synthetic vesicles, with potential applications in vesicle physiology, the pathobiology of cancer and other diseases, diagnostics using in vivo liquid biopsy, and drug delivery methods. PMID:28265065

  1. Removal of Vesicle Structures From Transmission Electron Microscope Images

    PubMed Central

    Jensen, Katrine Hommelhoff; Sigworth, Fred J.; Brandt, Sami Sebastian

    2016-01-01

    In this paper, we address the problem of imaging membrane proteins for single-particle cryo-electron microscopy reconstruction of the isolated protein structure. More precisely, we propose a method for learning and removing the interfering vesicle signals from the micrograph, prior to reconstruction. In our approach, we estimate the subspace of the vesicle structures and project the micrographs onto the orthogonal complement of this subspace. We construct a 2d statistical model of the vesicle structure, based on higher order singular value decomposition (HOSVD), by considering the structural symmetries of the vesicles in the polar coordinate plane. We then propose to lift the HOSVD model to a novel hierarchical model by summarizing the multidimensional HOSVD coefficients by their principal components. Along with the model, a solid vesicle normalization scheme and model selection criterion are proposed to make a compact and general model. The results show that the vesicle structures are accurately separated from the background by the HOSVD model that is also able to adapt to the asymmetries of the vesicles. This is a promising result and suggests even wider applicability of the proposed approach in learning and removal of statistical structures. PMID:26642456

  2. Dynamics of vesicles in a wall-bounded shear flow.

    PubMed

    Abkarian, M; Viallat, A

    2005-08-01

    We report a detailed study of the behavior (shapes, experienced forces, velocities) of giant lipid vesicles subjected to a shear flow close to a wall. Vesicle buoyancy, size, and reduced volume were separately varied. We show that vesicles are deformed by the flow and exhibit a tank-treading motion with steady orientation. Their shapes are characterized by two nondimensional parameters: the reduced volume and the ratio of the shear stress with the hydrostatic pressure. We confirm the existence of a force, able to lift away nonspherical buoyant vesicles from the substrate. We give the functional variation and the value of this lift force (up to 150 pN in our experimental conditions) as a function of the relevant physical parameters: vesicle-substrate distance, wall shear rate, viscosity of the solution, vesicle size, and reduced volume. Circulating deformable cells disclosing a nonspherical shape also experience this force of viscous origin, which contributes to take them away from the endothelium and should be taken into account in studies on cell adhesion in flow chambers, where cells membrane and the adhesive substrate are in relative motion. Finally, the kinematics of vesicles along the flow direction can be described in a first approximation with a model of rigid spheres.

  3. Characteristics of endoplasmic reticulum-derived transport vesicles

    PubMed Central

    1994-01-01

    We have isolated vesicles that mediate protein transport from the ER to Golgi membranes in perforated yeast. These vesicles, which form de novo during in vitro incubations, carry lumenal and membrane proteins that include core-glycosylated pro-alpha-factor, Bet1, Sec22, and Bos1, but not ER-resident Kar2 or Sec61 proteins. Thus, lumenal and membrane proteins in the ER are sorted prior to transport vesicle scission. Inhibition of Ypt1p-function, which prevents newly formed vesicles from docking to cis-Golgi membranes, was used to block transport. Vesicles that accumulate are competent for fusion with cis-Golgi membranes, but not with ER membranes, and thus are functionally committed to vectorial transport. A 900-fold enrichment was developed using differential centrifugation and a series of velocity and equilibrium density gradients. Electron microscopic analysis shows a uniform population of 60 nm vesicles that lack peripheral protein coats. Quantitative Western blot analysis indicates that protein markers of cytosol and cellular membranes are depleted throughout the purification, whereas the synaptobrevin-like Bet1, Sec22, and Bos1 proteins are highly enriched. Uncoated ER-derived transport vesicles (ERV) contain twelve major proteins that associate tightly with the membrane. The ERV proteins may represent abundant cargo and additional targeting molecules. PMID:8063853

  4. Photosynthetic vesicle architecture and constraints on efficient energy harvesting.

    PubMed

    Sener, Melih; Strümpfer, Johan; Timney, John A; Freiberg, Arvi; Hunter, C Neil; Schulten, Klaus

    2010-07-07

    Photosynthetic chromatophore vesicles found in some purple bacteria constitute one of the simplest light-harvesting systems in nature. The overall architecture of chromatophore vesicles and the structural integration of vesicle function remain poorly understood despite structural information being available on individual constituent proteins. An all-atom structural model for an entire chromatophore vesicle is presented, which improves upon earlier models by taking into account the stoichiometry of core and antenna complexes determined by the absorption spectrum of intact vesicles in Rhodobacter sphaeroides, as well as the well-established curvature-inducing properties of the dimeric core complex. The absorption spectrum of low-light-adapted vesicles is shown to correspond to a light-harvesting-complex 2 to reaction center ratio of 3:1. A structural model for a vesicle consistent with this stoichiometry is developed and used in the computation of excitonic properties. Considered also is the packing density of antenna and core complexes that is high enough for efficient energy transfer and low enough for quinone diffusion from reaction centers to cytochrome bc(1) complexes.

  5. Glucocorticoid affects dendritic transport of BDNF-containing vesicles.

    PubMed

    Adachi, Naoki; Numakawa, Tadahiro; Nakajima, Shingo; Fukuoka, Masashi; Odaka, Haruki; Katanuma, Yusuke; Ooshima, Yoshiko; Hohjoh, Hirohiko; Kunugi, Hiroshi

    2015-08-04

    Brain-derived neurotrophic factor (BDNF) is essential for neuronal survival, differentiation, and functions in the central nervous system (CNS). Because BDNF protein is sorted into secretory vesicles at the trans-Golgi network in the cell body after translation, transport of BDNF-containing vesicles to the secretion sites is an important process for its function. Here we examined the effect of dexamethasone (DEX), a synthetic glucocorticoid, on BDNF-containing vesicle transport and found that DEX decreased the proportion of stationary vesicles and increased velocity of the microtubule-based vesicle transport in dendrites of cortical neurons. Furthermore, DEX increased huntingtin (Htt) protein levels via glucocorticoid receptor (GR) activation, and reduction in the amount of Htt by a specific shRNA reversed the action of DEX on BDNF vesicle transport. Given that Htt protein is a positive regulator for the microtubule-dependent vesicular transport in neurons, our data suggest that glucocorticoid stimulates BDNF vesicle transport through upregulation of Htt protein levels.

  6. Priming addition facts with semantic relations.

    PubMed

    Bassok, Miriam; Pedigo, Samuel F; Oskarsson, An T

    2008-03-01

    Results from 2 relational-priming experiments suggest the existence of an automatic analogical coordination between semantic and arithmetic relations. Word pairs denoting object sets served as primes in a task that elicits "obligatory" activation of addition facts (5 + 3 activates 8; J. LeFevre, J. Bisanz, & L. Mrkonjic, 1988). Semantic relations between the priming words were either aligned or misaligned with the structure of addition (M. Bassok, V. M. Chase, & S. A. Martin, 1998). Obligatory activation of addition facts occurred when the digits were primed by categorically related words (tulips-daisies), which are aligned with addition, but did not occur when the digits were primed by unrelated words (hens-radios, Experiment 1) or by functionally related words (records-songs, Experiment 2), which are misaligned with addition. These findings lend support to the viability of automatic analogical priming (B. A. Spellman, K. J. Holyoak, & R. G. Morrison, 2001) and highlight the relevance of arithmetic applications to theoretical accounts of mental arithmetic.

  7. Conditional discriminations, symmetry, and semantic priming.

    PubMed

    Vaidya, Manish; Hudgins, Caleb D; Ortu, Daniele

    2015-09-01

    Psychologists interested in the study of symbolic behavior have found that people are faster at reporting that two words are related to one another than they are in reporting that two words are not related - an effect called semantic priming. This phenomenon has largely been documented in the context of natural languages using real words as stimuli. The current study asked whether laboratory-generated stimulus-stimulus relations established between arbitrary geometrical shapes would also show the semantic priming effect. Participants learned six conditional relations using a one-to-many training structure (A1-B1, A1-C1, A1-D1, A2-B2, A2-C2, A2-D2) and demonstrated, via accurate performance on tests of derived symmetry, that the trained stimulus functions had become reversible. In a lexical decision task, subjects also demonstrated a priming effect as they displayed faster reaction times to target stimuli when the prime and target came from the same trained or derived conditional relations, compared to the condition in which the prime and target came from different trained or derived conditional relations. These data suggest that laboratory-generated equivalence relations may serve as useful analogues of symbolic behavior. However, the fact that conditional relations training and symmetry alone were sufficient to produce the effect suggests that semantic priming like effects may be the byproduct of simpler stimulus-stimulus relations.

  8. An Electrophysiological Investigation of Early Effects of Masked Morphological Priming

    ERIC Educational Resources Information Center

    Morris, Joanna; Grainger, Jonathan; Holcomb, Phillip J.

    2008-01-01

    This experiment examined event-related responses to targets preceded by semantically transparent morphologically related primes (e.g., farmer-farm), semantically opaque primes with an apparent morphological relation (corner-corn), and orthographically, but not morphologically, related primes (scandal-scan) using the masked priming technique…

  9. Masked Priming from Orthographic Neighbors: An ERP Investigation

    ERIC Educational Resources Information Center

    Massol, Stephanie; Grainger, Jonathan; Dufau, Stephane; Holcomb, Phillip

    2010-01-01

    Two experiments combined masked priming with event-related potential (ERP) recordings to examine effects of primes that are orthographic neighbors of target words. Experiment 1 compared effects of repetition primes with effects of primes that were high-frequency orthographic neighbors of low-frequency targets (e.g., faute-faune [error-wildlife]),…

  10. Priming Ability-Relevant Social Categories Improves Intellectual Test Performance

    ERIC Educational Resources Information Center

    Lin, Phoebe S.; Kennette, Lynne N.; Van Havermaet, Lisa R.; Frank, Nichole M.; McIntyre, Rusty B.

    2012-01-01

    Research shows that priming affects behavioral tasks; fewer studies, however, have been conducted on how social category primes affect cognitive tasks. The present study aimed to examine the effects of social category primes on math performance and word recall. It was hypothesized that Asian prime words would improve math performance and word…

  11. Semantic Priming for Coordinate Distant Concepts in Alzheimer's Disease Patients

    ERIC Educational Resources Information Center

    Perri, R.; Zannino, G. D.; Caltagirone, C.; Carlesimo, G. A.

    2011-01-01

    Semantic priming paradigms have been used to investigate semantic knowledge in patients with Alzheimer's disease (AD). While priming effects produced by prime-target pairs with associative relatedness reflect processes at both lexical and semantic levels, priming effects produced by words that are semantically related but not associated should…

  12. Processing Speaker Variability in Repetition and Semantic/Associative Priming

    ERIC Educational Resources Information Center

    Lee, Chao-Yang; Zhang, Yu

    2015-01-01

    The effect of speaker variability on accessing the form and meaning of spoken words was evaluated in two short-term priming experiments. In the repetition priming experiment, participants listened to repeated or unrelated prime-target pairs, in which the prime and target were produced by the same speaker or different speakers. The results showed…

  13. Cytokinin primes plant responses to wounding and reduces insect performance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report a potential role of endogenous cytokinin supply in priming plant defense against herbivory. Cytokinin priming significantly reduced weight gain by insect larvae. Unlike previously described priming by volatile compounds, priming by cytokinin did not overcome vascular restrictions on system...

  14. Masked Inhibitory Priming in English: Evidence for Lexical Inhibition

    ERIC Educational Resources Information Center

    Davis, Colin J.; Lupker, Stephen J.

    2006-01-01

    Predictions derived from the interactive activation (IA) model were tested in 3 experiments using the masked priming technique in the lexical decision task. Experiment 1 showed a strong effect of prime lexicality: Classifications of target words were facilitated by orthographically related nonword primes (relative to unrelated nonword primes) but…

  15. Pure Mediated Priming: A Retrospective Semantic Matching Model

    ERIC Educational Resources Information Center

    Jones, Lara L.

    2010-01-01

    Mediated priming refers to the activation of a target (e.g., "stripes") by a prime (e.g., "lion") that is related indirectly via a connecting mediator (e.g., tiger). In previous mediated priming studies (e.g., McNamara & Altarriba, 1988), the mediator was associatively related to the prime. In contrast, pure mediated…

  16. Decomposition of Repetition Priming Components in Picture Naming

    ERIC Educational Resources Information Center

    Francis, Wendy S.; Corral, Nuvia I.; Jones, Mary L.; Saenz, Silvia P.

    2008-01-01

    Cognitive mechanisms underlying repetition priming in picture naming were decomposed in several experiments. Sets of encoding manipulations meant to selectively prime or reduce priming in object identification or word production components of picture naming were combined factorially to dissociate processes underlying priming in picture naming.…

  17. 46 CFR 111.12-1 - Prime movers.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... REQUIREMENTS Generator Construction and Circuits § 111.12-1 Prime movers. (a) Prime movers must meet section 58...-1). Further requirements for emergency generator prime movers are in 46 CFR subpart 112.50. (b) Each generator prime mover must have an overspeed device that is independent of the normal operating governor...

  18. 46 CFR 111.12-1 - Prime movers.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... REQUIREMENTS Generator Construction and Circuits § 111.12-1 Prime movers. (a) Prime movers must meet section 58...-1). Further requirements for emergency generator prime movers are in 46 CFR subpart 112.50. (b) Each generator prime mover must have an overspeed device that is independent of the normal operating governor...

  19. 46 CFR 111.12-1 - Prime movers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... REQUIREMENTS Generator Construction and Circuits § 111.12-1 Prime movers. (a) Prime movers must meet section 58...-1). Further requirements for emergency generator prime movers are in 46 CFR subpart 112.50. (b) Each generator prime mover must have an overspeed device that is independent of the normal operating governor...

  20. 46 CFR 111.12-1 - Prime movers.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... REQUIREMENTS Generator Construction and Circuits § 111.12-1 Prime movers. (a) Prime movers must meet section 58...-1). Further requirements for emergency generator prime movers are in 46 CFR subpart 112.50. (b) Each generator prime mover must have an overspeed device that is independent of the normal operating governor...

  1. 46 CFR 111.12-1 - Prime movers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... REQUIREMENTS Generator Construction and Circuits § 111.12-1 Prime movers. (a) Prime movers must meet section 58...-1). Further requirements for emergency generator prime movers are in 46 CFR subpart 112.50. (b) Each generator prime mover must have an overspeed device that is independent of the normal operating governor...

  2. Quantitative two-dimensional EXSY spectroscopy and dynamic behavior of a paramagnetic lanthanide macrocyclic chelate: YbDOTA (DOTA = 1,4,7,10-tetraazacyclododecane-N,N{prime},N{double_prime},N{double_prime}{prime}-tetraacetic acid)

    SciTech Connect

    Jacques, V.; Desreux, J.F.

    1994-08-31

    The chelate, YbDOTA (DOTA = 1,4,7,10-tetraazacyclododecane-N,N{prime},N{double_prime},N{double_prime}{prime}-tetraacetic acid) has two possible conformations. Interconversion between isomers was studied by EXSY NMR spectroscopy. The activation energy for conversion was found to be 65.6 kJ/mol for conversion from the major to the minor species.

  3. Cholinergic synaptic vesicle heterogeneity: evidence for regulation of acetylcholine transport

    SciTech Connect

    Gracz, L.M.; Wang, W.; Parsons, S.M.

    1988-07-12

    Crude cholinergic synaptic vesicles from a homogenate of the electric organ of Torpedo californica were centrifuged to equilibrium in an isosmotic sucrose density gradient. The classical VP/sub 1/ synaptic vesicles banding at 1.055 g/mL actively transported (/sup 3/H)acetylcholine (AcCh). An organelle banding at about 1.071 g/mL transported even more (/sup 3/H)AcCh. Transport by both organelles was inhibited by the known AcCh storage blockers trans-2-(4-phenylpiperidino)cyclohexanol (vesamicol, formerly AH5183) and nigericin. Relative to VP/sub 1/ vesicles the denser organelle was slightly smaller as shown by size-exclusion chromatography. It is concluded that the denser organelle corresponds to the recycling VP/sub 2/ synaptic vesicle originally described in intact Torpedo marmorata electric organ. The properties of the receptor for vesamicol were studied by measuring binding of (/sup 3/H)vesamicol, and the amount of SV2 antigen characteristic of secretory vesicles was assayed with a monoclonal antibody directed against it. Relative to VP/sub 1/ vesicles the VP/sub 2/ vesicles had a ratio of (/sup 3/H)AcCh transport activity to vesamicol receptor concentration that typically was 4-7-fold higher, whereas the ratio of SV2 antigen concentration to vesamicol receptor concentration was about 2-fold higher. The Hill coefficients ..cap alpha../sub H/ and equilibrium dissociation constants K for vesamicol binding to VP/sub 1/ and VP/sub 2/ vesicles were essentially the same. The positive Hill coefficient suggests that the vesamicol receptor exists as a homotropic oligomeric complex. The results demonstrate that VP/sub 1/ and VP/sub 2/ synaptic vesicles exhibit functional differences in the AcCh transport system, presumably as a result of regulatory phenomena.

  4. 1-O-Alkylglycerol vesicles (Algosomes): their formation and characterization.

    PubMed

    Gopinath, D; Ravi, D; Rao, B R; Apte, S S; Rambhau, D

    2002-10-10

    1-O-alkylglycerols (ALKG) have exhibited several biological activities and a prominent effect on blood-brain barrier permeability. They have markedly improved brain uptake of cancerostatic agents. Since ALKG are amphiphilic, we explored their tendency to assemble into bilayer vesicles, which can be applied as carriers for drugs. Vesicles (Algosomes) were formed by film hydration method using ALKG (tetra-, penta-, hexa-, hepta-, octa- or nona-decylglycerols) in combination with cholesterol (CHOL) and dicetyl phosphate (DCP) (1-O-alkylglycerol:CHOL:DCP in 45:45:10 molar ratio). On microscopic examination, the algosomes were found to be conspicuously spherical and the dispersion was a mixture of multi-lamellar and small-unilamellar vesicles. Phase transition temperatures of 1-O-hexadecylglycerol (HXDG) and CHOL mixtures were tested by differential scanning calorimetry (DSC). The changes in phase transition temperatures indicate the vesicle forming tendency of ALKG in presence of CHOL. Alkyl chain length dependent variations in vesicle size, zeta-potential (ZP) and capture volume (CV) could not be observed. Vesicles of 1-O-tetradecylglycerol (TTDG) showed improvement in CV with increase in CHOL content from 15 to 55 mol%. However the vesicle size decreased. On challenging algosomes with hypertonic salt solution [potassium iodide (KI) in water], vesicle size decreased and thus algosomes were found to be osmotically sensitive. Algosome dispersions on addition of higher concentrations of KI (40-100 mM) brought about increases in vesicle size and at concentrations 60 mM and above showed aggregation. All vesicular dispersions were stable for only a few days.

  5. Photoinduced bimolecular electron transfer kinetics in small unilamellar vesicles

    SciTech Connect

    Choudhury, Sharmistha Dutta; Kumbhakar, Manoj; Nath, Sukhendu; Pal, Haridas

    2007-11-21

    Photoinduced electron transfer (ET) from N,N-dimethylaniline to some coumarin derivatives has been studied in small unilamellar vesicles (SUVs) of the phospholipid, DL-{alpha}-dimyristoyl-phosphatidylcholine, using steady-state and time-resolved fluorescence quenching, both below and above the phase transition temperature of the vesicles. The primary interest was to examine whether Marcus inversion [H. Sumi and R. A. Marcus, J. Chem. Phys. 84, 4894 (1986)] could be observed for the present ET systems in these organized assemblies. The influence of the topology of SUVs on the photophysical properties of the reactants and consequently on their ET kinetics has also been investigated. Absorption and fluorescence spectral data of the coumarins in SUVs and the variation of their fluorescence decays with temperature indicate that the dyes are localized in the bilayer of the SUVs. Time-resolved area normalized emission spectra analysis, however, reveals that the dyes are distributed in two different microenvironments in the SUVs, which we attribute to the two leaflets of the bilayer, one toward bulk water and the other toward the inner water pool. The microenvironments in the two leaflets are, however, not indicated to be that significantly different. Time-resolved anisotropy decays were biexponential for all the dyes in SUVs, and this has been interpreted in terms of the compound motion model according to which the dye molecules can experience a fast wobbling-in-cone type of motion as well as a slow overall rotating motion of the cone containing the molecule. The expected bimolecular diffusion-controlled rates in SUVs, as estimated by comparing the microviscosities in SUVs (determined from rotational correlation times) and that in acetonitrile solution, are much slower than the observed fluorescence quenching rates, suggesting that reactant diffusion (translational) does not play any role in the quenching kinetics in the present systems. Accordingly, clear inversions are

  6. Onsager's irreversible thermodynamics of the dynamics of transient pores in spherical lipid vesicles.

    PubMed

    Martínez-Balbuena, L; Hernández-Zapata, E; Santamaría-Holek, I

    2015-09-01

    Onsager's irreversible thermodynamics is used to perform a systematic deduction of the kinetic equations governing the opening and collapse of transient pores in spherical vesicles. We show that the edge tension has to be determined from the initial stage of the pore relaxation and that in the final state the vesicle membrane is not completely relaxed, since the surface tension and the pressure difference are about 25% of its initial value. We also show that the pore life-time is controlled by the solution viscosity and its opening is driven by the solution leak-out and the surface tension drop. The final collapse is due to a non-linear interplay between the edge and the surface tensions together with the pressure difference. We also discuss the connection with previous models.

  7. Extracellular vesicles round off communication in the nervous system

    PubMed Central

    Budnik, Vivian; Ruiz-Cañada, Catalina; Wendler, Franz

    2016-01-01

    Functional neural competence and integrity require interactive exchanges among sensory and motor neurons, interneurons and glial cells. Recent studies have attributed some of the tasks needed for these exchanges to extracellular vesicles (such as exosomes and microvesicles), which are most prominently involved in shuttling reciprocal signals between myelinating glia and neurons, thus promoting neuronal survival, the immune response mediated by microglia, and synapse assembly and plasticity. Such vesicles have also been identified as important factors in the spread of neurodegenerative disorders and brain cancer. These extracellular vesicle functions add a previously unrecognized level of complexity to transcellular interactions within the nervous system. PMID:26891626

  8. Transfection of mammalian cells using block copolypeptide vesicles.

    PubMed

    Sun, Victor Z; Choe, Uh-Joo; Rodriguez, April R; Dai, Howard; Deming, Timothy J; Kamei, Daniel T

    2013-05-01

    An arginine-leucine block copolypeptide (R60 L20 ) is synthesized, which is capable of forming vesicles with controllable sizes, able to transport hydrophilic cargo across the cell membrane, and exhibit relatively low cytotoxicity. The R60 L20 vesicles also possess the ability to deliver DNA into mammalian cells for transfection. Although the transfection efficiency is lower than that of the commercially available transfection agent Lipofectamine 2000, the R60 L20 vesicles are able to achieve transfection with significantly lower cytotoxicity and immunogenicity. This behavior is potentially due to its stronger interaction with DNA which subsequently provides better protection against anionic heparin.

  9. Kinetics of enzyme-mediated hydrolysis of lipid vesicles

    NASA Astrophysics Data System (ADS)

    Zhdanov, Vladimir P.; Agnarsson, Björn; Höök, Fredrik

    2016-10-01

    Membrane enzymatic reactions can now be experimentally studied at the level of single sub-100 nm lipid vesicles. To interpret such experiments, we scrutinize theoretically various aspects of the hydrolysis of vesicles by single enzyme molecules and enzyme (e.g., PLA2) supplied with the constant rate. Using the mean-field kinetic model, we illustrate the shape of the corresponding kinetics and the dependence of the time scale of the reaction on the vesicle radius. Stochastic effects are illustrated as well. In addition, we discuss the likely mechanisms of the reaction-induced pore formation and bilayer rupture.

  10. Modulation of 2{prime}-5{prime} oligoadenylate synthetase by environmental stress in the marine sponge Geodia cydonium

    SciTech Connect

    Schroeder, H.C.; Wiens, M.; Mueller, W.E.G.; Kuusksalu, A.; Kelve, M.

    1997-07-01

    Recently the authors established the presence of relatively high amounts of 2{prime}-5{prime} oligoadenylates (2{prime}-5{prime} A) and 2{prime}-5{prime} oligoadenylate synthetase (2{prime}-5{prime} A synthetase) in the marine sponge Geodia cydonium. Here they determined by applying radioimmunoassay and high-performance liquid chromatographical methods that the concentration of 2{prime}-5{prime} A synthetase change following exposure of G. cydonium tissue to environmental stress. The 2{prime}-5{prime} A content and the activity of 2{prime}-5{prime} A synthetase, present in crude sponge extract, increase by up to three-fold after treating sponge cubes for 2 h with natural stressors including heat shock (26 C), cold shock (6 C), pH shock (pH 6), and hypertonic shock and subsequent incubation for 18 h under ambient conditions (16 C). No response was observed after exposure of sponges to an alkaline (pH 10) or hypotonic environment. Similar changes have been found for the expression of heat shock protein HSP70 in G. cydonium. These results show that 2{prime}-5{prime} A in sponges may be useful as a novel biomarker for environmental monitoring.

  11. Women's greater ability to perceive happy facial emotion automatically: gender differences in affective priming.

    PubMed

    Donges, Uta-Susan; Kersting, Anette; Suslow, Thomas

    2012-01-01

    There is evidence that women are better in recognizing their own and others' emotions. The female advantage in emotion recognition becomes even more apparent under conditions of rapid stimulus presentation. Affective priming paradigms have been developed to examine empirically whether facial emotion stimuli presented outside of conscious awareness color our impressions. It was observed that masked emotional facial expression has an affect congruent influence on subsequent judgments of neutral stimuli. The aim of the present study was to examine the effect of gender on affective priming based on negative and positive facial expression. In our priming experiment sad, happy, neutral, or no facial expression was briefly presented (for 33 ms) and masked by neutral faces which had to be evaluated. 81 young healthy volunteers (53 women) participated in the study. Subjects had no subjective awareness of emotional primes. Women did not differ from men with regard to age, education, intelligence, trait anxiety, or depressivity. In the whole sample, happy but not sad facial expression elicited valence congruent affective priming. Between-group analyses revealed that women manifested greater affective priming due to happy faces than men. Women seem to have a greater ability to perceive and respond to positive facial emotion at an automatic processing level compared to men. High perceptual sensitivity to minimal social-affective signals may contribute to women's advantage in understanding other persons' emotional states.

  12. Indole is an essential herbivore-induced volatile priming signal in maize

    PubMed Central

    Erb, Matthias; Veyrat, Nathalie; Robert, Christelle A. M.; Xu, Hao; Frey, Monika; Ton, Jurriaan; Turlings, Ted C. J.

    2015-01-01

    Herbivore-induced volatile organic compounds prime non-attacked plant tissues to respond more strongly to subsequent attacks. However, the key volatiles that trigger this primed state remain largely unidentified. In maize, the release of the aromatic compound indole is herbivore-specific and occurs earlier than other induced responses. We therefore hypothesized that indole may be involved in airborne priming. Using indole-deficient mutants and synthetic indole dispensers, we show that herbivore-induced indole enhances the induction of defensive volatiles in neighbouring maize plants in a species-specific manner. Furthermore, the release of indole is essential for priming of mono- and homoterpenes in systemic leaves of attacked plants. Indole exposure markedly increases the herbivore-induced production of the stress hormones jasmonate-isoleucine conjugate and abscisic acid, which represents a likely mechanism for indole-dependent priming. These results demonstrate that indole functions as a rapid and potent aerial priming agent that prepares systemic tissues and neighbouring plants for incoming attacks. PMID:25683900

  13. An ERP Investigation of Masked Cross-Script Translation Priming

    PubMed Central

    Hoshino, Noriko; Midgley, Katherine J.; Holcomb, Phillip J.; Grainger, Jonathan

    2010-01-01

    The time course of cross-script translation priming and repetition priming was examined in two different scripts using a combination of the masked priming paradigm with the recording of event-related potentials (ERPs). Japanese-English bilinguals performed a semantic categorization task in their second language (L2) English and in their first language (L1) Japanese. Targets were preceded by a visually presented related (translation equivalent/repeated) or unrelated prime. The results showed that the amplitudes of the N250 and N400 ERP components were significantly modulated for L2-L2 repetition priming, L1-L2 translation priming, and L1-L1 repetition priming, but not for L2-L1 translation priming. There was also evidence for priming effects in an earlier 100-200 ms time window for L1-L1 repetition priming and L1-L2 translation priming. We argue that a change in script across primes and targets provides optimal conditions for prime word processing, hence generating very fast-acting translation priming effects when primes are in L1. PMID:20478274

  14. [Priming effects in picture problems: preliminary solutions].

    PubMed

    Wippich, W; Mecklenbräuker, S; Weidmann, K; Reichert, A

    1995-01-01

    Two experiments explored whether picture puzzles are an adequate instrument to investigate implicit memory for pictorial information. During the testing phase, the subjects had to identify hidden figures in picture puzzles. In a preceding learning phase, the priming conditions were varied systematically. In the first experiment, some subjects had to solve picture puzzles in the learning phase, whereas others made esthetic judgments (global processing) or estimated the number of triangles in the picture puzzles (local processing). In the second experiment, the subjects inspected copies of figures that were hidden at testing, modified versions of these figures, or their names in the learning phase. In the first experiment, the subjects of the different encoding conditions showed comparable priming effects. Picture puzzles that had already been processed or seen during learning were solved more often than new ones. Interview data revealed that subjects in the local or global processing conditions did not identify hidden figures at encoding. Furthermore, these subjects could not discriminate between old and new picture puzzles in a final explicit test of recognition. Thus, nonconscious storage of perceptual information that is not semantically interpreted may be sufficient to evoke priming effects. In the second experiment, the subjects in the different encoding conditions showed reliable priming effects, too. The presentation of the duplicates at encoding produced the greatest amount of priming. Effects of verbal priming, however, indicate that the solution of picture puzzles is not based solely on perceptual information. Depending on the priming conditions at learning, the solution of picture puzzles may be based primarily on data-driven processing or may be guided more heavily by conceptual information. It is concluded that perceptual, lexical, and/or conceptual information can contribute to the solution of picture puzzles.

  15. Implicit phonological priming during visual word recognition.

    PubMed

    Wilson, Lisa B; Tregellas, Jason R; Slason, Erin; Pasko, Bryce E; Rojas, Donald C

    2011-03-15

    Phonology is a lower-level structural aspect of language involving the sounds of a language and their organization in that language. Numerous behavioral studies utilizing priming, which refers to an increased sensitivity to a stimulus following prior experience with that or a related stimulus, have provided evidence for the role of phonology in visual word recognition. However, most language studies utilizing priming in conjunction with functional magnetic resonance imaging (fMRI) have focused on lexical-semantic aspects of language processing. The aim of the present study was to investigate the neurobiological substrates of the automatic, implicit stages of phonological processing. While undergoing fMRI, eighteen individuals performed a lexical decision task (LDT) on prime-target pairs including word-word homophone and pseudoword-word pseudohomophone pairs with a prime presentation below perceptual threshold. Whole-brain analyses revealed several cortical regions exhibiting hemodynamic response suppression due to phonological priming including bilateral superior temporal gyri (STG), middle temporal gyri (MTG), and angular gyri (AG) with additional region of interest (ROI) analyses revealing response suppression in the left lateralized supramarginal gyrus (SMG). Homophone and pseudohomophone priming also resulted in different patterns of hemodynamic responses relative to one another. These results suggest that phonological processing plays a key role in visual word recognition. Furthermore, enhanced hemodynamic responses for unrelated stimuli relative to primed stimuli were observed in midline cortical regions corresponding to the default-mode network (DMN) suggesting that DMN activity can be modulated by task requirements within the context of an implicit task.

  16. Subliminal presentation of emotionally negative vs positive primes increases the perceived beauty of target stimuli.

    PubMed

    Era, Vanessa; Candidi, Matteo; Aglioti, Salvatore Maria

    2015-11-01

    Emotions have a profound influence on aesthetic experiences. Studies using affective priming procedures demonstrate, for example, that inducing a conscious negative emotional state biases the perception of abstract stimuli towards the sublime (Eskine et al. Emotion 12:1071-1074, 2012. doi: 10.1037/a0027200). Moreover, subliminal happy facial expressions have a positive impact on the aesthetic evaluation of abstract art (Flexas et al. PLoS ONE 8:e80154, 2013). Little is known about how emotion influences aesthetic perception of non-abstract, representational stimuli, especially those that are particularly relevant for social behaviour, like human bodies. Here, we explore whether the subliminal presentation of emotionally charged visual primes modulates the explicit subjective aesthetic judgment of body images. Using a forward/backward masking procedure, we presented subliminally positive and negative, arousal-matched, emotional or neutral primes and measured their effect on the explicit evaluation of perceived beauty (high vs low) and emotion (positive vs negative) evoked by abstract and body images. We found that negative primes increased subjective aesthetic evaluations of target bodies or abstract images in comparison with positive primes. No influence of primes on the emotional dimension of the targets was found, thus ruling out an unspecific arousal effect and strengthening the link between emotional valence and aesthetic appreciation. More specifically, that subliminal negative primes increase beauty ratings compared to subliminal positive primes indicates a clear link between negative emotions and positive aesthetic evaluations and vice versa, suggesting a possible link between negative emotion and the experience of sublime in art. The study expands previous research by showing the effect of subliminal negative emotions on the subjective aesthetic evaluation not only of abstract but also of body images.

  17. Chromatin assembly factor CAF-1 represses priming of plant defence response genes.

    PubMed

    Mozgová, Iva; Wildhaber, Thomas; Liu, Qinsong; Abou-Mansour, Eliane; L'Haridon, Floriane; Métraux, Jean-Pierre; Gruissem, Wilhelm; Hofius, Daniel; Hennig, Lars

    2015-09-01

    Plants have evolved efficient defence systems against pathogens that often rely on specific transcriptional responses. Priming is part of the defence syndrome, by establishing a hypersensitive state of defence genes such as after a first encounter with a pathogen. Because activation of defence responses has a fitness cost, priming must be tightly controlled to prevent spurious activation of defence. However, mechanisms that repress defence gene priming are poorly understood. Here, we show that the histone chaperone CAF-1 is required to establish a repressed chromatin state at defence genes. Absence of CAF-1 results in spurious activation of a salicylic acid-dependent pathogen defence response in plants grown under non-sterile conditions. Chromatin at defence response genes in CAF-1 mutants under non-inductive (sterile) conditions is marked by low nucleosome occupancy and high H3K4me3 at transcription start sites, resembling chromatin in primed wild-type plants. We conclude that CAF-1-mediated chromatin assembly prevents the establishment of a primed state that may under standard non-sterile growth conditions result in spurious activation of SA-dependent defence responses and consequential reduction of plant vigour.

  18. Refined contour analysis of giant unilamellar vesicles

    NASA Astrophysics Data System (ADS)

    Pécréaux, J.; Döbereiner, H.-G.; Prost, J.; Joanny, J.-F.; Bassereau, P.

    2004-03-01

    The fluctuation spectrum of giant unilamellar vesicles is measured using a high-resolution contour detection technique. An analysis at higher q vectors than previously achievable is now possible due to technical improvements of the experimental setup and of the detection algorithm. The global fluctuation spectrum is directly fitted to deduce the membrane tension and the bending modulus of lipid membranes. Moreover, we show that the planar analysis of fluctuations is valid for spherical objects, even at low wave vectors. Corrections due to the integration time of the video camera and to the section of a 3D object by the observation plane are introduced. A precise calculation of the error bars has been done in order to provide reliable error estimate. Eventually, using this technique, we have measured bending moduli for EPC, SOPC and \\chem{SOPC:CHOL} membranes confirming previously published values. An interesting application of this technique can be the measurement of the fluctuation spectra for non-equilibrium membranes, such as “active membranes”.

  19. Outer membrane vesicles as platform vaccine technology

    PubMed Central

    Stork, Michiel; van der Ley, Peter

    2015-01-01

    Abstract Outer membrane vesicles (OMVs) are released spontaneously during growth by many Gram‐negative bacteria. They present a range of surface antigens in a native conformation and have natural properties like immunogenicity, self‐adjuvation and uptake by immune cells which make them attractive for application as vaccines against pathogenic bacteria. In particular with Neisseria meningitidis, they have been investigated extensively and an OMV‐containing meningococcal vaccine has recently been approved by regulatory agencies. Genetic engineering of the OMV‐producing bacteria can be used to improve and expand their usefulness as vaccines. Recent work on meningitis B vaccines shows that OMVs can be modified, such as for lipopolysaccharide reactogenicity, to yield an OMV product that is safe and effective. The overexpression of crucial antigens or simultaneous expression of multiple antigenic variants as well as the expression of heterologous antigens enable expansion of their range of applications. In addition, modifications may increase the yield of OMV production and can be combined with specific production processes to obtain high amounts of well‐defined, stable and uniform OMV particle vaccine products. Further improvement can facilitate the development of OMVs as platform vaccine product for multiple applications. PMID:26912077

  20. Dysregulations of Synaptic Vesicle Trafficking in Schizophrenia.

    PubMed

    Egbujo, Chijioke N; Sinclair, Duncan; Hahn, Chang-Gyu

    2016-08-01

    Schizophrenia is a serious psychiatric illness which is experienced by about 1 % of individuals worldwide and has a debilitating impact on perception, cognition, and social function. Over the years, several models/hypotheses have been developed which link schizophrenia to dysregulations of the dopamine, glutamate, and serotonin receptor pathways. An important segment of these pathways that have been extensively studied for the pathophysiology of schizophrenia is the presynaptic neurotransmitter release mechanism. This set of molecular events is an evolutionarily well-conserved process that involves vesicle recruitment, docking, membrane fusion, and recycling, leading to efficient neurotransmitter delivery at the synapse. Accumulated evidence indicate dysregulation of this mechanism impacting postsynaptic signal transduction via different neurotransmitters in key brain regions implicated in schizophrenia. In recent years, after ground-breaking work that elucidated the operations of this mechanism, research efforts have focused on the alterations in the messenger RNA (mRNA) and protein expression of presynaptic neurotransmitter release molecules in schizophrenia and other neuropsychiatric conditions. In this review article, we present recent evidence from schizophrenia human postmortem studies that key proteins involved in the presynaptic release mechanism are dysregulated in the disorder. We also discuss the potential impact of dysfunctional presynaptic neurotransmitter release on the various neurotransmitter systems implicated in schizophrenia.

  1. Light gradients in spherical photosynthetic vesicles.

    PubMed

    Paillotin, G; Leibl, W; Gapiński, J; Breton, J; Dobek, A

    1998-07-01

    Light-gradient photovoltage measurements were performed on EDTA-treated thylakoids and on osmotically swollen thylakoids (blebs), both of spherical symmetry but of different sizes. In the case of EDTA vesicles, a negative polarity (due to the normal light gradient) was observed in the blue range of the absorption spectrum, and a positive polarity, corresponding to an inverse light gradient, was observed at lambda = 530 and lambda = 682 nm. The sign of the photovoltage polarity measured in large blebs (swollen thylakoids) is the same as that obtained for whole chloroplasts, although differences in the amplitudes are observed. An approach based on the use of polar coordinates was adapted for a theoretical description of these membrane systems of spherical symmetry. The light intensity distribution and the photovoltage in such systems were calculated. Fits to the photovoltage amplitudes, measured as a function of light wavelength, made it possible to derive the values of the dielectric constant of the protein, epsilons = 3, and the refractive index of the photosynthetic membrane for light propagating perpendicular and parallel to the membrane surface, nt = 1.42 and nn = 1.60, respectively. The latter two values determine the birefringence of the biological membrane, Deltan = nn - nt = 0.18.

  2. Extracellular Vesicles and Autophagy in Osteoarthritis

    PubMed Central

    Guo, Weimin; Chen, Mingxue; Huang, Jingxiang; Yuan, Zhiguo; Zhang, Yu; Wang, Mingjie; Li, Penghao; Wang, Aiyuan; Wang, Yu; Sui, Xiang; Zhang, Li; Xu, Wenjing; Lu, Shibi

    2016-01-01

    Osteoarthritis (OA) is a type of chronic joint disease that is characterized by the degeneration and loss of articular cartilage and hyperplasia of the synovium and subchondral bone. There is reasonable knowledge about articular cartilage physiology, biochemistry, and chondrocyte metabolism. However, the etiology and pathogenesis of OA remain unclear and need urgent clarification to guide the early diagnosis and treatment of OA. Extracellular vesicles (EVs) are small membrane-linking particles that are released from cells. In recent decades, several special biological properties have been found in EV, especially in terms of cartilage. Autophagy plays a critical role in the regulation of cellular homeostasis. Likewise, more and more research has gradually focused on the effect of autophagy on chondrocyte proliferation and function in OA. The synthesis and release of EV are closely associated with autophagy. At the same time, both EV and autophagy play a role in OA development. Based on the mechanism of EV and autophagy in OA development, EV may be beneficial in the early diagnosis of OA; on the other hand, the combination of EV and autophagy-related regulatory drugs may provide insight into possible OA therapeutic strategies. PMID:28078284

  3. Carotenoids in DPPC vesicles: membrane dynamics

    NASA Astrophysics Data System (ADS)

    Socaciu, Carmen; Lausch, Carsten; Diehl, Horst A.

    1999-09-01

    Incorporation of carotenoids into membranes is supposed to change their physical properties with consequences to signal transduction and membrane protein activities. Here the physical parameters membrane fluidity, micropolarity and anisotropy are considered and measured in multilamellar and unilamellar vesicles of dipalmitoylphosphatidyicholine (DPPC) after incorporation of 1, 2.5 and 5 mol% β-carotene, lutein, zeaxanthin, canthaxanthin, or astaxhanthin using 4 mol% pyrene or 1 μM 1,6-diphenyl-1,3,5-hexatriene (DPH) as fluorescent labels. Contrary to other investigations, no significant change in membrane fluidity (as evaluated by the pyrene excimer method) can be found. But a change of micropolarity in the pyrene label environment is obtained from the pyrene monomer fluorescence emission fine structure after incorporation of carotenoids. The membrane anisotropy is enhanced significantly only by those carotenoids which incorporate worst into the membrane. This leads to the hypothesis that carotenoid incorporation into membranes is governed not only by carotenoid polarity hut also by their ability to change membrane anisotropy.

  4. Early Morphological Decomposition of Suffixed Words: Masked Priming Evidence with Transposed-Letter Nonword Primes

    ERIC Educational Resources Information Center

    Beyersmann, Elisabeth; Dunabeitia, Jon Andoni; Carreiras, Manuel; Coltheart, Max; Castles, Anne

    2013-01-01

    Many studies have previously reported that the recognition of a stem target (e.g., "teach") is facilitated by the prior masked presentation of a prime consisting of a derived form of it (e.g., "teacher"). We conducted two lexical decision experiments to investigate masked morphological priming in Spanish. Experiment 1 showed…

  5. Smelly primes – when olfactory primes do or do not work

    PubMed Central

    Smeets, M. A. M.; Dijksterhuis, G. B.

    2014-01-01

    In applied olfactory cognition the effects that olfactory stimulation can have on (human) behavior are investigated. To enable an efficient application of olfactory stimuli a model of how they may lead to a change in behavior is proposed. To this end we use the concept of olfactory priming. Olfactory priming may prompt a special view on priming as the olfactory sense has some unique properties which make odors special types of primes. Examples of such properties are the ability of odors to influence our behavior outside of awareness, to lead to strong affective evaluations, to evoke specific memories, and to associate easily and quickly to other environmental stimuli. Opportunities and limitations for using odors as primes are related to these properties, and alternative explanations for reported findings are offered. Implications for olfactory semantic, construal, behavior and goal priming are given based on a brief overview of the priming literature from social psychology and from olfactory perception science. We end by formulating recommendations and ideas for a future research agenda and applications for olfactory priming. PMID:24575071

  6. Priming tool actions: Are real objects more effective primes than pictures?

    PubMed

    Squires, Scott D; Macdonald, Scott N; Culham, Jody C; Snow, Jacqueline C

    2016-04-01

    Humans are faster to grasp an object such as a tool if they have previewed the same object beforehand. This priming effect is strongest when actors gesture the use of the tool rather than simply move it, possibly because the previewed tool activates action-specific routines in dorsal-stream motor networks. Here, we examined whether real tools, which observers could physically act upon, serve as more potent primes than two-dimensional images of tools, which do not afford physical action. Participants were presented with a prime stimulus that could be either a real tool or a visually matched photograph of a tool. After a brief delay, participants interacted with a real tool target, either by 'grasping to move,' or 'grasping to use' it. The identities of the prime and target tools were either the same (congruent trials; e.g., spatula-spatula) or different (incongruent trials; e.g., whisk-spatula). As expected, participants were faster to initiate grasps during trials when they had to move the tool rather than gesture its use. Priming effects were observed for grasp-to-use, but not grasp-to-move, responses. Surprisingly, however, both pictures of tools and real tools primed action responses equally. Our results indicate that tool priming effects are driven by pictorial cues and their implied actions, even in the absence of volumetric cues that reflect the tangibility and affordances of the prime.

  7. Challenges posed by extracellular vesicles from eukaryotic microbes

    PubMed Central

    Wolf, Julie M.; Casadevall, Arturo

    2014-01-01

    Extracellular vesicles (EV) produced by eukaryotic microbes play an important role during infection. EV release is thought to benefit microbial invasion by delivering a high concentration of virulence factors to distal host cells or to the cytoplasm of a host cell. EV can significantly impact the outcome of host-pathogen interaction in a cargo-dependent manner. Release of EV from eukaryotic microbes poses unique challenges when compared to their bacterial or archaeal counterparts. Firstly, the membrane-bound organelles within eukaryotes facilitate multiple mechanisms of vesicle generation. Secondly, the fungal cell wall poses a unique barrier between the vesicle release site at the plasma membrane and its destined extracellular environment. This review focuses on these eukaryotic-specific aspects of vesicle synthesis and release. PMID:25460799

  8. Magnetic vesicles as MRI-trackable biogenic nanovectors

    NASA Astrophysics Data System (ADS)

    Andriola Silva, Amanda K.; Luciani, Nathalie; Gazeau, Florence; Wilhelm, Claire

    2012-03-01

    Magnetic labeling renders cells MRI-detectable which provides attractive solutions for tracking the fate of a transplanted cell population. Understanding the interplay of magnetic nanoparticles and cells is then an important point that should not be neglected. Here we show that in the condition of food starvation, macrophage cells emit vesicles containing nanoparticles. First, we inferred the intracellular iron oxide load from the magnetophoretic velocity of cells at a calibrated magnetic field gradient. After magnetic labeling and culture in stress conditions, the intracellular iron oxide load was once more determined and a detectable difference was observed before and after stress. Moreover, we identified in the stress conditioned medium membrane vesicle structures carrying magnetic particles. Besides pointing out the role of cell-derived vesicles in the sequestration of the intracellular magnetic label, experiments also demonstrated that vesicles were able to chaperone the magnetic cargo into naïve cells.

  9. Integral equation methods for vesicle electrohydrodynamics in three dimensions

    NASA Astrophysics Data System (ADS)

    Veerapaneni, Shravan

    2016-12-01

    In this paper, we develop a new boundary integral equation formulation that describes the coupled electro- and hydro-dynamics of a vesicle suspended in a viscous fluid and subjected to external flow and electric fields. The dynamics of the vesicle are characterized by a competition between the elastic, electric and viscous forces on its membrane. The classical Taylor-Melcher leaky-dielectric model is employed for the electric response of the vesicle and the Helfrich energy model combined with local inextensibility is employed for its elastic response. The coupled governing equations for the vesicle position and its transmembrane electric potential are solved using a numerical method that is spectrally accurate in space and first-order in time. The method uses a semi-implicit time-stepping scheme to overcome the numerical stiffness associated with the governing equations.

  10. The development and differentiation of human seminal vesicles.

    PubMed Central

    Brewster, S F

    1985-01-01

    The development and differentiation of the human seminal vesicles has been studied using 14 human male fetuses and 4 postnatal specimens. Earlier accounts of the morphogenesis of the seminal vesicle have been reviewed and commented upon in the light of the present findings. Previously undocumented observations include the movements of the mesenchyme, the formation of epithelial folds and the development of the lamina propria and tunica muscularis. Epithelial differentiation is also described. The function of the basal cells and the possible prepubertal secretory activity of the seminal vesicle are discussed. More work is required on the differentiation of the vesicle between birth and puberty. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 PMID:3870731

  11. Improved Methods of Producing and Administering Extracellular Vesicles | Poster

    Cancer.gov

    An efficient method of producing purified extracellular vesicles (EVs), in conjunction with a method that blocks liver macrophages from clearing EVs from the body, has produced promising results for the use of EVs in cancer therapy.

  12. Rho-GTPase-regulated vesicle trafficking in plant cell polarity.

    PubMed

    Chen, Xu; Friml, Jiří

    2014-02-01

    ROPs (Rho of plants) belong to a large family of plant-specific Rho-like small GTPases that function as essential molecular switches to control diverse cellular processes including cytoskeleton organization, cell polarization, cytokinesis, cell differentiation and vesicle trafficking. Although the machineries of vesicle trafficking and cell polarity in plants have been individually well addressed, how ROPs co-ordinate those processes is still largely unclear. Recent progress has been made towards an understanding of the co-ordination of ROP signalling and trafficking of PIN (PINFORMED) transporters for the plant hormone auxin in both root and leaf pavement cells. PIN transporters constantly shuttle between the endosomal compartments and the polar plasma membrane domains, therefore the modulation of PIN-dependent auxin transport between cells is a main developmental output of ROP-regulated vesicle trafficking. The present review focuses on these cellular mechanisms, especially the integration of ROP-based vesicle trafficking and plant cell polarity.

  13. Lipid-targeting peptide probes for extracellular vesicles

    PubMed Central

    Flynn, Aaron D.; Yin, Hang

    2017-01-01

    Extracellular vesicles released from cells are under intense investigation for their roles in cell-cell communication and cancer progression. However, individual vesicles have been difficult to probe as their small size renders them invisible by conventional light microscopy. However, as a consequence of their small size these vesicles possess highly curved lipid membranes that offer an unconventional target for curvature-sensing probes. In this article, we present a strategy for using peptide-based biosensors to detect highly curved membranes and the negatively charged membrane lipid phosphatidylserine, we delineate several assays used to validate curvature- and lipid-targeting mechanisms, and we explore potential applications in probing extracellular vesicles released from sources such as apoptotic cells, cancer cells, or activated platelets. PMID:26909741

  14. Assembly of cells and vesicles for organ engineering

    NASA Astrophysics Data System (ADS)

    Taguchi, Tetsushi

    2011-12-01

    The development of materials and technologies for the assembly of cells and/or vesicles is a key for the next generation of tissue engineering. Since the introduction of the tissue engineering concept in 1993, various types of scaffolds have been developed for the regeneration of connective tissues in vitro and in vivo. Cartilage, bone and skin have been successfully regenerated in vitro, and these regenerated tissues have been applied clinically. However, organs such as the liver and pancreas constitute numerous cell types, contain small amounts of extracellular matrix, and are highly vascularized. Therefore, organ engineering will require the assembly of cells and/or vesicles. In particular, adhesion between cells/vesicles will be required for regeneration of organs in vitro. This review introduces and discusses the key technologies and materials for the assembly of cells/vesicles for organ regeneration.

  15. Yeast Membrane Vesicles: Isolation and General Characteristics1

    PubMed Central

    Christensen, Michael S.; Cirillo, Vincent P.

    1972-01-01

    Yeast membrane vesicles are formed when packed yeast are ground manually in a porcelain mortar and pestle with glass beads (0.2 mm diameter). These vesicles can be separated from the other components of the grinding mixture by a combination of centrifugation steps and elution from a column of the same glass beads (0.2 mm diameter). Isolated vesicles are osmotically sensitive, contain cytoplasmic components, and have energy-independent transport function. They are unable to metabolize glucose, but have respiratory function which is thought to be associated with intravesicular mitochondria. Invertase and oligomycin-insensitive adenosine triphosphatase are present in lysed vesicle preparations, and the appropriateness of these enzyme activities as membrane markers is discussed. Images PMID:4337848

  16. Electro-hydrodynamic effects on lipid membranes in giant vesicles

    NASA Astrophysics Data System (ADS)

    Staykova, Margarita; Yamamoto, Tetsuya; Lipowsky, Reinhard; Dimova, Rumiana

    2009-11-01

    Electric fields are widely applied for cell manipulation in numerous micron-scale systems. Here, we show for the first time that alternating electric fields may cause pronounced flows in the membrane of giant lipid vesicles as well as in the surrounding fluid media.^ The lipid vesicles are not only biomimetic model for the cell membrane but also have many potential biotechnological applications, e.g. as drug-delivery systems and micro-reactors. The reported effects should be considered in electric micro-manipulation procedures on cells and vesicles. They might be useful for applications in microfluidic technologies, for lipid mixing, trapping and displacement, as will be demonstrated. We also believe that our method for visualization of the lipid flows by fluorescently labeled intra-membrane domains will be helpful for studies on membrane behavior in vesicles subjected to shear or mechanical stresses.

  17. Homologous prime-boost strategy in neonate mice using Neisseria lactamica.

    PubMed

    Ito, André Y; Néri, Simone; Machado, Marta S S; Tunes, Claudia F; De Gaspari, Elizabeth N

    2009-05-26

    The aim of the present study was to investigate the immune response to native outer membrane vesicles (NOMVs) of Neisseria lactamica with and without Bordetella pertussis (BP) as adjuvant in intranasal (i.n./i.m) immunization. N. lactamica NOMVs delivered intranasally (i.n) to BALB/c mice in a final volume of 5microl that was gradually introduced with a micropipette, Animals received 1, 2, 3, or 4 doses of antigens at 3, 7, 9 and 12 days after birth. On the 35th day, the animals were immunized intramuscularly (i.m.) with (NOMV) of N. lactamica. The prime-booster strategy using NOMV of N. lactamica with BP as adjuvant in the primer (i.n.) and booster (i.m.) is an effective immunization protocol for inducing humoral immune responses producing IgG antibodies of intermediate to high avidity.

  18. Extracellular Vesicles in Alzheimer’s Disease: Friends or Foes? Focus on Aβ-Vesicle Interaction

    PubMed Central

    Joshi, Pooja; Benussi, Luisa; Furlan, Roberto; Ghidoni, Roberta; Verderio, Claudia

    2015-01-01

    The intercellular transfer of amyloid-β (Aβ) and tau proteins has received increasing attention in Alzheimer’s disease (AD). Among other transfer modes, Aβ and tau dissemination has been suggested to occur through release of Extracellular Vesicles (EVs), which may facilitate delivery of pathogenic proteins over large distances. Recent evidence indicates that EVs carry on their surface, specific molecules which bind to extracellular Aβ, opening the possibility that EVs may also influence Aβ assembly and synaptotoxicity. In this review we focus on studies which investigated the impact of EVs in Aβ-mediated neurodegeneration and showed either detrimental or protective role for EVs in the pathology. PMID:25741766

  19. P2X7 receptors trigger ATP exocytosis and modify secretory vesicle dynamics in neuroblastoma cells.

    PubMed

    Gutiérrez-Martín, Yolanda; Bustillo, Diego; Gómez-Villafuertes, Rosa; Sánchez-Nogueiro, Jesús; Torregrosa-Hetland, Cristina; Binz, Thomas; Gutiérrez, Luis Miguel; Miras-Portugal, María Teresa; Artalejo, Antonio R

    2011-04-01

    Previously, we reported that purinergic ionotropic P2X7 receptors negatively regulate neurite formation in Neuro-2a (N2a) mouse neuroblastoma cells through a Ca(2+)/calmodulin-dependent kinase II-related mechanism. In the present study we used this cell line to investigate a parallel though faster P2X7 receptor-mediated signaling pathway, namely Ca(2+)-regulated exocytosis. Selective activation of P2X7 receptors evoked exocytosis as assayed by high resolution membrane capacitance measurements. Using dual-wavelength total internal reflection microscopy, we have observed both the increase in near-membrane Ca(2+) concentration and the exocytosis of fluorescently labeled vesicles in response to P2X7 receptor stimulation. Moreover, activation of P2X7 receptors also affects vesicle motion in the vertical and horizontal directions, thus, involving this receptor type in the control of early steps (docking and priming) of the secretory pathway. Immunocytochemical and RT-PCR experiments evidenced that N2a cells express the three neuronal SNAREs as well as vesicular nucleotide and monoamine (VMAT-1 and VMAT-2) transporters. Biochemical measurements indicated that ionomycin induced a significant release of ATP from N2a cells. Finally, P2X7 receptor stimulation and ionomycin increased the incidence of small transient inward currents, reminiscent of postsynaptic quantal events observed at synapses. Small transient inward currents were dependent on extracellular Ca(2+) and were abolished by Brilliant Blue G, suggesting they were mediated by P2X7 receptors. Altogether, these results suggest the existence of a positive feedback mechanism mediated by P2X7 receptor-stimulated exocytotic release of ATP that would act on P2X7 receptors on the same or neighbor cells to further stimulate its own release and negatively control N2a cell differentiation.

  20. P2X7 Receptors Trigger ATP Exocytosis and Modify Secretory Vesicle Dynamics in Neuroblastoma Cells*

    PubMed Central

    Gutiérrez-Martín, Yolanda; Bustillo, Diego; Gómez-Villafuertes, Rosa; Sánchez-Nogueiro, Jesús; Torregrosa-Hetland, Cristina; Binz, Thomas; Gutiérrez, Luis Miguel; Miras-Portugal, María Teresa; Artalejo, Antonio R.

    2011-01-01

    Previously, we reported that purinergic ionotropic P2X7 receptors negatively regulate neurite formation in Neuro-2a (N2a) mouse neuroblastoma cells through a Ca2+/calmodulin-dependent kinase II-related mechanism. In the present study we used this cell line to investigate a parallel though faster P2X7 receptor-mediated signaling pathway, namely Ca2+-regulated exocytosis. Selective activation of P2X7 receptors evoked exocytosis as assayed by high resolution membrane capacitance measurements. Using dual-wavelength total internal reflection microscopy, we have observed both the increase in near-membrane Ca2+ concentration and the exocytosis of fluorescently labeled vesicles in response to P2X7 receptor stimulation. Moreover, activation of P2X7 receptors also affects vesicle motion in the vertical and horizontal directions, thus, involving this receptor type in the control of early steps (docking and priming) of the secretory pathway. Immunocytochemical and RT-PCR experiments evidenced that N2a cells express the three neuronal SNAREs as well as vesicular nucleotide and monoamine (VMAT-1 and VMAT-2) transporters. Biochemical measurements indicated that ionomycin induced a significant release of ATP from N2a cells. Finally, P2X7 receptor stimulation and ionomycin increased the incidence of small transient inward currents, reminiscent of postsynaptic quantal events observed at synapses. Small transient inward currents were dependent on extracellular Ca2+ and were abolished by Brilliant Blue G, suggesting they were mediated by P2X7 receptors. Altogether, these results suggest the existence of a positive feedback mechanism mediated by P2X7 receptor-stimulated exocytotic release of ATP that would act on P2X7 receptors on the same or neighbor cells to further stimulate its own release and negatively control N2a cell differentiation. PMID:21292765