Sample records for mutant clc-0 pore

  1. Proton Sensing of CLC-0 Mutant E166D (United States)

    Traverso, Sonia; Zifarelli, Giovanni; Aiello, Rita; Pusch, Michael


    CLC Cl− channels are homodimers in which each subunit has a proper pore and a (fast) gate. An additional slow gate acts on both pores. A conserved glutamate (E166 in CLC-0) is a major determinant of gating in CLC-0 and is crucially involved in Cl−/H+ antiport of CLC-ec1, a CLC of known structure. We constructed tandem dimers with one wild-type (WT) and one mutant subunit (E166A or E166D) to show that these mutations of E166 specifically alter the fast gate of the pore to which they belong without effect on the fast gate of the neighboring pore. In addition both mutations activate the common slow gate. E166A pores have a large, voltage-independent open probability of the fast gate (popen), whereas popen of E166D pores is dramatically reduced. Similar to WT, popen of E166D was increased by lowering pHint. At negative voltages, E166D presents a persistent inward current that is blocked by p-chlorophenoxy-acetic acid (CPA) and increased at low pHext. The pHext dependence of the persistent current is analogous to a similar steady inward current in WT CLC-0. Surprisingly, however, the underlying unitary conductance of the persistent current in E166D is about an order of magnitude smaller than that of the transient deactivating inward Cl− current. Collectively, our data support the possibility that the mutated CLC-0 channel E166D can assume two distinct open states. Voltage-independent protonation of D166 from the outside favors a low conductance state, whereas protonation from the inside favors the high conductance state. PMID:16380443

  2. Gating Competence of Constitutively Open CLC-0 Mutants Revealed by the Interaction with a Small Organic Inhibitor (United States)

    Traverso, Sonia; Elia, Laura; Pusch, Michael


    Opening of CLC chloride channels is coupled to the translocation of the permeant anion. From the recent structure determination of bacterial CLC proteins in the closed and open configuration, a glutamate residue was hypothesized to form part of the Cl−-sensitive gate. The negatively charged side-chain of the glutamate was suggested to occlude the permeation pathway in the closed state, while opening of a single protopore of the double-pore channel would reflect mainly a movement of this side-chain toward the extracellular pore vestibule, with little rearrangement of the rest of the channel. Here we show that mutating this critical residue (Glu166) in the prototype Torpedo CLC-0 to alanine, serine, or lysine leads to constitutively open channels, whereas a mutation to aspartate strongly slowed down opening. Furthermore, we investigated the interaction of the small organic channel blocker p-chlorophenoxy-acetic acid (CPA) with the mutants E166A and E166S. Both mutants were strongly inhibited by CPA at negative voltages with a >200-fold larger affinity than for wild-type CLC-0 (apparent KD at −140 mV ∼4 μM). A three-state linear model with an open state, a low-affinity and a high-affinity CPA-bound state can quantitatively describe steady-state and kinetic properties of the CPA block. The parameters of the model and additional mutagenesis suggest that the high-affinity CPA-bound state is similar to the closed configuration of the protopore gate of wild-type CLC-0. In the E166A mutant the glutamate side chain that occludes the permeation pathway is absent. Thus, if gating consists only in movement of this side-chain the mutant E166A should not be able to assume a closed conformation. It may thus be that fast gating in CLC-0 is more complex than anticipated from the bacterial structures. PMID:12913089

  3. Mechanism of block of single protopores of the Torpedo chloride channel ClC-0 by 2-(p-chlorophenoxy)butyric acid (CPB). (United States)

    Pusch, M; Accardi, A; Liantonio, A; Ferrera, L; De Luca, A; Camerino, D C; Conti, F


    We investigated in detail the mechanism of inhibition by the S(-) enantiomer of 2-(p-chlorophenoxy)butyric acid (CPB) of the Torpedo Cl(-)channel, ClC-0. The substance has been previously shown to inhibit the homologous skeletal muscle channel, CLC-1. ClC-0 is a homodimer with probably two independently gated protopores that are conductive only if an additional common gate is open. As a simplification, we used a mutant of ClC-0 (C212S) that has the common gate "locked open" (Lin, Y.W., C.W. Lin, and T.Y. Chen. 1999. J. Gen. Physiol. 114:1-12). CPB inhibits C212S currents only when applied to the cytoplasmic side, and single-channel recordings at voltages (V) between -120 and -80 mV demonstrate that it acts independently on individual protopores by introducing a long-lived nonconductive state with no effect on the conductance and little effect on the lifetime of the open state. Steady-state macroscopic currents at -140 mV are half-inhibited by approximately 0.5 mM CPB, but the inhibition decreases with V and vanishes for V > or = 40 mV. Relaxations of CPB inhibition after voltage steps are seen in the current responses as an additional exponential component that is much slower than the gating of drug-free protopores. For V = 60 mV) with an IC50 of approximately 30-40 mM. Altogether, these findings support a model for the mechanism of CPB inhibition in which the drug competes with Cl(-) for binding to a site of the pore where it blocks permeation. CPB binds preferentially to closed channels, and thereby also strongly alters the gating of the single protopore. Since the affinity of CPB for open WT pores is extremely low, we cannot decide in this case if it acts also as an open pore blocker. However, the experiments with the mutant K519E strongly support this interpretation. CPB block may become a useful tool to study the pore of ClC channels. As a first application, our results provide additional evidence for a double-barreled structure of ClC-0 and ClC-1.

  4. Mechanism of accelerated current decay caused by an episodic ataxia type-1-associated mutant in a potassium channel pore. (United States)

    Peters, Christian J; Werry, Daniel; Gill, Hira S; Accili, Eric A; Fedida, David


    In Kv1.1, single point mutants found below the channel activation gate at residue V408 are associated with human episodic ataxia type-1, and impair channel function by accelerating decay of outward current during periods of membrane depolarization and channel opening. This decay is usually attributed to C-type inactivation, but here we provide evidence that this is not the case. Using voltage-clamp fluorimetry in Xenopus oocytes, and single-channel patch clamp in mouse ltk- cells, of the homologous Shaker channel (with the equivalent mutation V478A), we have determined that the mutation may cause current decay through a local effect at the activation gate, by destabilizing channel opening. We demonstrate that the effect of the mutant is similar to that of trapped 4-aminopyridine in antagonizing channel opening, as the mutation and 10 mm 4-AP had similar, nonadditive effects on fluorescence recorded from the voltage-sensitive S4 helix. We propose a model where the Kv1.1 activation gate fails to enter a stabilized open conformation, from which the channel would normally C-type inactivate. Instead, the lower pore lining helix is able to enter an activated-not-open conformation during depolarization. These results provide an understanding of the molecular etiology underlying episodic ataxia type-1 due to V408A, as well as biophysical insights into the links between the potassium channel activation gate, the voltage sensor and the selectivity filter.

  5. Validation of a mutant of the pore-forming toxin sticholysin-I for the construction of proteinase-activated immunotoxins. (United States)

    Pentón, David; Pérez-Barzaga, Victor; Díaz, Iscel; Reytor, Mey L; Campos, Javier; Fando, Rafael; Calvo, Loany; Cilli, Eduardo M; Morera, Vivian; Castellanos-Serra, Lila R; Pazos, Fabiola; Lanio, María E; Alvarez, Carlos; Pons, Tirso; Tejuca, Mayra


    The use of pore-forming toxins from sea anemones (actinoporins) in the construction of immunotoxins (ITs) against tumour cells is an alternative for cancer therapy. However, the main disadvantage of actinoporin-based ITs obtained so far has been the poor cellular specificity associated with the toxin's ability to bind and exert its activity in almost any cell membrane. Our final goal is the construction of tumour proteinase-activated ITs using a cysteine mutant at the membrane binding region of sticholysin-I (StI), a cytolysin isolated from the sea anemone Stichodactyla helianthus. The mutant and the ligand moiety would be linked by proteinase-sensitive peptides through the StI cysteine residue blocking the toxin binding region and hence the IT non-specific killing activity. To accomplish this objective the first step was to obtain the mutant StI W111C, and to evaluate the impact of mutating tryptophan 111 by cysteine on the toxin pore-forming capacity. After proteolysis of the cleavage sequence, a short peptide would remain attached to the toxin. The next step was to evaluate whether this mutant is able to form pores even with a residual peptide linked to cysteine 111. In this work we demonstrated that (i) StI W111C shows pore-forming capacity in a nanomolar range, although it is 8-fold less active than the wild-type recombinant StI, corroborating the previously reported importance of residue 111 for the binding of StI to membranes, and (ii) the mutant is able to form pores even with a residual seven-residue peptide linked to cysteine 111. In addition, it was demonstrated that binding of a large molecule to cysteine 111 renders an inactive toxin that is no longer able to bind to the membrane. These results validate the mutant StI W111C for its use in the construction of tumour proteinase-activated ITs.

  6. Functional characterization of sticholysin I and W111C mutant reveals the sequence of the actinoporin's pore assembly.

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    Valeria Antonini

    Full Text Available The use of pore-forming toxins in the construction of immunotoxins against tumour cells is an alternative for cancer therapy. In this protein family one of the most potent toxins are the actinoporins, cytolysins from sea anemones. We work on the construction of tumour proteinase-activated immunotoxins using sticholysin I (StI, an actinoporin isolated from the sea anemone Stichodactyla helianthus. To accomplish this objective, recombinant StI (StIr with a mutation in the membrane binding region has been employed. In this work, it was evaluated the impact of mutating tryptophan 111 to cysteine on the toxin pore forming capability. StI W111C is still able to permeabilize erythrocytes and liposomes, but at ten-fold higher concentration than StI. This is due to its lower affinity for the membrane, which corroborates the importance of residue 111 for the binding of actinoporins to the lipid bilayer. In agreement, other functional characteristics not directly associated to the binding, are essentially the same for both variants, that is, pores have oligomeric structures with similar radii, conductance, cation-selectivity, and instantaneous current-voltage behavior. In addition, this work provides experimental evidence sustaining the toroidal protein-lipid actinoporins lytic structures, since the toxins provoke the trans-bilayer movement (flip-flop of a pyrene-labeled analogue of phosphatidylcholine in liposomes, indicating the existence of continuity between the outer and the inner membrane leaflet. Finally, our planar lipid membranes results have also contributed to a better understanding of the actinoporin's pore assembly mechanism. After the toxin binding and the N-terminal insertion in the lipid membrane, the pore assembly occurs by passing through different transient sub-conductance states. These states, usually 3 or 4, are due to the successive incorporation of N-terminal α-helices and lipid heads to the growing pores until a stable toroidal

  7. Functional analysis of a frame-shift mutant of the dihydropyridine receptor pore subunit (α1S expressing two complementary protein fragments

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    Mortenson Lindsay


    Full Text Available Abstract Background The L-type Ca2+ channel formed by the dihydropyridine receptor (DHPR of skeletal muscle senses the membrane voltage and opens the ryanodine receptor (RyR1. This channel-to-channel coupling is essential for Ca2+ signaling but poorly understood. We characterized a single-base frame-shift mutant of α1S, the pore subunit of the DHPR, that has the unusual ability to function voltage sensor for excitation-contraction (EC coupling by virtue of expressing two complementary hemi-Ca2+ channel fragments. Results Functional analysis of cDNA transfected dysgenic myotubes lacking α1S were carried out using voltage-clamp, confocal Ca2+ indicator fluoresence, epitope immunofluorescence and immunoblots of expressed proteins. The frame-shift mutant (fs-α1S expressed the N-terminal half of α1S (M1 to L670 and the C-terminal half starting at M701 separately. The C-terminal fragment was generated by an unexpected restart of translation of the fs-α1S message at M701 and was eliminated by a M701I mutation. Protein-protein complementation between the two fragments produced recovery of skeletal-type EC coupling but not L-type Ca2+ current. Discussion A premature stop codon in the II-III loop may not necessarily cause a loss of DHPR function due to a restart of translation within the II-III loop, presumably by a mechanism involving leaky ribosomal scanning. In these cases, function is recovered by expression of complementary protein fragments from the same cDNA. DHPR-RyR1 interactions can be achieved via protein-protein complementation between hemi-Ca2+ channel proteins, hence an intact II-III loop is not essential for coupling the DHPR voltage sensor to the opening of RyR1 channel.

  8. An excretory function for the Escherichia coli outer membrane pore TolC: upregulation of marA and soxS transcription and Rob activity due to metabolites accumulated in tolC mutants. (United States)

    Rosner, Judah L; Martin, Robert G


    Efflux pumps function to rid bacteria of xenobiotics, including antibiotics, bile salts, and organic solvents. TolC, which forms an outer membrane channel, is an essential component of several efflux pumps in Escherichia coli. We asked whether TolC has a role during growth in the absence of xenobiotics. Because tolC transcription is activated by three paralogous activators, MarA, SoxS, and Rob, we examined the regulation of these activators in tolC mutants. Using transcriptional fusions, we detected significant upregulation of marRAB and soxS transcription and Rob protein activity in tolC mutants. Three mechanisms could be distinguished: (i) activation of marRAB transcription was independent of marRAB, soxR, and rob functions; (ii) activation of soxS transcription required SoxR, a sensor of oxidants; and (iii) Rob protein was activated posttranscriptionally. This mechanism is similar to the mechanisms of upregulation of marRAB, soxS, and Rob by treatment with certain phenolics, superoxides, and bile salts, respectively. The transcription of other marA/soxS/rob regulon promoters, including tolC itself, was also elevated in tolC mutants. We propose that TolC is involved in the efflux of certain cellular metabolites, not only xenobiotics. As these metabolites accumulate during growth, they trigger the upregulation of MarA, SoxS, and Rob, which in turn upregulate tolC and help rid the bacteria of these metabolites, thereby restoring homeostasis.

  9. Dilated pore of winer

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    Mittal R


    Full Text Available Two cases of dilated pore of Winer were observed. First case had single defined black papule with well defined margin, central pore and discharge of black powdery material from nose since 3 years. The second case had one 9mm, black well-defined papule with central pore discharging black powdery material on right forearm since 9 months and 9 similar smaller papules were seen on forearm and lower abdomen. Histopathologically both revealed greatly dilated infundibulum lined by acanthotic epidermis and atrophic subinfundibular hair structures thus confirming diagnosis of dilated pore of Winer

  10. Fingerprint pores extractor

    CSIR Research Space (South Africa)

    Mngenge, NA


    Full Text Available alone. Sweat pores have been less utilized in the past due to constraints imposed by fingerprint scanning devices and resolution standards. Recently, progress has been made on both scanning devices and resolution standards to support the use of pores...

  11. Pore size distribution mapping


    Strange, John H.; J. Beau W. WEBBER; Schmidt, S.D.


    Pore size distribution mapping has been demonstrated using NMR cryoporometry\\ud in the presence of a magnetic field gradient, This novel method is extendable to 2D and 3D mapping. It offers a unique nondestructive method of obtaining full pore-size distributions in the range 3 to 100 nm at any point within a bulk sample. \\ud

  12. Velocities in Solar Pores (United States)

    Balasubramaniam, K. S.; Keil, S. L.; Smaldone, L. A.


    We investigate the three dimensional structure of solar pores and their surroundings using high spatial and spectral resolution data. We present evidence that surface velocities decrease around pores with a corresponding increase in the line-of-sight (LOS) velocities. LOS velocities in pores increase with the strength of the magnetic field. Surface velocities show convergence toward a weak downflow which appear to trace boundaries resembling meso-granular and super granular flows. The observed magnetic fields in the pores appear near these boundaries. We analyze the vertical velocity structure in pores and show that they generally have downflows decreasing exponentially with height, with a scale height of about 90 km. Evidence is also presented for the expanding nature of flux tubes. Finally we describe a phenomenological model for pores. This work was supported by AFOSR Task 2311G3. LAS was partially supported by the Progetto Nazionale Astrofisica e Fisica Cosmica of MURST and Scambi Internazionali of the Universita degli Studi di Napoli Frederico II. National Solar Observatory, NOAO, is operated for the National Science Foundation by AURA, Inc.

  13. Properties of solar pores

    NARCIS (Netherlands)

    Sütterlin, Peter


    We present the results of an extensive investigation of the properties of solar pores. Spectra of all 4 Stokes parameters of several magnetic sensitive absorption lines as well as Stokes I only spectra of lines with low or vanishing Landéfactor have been observed. An inversion code based on the Leve

  14. The pore space scramble (United States)

    Gormally, Alexandra; Bentham, Michelle; Vermeylen, Saskia; Markusson, Nils


    Climate change and energy security continue to be the context of the transition to a secure, affordable and low carbon energy future, both in the UK and beyond. This is reflected in for example, binding climate policy targets at the EU level, the introduction of renewable energy targets, and has also led to an increasing interest in Carbon Capture and Storage (CCS) technology with its potential to help mitigate against the effects of CO2 emissions from fossil fuel burning. The UK has proposed a three phase strategy to integrate CCS into its energy system in the long term focussing on off-shore subsurface storage (DECC, 2014). The potential of CCS therefore, raises a number of challenging questions and issues surrounding the long-term storage of CO2 captured and injected into underground spaces and, alongside other novel uses of the subsurface, contributes to opening a new field for discussion on the governance of the subsurface. Such 'novel' uses of the subsurface have lead to it becoming an increasingly contested space in terms of its governance, with issues emerging around the role of ownership, liability and property rights of subsurface pore space. For instance, questions over the legal ownership of pore space have arisen with ambiguity over the legal standpoint of the surface owner and those wanting to utilise the pore space for gas storage, and suggestions of whether there are depths at which legal 'ownership' becomes obsolete (Barton, 2014). Here we propose to discuss this 'pore space scramble' and provide examples of the competing trajectories of different stakeholders, particularly in the off-shore context given its priority in the UK. We also propose to highlight the current ambiguity around property law of pore space in the UK with reference to approaches currently taken in different national contexts. Ultimately we delineate contrasting models of governance to illustrate the choices we face and consider the ethics of these models for the common good

  15. Pore dynamics in lipid membranes (United States)

    Gozen, I.; Dommersnes, P.


    Transient circular pores can open in plasma membrane of cells due to mechanical stress, and failure to repair such pores lead to cell death. Similar pores in the form of defects also exist among smectic membranes, such as in myelin sheaths or mitochondrial membranes. The formation and growth of membrane defects are associated with diseases, for example multiple sclerosis. A deeper understanding of membrane pore dynamics can provide a more refined picture of membrane integrity-related disease development, and possibly also treatment options and strategies. Pore dynamics is also of great importance regarding healthcare applications such as drug delivery, gene or as recently been implied, cancer therapy. The dynamics of pores significantly differ in stacks which are confined in 2D compared to those in cells or vesicles. In this short review, we will summarize the dynamics of different types of pores that can be observed in biological membranes, which include circular transient, fusion and hemi-fusion pores. We will dedicate a section to floral and fractal pores which were discovered a few years ago and have highly peculiar characteristics. Finally, we will discuss the repair mechanisms of large area pores in conjunction with the current cell membrane repair hypotheses.

  16. Soils, Pores, and NMR (United States)

    Pohlmeier, Andreas; Haber-Pohlmeier, Sabina; Haber, Agnes; Sucre, Oscar; Stingaciu, Laura; Stapf, Siegfried; Blümich, Bernhard


    Within Cluster A, Partial Project A1, the pore space exploration by means of Nuclear Magnetic Resonance (NMR) plays a central role. NMR is especially convenient since it probes directly the state and dynamics of the substance of interest: water. First, NMR is applied as relaxometry, where the degree of saturation but also the pore geometry controls the NMR signature of natural porous systems. Examples are presented where soil samples from the Selhausen, Merzenhausen (silt loams), and Kaldenkirchen (sandy loam) test sites are investigated by means of Fast Field Cycling Relaxometry at different degrees of saturation. From the change of the relaxation time distributions with decreasing water content and by comparison with conventional water retention curves we conclude that the fraction of immobile water is characterized by T1 samples (Haber-Pohlmeier et al. 2010). Third, relaxometric information forms the basis of understanding magnetic resonance imaging (MRI) results. The general difficulty of imaging in soils are the inherent fast T2 relaxation times due to i) the small pore sizes, ii) presence of paramagnetic ions in the solid matrix, and iii) diffusion in internal gradients. The last point is important, since echo times can not set shorter than about 1ms for imaging purposes. The way out is either the usage of low fields for imaging in soils or special ultra-short pulse sequences, which do not create echoes. In this presentation we will give examples on conventional imaging of macropore fluxes in soil cores (Haber-Pohlmeier et al. 2010), and the combination with relaxometric imaging, as well as the advantages and drawbacks of low-field and ultra-fast pulse imaging. Also first results on the imaging of soil columns measured by SIP in Project A3 are given. Haber-Pohlmeier, S., S. Stapf, et al. (2010). "Waterflow Monitored by Tracer Transport in Natural Porous Media Using MRI." Vadose Zone J.: submitted. Haber-Pohlmeier, S., S. Stapf, et al. (2010). "Relaxation in a

  17. A pore water conductivity sensor

    NARCIS (Netherlands)

    Hilhorst, M.A.


    The electrical permittivity and conductivity of the bulk soil are a function of the permittivity and conductivity of the pore water. For soil water contents higher than 0.10 both functions are equal, facilitating in situ conductivity measurements of the pore water. A novel method is described, based

  18. Ion transport across transmembrane pores

    NARCIS (Netherlands)

    Leontiadou, Hari; Mark, Alan E.; Marrink, Siewert-Jan


    To study the pore-mediated transport of ionic species across a lipid membrane, a series of molecular dynamics simulations have been performed of a dipalmitoyl-phosphatidyl-choline bilayer containing a preformed water pore in the presence of sodium and chloride ions. It is found that the stability of

  19. Crystal structure of listeriolysin O reveals molecular details of oligomerization and pore formation (United States)

    Köster, Stefan; van Pee, Katharina; Hudel, Martina; Leustik, Martin; Rhinow, Daniel; Kühlbrandt, Werner; Chakraborty, Trinad; Yildiz, Özkan


    Listeriolysin O (LLO) is an essential virulence factor of Listeria monocytogenes that causes listeriosis. Listeria monocytogenes owes its ability to live within cells to the pH- and temperature-dependent pore-forming activity of LLO, which is unique among cholesterol-dependent cytolysins. LLO enables the bacteria to cross the phagosomal membrane and is also involved in activation of cellular processes, including the modulation of gene expression or intracellular Ca2+ oscillations. Neither the pore-forming mechanism nor the mechanisms triggering the signalling processes in the host cell are known in detail. Here, we report the crystal structure of LLO, in which we identified regions important for oligomerization and pore formation. Mutants were characterized by determining their haemolytic and Ca2+ uptake activity. We analysed the pore formation of LLO and its variants on erythrocyte ghosts by electron microscopy and show that pore formation requires precise interface interactions during toxin oligomerization on the membrane.

  20. Reduction of Streptolysin O (SLO Pore-Forming Activity Enhances Inflammasome Activation

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    Peter A. Keyel


    Full Text Available Pore-forming toxins are utilized by bacterial and mammalian cells to exert pathogenic effects and induce cell lysis. In addition to rapid plasma membrane repair, macrophages respond to pore-forming toxins through activation of the NLRP3 inflammasome, leading to IL-1β secretion and pyroptosis. The structural determinants of pore-forming toxins required for NLRP3 activation remain unknown. Here, we demonstrate using streptolysin O (SLO that pore-formation controls IL-1β secretion and direct toxicity. An SLO mutant incapable of pore-formation did not promote direct killing, pyroptosis or IL-1β production. This indicated that pore formation is necessary for inflammasome activation. However, a partially active mutant (SLO N402C that was less toxic to macrophages than wild-type SLO, even at concentrations that directly lysed an equivalent number of red blood cells, enhanced IL-1β production but did not alter pyroptosis. This suggests that direct lysis may attenuate immune responses by preventing macrophages from successfully repairing their plasma membrane and elaborating more robust cytokine production. We suggest that mutagenesis of pore-forming toxins represents a strategy to enhance adjuvant activity.

  1. Lysenin: a sphingomyelin specific pore-forming toxin. (United States)

    Shogomori, Hidehiko; Kobayashi, Toshihide


    Sphingomyelin is a major sphingolipid in mammalian cells. Recent results indicate that sphingomyelin is a reservoir of lipid second messengers, ceramide and sphingosine-1-phosphate. Sphingomyelin is also a major component of sphingolipid and cholesterol-rich membrane domains (lipid rafts). Lysenin is a pore-forming toxin that specifically binds sphingomyelin. The binding of lysenin to sphingomyelin is dependent on the membrane distribution of the lipid, i.e. the toxin selectively binds sphingomyelin clusters. Development of a non-toxic lysenin mutant revealed the spatial and functional heterogeneity of sphingolipid-rich membrane domains.

  2. Structural basis for pore-forming mechanism of staphylococcal α-hemolysin. (United States)

    Sugawara, Takaki; Yamashita, Daichi; Kato, Koji; Peng, Zhao; Ueda, Junki; Kaneko, Jun; Kamio, Yoshiyuki; Tanaka, Yoshikazu; Yao, Min


    Staphylococcal α-hemolysin (α-HL) is a β-barrel pore-forming toxin (PFT) expressed by Staphylococcus aureus. α-HL is secreted as a water-soluble monomeric protein, which binds to target membranes and forms membrane-inserted heptameric pores. To explore the pore-forming mechanism of α-HL in detail, we determined the crystal structure of the α-HL monomer and prepore using H35A mutant and W179A/R200A mutant, respectively. Although the overall structure of the monomer was similar to that of other staphylococcal PFTs, a marked difference was observed in the N-terminal amino latch, which bent toward the prestem. Moreover, the prestem was fastened by the cap domain with a key hydrogen bond between Asp45 and Tyr118. Prepore structure showed that the transmembrane region is roughly formed with flexibility, although the upper half of the β-barrel is formed appropriately. Structure comparison among monomer, prepore and pore revealed a series of motions, in which the N-terminal amino latch released upon oligomerization destroys its own key hydrogen bond between Asp45-Tyr118. This action initiated the protrusion of the prestem. Y118F mutant and the N-terminal truncated mutant markedly decreased in the hemolytic activity, indicating the importance of the key hydrogen bond and the N-terminal amino latch on the pore formation. Based on these observations, we proposed a dynamic molecular mechanism of pore formation for α-HL.


    Field studies were conducted over several years at municipal wastewater treatment plants employing line pore diffused aeration systems. These studies were designed to produce reliable information on the performance and operational requirements of fine pore devices under process ...

  4. Pore formation by Cry toxins. (United States)

    Soberón, Mario; Pardo, Liliana; Muñóz-Garay, Carlos; Sánchez, Jorge; Gómez, Isabel; Porta, Helena; Bravo, Alejandra


    Bacillus thuringiensis (Bt) bacteria produce insecticidal Cry and Cyt proteins used in the biological control of different insect pests. In this review, we will focus on the 3d-Cry toxins that represent the biggest group of Cry proteins and also on Cyt toxins. The 3d-Cry toxins are pore-forming toxins that induce cell death by forming ionic pores into the membrane of the midgut epithelial cells in their target insect. The initial steps in the mode of action include ingestion of the protoxin, activation by midgut proteases to produce the toxin fragment and the interaction with the primary cadherin receptor. The interaction of the monomeric CrylA toxin with the cadherin receptor promotes an extra proteolytic cleavage, where helix alpha-1 of domain I is eliminated and the toxin oligomerization is induced, forming a structure of 250 kDa. The oligomeric structure binds to a secondary receptor, aminopeptidase N or alkaline phosphatase. The secondary receptor drives the toxin into detergent resistant membrane microdomains formingpores that cause osmotic shock, burst of the midgut cells and insect death. Regarding to Cyt toxins, these proteins have a synergistic effect on the toxicity of some Cry toxins. Cyt proteins are also proteolytic activated in the midgut lumen of their target, they bind to some phospholipids present in the mosquito midgut cells. The proposed mechanism of synergism between Cry and Cyt toxins is that Cyt1Aa function as a receptor for Cry toxins. The Cyt1A inserts into midgut epithelium membrane and exposes protein regions that are recognized by Cry11Aa. It was demonstrated that this interaction facilitates the oligomerization of Cry11Aa and also its pore formation activity.

  5. The C-terminus of IcmT is essential for pore formation and for intracellular trafficking of Legionella pneumophila within Acanthamoeba polyphaga. (United States)

    Molmeret, Maëlle; Alli, O A Terry; Radulic, Marina; Susa, Milorad; Doric, Miljenko; Kwaik, Yousef Abu


    We have shown previously that the five rib (release of intracellular bacteria) mutants of Legionella pneumophila are competent for intracellular replication but defective in pore formation-mediated cytolysis and egress from protozoan and mammalian cells. The rib phenotype results from a point mutation (deletion) DeltaG544 in icmT that is predicted to result in the expression of a protein truncated by 32 amino acids from the C-terminus. In contrast to the rib mutants that are capable of intracellular replication, an icmT null mutant was completely defective in intracellular replication within mammalian and protozoan cells, in addition to its defect in pore formation-mediated cytolysis. The icmT wild-type allele complemented the icmT null mutant for both defects of intracellular replication and pore formation-mediated cytolysis and egress from mammalian cells. In contrast, the icmTDeltaG544 allele complemented the icmT null mutant for intracellular growth, but not for the pore-forming activity. Consistent with their defect in pore formation-mediated cytotoxicity in vitro, both mutants failed to cause pulmonary inflammation in A/J mice. Interestingly, the rib mutant was severely defective in intracellular growth within Acanthamoeba polyphaga. Confocal laser scanning and electron microscopy confirmed that the rib mutant and the icmT null mutant were severely and completely defective, respectively, in intracellular growth in A. polyphaga, and the respective defects correlated with fusion of the bacterial phagosomes to lysosomes. Taken together, the data showed that the C-terminus domain of IcmT is essential for the pore-forming activity and is required for intracellular trafficking and replication within A. polyphaga, but not within mammalian cells.


    Energy Technology Data Exchange (ETDEWEB)

    Jagiello, Jacek [Micromeritics Instrument Corporation; Kenvin, Jeffrey [Micromeritics Instrument Corporation; Oliver, James P [Micromeritics Instrument Corporation; Lupini, Andrew R [ORNL; Contescu, Cristian I [ORNL


    In this work, we present a model for analyzing activated carbon micropore structures based on graphene sheet walls of finite thickness and extent. This is a two-dimensional modification of the widely used infinite slit pore model that assumes graphite-like infinitely extended pore walls. The proposed model has two versions: (1) a strip pore constructed with graphene strip walls that have finite length L in the x direction and are infinite in the y direction. Strip pores are open on both sides in the x direction. (2) A channel pore is a strip pore partially closed along one edge by a perpendicularly oriented graphene wall. This more realistic model allows pore termination via both physical pore entrances and pore blockage. The model consequently introduces heterogeneity of the adsorption potential that is reduced near pore entrances and enhanced near corners of pore walls. These energetically heterogeneous structures fill with adsorbate more gradually than homogeneous pores of the same width. As a result, the calculated adsorption isotherms are smoother and less steep for the finite versus the infinite pore model. In the application of this model for carbon characterization it is necessary to make an assumption about the pore length. In this work we made this assumption based on the high resolution scanning transmission electron microscopy (STEM) results. We find the agreement between the experiment and the model significantly better for the finite than for the infinite pore model.

  7. Characterization of a nuclear pore protein sheds light on the roles and composition of the Toxoplasma gondii nuclear pore complex. (United States)

    Courjol, Flavie; Mouveaux, Thomas; Lesage, Kevin; Saliou, Jean-Michel; Werkmeister, Elisabeth; Bonabaud, Maurine; Rohmer, Marine; Slomianny, Christian; Lafont, Franck; Gissot, Mathieu


    The nuclear pore is a key structure in eukaryotes regulating nuclear-cytoplasmic transport as well as a wide range of cellular processes. Here, we report the characterization of the first Toxoplasma gondii nuclear pore protein, named TgNup302, which appears to be the orthologue of the mammalian Nup98-96 protein. We produced a conditional knock-down mutant that expresses TgNup302 under the control of an inducible tetracycline-regulated promoter. Under ATc treatment, a substantial decrease of TgNup302 protein in inducible knock-down (iKD) parasites was observed, causing a delay in parasite proliferation. Moreover, the nuclear protein TgENO2 was trapped in the cytoplasm of ATc-treated mutants, suggesting that TgNup302 is involved in nuclear transport. Fluorescence in situ hybridization revealed that TgNup302 is essential for 18S RNA export from the nucleus to the cytoplasm, while global mRNA export remains unchanged. Using an affinity tag purification combined with mass spectrometry, we identified additional components of the nuclear pore complex, including proteins potentially interacting with chromatin. Furthermore, reverse immunoprecipitation confirmed their interaction with TgNup302, and structured illuminated microscopy confirmed the NPC localization of some of the TgNup302-interacting proteins. Intriguingly, facilitates chromatin transcription complex (FACT) components were identified, suggesting the existence of an NPC-chromatin interaction in T. gondii. Identification of TgNup302-interacting proteins also provides the first glimpse at the NPC structure in Apicomplexa, suggesting a structural conservation of the NPC components between distant eukaryotes.

  8. Importance of polarity of the α4-α5 loop residue-Asn(166) in the pore-forming domain of the Bacillus thuringiensis Cry4Ba toxin: implications for ion permeation and pore opening. (United States)

    Juntadech, Thanate; Kanintronkul, Yodsoi; Kanchanawarin, Chalermpol; Katzenmeier, Gerd; Angsuthanasombat, Chanan


    Bacillus thuringiensis Cry4Ba toxin is lethal to mosquito-larvae by forming ion-permeable pores in the target midgut cell membrane. Previously, the polarity of Asn(166) located within the α4-α5 loop composing the Cry4Ba pore-forming domain was shown to be crucial for larvicidal activity. Here, structurally stable-mutant toxins of both larvicidal-active (N166D) and inactive (N166A and N166I) mutants were FPLC-purified and characterized for their relative activities in liposomal-membrane permeation and single-channel formation. Similar to the 65-kDa trypsin-activated wild-type toxin, the N166D bio-active mutant toxin was still capable of releasing entrapped calcein from lipid vesicles. Conversely, the two other bio-inactive mutants showed a dramatic decrease in causing membrane permeation. When the N166D mutant was incorporated into planar lipid bilayers (under symmetrical conditions at 150mM KCl, pH8.5), it produced single-channel currents with a maximum conductance of about 425pS comparable to the wild-type toxin. However, maximum conductances for single K(+)-channels formed by both bio-inactive mutants (N166I and N166A) were reduced to approximately 165-205pS. Structural dynamics of 60-ns simulations of a trimeric α4-α5 pore model in a fully hydrated-DMPC system revealed that an open-pore structure could be observed only for the simulated pores of the wild type and N166D. Additionally, the number of lipid molecules interacting with both wild-type and N166D pores is relatively higher than those of N166A and N166I pores. Altogether, our results further signify that the polarity at the α4-α5 loop residue-Asn(166) is directly involved in ion permeation through the Cry4Ba toxin-induced ionic pore and pore opening at the membrane-water interface.

  9. Engineering a pH responsive pore forming protein (United States)

    Kisovec, Matic; Rezelj, Saša; Knap, Primož; Cajnko, Miša Mojca; Caserman, Simon; Flašker, Ajda; Žnidaršič, Nada; Repič, Matej; Mavri, Janez; Ruan, Yi; Scheuring, Simon; Podobnik, Marjetka; Anderluh, Gregor


    Listeriolysin O (LLO) is a cytolysin capable of forming pores in cholesterol-rich lipid membranes of host cells. It is conveniently suited for engineering a pH-governed responsiveness, due to a pH sensor identified in its structure that was shown before to affect its stability. Here we introduced a new level of control of its hemolytic activity by making a variant with hemolytic activity that was pH-dependent. Based on detailed structural analysis coupled with molecular dynamics and mutational analysis, we found that the bulky side chain of Tyr406 allosterically affects the pH sensor. Molecular dynamics simulation further suggested which other amino acid residues may also allosterically influence the pH-sensor. LLO was engineered to the point where it can, in a pH-regulated manner, perforate artificial and cellular membranes. The single mutant Tyr406Ala bound to membranes and oligomerized similarly to the wild-type LLO, however, the final membrane insertion step was pH-affected by the introduced mutation. We show that the mutant toxin can be activated at the surface of artificial membranes or living cells by a single wash with slightly acidic pH buffer. Y406A mutant has a high potential in development of novel nanobiotechnological applications such as controlled release of substances or as a sensor of environmental pH. PMID:28176876

  10. Engineering a pH responsive pore forming protein (United States)

    Kisovec, Matic; Rezelj, Saša; Knap, Primož; Cajnko, Miša Mojca; Caserman, Simon; Flašker, Ajda; Žnidaršič, Nada; Repič, Matej; Mavri, Janez; Ruan, Yi; Scheuring, Simon; Podobnik, Marjetka; Anderluh, Gregor


    Listeriolysin O (LLO) is a cytolysin capable of forming pores in cholesterol-rich lipid membranes of host cells. It is conveniently suited for engineering a pH-governed responsiveness, due to a pH sensor identified in its structure that was shown before to affect its stability. Here we introduced a new level of control of its hemolytic activity by making a variant with hemolytic activity that was pH-dependent. Based on detailed structural analysis coupled with molecular dynamics and mutational analysis, we found that the bulky side chain of Tyr406 allosterically affects the pH sensor. Molecular dynamics simulation further suggested which other amino acid residues may also allosterically influence the pH-sensor. LLO was engineered to the point where it can, in a pH-regulated manner, perforate artificial and cellular membranes. The single mutant Tyr406Ala bound to membranes and oligomerized similarly to the wild-type LLO, however, the final membrane insertion step was pH-affected by the introduced mutation. We show that the mutant toxin can be activated at the surface of artificial membranes or living cells by a single wash with slightly acidic pH buffer. Y406A mutant has a high potential in development of novel nanobiotechnological applications such as controlled release of substances or as a sensor of environmental pH.

  11. Microlens arrays with integrated pores

    Directory of Open Access Journals (Sweden)

    Shu Yang


    Full Text Available Microlenses are important optical components that image, detect, and couple light. But most synthetic microlenses have fixed position and shape once they are fabricated, so their possible range of tunability and complexity is rather limited. By comparison, biology provides many varied, new paradigms for the development of adaptive optical networks. Here, we discuss inspirational examples of biological lenses and their synthetic analogs. We focus on the fabrication and characterization of biomimetic microlens arrays with integrated pores, whose appearance and function are similar to highly efficient optical elements formed by brittlestars. The complex design can be created by three-beam interference lithography. The synthetic lens has strong focusing ability for use as an adjustable lithographic mask and a tunable optical device coupled with the microfluidic system. Replacing rigid microlenses with soft hydrogels provides a way of changing the lens geometry and refractive index continuously in response to external stimuli, resulting in intelligent, multifunctional, tunable optics.

  12. Atomic Structure of Graphene Subnanometer Pores. (United States)

    Robertson, Alex W; Lee, Gun-Do; He, Kuang; Gong, Chuncheng; Chen, Qu; Yoon, Euijoon; Kirkland, Angus I; Warner, Jamie H


    The atomic structure of subnanometer pores in graphene, of interest due to graphene's potential as a desalination and gas filtration membrane, is demonstrated by atomic resolution aberration corrected transmission electron microscopy. High temperatures of 500 °C and over are used to prevent self-healing of the pores, permitting the successful imaging of open pore geometries consisting of between -4 to -13 atoms, all exhibiting subnanometer diameters. Picometer resolution bond length measurements are used to confirm reconstruction of five-membered ring projections that often decorate the pore perimeter, knowledge which is used to explore the viability of completely self-passivated subnanometer pore structures; bonding configurations where the pore would not require external passivation by, for example, hydrogen to be chemically inert.

  13. Connexin mutants and cataracts

    Directory of Open Access Journals (Sweden)

    Eric C Beyer


    Full Text Available The lens is a multicellular, but avascular tissue that must stay transparent to allow normal transmission of light and focusing of it on the retina. Damage to lens cells and/or proteins can cause cataracts, opacities that disrupt these processes. The normal survival of the lens is facilitated by an extensive network of gap junctions formed predominantly of connexin46 and connexin50. Mutations of the genes that encode these connexins (GJA3 and GJA8 have been identified and linked to inheritance of cataracts in human families and mouse lines. In vitro expression studies of several of these mutants have shown that they exhibit abnormalities that may lead to disease. Many of the mutants reduce or modify intercellular communication due to channel alterations (including loss of function or altered gating or due to impaired cellular trafficking which reduces the number of gap junction channels within the plasma membrane. However, the abnormalities detected in studies of other mutants suggest that they cause cataracts through other mechanisms including gain of hemichannel function (leading to cell injury and death and formation of cytoplasmic accumulations (that may act as light scattering particles. These observations and the anticipated results of ongoing studies should elucidate the mechanisms of cataract development due to mutations of lens connexins and abnormalities of other lens proteins. They may also contribute to our understanding of the mechanisms of disease due to connexin mutations in other tissues.

  14. Secretos de Mutantes


    Marín, Martha; Muñoz, Germán; Serrano, Rafael


    Apartándose de enfoques que consideran las culturas juveniles como ‘desviaciones sociales', ‘tribus urbanas' o ‘nuevos movimientos políticos', Secretos de mutantes bucea en culturas juveniles urbanas como la Skinhead, el Punk, el Metal, el Hardcore, el Grunge y el Hip Hop, explorándolas desde un punto de vista inédito: su dimensión de creación, para percibir los cruciales y casi desconocidos procesos que sus miembros llevan a cabo en estos vastos universos de experimentación. Esta obra se nut...

  15. Gas transport and subsoil pore characteristics

    DEFF Research Database (Denmark)

    Berisso, Feto Esimo; Schjønning, Per; Keller, Thomas


    Arrangements of elementary soil particles during soil deposition and subsequent biological and physical processes in long-term pedogenesis are expected to lead to anisotropy of the non-tilled subsoil pore system. Soil compaction by agricultural machinery is known to affect soil pore characteristi...

  16. Cavitation and pore blocking in nanoporous glasses. (United States)

    Reichenbach, C; Kalies, G; Enke, D; Klank, D


    In gas adsorption studies, porous glasses are frequently referred to as model materials for highly disordered mesopore systems. Numerous works suggest that an accurate interpretation of physisorption isotherms requires a complete understanding of network effects upon adsorption and desorption, respectively. The present article deals with nitrogen and argon adsorption at different temperatures (77 and 87 K) performed on a series of novel nanoporous glasses (NPG) with different mean pore widths. NPG samples contain smaller mesopores and significantly higher microporosity than porous Vycor glass or controlled pore glass. Since the mean pore width of NPG can be tuned sensitively, the evolution of adsorption characteristics with respect to a broadening pore network can be investigated starting from the narrowest nanopore width. With an increasing mean pore width, a H2-type hysteresis develops gradually which finally transforms into a H1-type. In this connection, a transition from a cavitation-induced desorption toward desorption controlled by pore blocking can be observed. Furthermore, we find concrete hints for a pore size dependence of the relative pressure of cavitation in highly disordered pore systems. By comparing nitrogen and argon adsorption, a comprehensive insight into adsorption mechanisms in novel disordered materials is provided.

  17. Coating of silicon pore optics

    DEFF Research Database (Denmark)

    Cooper-Jensen, Carsten P.; Ackermann, M.; Christensen, Finn Erland


    For the International X-ray observatory (IXO), a mirror module with an effective area of 3 m2 at 1.25 keV and at least 0.65 m2 at 6 keV has to be realized. To achieve this goal, coated silicon pore optics has been developed over the last years. One of the challenges is to coat the Si plates...... and still to realize Si-Si bonding. It has been demonstrated that ribbed silicon plates can be produced and assembled into stacks. All previously work has been done using uncoated Si plates. In this paper we describe how to coat the ribbed Si plates with an Ir coating and a top C coating through a mask so...... that there will be coating only between the ribs and not in the area where bonding takes place. The paper includes description of the mounting jig and how to align the mask on top of the plate. We will also present energy scans from Si plates coated through a mask....

  18. Particle diffusion in complex nanoscale pore networks

    DEFF Research Database (Denmark)

    Müter, Dirk; Sørensen, Henning Osholm; Bock, H.


    decreased to as much as 60% when particle size increased from 1% to 35% of the average pore diameter. When particles were attracted to the pore surfaces, even very small particles, diffusion was drastically inhibited, by as much as a factor of 100. Thus, the size of particles and their interaction......We studied the diffusion of particles in the highly irregular pore networks of chalk, a very fine-grained rock, by combining three-dimensional X-ray imaging and dissipative particle dynamics (DPD) simulations. X-ray imaging data were collected at 25 nm voxel dimension for two chalk samples...... with very different porosities (4% and 26%). The three-dimensional pore systems derived from the tomograms were imported into DPD simulations and filled with spherical particles of variable diameter and with an optional attractive interaction to the pore surfaces. We found that diffusion significantly...


    Directory of Open Access Journals (Sweden)

    S. Malathi


    Full Text Available In recent years there has been exponential growth in the use of bio- metrics for user authentication applications. Automated Fingerprint Identification systems have become popular tool in many security and law enforcement applications. Most of these systems rely on minutiae (ridge ending and bifurcation features. With the advancement in sensor technology, high resolution fingerprint images (1000 dpi pro- vide micro level of features (pores that have proven to be useful fea- tures for identification. In this paper, we propose a new strategy for fingerprint matching based on pores by reliably extracting the pore features The extraction of pores is done by Marker Controlled Wa- tershed segmentation method and the centroids of each pore are con- sidered as feature vectors for matching of two fingerprint images. Experimental results shows that the proposed method has better per- formance with lower false rates and higher accuracy.

  20. The E1 protein is mandatory for pore formation by Semliki Forest virus spikes. (United States)

    Dick, M; Barth, B U; Kempf, C


    Insect cells (Aedes albopictus, clone C6/36) were infected with various variants of Semliki Forest virus including the wild type using the SFV replicon system. The variants included deletion mutants lacking one of the structural proteins and a mutant with a point mutation in p62 (SQL). The latter mutation results in a failure to process p62 to E2 and E3. After infection of the cells with different variant viruses and subsequent expression of viral proteins in the host cell plasma membrane low pH-induced pore formation was detected by measuring the efflux of a radiolabeled compound. The results of these experiments clearly showed that the E1 protein is mandatory for the acid-induced pore formation. A participation of the 6K or C-protein could be excluded. Furthermore, results obtained with the SQL mutant suggest that dissociation of the E1/E2 heterodimer and subsequent homooligomerization of E1 are required for pore formation.

  1. Concerted Motions Networking Pores and Distant Ferroxidase Centers Enable Bacterioferritin Function and Iron Traffic£ξ (United States)

    Yao, Huili; Rui, Huan; Kumar, Ritesh; Eshelman, Kate; Lovell, Scott; Battaile, Kevin P.; Im, Wonpil; Rivera, Mario


    X-ray crystallography, molecular dynamics (MD) simulations and biochemistry were utilized to investigate the effect of introducing hydrophobic interactions in the 4-fold (N148L and Q151L) and B-pores (D34F) of Pseudomonas aeruginosa bacterioferritin B (BfrB) on BfrB function. The structures show only local structural perturbations and confirm the anticipated hydrophobic interactions. Surprisingly, structures obtained after soaking crystals in Fe2+-containing crystallization solution revealed that although iron loads into the ferroxidase centers of the mutants, the side chains of ferroxidase ligands E51 and H130 do not reorganize to bind the iron ions, as is seen in the wt BfrB structures. Similar experiments with a double mutant (C89S/K96C) prepared to introduce changes outside the pores show competent ferroxidase centers that function akin to those in wt BfrB. MD simulations comparing wt BfrB with the D34F and N148L mutants show that the mutants exhibit significantly reduced flexibility, and reveal a network of concerted motions linking ferroxidase centers and 4-fold and B-pores, which are important for imparting ferroxidase centers in BfrB with the required flexibility to function efficiently. In agreement, the efficiency of Fe2+ oxidation and uptake of the 4-fold and B-pore mutants in solution is significantly compromised relative to wt or C89S/K96C BfrB. Finally, our structures show a large number of previously unknown iron binding sites in the interior cavity and B-pores of BfrB, which reveal in unprecedented detail conduits followed by iron and phosphate ions across the BfrB shell, as well as paths in the interior cavity that may facilitate nucleation of the iron phosphate mineral. PMID:25640193

  2. Designing Nonwovens to Meet Pore Size Specifications

    Directory of Open Access Journals (Sweden)

    Glen E. Simmonds


    Full Text Available New nonwovens applications in areas such as filtration require a media designed to particular pore size specifications in the 3 to 20 micron range. The purpose of this work was to develop a basis by which to design and construct a fabric with given pore size specifications. While doing so we have provided a validation for two different mathematical models. We have also found that bicomponent spunbonded islands-in-the-sea nonwoven fabrics can be designed very precisely to achieve target pore diameters and porosity. Mathematical models can be used to develop fabric specifications in the standard manufacturing terms of basis weight and fiber diameter. Measured mean flow pore diameters for the test fabrics showed excellent correlation to targeted mean flow pore diameters for both models. The experimental fit to the Bryner model is the better of the two, but requires specification of fabric thickness in addition to basis weight and fiber diameter to achieve actual mean pore diameters that closely match target values. Experimental validation of the influence of fabric thickness on the mean flow pore diameter at constant basis weight and fiber diameter remains open for further investigation. In addition, achieving complete separation of the island and sea polymers along with unbundling of the island fibers remain areas for improvement.

  3. ECB deacylase mutants (United States)

    Arnold, Frances H.; Shao, Zhixin; Zhao, Huimin; Giver, Lorraine J.


    A method for in vitro mutagenesis and recombination of polynucleotide sequences based on polymerase-catalyzed extension of primer oligonucleotides is disclosed. The method involves priming template polynucleotide(s) with random-sequences or defined-sequence primers to generate a pool of short DNA fragments with a low level of point mutations. The DNA fragments are subjected to denaturization followed by annealing and further enzyme-catalyzed DNA polymerization. This procedure is repeated a sufficient number of times to produce full-length genes which comprise mutants of the original template polynucleotides. These genes can be further amplified by the polymerase chain reaction and cloned into a vector for expression of the encoded proteins.

  4. Control of pore size in epoxy systems.

    Energy Technology Data Exchange (ETDEWEB)

    Sawyer, Patricia Sue; Lenhart, Joseph Ludlow (North Dakota State University, Fargo, ND); Lee, Elizabeth (North Dakota State University, Fargo, ND); Kallam, Alekhya (North Dakota State University, Fargo, ND); Majumdar, Partha (North Dakota State University, Fargo, ND); Dirk, Shawn M.; Gubbins, Nathan; Chisholm, Bret J. (North Dakota State University, Fargo, ND); Celina, Mathias Christopher; Bahr, James (North Dakota State University, Fargo, ND); Klein, Robert J.


    Both conventional and combinatorial approaches were used to study the pore formation process in epoxy based polymer systems. Sandia National Laboratories conducted the initial work and collaborated with North Dakota State University (NDSU) using a combinatorial research approach to produce a library of novel monomers and crosslinkers capable of forming porous polymers. The library was screened to determine the physical factors that control porosity, such as porogen loading, polymer-porogen interactions, and polymer crosslink density. We have identified the physical and chemical factors that control the average porosity, pore size, and pore size distribution within epoxy based systems.

  5. Analytical applications for pore-forming proteins. (United States)

    Kasianowicz, John J; Balijepalli, Arvind K; Ettedgui, Jessica; Forstater, Jacob H; Wang, Haiyan; Zhang, Huisheng; Robertson, Joseph W F


    Proteinaceous nanometer-scale pores are ubiquitous in biology. The canonical ionic channels (e.g., those that transport Na(+), K(+), Ca(2+), and Cl(-) across cell membranes) play key roles in many cellular processes, including nerve and muscle activity. Another class of channels includes bacterial pore-forming toxins, which disrupt cell function, and can lead to cell death. We describe here the recent development of these toxins for a wide range of biological sensing applications. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale.

  6. Straight Pore Microfilter with Efficient Regeneration Project (United States)

    National Aeronautics and Space Administration — This Small Business Innovation Research Phase II project is directed toward development of a novel microfiltration filter that has distinctively narrow pore size...

  7. Straight Pore Microfilter with Efficient Regeneration Project (United States)

    National Aeronautics and Space Administration — This Small Business Innovation Research Phase I project is directed toward development of a novel microfiltration filter that has distinctively narrow pore size...

  8. A Potential Model for Cylindrical Pores

    Institute of Scientific and Technical Information of China (English)

    张现仁; 汪文川


    An analytical potential for cylindrical pores has been derived by introducing a variational method into the integration for the calculation of the interaction energy between the wall molecules and a test molecule, all of which are represented by Lennard-Jones potential. The model proposed gives good fit to the results from the cylindrical surface model and the pseudoatom model. To test the potential proposed rigorously, we have carried out grand canonical ensemble Monte Carlo(GCMC) simulation of nitrogen in the MCM-41 pore at 77 K, and compared the simulated adsorption isotherm with the experimental data reported in the literature. The simulated isotherm from our model is in almost qualitative agreement with experiment. Consequently, the model proposed provides an explicit and accurate description of cylindrical pores represented by the Lennard-Jones potential. Moreover, the model can be easily applied to a variety of cylindrical pores, ranging from cylindrical surface to finite thickness walls, in both theoretical studies and computer simulations.

  9. Pore structure in blended cement pastes

    DEFF Research Database (Denmark)

    Canut, Mariana Moreira Cavalcanti

    Supplementary cementitious materials (SCMs), such as slag and fly ash, are increasingly used as a substitute for Portland cement in the interests of improvement of engineering properties and sustainability of concrete. According to studies improvement of engineering properties can be explained...... supplement each other. Cement pastes (w/b=0.4) with and without slag and fly ash cured at two moisture (sealed and saturated) and temperature (20 and 55ºC) conditions were used to investigate the combined impact of SCMs addition and curing on the pore structure of pastes cured up to two years. Also...... volume and threshold pore size were found when comparing with plain cement paste at the same curing conditions. The porosity methods MIP, LTC and SEM have been shown to be suitable to characterise pore parameters of the pastes. MIP is a simple and fast method which covers a large range of pore sizes...

  10. Analysis of a spatially deconvolved solar pore

    CERN Document Server

    Noda, C Quintero; Cobo, B Ruiz; Suematsu, Y; Katsukawa, Y; Ichimoto, K


    Solar pores are active regions with large magnetic field strengths and apparent simple magnetic configurations. Their properties resemble the ones found for the sunspot umbra although pores do not show penumbra. Therefore, solar pores present themselves as an intriguing phenomenon that is not completely understood. We examine in this work a solar pore observed with Hinode/SP using two state of the art techniques. The first one is the spatial deconvolution of the spectropolarimetric data that allows removing the stray light contamination induced by the spatial point spread function of the telescope. The second one is the inversion of the Stokes profiles assuming local thermodynamic equilibrium that let us to infer the atmospheric physical parameters. After applying these techniques, we found that the spatial deconvolution method does not introduce artefacts, even at the edges of the magnetic structure, where large horizontal gradients are detected on the atmospheric parameters. Moreover, we also describe the p...

  11. Ion permeation and block of the gating pore in the voltage sensor of NaV1.4 channels with hypokalemic periodic paralysis mutations. (United States)

    Sokolov, Stanislav; Scheuer, Todd; Catterall, William A


    Hypokalemic periodic paralysis and normokalemic periodic paralysis are caused by mutations of the gating charge-carrying arginine residues in skeletal muscle Na(V)1.4 channels, which induce gating pore current through the mutant voltage sensor domains. Inward sodium currents through the gating pore of mutant R666G are only approximately 1% of central pore current, but substitution of guanidine for sodium in the extracellular solution increases their size by 13- +/- 2-fold. Ethylguanidine is permeant through the R666G gating pore at physiological membrane potentials but blocks the gating pore at hyperpolarized potentials. Guanidine is also highly permeant through the proton-selective gating pore formed by the mutant R666H. Gating pore current conducted by the R666G mutant is blocked by divalent cations such as Ba(2+) and Zn(2+) in a voltage-dependent manner. The affinity for voltage-dependent block of gating pore current by Ba(2+) and Zn(2+) is increased at more negative holding potentials. The apparent dissociation constant (K(d)) values for Zn(2+) block for test pulses to -160 mV are 650 +/- 150 microM, 360 +/- 70 microM, and 95.6 +/- 11 microM at holding potentials of 0 mV, -80 mV, and -120 mV, respectively. Gating pore current is blocked by trivalent cations, but in a nearly voltage-independent manner, with an apparent K(d) for Gd(3+) of 238 +/- 14 microM at -80 mV. To test whether these periodic paralyses might be treated by blocking gating pore current, we screened several aromatic and aliphatic guanidine derivatives and found that 1-(2,4-xylyl)guanidinium can block gating pore current in the millimolar concentration range without affecting normal Na(V)1.4 channel function. Together, our results demonstrate unique permeability of guanidine through Na(V)1.4 gating pores, define voltage-dependent and voltage-independent block by divalent and trivalent cations, respectively, and provide initial support for the concept that guanidine-based gating pore blockers

  12. Visualization of enzyme activities inside earthworm pores (United States)

    Hoang, Duyen; Razavi, Bahar S.


    In extremely dynamic microhabitats as bio-pores made by earthworm, the in situ enzyme activities are assumed as a footprint of complex biotic interactions. Our study focused on the effect of earthworm on the enzyme activities inside bio-pores and visualizing the differences between bio-pores and earthworm-free soil by zymography technique (Spohn and Kuzyakov, 2013). For the first time, we aimed at quantitative imaging of enzyme activities in bio-pores. Lumbricus terrestris L. was placed into transparent box (15×20×15cm). After two weeks when bio-pore systems were formed by earthworms, we visualized in situ enzyme activities of five hydrolytic enzymes (β-glucosidase, cellobiohydrolase, chitinase, xylanase, leucine-aminopeptidase, and phosphatase. Zymography showed higher activity of β-glucosidase, chitinase, xylanase and phosphatase in biopores comparing to bulk soil. However, the differences in activity of cellobiohydrolase and leucine aminopeptidase between bio-pore and bulk soil were less pronounced. This demonstrated an applicability of zymography approach to monitor and to distinguish the in situ activity of hydrolytic enzymes in soil biopores.

  13. Pore REconstruction and Segmentation (PORES) method for improved porosity quantification of nanoporous materials

    Energy Technology Data Exchange (ETDEWEB)

    Van Eyndhoven, G., E-mail: [iMinds-Vision Lab, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk (Belgium); Kurttepeli, M. [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Van Oers, C.J.; Cool, P. [Laboratory of Adsorption and Catalysis, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk (Belgium); Bals, S. [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Batenburg, K.J. [iMinds-Vision Lab, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk (Belgium); Centrum Wiskunde and Informatica, Science Park 123, NL-1090 GB Amsterdam (Netherlands); Mathematical Institute, Universiteit Leiden, Niels Bohrweg 1, NL-2333 CA Leiden (Netherlands); Sijbers, J. [iMinds-Vision Lab, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk (Belgium)


    Electron tomography is currently a versatile tool to investigate the connection between the structure and properties of nanomaterials. However, a quantitative interpretation of electron tomography results is still far from straightforward. Especially accurate quantification of pore-space is hampered by artifacts introduced in all steps of the processing chain, i.e., acquisition, reconstruction, segmentation and quantification. Furthermore, most common approaches require subjective manual user input. In this paper, the PORES algorithm “POre REconstruction and Segmentation” is introduced; it is a tailor-made, integral approach, for the reconstruction, segmentation, and quantification of porous nanomaterials. The PORES processing chain starts by calculating a reconstruction with a nanoporous-specific reconstruction algorithm: the Simultaneous Update of Pore Pixels by iterative REconstruction and Simple Segmentation algorithm (SUPPRESS). It classifies the interior region to the pores during reconstruction, while reconstructing the remaining region by reducing the error with respect to the acquired electron microscopy data. The SUPPRESS reconstruction can be directly plugged into the remaining processing chain of the PORES algorithm, resulting in accurate individual pore quantification and full sample pore statistics. The proposed approach was extensively validated on both simulated and experimental data, indicating its ability to generate accurate statistics of nanoporous materials. - Highlights: • An electron tomography reconstruction/segmentation method for nanoporous materials. • The method exploits the porous nature of the scanned material. • Validated extensively on both simulation and real data experiments. • Results in increased image resolution and improved porosity quantification.

  14. Membrane damage by an α-helical pore-forming protein, Equinatoxin II, proceeds through a succession of ordered steps. (United States)

    Rojko, Nejc; Kristan, Katarina Č; Viero, Gabriella; Žerovnik, Eva; Maček, Peter; Dalla Serra, Mauro; Anderluh, Gregor


    Actinoporin equinatoxin II (EqtII) is an archetypal example of α-helical pore-forming toxins that porate cellular membranes by the use of α-helices. Previous studies proposed several steps in the pore formation: binding of monomeric protein onto the membrane, followed by oligomerization and insertion of the N-terminal α-helix into the lipid bilayer. We studied these separate steps with an EqtII triple cysteine mutant. The mutant was engineered to monitor the insertion of the N terminus into the lipid bilayer by labeling Cys-18 with a fluorescence probe and at the same time to control the flexibility of the N-terminal region by the disulfide bond formed between cysteines introduced at positions 8 and 69. The insertion of the N terminus into the membrane proceeded shortly after the toxin binding and was followed by oligomerization. The oxidized, non-lytic, form of the mutant was still able to bind to membranes and oligomerize at the same level as the wild-type or the reduced form. However, the kinetics of the N-terminal helix insertion, the release of calcein from erythrocyte ghosts, and hemolysis of erythrocytes was much slower when membrane-bound oxidized mutant was reduced by the addition of the reductant. Results show that the N-terminal region needs to be inserted in the lipid membrane before the oligomerization into the final pore and imply that there is no need for a stable prepore formation. This is different from β-pore-forming toxins that often form β-barrel pores via a stable prepore complex.

  15. Low Pore Connectivity in Natural Rock

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Qinhong; Ewing, Robert P.; Dultz, Stefan


    As repositories for CO₂ and radioactive waste, as oil and gas reservoirs, and as contaminated sites needing remediation, rock formations play a central role in energy and environmental management. The connectivity of the rock's porespace strongly affects fluid flow and solute transport. This work examines pore connectivity and its implications for fluid flow and chemical transport. Three experimental approaches (imbibition, tracer concentration profiles, and imaging) were used in combination with network modeling. In the imbibition results, three types of imbibition slope [log (cumulative imbibition) vs. log (imbibition time)] were found: the classical 0.5, plus 0.26, and 0.26 transitioning to 0.5. The imbibition slope of 0.26 seen in Indiana sandstone, metagraywacke, and Barnett shale indicates low pore connectivity, in contrast to the slope of 0.5 seen in the well-connected Berea sandstone. In the tracer profile work, rocks exhibited different distances to the plateau porosity, consistent with the pore connectivity from the imbibition tests. Injection of a molten metal into connected pore spaces, followed by 2-D imaging of the solidified alloy in polished thin sections, allowed direct assessment of pore structure and lateral connection in the rock samples. Pore-scale network modeling gave results consistent with measurements, confirming pore connectivity as the underlying cause of both anomalous behaviors: imbibition slope not having the classical value of 0.5, and accessible porosity being a function of distance from the edge. A poorly connected porespace will exhibit anomalous behavior in fluid flow and chemical transport, such as a lower imbibition slope (in air–water system) and diffusion rate than expected from classical behavior.

  16. Low pore connectivity in natural rock. (United States)

    Hu, Qinhong; Ewing, Robert P; Dultz, Stefan


    As repositories for CO(2) and radioactive waste, as oil and gas reservoirs, and as contaminated sites needing remediation, rock formations play a central role in energy and environmental management. The connectivity of the rock's porespace strongly affects fluid flow and solute transport. This work examines pore connectivity and its implications for fluid flow and chemical transport. Three experimental approaches (imbibition, tracer concentration profiles, and imaging) were used in combination with network modeling. In the imbibition results, three types of imbibition slope [log (cumulative imbibition) vs. log (imbibition time)] were found: the classical 0.5, plus 0.26, and 0.26 transitioning to 0.5. The imbibition slope of 0.26 seen in Indiana sandstone, metagraywacke, and Barnett shale indicates low pore connectivity, in contrast to the slope of 0.5 seen in the well-connected Berea sandstone. In the tracer profile work, rocks exhibited different distances to the plateau porosity, consistent with the pore connectivity from the imbibition tests. Injection of a molten metal into connected pore spaces, followed by 2-D imaging of the solidified alloy in polished thin sections, allowed direct assessment of pore structure and lateral connection in the rock samples. Pore-scale network modeling gave results consistent with measurements, confirming pore connectivity as the underlying cause of both anomalous behaviors: imbibition slope not having the classical value of 0.5, and accessible porosity being a function of distance from the edge. A poorly connected porespace will exhibit anomalous behavior in fluid flow and chemical transport, such as a lower imbibition slope (in air-water system) and diffusion rate than expected from classical behavior.

  17. Pore Structure Characterization of Indiana Limestone and Pink Dolomite from Pore Network Reconstructions

    Directory of Open Access Journals (Sweden)

    Freire-Gormaly Marina


    Full Text Available Carbon sequestration in deep underground saline aquifers holds significant promise for reducing atmospheric carbon dioxide emissions (CO2. However, challenges remain in predicting the long term migration of injected CO2. Addressing these challenges requires an understanding of pore-scale transport of CO2 within existing brine-filled geological reservoirs. Studies on the transport of fluids through geological porous media have predominantly focused on oil-bearing formations such as sandstone. However, few studies have considered pore-scale transport within limestone and other carbonate formations, which are found in potential storage sites. In this work, high-resolution micro-Computed Tomography (microCT was used to obtain pore-scale structural information of two model carbonates: Indiana Limestone and Pink Dolomite. A modified watershed algorithm was applied to extract pore network from the reconstructed microCT volumetric images of rock samples and compile a list of pore-scale characteristics from the extracted networks. These include statistical distributions of pore size and radius, pore-pore separation, throat radius, and network coordination. Finally, invasion percolation algorithms were applied to determine saturation-pressure curves for the rock samples. The statistical distributions were comparable to literature values for the Indiana Limestone. This served as validation for the network extraction approach for Pink Dolomite, which has not been considered previously. Based on the connectivity and the pore-pore separation, formations such as Pink Dolomite may present suitable storage sites for carbon storage. The pore structural distributions and saturation curves obtained in this study can be used to inform core- and reservoir-scale modeling and experimental studies of sequestration feasibility.

  18. Modeling branching pore structures in membrane filters (United States)

    Sanaei, Pejman; Cummings, Linda J.


    Membrane filters are in widespread industrial use, and mathematical models to predict their efficacy are potentially very useful, as such models can suggest design modifications to improve filter performance and lifetime. Many models have been proposed to describe particle capture by membrane filters and the associated fluid dynamics, but most such models are based on a very simple structure in which the pores of the membrane are assumed to be simple circularly-cylindrical tubes spanning the depth of the membrane. Real membranes used in applications usually have much more complex geometry, with interconnected pores which may branch and bifurcate. Pores are also typically larger on the upstream side of the membrane than on the downstream side. We present an idealized mathematical model, in which a membrane consists of a series of bifurcating pores, which decrease in size as the membrane is traversed. Feed solution is forced through the membrane by applied pressure, and particles are removed from the feed either by sieving, or by particle adsorption within pores (which shrinks them). Thus the membrane's permeability decreases as the filtration progresses, ultimately falling to zero. We discuss how filtration efficiency depends on the characteristics of the branching structure. Partial support from NSF DMS 1261596 is gratefully acknowledged.

  19. Performance of Small Pore Microchannel Plates (United States)

    Siegmund, O. H. W.; Gummin, M. A.; Ravinett, T.; Jelinsky, S. R.; Edgar, M.


    Small pore size microchannel plates (MCP's) are needed to satisfy the requirements for future high resolution small and large format detectors for astronomy. MCP's with pore sizes in the range 5 micron to 8 micron are now being manufactured, but they are of limited availability and are of small size. We have obtained sets of Galileo 8 micron and 6.5 micron MCP's, and Philips 6 micron and 7 micron pore MCP's, and compared them to our larger pore MCP Z stacks. We have tested back to back MCP stacks of four of these MCP's and achieved gains greater than 2 x 1O(exp 7) with pulse height distributions of less than 40% FWHM, and background rates of less than 0.3 events sec(exp -1) cm(exp -2). Local counting rates up to approx. 100 events/pore/sec have been attained with little drop of the MCP gain. The bare MCP quantum efficiencies are somewhat lower than those expected, however. Flat field images are characterized by an absence of MCP fixed pattern noise.

  20. Modeling Tissue Growth Within Nonwoven Scaffolds Pores (United States)

    Church, Jeffrey S.; Alexander, David L.J.; Russell, Stephen J.; Ingham, Eileen; Ramshaw, John A.M.; Werkmeister, Jerome A.


    In this study we present a novel approach for predicting tissue growth within the pores of fibrous tissue engineering scaffolds. Thin nonwoven polyethylene terephthalate scaffolds were prepared to characterize tissue growth within scaffold pores, by mouse NR6 fibroblast cells. On the basis of measurements of tissue lengths at fiber crossovers and along fiber segments, mathematical models were determined during the proliferative phase of cell growth. Tissue growth at fiber crossovers decreased with increasing interfiber angle, with exponential relationships determined on day 6 and 10 of culture. Analysis of tissue growth along fiber segments determined two growth profiles, one with enhanced growth as a result of increased tissue lengths near the fiber crossover, achieved in the latter stage of culture. Derived mathematical models were used in the development of a software program to visualize predicted tissue growth within a pore. This study identifies key pore parameters that contribute toward tissue growth, and suggests models for predicting this growth, based on fibroblast cells. Such models may be used in aiding scaffold design, for optimum pore infiltration during the tissue engineering process. PMID:20687775

  1. Analysis of a spatially deconvolved solar pore (United States)

    Quintero Noda, C.; Shimizu, T.; Ruiz Cobo, B.; Suematsu, Y.; Katsukawa, Y.; Ichimoto, K.


    Solar pores are active regions with large magnetic field strengths and apparent simple magnetic configurations. Their properties resemble the ones found for the sunspot umbra although pores do not show penumbra. Therefore, solar pores present themselves as an intriguing phenomenon that is not completely understood. We examine in this work a solar pore observed with Hinode/SP using two state of the art techniques. The first one is the spatial deconvolution of the spectropolarimetric data that allows removing the stray light contamination induced by the spatial point spread function of the telescope. The second one is the inversion of the Stokes profiles assuming local thermodynamic equilibrium that let us to infer the atmospheric physical parameters. After applying these techniques, we found that the spatial deconvolution method does not introduce artefacts, even at the edges of the magnetic structure, where large horizontal gradients are detected on the atmospheric parameters. Moreover, we also describe the physical properties of the magnetic structure at different heights finding that, in the inner part of the solar pore, the temperature is lower than outside, the magnetic field strength is larger than 2 kG and unipolar, and the line-of-sight velocity is almost null. At neighbouring pixels, we found low magnetic field strengths of same polarity and strong downward motions that only occur at the low photosphere, below the continuum optical depth log τ = -1. Finally, we studied the spatial relation between different atmospheric parameters at different heights corroborating the physical properties described before.

  2. Reversal of charge selectivity in transmembrane protein pores by using noncovalent molecular adapters (United States)

    Gu, Li-Qun; Dalla Serra, Mauro; Vincent, J. Bryan; Vigh, Gyula; Cheley, Stephen; Braha, Orit; Bayley, Hagan


    In this study, the charge selectivity of staphylococcal α-hemolysin (αHL), a bacterial pore-forming toxin, is manipulated by using cyclodextrins as noncovalent molecular adapters. Anion-selective versions of αHL, including the wild-type pore and various mutants, become more anion selective when β-cyclodextrin (βCD) is lodged within the channel lumen. By contrast, the negatively charged adapter, hepta-6-sulfato-β-cyclodextrin (s7βCD), produces cation selectivity. The cyclodextrin adapters have similar effects when placed in cation-selective mutant αHL pores. Most probably, hydrated Cl− ions partition into the central cavity of βCD more readily than K+ ions, whereas s7βCD introduces a charged ring near the midpoint of the channel lumen and confers cation selectivity through electrostatic interactions. The molecular adapters generate permeability ratios (PK+/PCl−) over a 200-fold range and should be useful in the de novo design of membrane channels both for basic studies of ion permeation and for applications in biotechnology. PMID:10760267

  3. TIG Dressing Effects on Weld Pores and Pore Cracking of Titanium Weldments

    Directory of Open Access Journals (Sweden)

    Hui-Jun Yi


    Full Text Available Weld pores redistribution, the effectiveness of using tungsten inert gas (TIG dressing to remove weld pores, and changes in the mechanical properties due to the TIG dressing of Ti-3Al-2.5V weldments were studied. Moreover, weld cracks due to pores were investigated. The results show that weld pores less than 300 μm in size are redistributed or removed via remelting due to TIG dressing. Regardless of the temperature condition, TIG dressing welding showed ductility, and there was a loss of 7% tensile strength of the weldments. Additionally, it was considered that porosity redistribution by TIG dressing was due to fluid flow during the remelting of the weld pool. Weld cracks in titanium weldment create branch cracks around pores that propagate via the intragranular fracture, and oxygen is dispersed around the pores. It is suggested that the pore locations around the LBZ (local brittle zone and stress concentration due to the pores have significant effects on crack initiation and propagation.

  4. Chromatographic performance of large-pore versus small-pore columns in micellar liquid chromatography. (United States)

    McCormick, Timothy J; Foley, Joe P; Lloyd, David K


    Micellar liquid chromatography (MLC) is useful in bioanalysis because proteinaceous biofluids can be directly injected onto the column. The technique has been limited in part because of the apparently weak eluting power of micellar mobile phases. It has recently been shown [Anal. Chem. 72 (2000) 294] that this may be overcome by the use of large pore size stationary phases. In this work, large-pore (1000 A) C(18) stationary phases were evaluated relative to conventional small-pore (100 A) C(18) stationary phases for the direct sample injection of drugs in plasma. Furthermore, the difference between the large and small pore phases in gradient elution separations of mixtures of widely varying hydrophobicities was investigated. Large-pore stationary phases were found to be very effective for eluting moderately to highly hydrophobic compounds such as ibuprofen, crotamiton, propranolol, and dodecanophenone, which were highly retained on the small-pore stationary phases typically used in MLC. The advantages of direct introduction of biological samples (drugs in plasma) and rapid column re-equilibration after gradient elution in MLC were maintained with large-pore phases. Finally, recoveries, precision, linearity, and detection limits for the determination of quinidine and DPC 961 in spiked bovine plasma were somewhat better using MLC with wide pore phases.

  5. Modeling of N2 adsorption in MCM-41 materials: hexagonal pores versus cylindrical pores. (United States)

    Ustinov, Eugene A


    Low-temperature nitrogen adsorption in hexagonal pores and equivalent cylindrical pores is analyzed using nonlocal density functional theory extended to amorphous solids (NLDFT-AS). It is found that, despite significant difference of the density distribution over the cross-section of the pore, the capillary condensation/evaporation pressure is not considerably affected by the pore shape being slightly lower in the case of hexagonal geometry. However, the condensation/evaporation step in the hexagonal pore is slightly larger than that in the equivalent cylindrical pore because in the latter case the pore wall surface area and, hence, the amount adsorbed at pressures below the evaporation pressure are underestimated by 5%. We show that a dimensionless parameter defined as the ratio of the condensation/evaporation step and the upper value of the amount adsorbed at the condensation/evaporation pressure can be used as an additional criterion of the correct choice of the gas-solid molecular parameters along with the dependence of condensation/evaporation pressure on the pore diameter. Application of the criteria to experimental data on nitrogen adsorption on a series of MCM-41 silica at 77 K corroborates some evidence that the capillary condensation occurs at equilibrium conditions.

  6. Moving Magnetic Features around a Pore

    CERN Document Server

    Kaithakkal, A J; Solanki, S K; Lagg, A; Barthol, P; Gandorfer, A; Gizon, L; Hirzberger, J; vanNoort, M; Rodríguez, J Blanco; Iniesta, J C Del Toro; Suárez, D Orozco; Schmidt, W; Pillet, V Martínez; Knölker, M


    Spectropolarimetric observations from Sunrise II/IMaX obtained in June 2013 are used for a statistical analysis to determine the physical properties of moving magnetic features (MMFs) observed near a pore. MMFs of the same and opposite polarity with respect to the pore are found to stream from its border at an average speed of 1.3 km s$^{-1}$ and 1.2 km s$^{-1}$ respectively, with mainly same-polarity MMFs found further away from the pore. MMFs of both polarities are found to harbor rather weak, inclined magnetic fields. Opposite-polarity MMFs are blue-shifted, while same-polarity MMFs do not show any preference for up- or downflows. Most of the MMFs are found to be of sub-arcsecond size and carry a mean flux of $\\sim$ 1.2$\\times 10^{17}$ Mx.

  7. Optical detection of pores in adipocyte membrane (United States)

    Yanina, I. Yu.; Doubrovski, V. A.; Tuchin, V. V.


    Structures that can be interpreted as cytoplasm droplets leaking through the membrane are experimentally detected on the membranes of adipocytes using optical digital microscopy. The effect of an aqueous alcohol solution of brilliant green on the amount and sizes of structures is studied. It is demonstrated that the optical irradiation of the adipocytes that are sensitized with the aid of the brilliant green leads to an increase in the amount of structures (pores) after the irradiation. The experimental results confirm the existence of an earlier-proposed effect of photochemical action on the sensitized cells of adipose tissue that involves additional formation of pores in the membrane of the sensitized cell under selective optical irradiation. The proposed method for the detection of micropores in the membrane of adipose tissue based on the detection of the cytoplasm droplets leaking from the cell can be considered as a method for the optical detection of nanosized pores.

  8. Pore Pressure Measurements Inside Rubble Mound Breakwaters

    DEFF Research Database (Denmark)

    Helgason, Einar; Burcharth, H. F.; Grüne, Joachim


    The present paper presents pore pressure measurements from large scale model tests performed at the Large Wave Channel, Hannover, Germany and small scale model test performed at the Hydraulic & Coastal Engineering Laboratory, Aalborg University, Denmark. Information on pore pressure attenuation......, and compared to a damping model presented by Burcharth et al. (1999). Reasonable agreement is found when considering the difference in the grading and uniformity of the model core materials. Comparison between results obtained from small and large scale model tests showed no clear evidence of scale effects....

  9. Porous media fluid transport and pore structure

    CERN Document Server

    Dullien, F A L


    This book examines the relationship between transport properties and pore structure of porous material. Models of pore structure are presented with a discussion of how such models can be used to predict the transport properties of porous media. Portions of the book are devoted to interpretations of experimental results in this area and directions for future research. Practical applications are given where applicable, and are expected to be useful for a large number of different fields, including reservoir engineering, geology, hydrogeology, soil science, chemical process engineering, biomedica

  10. Active Pore Volume in Danish Peat Soils

    DEFF Research Database (Denmark)

    Forsmann, Ditte M.; Kjærgaard, Charlotte


    Phosphorus release within the soil matrix caused by the changed redox conditions due to re-establishment of a riparian wetland can be critical for the aquatic environment. However, phosphorous released in the soil will not always result in an immediate contribution to this loss to the aquatic...... is not actively transported out of the system, but is only transported via diffusion, which is a very slow process. Thus it is interesting to investigate the size of the active pore volume in peat soils. The hypothesis of this study is that the active pores volume of a peat soil can be expressed using bulk...

  11. Conservation agriculture effects on soil pore characteristics

    DEFF Research Database (Denmark)

    Munkholm, Lars Juhl; Abdollahi, Lotfollah

    of quantitative knowledge to support this statement. This study examines the long-term effects of crop rotations, residue management and tillage on soil pore characteristics of two sandy loam soils in Denmark. Results are reported from a split plot field experiment rotation as main plot factor and tillage...... air permeability and pore continuity index. Generally, residue input, especially when combined with direct drilling at the Foulum site, decreased bulk density and the volume of blocked air porosity, and increased air-filled porosity, volumetric water content, air permeability and gas diffusivity. Our...

  12. Control of ionic selectivity by a pore helix residue in the Kv1.2 channel. (United States)

    Chao, Chia-Chia; Huang, Chieh-Chen; Kuo, Chang-Shin; Leung, Yuk-Man


    Interaction between the selectivity filter and the adjacent pore helix of voltage-gated K(+) (Kv) channels controls pore stability during K(+) conduction. Kv channels, having their selectivity filter destabilized during depolarization, are said to undergo C-type inactivation. We examined the functionality of a residue at the pore helix of the Kv1.2 channel (V370), which reportedly affects C-type inactivation. A mutation into glycine (V370G) caused a shift in reversal potential from around -72 to -9 mV. The permeability ratios (P(Na)/P(K)) of the wild type and V370G mutant are 0.04 and 0.76, respectively. In the wild-type, P(Rb)/P(K), P(Cs)/P(K) and P(Li)/P(K) are 0.78, 0.10 and 0.05, respectively. Kv1.2 V370G channels had enhanced permeability to Rb(+) and Cs(+) (P(Rb)/P(K) and P(Cs)/P(K) are 1.63 and 1.18, respectively); however, Li(+) permeability was not significantly augmented (P(Li)/P(K) is 0.13). Therefore, in addition to its known effect on pore stability, V370 of Kv1.2 is also crucial in controlling ion selectivity.

  13. Mutagenesis and functional analysis of the pore-forming toxin HALT-1 from Hydra magnipapillata. (United States)

    Liew, Yvonne Jing Mei; Soh, Wai Tuck; Jiemy, William Febry; Hwang, Jung Shan


    Actinoporins are small 18.5 kDa pore-forming toxins. A family of six actinoporin genes has been identified in the genome of Hydra magnipapillata, and HALT-1 (Hydra actinoporin-like toxin-1) has been shown to have haemolytic activity. In this study, we have used site-directed mutagenesis to investigate the role of amino acids in the pore-forming N-terminal region and the conserved aromatic cluster required for cell membrane binding. A total of 10 mutants of HALT-1 were constructed and tested for their haemolytic and cytolytic activity on human erythrocytes and HeLa cells, respectively. Insertion of 1-4 negatively charged residues in the N-terminal region of HALT-1 strongly reduced haemolytic and cytolytic activity, suggesting that the length or charge of the N-terminal region is critical for pore-forming activity. Moreover, substitution of amino acids in the conserved aromatic cluster reduced haemolytic and cytolytic activity by more than 80%, suggesting that these aromatic amino acids are important for attachment to the lipid membrane as shown for other actinoporins. The results suggest that HALT-1 and other actinoporins share similar mechanisms of pore formation and that it is critical for HALT-1 to maintain an amphipathic helix at the N-terminus and an aromatic amino acid-rich segment at the site of membrane binding.

  14. Mutagenesis and Functional Analysis of the Pore-Forming Toxin HALT-1 from Hydra magnipapillata

    Directory of Open Access Journals (Sweden)

    Yvonne Jing Mei Liew


    Full Text Available Actinoporins are small 18.5 kDa pore-forming toxins. A family of six actinoporin genes has been identified in the genome of Hydra magnipapillata, and HALT-1 (Hydra actinoporin-like toxin-1 has been shown to have haemolytic activity. In this study, we have used site-directed mutagenesis to investigate the role of amino acids in the pore-forming N-terminal region and the conserved aromatic cluster required for cell membrane binding. A total of 10 mutants of HALT-1 were constructed and tested for their haemolytic and cytolytic activity on human erythrocytes and HeLa cells, respectively. Insertion of 1–4 negatively charged residues in the N-terminal region of HALT-1 strongly reduced haemolytic and cytolytic activity, suggesting that the length or charge of the N-terminal region is critical for pore-forming activity. Moreover, substitution of amino acids in the conserved aromatic cluster reduced haemolytic and cytolytic activity by more than 80%, suggesting that these aromatic amino acids are important for attachment to the lipid membrane as shown for other actinoporins. The results suggest that HALT-1 and other actinoporins share similar mechanisms of pore formation and that it is critical for HALT-1 to maintain an amphipathic helix at the N-terminus and an aromatic amino acid-rich segment at the site of membrane binding.

  15. Fluctuations of a fluid inside a pore

    NARCIS (Netherlands)

    Zvelindovsky, AV; Zatovsky, AV

    The correlation theory of the thermal hydrodynamic fluctuations of compressible viscous fluids within a spherical pore has been developed. The fluctuation motions ape described by the linearized Navier-Stokes and heat transfer equations, containing spontaneous viscous stresses and heat fluxes. The

  16. Induction of nano pore in Agrobacterial hemoglobin

    Directory of Open Access Journals (Sweden)

    Mojtaba Tousheh


    Full Text Available Introduction: A variety of oxygen-transport and -binding proteins exist in organisms including bacteria, protozoans, and fungi all have hemoglobin-like proteins. In addition to dealing with transport and sensing of oxygen, they may also deal with NO2, CO2, sulfide compounds, and even O2 scavenging in environments. Also they detoxified chlorinated materials like P450 enzymes and peroxidases and use as a detector of nitrate and hydrogen peroxide. Pore-forming bacterial globins are interested for filtration. Materials and methods: Although there are data for bacterial toxin as a filter, here we used Agrobacterial hem to induce nano pore in the heme structure using point mutation. Results: Investigations showed that three amino acids leucine 76, alanine 83 and histidine 80 are important for pore formation in Agrobacterium hemoglobin. A point mutation on leucine 76 to glycine, histidine 80 to asparagine and alanine 83 to lysine step by step led to create the nano pore 0.7- 0.8 nm in the globin. Discussion and conclusion: These mutations in bacterial hemoglobin increase the stability when mutation is with it’s at pH7. This mutation decreases the aliphatic index however increase the stability index.

  17. Particle diffusion in complex nanoscale pore networks

    DEFF Research Database (Denmark)

    Müter, Dirk; Sørensen, Henning Osholm; Bock, H.;


    We studied the diffusion of particles in the highly irregular pore networks of chalk, a very fine-grained rock, by combining three-dimensional X-ray imaging and dissipative particle dynamics (DPD) simulations. X-ray imaging data were collected at 25 nm voxel dimension for two chalk samples with v...

  18. Maximal pore size in UF membranes

    NARCIS (Netherlands)

    Arkhangelsky, E.; Duek, A.; Gitis, V.


    The ultrafiltration membrane rejection capability is most often characterized by molecular weight cutoff (MWCO). The value is found by rejection of organic solutes and the evaluation of particle retention requires a conversion of either MWCO to pore size or particle diameter to molecular weight. The

  19. Silicon Pore Optics development for ATHENA

    DEFF Research Database (Denmark)

    Collon, Maximilien J.; Vacanti, Giuseppe; Guenther, Ramses


    ) to meet the science requirements of large effective area (1-2 m(2) at a few keV) at a focal length of 12 m. To meet the high angular resolution (5 arc seconds) requirement the X-ray lens will also need to be very accurate. Silicon Pore Optics (SPO) technology has been invented to enable building...

  20. Observations of sausage modes in magnetic pores

    CERN Document Server

    Morton, R J; Jess, D B; Mathioudakis, M


    We present here evidence for the observation of the magneto-hydrodynamic (MHD) sausage modes in magnetic pores in the solar photosphere. Further evidence for the omnipresent nature of acoustic global modes is also found. The empirical decomposition method of wave analysis is used to identify the oscillations detected through a 4170 {\\AA} 'blue continuum' filter observed with the Rapid Oscillations in the Solar Atmosphere (ROSA) instrument. Out of phase, periodic behavior in pore size and intensity is used as an indicator of the presence of magneto-acoustic sausage oscillations. Multiple signatures of the magneto-acoustic sausage mode are found in a number of pores. The periods range from as short as 30 s up to 450 s. A number of the magneto-acoustic sausage mode oscillations found have periods of 3 and 5 minutes, similar to the acoustic global modes of the solar interior. It is proposed that these global oscillations could be the driver of the sausage type magneto-acoustic MHD wave modes in pores.

  1. Fluctuations of a fluid inside a pore

    NARCIS (Netherlands)

    Zvelindovsky, AV; Zatovsky, AV


    The correlation theory of the thermal hydrodynamic fluctuations of compressible viscous fluids within a spherical pore has been developed. The fluctuation motions ape described by the linearized Navier-Stokes and heat transfer equations, containing spontaneous viscous stresses and heat fluxes. The e

  2. Fluctuations of a fluid inside a pore

    NARCIS (Netherlands)

    Zvelindovsky, AV; Zatovsky, AV


    The correlation theory of the thermal hydrodynamic fluctuations of compressible viscous fluids within a spherical pore has been developed. The fluctuation motions ape described by the linearized Navier-Stokes and heat transfer equations, containing spontaneous viscous stresses and heat fluxes. The e

  3. Pore-Forming Toxins Trigger the Purge. (United States)

    Bonfini, Alessandro; Buchon, Nicolas


    The intestinal epithelium responds to pathogens by coordinating microbial elimination with tissue repair, both required to survive an infection. In this issue of Cell Host & Microbe, Lee et al. (2016) discover a rapid and evolutionarily conserved response to pore-forming toxins in the gut, involving cytoplasm ejection and enterocyte regrowth.

  4. Silicon Pore Optics development for ATHENA

    DEFF Research Database (Denmark)

    Collon, Maximilien J.; Vacanti, Giuseppe; Guenther, Ramses;


    ) to meet the science requirements of large effective area (1-2 m(2) at a few keV) at a focal length of 12 m. To meet the high angular resolution (5 arc seconds) requirement the X-ray lens will also need to be very accurate. Silicon Pore Optics (SPO) technology has been invented to enable building...

  5. Pore mutations in ammonium transporter AMT1 with increased electrogenic ammonium transport activity. (United States)

    Loqué, Dominique; Mora, Silvia I; Andrade, Susana L A; Pantoja, Omar; Frommer, Wolf B


    AMT/Mep ammonium transporters mediate high affinity ammonium/ammonia uptake in bacteria, fungi, and plants. The Arabidopsis AMT1 proteins mediate uptake of the ionic form of ammonium. AMT transport activity is controlled allosterically via a highly conserved cytosolic C terminus that interacts with neighboring subunits in a trimer. The C terminus is thus capable of modulating the conductivity of the pore. To gain insight into the underlying mechanism, pore mutants suppressing the inhibitory effect of mutations in the C-terminal trans-activation domain were characterized. AMT1;1 carrying the mutation Q57H in transmembrane helix I (TMH I) showed increased ammonium uptake but reduced capacity to take up methylammonium. To explore whether the transport mechanism was altered, the AMT1;1-Q57H mutant was expressed in Xenopus oocytes and analyzed electrophysiologically. AMT1;1-Q57H was characterized by increased ammonium-induced and reduced methylammonium-induced currents. AMT1;1-Q57H possesses a 100x lower affinity for ammonium (K(m)) and a 10-fold higher V(max) as compared with the wild type form. To test whether the trans-regulatory mechanism is conserved in archaeal homologs, AfAmt-2 from Archaeoglobus fulgidus was expressed in yeast. The transport function of AfAmt-2 also depends on trans-activation by the C terminus, and mutations in pore-residues corresponding to Q57H of AMT1;1 suppress nonfunctional AfAmt-2 mutants lacking the activating C terminus. Altogether, our data suggest that bacterial and plant AMTs use a conserved allosteric mechanism to control ammonium flux, potentially using a gating mechanism that limits flux to protect against ammonium toxicity.

  6. Facial skin pores: a multiethnic study (United States)

    Flament, Frederic; Francois, Ghislain; Qiu, Huixia; Ye, Chengda; Hanaya, Tomoo; Batisse, Dominique; Cointereau-Chardon, Suzy; Seixas, Mirela Donato Gianeti; Dal Belo, Susi Elaine; Bazin, Roland


    Skin pores (SP), as they are called by laymen, are common and benign features mostly located on the face (nose, cheeks, etc) that generate many aesthetic concerns or complaints. Despite the prevalence of skin pores, related literature is scarce. With the aim of describing the prevalence of skin pores and anatomic features among ethnic groups, a dermatoscopic instrument, using polarized lighting, coupled to a digital camera recorded the major features of skin pores (size, density, coverage) on the cheeks of 2,585 women in different countries and continents. A detection threshold of 250 μm, correlated to clinical scorings by experts, was input into a specific software to further allow for automatic counting of the SP density (N/cm2) and determination of their respective sizes in mm2. Integrating both criteria also led to establishing the relative part of the skin surface (as a percentage) that is actually covered by SP on cheeks. The results showed that the values of respective sizes, densities, and skin coverage: 1) were recorded in all studied subjects; 2) varied greatly with ethnicity; 3) plateaued with age in most cases; and 4) globally refected self-assessment by subjects, in particular those who self-declare having “enlarged pores” like Brazilian women. Inversely, Chinese women were clearly distinct from other ethnicities in having very low density and sizes. Analyzing the present results suggests that facial skin pore’s morphology as perceived by human eye less result from functional criteria of associated appendages such as sebaceous glands. To what extent skin pores may be viewed as additional criteria of a photo-altered skin is an issue to be further addressed. PMID:25733918

  7. Pore Structure of Cement Pastes Blended with Volcanic Rock

    Institute of Scientific and Technical Information of China (English)

    YU Lehua; ZHOU Shuangxi; LI Liling


    The pore parameters of cement pastes blended with volcanic rock at the curing age of 1, 28 and 90 d were de-termined by a mercury intrusion porosimetry. The pore structure of the pastes was characterized through the analysis of porosity, average pore diameter, the most probable pore aperture, pore size distribution, as well as total pore volume. For the improvement of mechanical property and durability of cement-based material, the correlation of the formed pore structure with hydration time and replacement level of volcanic rock for cement was revealed. The results indicate that volcanic rock can diminish porosity and reduce pore size in cement paste when curing time prolongs, which is particu-larly prominent with replacement level of less than 20% in late period. The more harmful pores (i.e., capillary pore) are gradually transformed into harmless pore (i.e., gel pores or micropore), even fully filled and disappeared when hydration products increase. The pore structure of the cement paste is thus refined. The beneficial effect of volcanic rock on the pore structure of cement paste could enhance the mechanical property and durability of cement-based material.

  8. Tuning the ion selectivity of two-pore channels

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Jiangtao; Zeng, Weizhong; Jiang, Youxing (UTSMC)


    Organellar two-pore channels (TPCs) contain two copies of a Shaker-like six-transmembrane (6-TM) domain in each subunit and are ubiquitously expressed in plants and animals. Interestingly, plant and animal TPCs share high sequence similarity in the filter region, yet exhibit drastically different ion selectivity. Plant TPC1 functions as a nonselective cation channel on the vacuole membrane, whereas mammalian TPC channels have been shown to be endo/lysosomal Na+-selective or Ca2+-release channels. In this study, we performed systematic characterization of the ion selectivity of TPC1 from Arabidopsis thaliana (AtTPC1) and compared its selectivity with the selectivity of human TPC2 (HsTPC2). We demonstrate that AtTPC1 is selective for Ca2+ over Na+, but nonselective among monovalent cations (Li+, Na+, and K+). Our results also confirm that HsTPC2 is a Na+-selective channel activated by phosphatidylinositol 3,5-bisphosphate. Guided by our recent structure of AtTPC1, we converted AtTPC1 to a Na+-selective channel by mimicking the selectivity filter of HsTPC2 and identified key residues in the TPC filters that differentiate the selectivity between AtTPC1 and HsTPC2. Furthermore, the structure of the Na+-selective AtTPC1 mutant elucidates the structural basis for Na+ selectivity in mammalian TPCs.

  9. Study of pores produced in underwater wet welding

    Institute of Scientific and Technical Information of China (English)

    Shen Xiaoqin; Liu Shiming


    This paper deals with the effect of water depth in the range of 10 m to 80 m upon the formation of pores produced during underwater wet welding. The results show that it is easy for the inner pores to occur owing to the particularity of the molten metal solidification that the outer pores begin to appear when the water depth increases to about 60 m, that the porosity increases and pore grows up as the water depth increases, and that pores are all hydrogen-cont aining ones through the examination of the variation of number of pores with the residual hydrogen and oxygen content in the weld metal.

  10. Mechanism for attenuated outward conductance induced by mutations in the cytoplasmic pore of Kir2.1 channels (United States)

    Chang, Hsueh-Kai; Iwamoto, Masayuki; Oiki, Shigetoshi; Shieh, Ru-Chi


    Outward currents through Kir2.1 channels regulate the electrical properties of excitable cells. These currents are subject to voltage-dependent attenuation by the binding of polyamines to high- and low-affinity sites, which leads to inward rectification, thereby controlling cell excitability. To examine the effects of positive charges at the low-affinity site in the cytoplasmic pore on inward rectification, we studied a mutant Kir channel (E224K/H226E) and measured single-channel currents and streaming potentials (Vstream), the latter provide the ratio of water to ions queued in a single-file permeation process in the selectivity filter. The water-ion coupling ratio was near one at a high K+ concentration ([K+]) for the wild-type channel and increased substantially as [K+] decreased. On the other hand, fewer ions occupied the selectivity filter in the mutant at all [K+]. A model for the Kir channel involving a K+ binding site in the wide pore was introduced. Model analyses revealed that the rate constants associated with the binding and release to and from the wide-pore K+ binding site was modified in the mutant. These effects lead to the reduced contribution of a conventional two-ion permeation mode to total conductance, especially at positive potentials, thereby inward rectification.

  11. Mineral dissolution kinetics at the pore scale

    Energy Technology Data Exchange (ETDEWEB)

    Li, L.; Steefel, C.I.; Yang, L.


    Mineral dissolution rates in the field have been reported to be orders of magnitude slower than those measured in the laboratory, an unresolved discrepancy that severely limits our ability to develop scientifically defensible predictive or even interpretive models for many geochemical processes in the earth and environmental sciences. One suggestion links this discrepancy to the role of physical and chemical heterogeneities typically found in subsurface soils and aquifers in producing scale-dependent rates where concentration gradients develop. In this paper, we examine the possibility that scale-dependent mineral dissolution rates can develop even at the single pore and fracture scale, the smallest and most fundamental building block of porous media. To do so, we develop two models to analyze mineral dissolution kinetics at the single pore scale: (1) a Poiseuille Flow model that applies laboratory-measured dissolution kinetics at the pore or fracture wall and couples this to a rigorous treatment of both advective and diffusive transport, and (2) a Well-Mixed Reactor model that assumes complete mixing within the pore, while maintaining the same reactive surface area, average flow rate, and geometry as the Poiseuille Flow model. For a fracture, a 1D Plug Flow Reactor model is considered in addition to quantify the effects of longitudinal versus transverse mixing. The comparison of averaged dissolution rates under various conditions of flow, pore size, and fracture length from the three models is used as a means to quantify the extent to which concentration gradients at the single pore and fracture scale can develop and render rates scale-dependent. Three important minerals that dissolve at widely different rates, calcite, plagioclase, and iron hydroxide, are considered. The modeling indicates that rate discrepancies arise primarily where concentration gradients develop due to comparable rates of reaction and advective transport, and incomplete mixing via molecular

  12. Evolution of Pore Size Distribution and Mean Pore Size in Lotus-type Porous Magnesium Fabricated with Gasar Process

    Institute of Scientific and Technical Information of China (English)

    Yuan LIU; Yanxiang LI; Huawei ZHANG; Jiang WAN


    The effect of gas pressures on the mean pore size, the porosity and the pore size distribution of lotus-type porous magnesium fabricated with Gasar process were investigated. The theoretical analysis and the experimental results all indicate that there exists an optimal ratio of the partial pressures of hydrogen pH2 to argon pAr for producing lotus-type structures with narrower pore size distribution and smaller pore size. The effect of solidification mode on the pore size distribution and pore size was also discussed.

  13. Nif- Hup- mutants of Rhizobium japonicum.


    Moshiri, F; Stults, L; Novak, P.; Maier, R J


    Two H2 uptake-negative (Hup-) Rhizobium japonicum mutants were obtained that also lacked symbiotic N2 fixation (acetylene reduction) activity. One of the mutants formed green nodules and was deficient in heme. Hydrogen oxidation activity in this mutant could be restored by the addition of heme plus ATP to crude extracts. Bacteroid extracts from the other mutant strain lacked hydrogenase activity and activity for both of the nitrogenase component proteins. Hup+ revertants of the mutant strains...

  14. Induction of nano pore in Agrobacterial hemoglobin


    Mojtaba Tousheh; Giti Emtiazi; Peyman Derikvand


    Introduction: A variety of oxygen-transport and -binding proteins exist in organisms including bacteria, protozoans, and fungi all have hemoglobin-like proteins. In addition to dealing with transport and sensing of oxygen, they may also deal with NO2, CO2, sulfide compounds, and even O2 scavenging in environments. Also they detoxified chlorinated materials like P450 enzymes and peroxidases and use as a detector of nitrate and hydrogen peroxide. Pore-forming bacterial globins are interested fo...

  15. Biomimetic collagen scaffolds with anisotropic pore architecture. (United States)

    Davidenko, N; Gibb, T; Schuster, C; Best, S M; Campbell, J J; Watson, C J; Cameron, R E


    Sponge-like matrices with a specific three-dimensional structural design resembling the actual extracellular matrix of a particular tissue show significant potential for the regeneration and repair of a broad range of damaged anisotropic tissues. The manipulation of the structure of collagen scaffolds using a freeze-drying technique was explored in this work as an intrinsically biocompatible way of tailoring the inner architecture of the scaffold. The research focused on the influence of temperature gradients, imposed during the phase of crystallisation of collagen suspensions, upon the degree of anisotropy in the microstructures of the scaffolds produced. Moulding technology was employed to achieve differences in heat transfer rates during the freezing processes. For this purpose various moulds with different configurations were developed with a view to producing uniaxial and multi-directional temperature gradients across the sample during this process. Scanning electron microscopy analysis of different cross-sections (longitudinal and horizontal) of scaffolds revealed that highly aligned matrices with axially directed pore architectures were obtained where single unidirectional temperature gradients were induced. Altering the freezing conditions by the introduction of multiple temperature gradients allowed collagen scaffolds to be produced with complex pore orientations, and anisotropy in pore size and alignment.

  16. Pore morphology study of silica aerogels

    Energy Technology Data Exchange (ETDEWEB)

    Hua, D.W.; Anderson, J.; Haereid, S.; Smith, D.M. [UNM/NSF Center for Micro-Engineered Ceramics, Albuquerque, NM (United States); Beaucage, G. [Sandia National Labs., Albuquerque, NM (United States)


    Silica aerogels have numerous properties which suggest applications such as ultra high efficiency thermal insulation. These properties relate directly to the aerogel`s pore size distribution. The micro and meso pore size ranges can be investigated by normal small angle x-ray scattering and possibly, nitrogen adsorption. However, the measurement of larger pores (> 250 {angstrom}) is more difficult. Due to their limited mechanical strength, mercury porosimetry and nitrogen condensation can disrupt the gel structure and electron microscopy provides only limited large scale structure information. The use of small angle light scattering techniques seems to have promise, the only hurdle is that aerogels exhibit significant multiple scattering. This can be avoided if one observes the gels in the wet stage since the structure of the aerogel should be very similar to the wet gel (as the result of supercritical drying). Thus, if one can match the refractive index, the morphology can be probed. The combination of certain alcoholic solvents fit this index matching criteria. Preliminary results for the gel network (micron range) and primary particle structure (manometer) are reported by using small angle light scattering and ultra-small angle x-ray scattering. The effects on structure over the length scale range of <1 nm to >5 {mu}m under different conditions (precursors, pH, etc.) are presented. The change in structure of an aerogel during isostatic compaction to 228 MPa (to simulate drying from wetting solvents) are also discussed.

  17. Characterization of antimicrobial activity against Listeria and cytotoxicity of native melittin and its mutant variants. (United States)

    Wu, Xi; Singh, Atul K; Wu, Xiaoyu; Lyu, Yuan; Bhunia, Arun K; Narsimhan, Ganesan


    Antimicrobial peptides (AMPs) are relatively short peptides that have the ability to penetrate the cell membrane, form pores leading to cell death. This study compares both antimicrobial activity and cytotoxicity of native melittin and its two mutants, namely, melittin I17K (GIGAVLKVLTTGLPALKSWIKRKRQQ) with a higher charge and lower hydrophobicity and mutant G1I (IIGAVLKVLTTGLPALISWIKRKRQQ) of higher hydrophobicity. The antimicrobial activity against different strains of Listeria was investigated by bioassay, viability studies, fluorescence and transmission electron microscopy. Cytotoxicity was examined by lactate dehydrogenase (LDH) assay on mammalian Caco-2 cells. The minimum inhibitory concentration of native, mutant I17K, mutant G1I against Listeria monocytogenes F4244 was 0.315±0.008, 0.814±0.006 and 0.494±0.037μg/ml respectively, whereas the minimum bactericidal concentration values were 3.263±0.0034, 7.412±0.017 and 5.366±0.019μg/ml respectively. Lag time for inactivation of L. monocytogenes F4244 was observed at concentrations below 0.20 and 0.78μg/ml for native and mutant melittin I17K respectively. The antimicrobial activity against L. monocytogenes F4244 was in the order native>G1I>I17K. Native melittin was cytotoxic to mammalian Caco-2 cells above concentration of 2μg/ml, whereas the two mutants exhibited negligible cytotoxicity up to a concentration of 8μg/ml. Pore formation in cell wall/membrane was observed by transmission electron microscopy. Molecular dynamics (MD) simulation of native and its mutants indicated that (i) surface native melittin and G1I exhibited higher tendency to penetrate a mimic of bacterial cell membrane and (ii) transmembrane native and I17K formed water channel in mimics of bacterial and mammalian cell membranes.


    Microbiologically-based procedures were used to describe biofouling phenomena on fine pore aeration devices and to determine whether biofilm characteristics could be related to diffuser process performance parameters. Fine pore diffusers were obtained from five municipal wastewa...

  19. Wild Accessions and Mutant Resources

    DEFF Research Database (Denmark)

    Kawaguchi, Masayoshi; Sandal, Niels Nørgaard


    Lotus japonicus, Lotus burttii, and Lotus filicaulis are species of Lotus genus that are utilized for molecular genetic analysis such as the construction of a linkage map and QTL analysis. Among them, a number of mutants have been isolated from two wild accessions: L. japonicus Gifu B-129...

  20. New ultrasonic technique for the study of the pore shape of track-etched pores in polymer films

    Energy Technology Data Exchange (ETDEWEB)

    Gomez Alvarez-Arenas, T.E., E-mail: tgomez@ia.cetef.csic.e [Instituto de Acustica, CSIC, Serrano 144, 28006 Madrid (Spain); Apel, P.Yu.; Orelovitch, O.L. [Flerov Lab. of Nuclear Reactions, JINR, Dubna (Russian Federation); Munoz, M. [Institute of Applied Physics, CSIC, Serrano 144, Madrid (Spain)


    A new technique for the study of the pore shape of track-etched pores in polymer films is presented. This technique is based on the use of air-coupled ultrasounds and phase and magnitude spectral analysis. Transmission of ultrasounds through these membranes is made up of two contributions: propagation through the solid part and propagation along the pore channels. A time-domain procedure to separate these to contributions is presented. Sensitivity of ultrasounds propagation in the pore channels to variations of pore shape is studied. Membranes with similar properties (gas flow rate values) but slight differences in the pore shape are studied. The proposed technique reveals to be sensitive to such differences; unlike other techniques, it is capable to provide information in a separate way about pore aperture at the surface and pore diameter inside the membrane, in addition the technique is non-destructive.

  1. Pore network and pore scale modeling of reactive transport in porous media (United States)

    Adler, P. M.; Vu, T. M.; Varloteaux, C.; Bekri, S.


    The study of the evolution of a porous medium where a reactive fluid flows is conditioned by the accurate determination of three macroscopic parameters governing the solute displacement, namely the solute velocity, dispersion and mean reaction rate. Of course, a possible application of such studies is CO2 sequestration. This presentation proposes to approach the determination of these parameters by two different ways and to compare them; both are on the pore scale. In the first one called PNM (for pore-network model), a pore-network is extracted from micro tomography images of a real porous medium. This network is composed of spherical pores joined by circular tubes; it is used to calculate transport macroscopic parameters and porosity-permeability evolution during the reactive transport flow as functions of dimensionless numbers representing the reaction and flow rate regimes. The flow is calculated by using Kirchhoff laws. Transport is determined in the asymptotic regime where the solute concentration undergoes an exponential evolution with time. In the second approach called PSM (for pore scale model), the pore-network model is used as a three dimensional medium which is discretized by the Level Set Method. The Stokes equations are solved in order to determine the local flow field and the corresponding permeability. The solute concentration is obtained by solving the local convection-diffusion equation in the 3D pore-network; numerical dispersion is reduced by a Flux Limiting Scheme. Two different geometries of porous media are addressed by both numerical codes. The first pore-network geometry is used to validate the PNM assumptions, whereas the second pore-network is defined for a better understanding of the dominant solute distribution. One of the main results obtained with the first pore-network is the dependence of the concentration profile on the Péclet number Pe in the pore-bodies. When this number increases, one has to switch from an assumption of

  2. Soil Pore Network Visualisation and Quantification using ImageJ

    DEFF Research Database (Denmark)

    Garbout, Amin; Pajor, Radoslaw; Otten, Wilfred

    Computed Tomography data. We used ImageJ to analyze images of pore geometries in soils generated by X-ray micro Computed Tomography. Soil samples were scanned at 30 μm resolution, and we produced replicated samples with different pore geometries by packing different sized soil aggregates at pre......-defined densities. First, scanned grayscale data of soil volumes were thresholded to separate solid and pore phases. Then, pore networks were extracted with the Skeletonize3D plug-in (Ignacio Arganda-Carreras), exploiting an ITK algorithm: binary thinning was used for finding the centerlines (”skeleton”) of pores...

  3. Energy conversion device with support member having pore channels (United States)

    Routkevitch, Dmitri [Longmont, CO; Wind, Rikard A [Johnstown, CO


    Energy devices such as energy conversion devices and energy storage devices and methods for the manufacture of such devices. The devices include a support member having an array of pore channels having a small average pore channel diameter and having a pore channel length. Material layers that may include energy conversion materials and conductive materials are coaxially disposed within the pore channels to form material rods having a relatively small cross-section and a relatively long length. By varying the structure of the materials in the pore channels, various energy devices can be fabricated, such as photovoltaic (PV) devices, radiation detectors, capacitors, batteries and the like.

  4. Single molecule atomic force microscopy of aerolysin pore complexes reveals unexpected star-shaped topography. (United States)

    He, Jianfeng; Wang, Jiabin; Hu, Jun; Sun, Jielin; Czajkowsky, Daniel Mark; Shao, Zhifeng


    Aerolysin is the paradigmatic member of a large family of toxins that convert from a water-soluble monomer/dimer into a membrane-spanning oligomeric pore. While there is x-ray crystallographic data of its water-soluble conformation, the most recent structural model of the membrane-inserted pore is based primarily on data of water-soluble tetradecamers of mutant protein, together with computational modeling ultimately performed in vacuum. Here we examine this pore model with atomic force microscopy (AFM) of membrane-associated wild-type complexes and all-atom molecular dynamics (MD) simulations in water. In striking contrast to a disc-shaped cap region predicted by the present model, the AFM images reveal a star-shaped complex, with a central ring surrounded by seven radial projections. Further, the MD simulations suggest that the locations of the receptor-binding (D1) domains in the present model are not correct. However, a modified model in which the D1 domains, rather than localized at fixed positions, adopt a wide range of configurations through fluctuations of an intervening linker is compatible with existing data. Thus our work not only demonstrates the importance of directly resolving such complexes in their native environment but also points to a dynamic receptor binding region, which may be critical for toxin assembly on the cell surface.

  5. Displacement of soil pore water by trichloroethylene (United States)

    Wershaw, R. L.; Aiken, G.R.; Imbrigiotta, T.E.; Goldberg, M.C.


    Dense nonaqueous phase liquids (DNAPLS) are important pollutants because of their widespread use as chemical and industrial solvents. An example of the pollution caused by the discharge of DNAPLs is found at the Picatinny Arsenal, New Jersey, where trichloroethylene (TCE) has been discharged directly into the unsaturated zone. This discharge has resulted in the formation of a plume of TCE-contaminated water in the aquifer downgradient of the discharge. A zone of dark-colored groundwater containing a high dissolved organic C content has been found near the point of discharge of the TCE. The colored-water plume extends from the point of discharge at least 30 m (100 feet) downgradient. Fulvic acids isolated from the colored-waters plume, from water from a background well that has not been affected by the discharge of chlorinated solvents, and from soil pore water collected in a lysimeter installed at an uncontaminated site upgradient of the study area have been compared. Nuclear magnetic resonance spectra of the fulvic acids from the colored waters and from the lysimeter are very similar, but are markedly different from the nuclear magnetic resonance spectrum of the fulvic acid from the background well. The three-dimensional fluorescence spectrum and the DOC fractionation profile of the colored groundwater and the soil pore water are very similar to each other, but quite different from those of the background water. It is proposed from these observations that this colored water is soil pore water that has been displaced by a separate DNAPL liquid phase downward to the saturated zone.

  6. Silicon pore optics developments and status

    DEFF Research Database (Denmark)

    Bavdaz, Marcos; Wille, Eric; Wallace, Kotska;


    Silicon Pore Optics (SPO) is a lightweight high performance X-ray optics technology being developed in Europe, driven by applications in observatory class high energy astrophysics missions. An example of such application is the former ESA science mission candidate ATHENA (Advanced Telescope...... of the SPO technology. The technology development programme has succeeded in maturing the SPO further and achieving important milestones, in each of the main activity streams: environmental compatibility, industrial production and optical performance. In order to accurately characterise the increasing...... performance of this innovative optical technology, the associated X-ray test facilities and beam-lines have been refined and upgraded. © 2012 SPIE....

  7. Viral Subversion of the Nuclear Pore Complex

    Directory of Open Access Journals (Sweden)

    Valerie Le Sage


    Full Text Available The nuclear pore complex (NPC acts as a selective barrier between the nucleus and the cytoplasm and is responsible for mediating communication by regulating the transport of RNA and proteins. Numerous viral pathogens have evolved different mechanisms to hijack the NPC in order to regulate trafficking of viral proteins, genomes and even capsids into and out of the nucleus thus promoting virus replication. The present review examines the different strategies and the specific nucleoporins utilized during viral infections as a means of promoting their life cycle and inhibiting host viral defenses.

  8. Analysis of quantitative pore features based on mathematical morphology

    Institute of Scientific and Technical Information of China (English)

    QI Heng-nian; CHEN Feng-nong; WANG Hang-jun


    Wood identification is a basic technique of wood science and industry. Pore features are among the most important identification features for hardwoods. We have used a method based on an analysis of quantitative pore feature, which differs from traditional qualitative methods. We applies mathematical morphology methods such as dilation and erosion, open and close transformation of wood cross-sections, image repairing, noise filtering and edge detection to segment the pores from their background. Then the mean square errors (MSE) of pores were computed to describe the distribution of pores. Our experiment shows that it is easy to classift the pore features into three basic types, just as in traditional qualitative methods, but with the use of MSE of pores. This quantitative method improves wood identification considerably.

  9. Quantifying similarity of pore-geometry in nanoporous materials (United States)

    Lee, Yongjin; Barthel, Senja D.; Dłotko, Paweł; Moosavi, S. Mohamad; Hess, Kathryn; Smit, Berend


    In most applications of nanoporous materials the pore structure is as important as the chemical composition as a determinant of performance. For example, one can alter performance in applications like carbon capture or methane storage by orders of magnitude by only modifying the pore structure. For these applications it is therefore important to identify the optimal pore geometry and use this information to find similar materials. However, the mathematical language and tools to identify materials with similar pore structures, but different composition, has been lacking. We develop a pore recognition approach to quantify similarity of pore structures and classify them using topological data analysis. This allows us to identify materials with similar pore geometries, and to screen for materials that are similar to given top-performing structures. Using methane storage as a case study, we also show that materials can be divided into topologically distinct classes requiring different optimization strategies.

  10. Nuclear protein import is reduced in cells expressing nuclear envelopathy-causing lamin A mutants

    Energy Technology Data Exchange (ETDEWEB)

    Busch, Albert; Kiel, Tilman; Heupel, Wolfgang-M. [University of Wuerzburg, Institute of Anatomy and Cell Biology, Koellikerstrasse 6, 97070 Wuerzburg (Germany); Wehnert, Manfred [Institute of Human Genetics, University of Greifswald, Greifswald (Germany); Huebner, Stefan, E-mail: [University of Wuerzburg, Institute of Anatomy and Cell Biology, Koellikerstrasse 6, 97070 Wuerzburg (Germany)


    Lamins, which form the nuclear lamina, not only constitute an important determinant of nuclear architecture, but additionally play essential roles in many nuclear functions. Mutations in A-type lamins cause a wide range of human genetic disorders (laminopathies). The importance of lamin A (LaA) in the spatial arrangement of nuclear pore complexes (NPCs) prompted us to study the role of LaA mutants in nuclear protein transport. Two mutants, causing prenatal skin disease restrictive dermopathy (RD) and the premature aging disease Hutchinson Gilford progeria syndrome, were used for expression in HeLa cells to investigate their impact on the subcellular localization of NPC-associated proteins and nuclear protein import. Furthermore, dynamics of the LaA mutants within the nuclear lamina were studied. We observed affected localization of NPC-associated proteins, diminished lamina dynamics for both LaA mutants and reduced nuclear import of representative cargo molecules. Intriguingly, both LaA mutants displayed similar effects on nuclear morphology and functions, despite their differences in disease severity. Reduced nuclear protein import was also seen in RD fibroblasts and impaired lamina dynamics for the nucleoporin Nup153. Our data thus represent the first study of a direct link between LaA mutant expression and reduced nuclear protein import.

  11. Inactivation of the pore-forming toxin Sticholysin I by peroxynitrite: protection by cys groups incorporated in the toxin. (United States)

    León, L; Lissi, E A; Celedón, G; Gonzalez, G; Pazos, F; Alvarez, C; Lanio, M E


    Sea anemones synthesize a variety of toxic peptides and proteins of biological interest. The Caribbean Sea anemone Stichodactyla helianthus, produces two pore-forming toxins, Sticholysin I (St I) and Stichloysin II (St II), with the ability to form oligomeric pores in cell and lipid bilayers characteristically lacking cysteine in their amino acid sequences. Recently, two mutants of a recombinant variant of Sticholysin I (rSt I) have been obtained with a Cys residue in functionally relevant regions for the pore-forming activity of the toxin: r St I F15C (in the amino terminal sequence) and r St I R52C (in the binding site). Aiming at characterizing the effects of oxidants in toxins devoid (r St I) or containing -SH moieties (r St I F15C and r St I R52C), we measured their hemolytic activity and pore forming capacity prior and after their incubation with peroxynitrite (ONOO(-)). At low ONOO(-)/Toxin ratios, nearly 0.8 Trp groups are modified by each added peroxynitrite molecule, and the toxin activity is reduced in ca. 20 %. On the other hand, in -SH bearing mutants only 0.5 Trp groups are modified by each peroxynitrite molecule and the toxin activity is only reduced in 10 %. The results indicated that Cys is the initial target of the oxidative damage and that Trp residues in Cys-containing toxins were less damaged than those in r St I. This relative protection of Trp groups correlates with a smaller loss of hemolytic activity and permeabilization ability in liposomes and emphasizes the relevance of Trp groups in the pore forming capacity of the toxins.

  12. Temperature induced pore fluid pressurization in geomaterials

    CERN Document Server

    Ghabezloo, Siavash


    The theoretical basis of the thermal response of the fluid-saturated porous materials in undrained condition is presented. It has been demonstrated that the thermal pressurization phenomenon is controlled by the discrepancy between the thermal expansion of the pore fluid and of the solid phase, the stress-dependency of the compressibility and the non-elastic volume changes of the porous material. For evaluation of the undrained thermo-poro-elastic properties of saturated porous materials in conventional triaxial cells, it is important to take into account the effect of the dead volume of the drainage system. A simple correction method is presented to correct the measured pore pressure change and also the measured volumetric strain during an undrained heating test. It is shown that the porosity of the tested material, its drained compressibility and the ratio of the volume of the drainage system to the one of the tested sample, are the key parameters which influence the most the error induced on the measuremen...

  13. Atmosphere above a large solar pore

    CERN Document Server

    Sobotka, M; Jurcak, J; Heinzel, P; Del Moro, D


    A large solar pore with a granular light bridge was observed on October 15, 2008 with the IBIS spectrometer at the Dunn Solar Telescope and a 69-min long time series of spectral scans in the lines Ca II 854.2 nm and Fe I 617.3 nm was obtained. The intensity and Doppler signals in the Ca II line were separated. This line samples the middle chromosphere in the core and the middle photosphere in the wings. Although no indication of a penumbra is seen in the photosphere, an extended filamentary structure, both in intensity and Doppler signals, is observed in the Ca II line core. An analysis of morphological and dynamical properties of the structure shows a close similarity to a superpenumbra of a sunspot with developed penumbra. A special attention is paid to the light bridge, which is the brightest feature in the pore seen in the Ca II line centre and shows an enhanced power of chromospheric oscillations at 3-5 mHz. Although the acoustic power flux in the light bridge is five times higher than in the "quiet" chr...

  14. Molecular dynamics simulation of flow in pores (United States)

    Blömer, Jan


    The gaseous flow in nano-scale pores is of wide interest for many today's industrial applications, e.g., in microelectronics, nano-mechanical devices (Knudsen compressor) and reaction and adsorption at porous surfaces. This can be seen from a variety of papers of recent RGD Symposia. Furthermore it is possible to separate gases by porous membranes. Although the fundamental problem of all these applications is same, namely the important role of the gas-surface interaction in such small structures, we will primarily concentrate on the separation of different gas species by porous membranes. These membranes are typically very robust (temperature, chemical resistance) because they are made from ceramics which offers new application fields. Porous flow can roughly be divided in several flow regimes by the Knudsen number: From viscous flow to Knudsen diffusion to surface diffusion and up to capillary condensation. A Molecular Dynamics (MD) model for the gas as well as the surface is formulated to investigate the interaction of gas atoms or molecules with internal degrees of freedom and the pore. The MD method seems to be well suited to study these phenomena because it can deal with the high density and the many-body-interactions, which occur during the multilayer adsorption and condensation at the surface, although it is clear that it is limited to a small physical space because of its high computational consumption.

  15. Functional significance of the highly conserved Glu(570) in the putative pore-forming helix 3 of the Bordetella pertussis haemolysin toxin. (United States)

    Kurehong, Chattip; Powthongchin, Busaba; Thamwiriyasati, Niramon; Angsuthanasombat, Chanan


    Adenylate cyclase-haemolysin toxin (CyaA) is a virulence factor secreted from the etiologic agent of whooping cough, Bordetella pertussis. Previously, the haemolysin or pore-forming domain (CyaA-PF) has been shown to cause cell lysis of sheep erythrocytes independently, and the predicted helix 3((570-593)) within the PF-hydrophobic stretch could be a pore-lining constituent. Here, a plausible involvement in haemolytic activity of polar or charged residues (Glu(570), Gln(574), Glu(581), Ser(584) and Ser(585)) lining the hydrophilic side of CyaA-PF helix 3 was investigated via single-alanine substitutions. All the 126-kDa mutant proteins over-expressed in Escherichia coli were verified for toxin acylation as the results are corresponding to the wild-type toxin. When haemolytic activity of E. coli lysates containing soluble mutant proteins was tested against sheep erythrocytes, the importance of Glu(570), which is highly conserved among the pore-forming RTX cytotoxin family, was revealed for pore formation, conceivably for a general pore-lining residue involved in ion conduction.

  16. Pore formation in lipid membrane I: Continuous reversible trajectory from intact bilayer through hydrophobic defect to transversal pore. (United States)

    Akimov, Sergey A; Volynsky, Pavel E; Galimzyanov, Timur R; Kuzmin, Peter I; Pavlov, Konstantin V; Batishchev, Oleg V


    Lipid membranes serve as effective barriers allowing cells to maintain internal composition differing from that of extracellular medium. Membrane permeation, both natural and artificial, can take place via appearance of transversal pores. The rearrangements of lipids leading to pore formation in the intact membrane are not yet understood in details. We applied continuum elasticity theory to obtain continuous trajectory of pore formation and closure, and analyzed molecular dynamics trajectories of pre-formed pore reseal. We hypothesized that a transversal pore is preceded by a hydrophobic defect: intermediate structure spanning through the membrane, the side walls of which are partially aligned by lipid tails. This prediction was confirmed by our molecular dynamics simulations. Conversion of the hydrophobic defect into the hydrophilic pore required surmounting some energy barrier. A metastable state was found for the hydrophilic pore at the radius of a few nanometers. The dependence of the energy on radius was approximately quadratic for hydrophobic defect and small hydrophilic pore, while for large radii it depended on the radius linearly. The pore energy related to its perimeter, line tension, thus depends of the pore radius. Calculated values of the line tension for large pores were in quantitative agreement with available experimental data.

  17. Bacterial mutants for enhanced succinate production

    NARCIS (Netherlands)

    Baart, G.J.E.; Beauprez, J.J.R.; Foulquie, M.M.R.; Heijnen, J.J.; Maertens, J.


    The present invention relates to a method for obtaining enhanced metabolite production in micro-organisms, and to mutants and/or transformants obtained with said method. More particularly, it relates to bacterial mutants and/or transformants for enhanced succinate production, especially mutants and/

  18. Problem-Solving Test: Tryptophan Operon Mutants (United States)

    Szeberenyi, Jozsef


    This paper presents a problem-solving test that deals with the regulation of the "trp" operon of "Escherichia coli." Two mutants of this operon are described: in mutant A, the operator region of the operon carries a point mutation so that it is unable to carry out its function; mutant B expresses a "trp" repressor protein unable to bind…

  19. An emerging pore-making strategy: confined swelling-induced pore generation in block copolymer materials. (United States)

    Wang, Yong; Li, Fengbin


    Block copolymers (BCPs) composed of two or more thermodynamically incompatible homopolymers self-assemble into periodic microdomains. Exposing self-assembled BCPs with solvents selective to one block causes a swelling of the domains composed of this block. Strong swelling in the confinement imposed by the matrix of the other glassy block leads to well-defined porous structures via morphology reconstruction. This confined swelling-induced pore-making process has emerged recently as a new strategy to produce porous materials due to synergic advantages that include extreme simplicity, high pore regularity, involvement of no chemical reactions, no weight loss, reversibility of the pore forming process, etc. The mechanism, kinetics, morphology, and governing parameters of the confined swelling-induced pore-making process in BCP thin films are discussed, and the main applications of nanoporous thin films in the fields of template synthesis, surface patterning, and guidance for the areal arrangements of nanomaterials and biomolecules are summarized. Recent, promising results of extending this mechanism to produce BCP nanofibers or nanotubes and bulk materials with well-defined porosity, which makes this strategy also attractive to researchers outside the nanocommunity, are also presented. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Computational modeling of electrokinetic transport in random networks of micro-pores and nano-pores (United States)

    Alizadeh, Shima; Mani, Ali


    A reduced order model has been developed to study the nonlinear electrokinetic behaviors emerging in the transport of ionic species through micro-scale and nano-scale porous media. In this approach a porous structure is modeled as a network of long and thin pores. By assuming transport equilibrium in the thin dimensions for each pore, a 1D transport equation is developed in the longitudinal direction covering a wide range of conditions including extreme limits of thick and thin electric double layers. This 1D model includes transport via diffusion, electromigration and wide range of advection mechanisms including pressure driven flow, electroosmosis, and diffusion osmosis. The area-averaged equations governing the axial transport from different pores are coupled at the pore intersections using the proper conservation laws. Moreover, an asymptotic treatment has been included in order to remove singularities in the limit of small concentration. The proposed method provides an efficient framework for insightful simulations of porous electrokinetic systems with applications in water desalination and energy storage. PhD student in Mechanical Engineering, Stanford University. She received her Master's degree in Mechanical Engineering from Stanford at 2013. Her research interests include CFD, high performance computing, and optimization.

  1. Structure and mechanism of peptide-induced membrane pores (United States)

    Qian, Shuo

    This thesis reports the studies of the structure and mechanism of peptide-induced membrane pores by antimicrobial peptide alamethicin and by a peptide named Baxalpha5, which is derived from Bax protein. Alamethicin is one of best known antimicrobial peptides, which are ubiquitous throughout the biological world. Bax-alpha5 peptide is the pore-forming domain of apoptosis regulator protein Bax, which activates pore formation on outer mitochondrial membrane to release cytochrome c to initiate programmed cell death. Both peptides as well as many other pore-forming peptides, induce pores in membrane, however the structure and mechanism of the pore formation were unknown. By utilizing grazing angle x-ray diffraction, I was able to reconstruct the electron density profile of the membrane pores induced by both peptides. The fully hydrated multiple bilayers of peptide-lipid mixture on solid substrate were prepared in the condition that pores were present, as established previously by neutron in-plane scattering and oriented circular dichroism. At dehydrated conditions, the inter bilayer distance of the sample shortened and the interactions between bilayers caused the membrane pores to become long-ranged correlated and formed a periodically ordered lattice of rhombohedral symmetry, so that x-ray diffraction can be applied. To help solving the phase problem of diffraction, a brominated lipid was used and multi-wavelength anomalous diffraction was performed below the bromine K-edge. The reconstructed electron density profiles unambiguously revealed that the alamethicin-induced membrane pore is of barrel-stave type, while the Bax-alpha5 induced pore is of lipidic toroidal (wormhole) type. The underlying mechanism of pore formation was resolved by observing the time-dependent process of pore formation in vesicles exposed to Bax-alpha5 solutions, as well as the membrane thinning experiment. This demonstrated that Bax-alpha5 exhibited the same sigmoidal concentration dependence as

  2. A Stereolithography Pore-Throat Model (United States)

    Crandall, D.; Ahmadi, G.; Ferer, M.; Smith, D. H.


    A new experimental, heterogeneous pore-throat model has been designed and fabricated using stereolithography (SL). In SL production, a laser cures a thin layer of photo-sensitive resin on the surface of a vat of liquid resin; a moveable platform then submerges the cured layer and a new layer is cured on top of the previous one, creating a physical model from a computer generated model. This layered fabrication of a computer generated model has enabled the production of an experimental porous medium with improved fluid resistance properties, as compared to previously studied, constant-height etched cells. A uniform distribution of throat widths was randomly placed throughout the pore-throat matrix and the throat height of each throat was assigned to increase the range of viscous and capillary resistances within the physical model. This variation in both throat height and width generated a porous medium with fairly low porosity (43%), permeability (~400 D), and wide range of geometric resistance properties. Experimental, two-phase immiscible drainage studies in the porous flowcell were performed. Analysis of the captured images was performed with open-source image processing software. These analysis techniques utilized the capability of both ImageJ and the Gnu Image Manipulation Program to be customized with ancillary codes. This enabled batch procedures to be created that converted the original grey-scale bitmaps to binary data sets, which were then analyzed with in-house codes. The fractal dimension, Df, (measured with box-counting) and percent saturation of these experiments were calculated and shown to compare favorably to fractal predictions and previous flowcell studies. Additionally, using the computer generated pore-throat geometry, a computational fluid dynamics model of two- phase flow through the porous medium was created. This model was created using FLUENT code and the Volume of Fluid method. The percent saturation of the less-viscous invading fluid

  3. Nif- Hup- mutants of Rhizobium japonicum. (United States)

    Moshiri, F; Stults, L; Novak, P; Maier, R J


    Two H2 uptake-negative (Hup-) Rhizobium japonicum mutants were obtained that also lacked symbiotic N2 fixation (acetylene reduction) activity. One of the mutants formed green nodules and was deficient in heme. Hydrogen oxidation activity in this mutant could be restored by the addition of heme plus ATP to crude extracts. Bacteroid extracts from the other mutant strain lacked hydrogenase activity and activity for both of the nitrogenase component proteins. Hup+ revertants of the mutant strains regained both H2 uptake ability and nitrogenase activity. Images PMID:6874648

  4. Pore size matters for potassium channel conductance (United States)

    Moldenhauer, Hans; Pincuntureo, Matías


    Ion channels are membrane proteins that mediate efficient ion transport across the hydrophobic core of cell membranes, an unlikely process in their absence. K+ channels discriminate K+ over cations with similar radii with extraordinary selectivity and display a wide diversity of ion transport rates, covering differences of two orders of magnitude in unitary conductance. The pore domains of large- and small-conductance K+ channels share a general architectural design comprising a conserved narrow selectivity filter, which forms intimate interactions with permeant ions, flanked by two wider vestibules toward the internal and external openings. In large-conductance K+ channels, the inner vestibule is wide, whereas in small-conductance channels it is narrow. Here we raise the idea that the physical dimensions of the hydrophobic internal vestibule limit ion transport in K+ channels, accounting for their diversity in unitary conductance. PMID:27619418

  5. Silicon pore optics development for ATHENA (United States)

    Collon, Maximilien J.; Vacanti, Giuseppe; Günther, Ramses; Yanson, Alex; Barrière, Nicolas; Landgraf, Boris; Vervest, Mark; Chatbi, Abdelhakim; Beijersbergen, Marco W.; Bavdaz, Marcos; Wille, Eric; Haneveld, Jeroen; Koelewijn, Arenda; Leenstra, Anne; Wijnperle, Maurice; van Baren, Coen; Müller, Peter; Krumrey, Michael; Burwitz, Vadim; Pareschi, Giovanni; Conconi, Paolo; Christensen, Finn E.


    The ATHENA mission, a European large (L) class X-ray observatory to be launched in 2028, will essentially consist of an X-ray lens and two focal plane instruments. The lens, based on a Wolter-I type double reflection grazing incidence angle design, will be very large (~ 3 m in diameter) to meet the science requirements of large effective area (1-2 m2 at a few keV) at a focal length of 12 m. To meet the high angular resolution (5 arc seconds) requirement the X-ray lens will also need to be very accurate. Silicon Pore Optics (SPO) technology has been invented to enable building such a lens and thus enabling the ATHENA mission. We will report in this paper on the latest status of the development, including details of X-ray test campaigns.

  6. Silicon pore optics for the ATHENA telescope (United States)

    Collon, Maximilien J.; Vacanti, Giuseppe; Günther, Ramses; Yanson, Alex; Barriere, Nicolas; Landgraf, Boris; Vervest, Mark; Chatbi, Abdelhakim; van der Hoeven, Roy; Beijersbergen, Marco W.; Bavdaz, Marcos; Wille, Eric; Shortt, Brian; Haneveld, Jeroen; Koelewijn, Arenda; van Baren, Coen; Eigenraam, Alexander; Müller, Peter; Krumrey, Michael; Burwitz, Vadim; Pareschi, Giovanni; Conconi, Paolo; Massahi, Sonny; Christensen, Finn E.; Valsecchi, Giuseppe


    Silicon Pore Optics is a high-energy optics technology, invented to enable the next generation of high-resolution, large area X-ray telescopes such as the ATHENA observatory, a European large (L) class mission with a launch date of 2028. The technology development is carried out by a consortium of industrial and academic partners and focuses on building an optics with a focal length of 12 m that shall achieve an angular resolution better than 5". So far we have built optics with a focal length of 50 m and 20 m. This paper presents details of the work carried out to build silicon stacks for a 12 m optics and to integrate them into mirror modules. It will also present results of x-ray tests taking place at PTB's XPBF with synchrotron radiation and the PANTER test facility.

  7. Distributed Pore Chemistry in Porous Organic Polymers (United States)

    Koontz, Steven L. (Inventor)


    A method for making a biocompatible polymer article using a uniform atomic oxygen treatment is disclosed. The substrate may be subsequently optionally grated with a compatibilizing compound. Compatibilizing compounds may include proteins, phosphorylcholine groups, platelet adhesion preventing polymers, albumin adhesion promoters, and the like. The compatibilized substrate may also have a living cell layer adhered thereto. The atomic oxygen is preferably produced by a flowing afterglow microwave discharge. wherein the substrate resides in a sidearm out of the plasma. Also, methods for culturing cells for various purposes using the various membranes are disclosed as well. Also disclosed are porous organic polymers having a distributed pore chemistry (DPC) comprising hydrophilic and hydrophobic regions. and a method for making the DPC by exposing the polymer to atomic oxygen wherein the rate of hydrophilization is greater than the rate of mass loss.

  8. Evaluating transport in irregular pore networks

    CERN Document Server

    Klimenko, Dimitri A; Klimenko, Alexander Y; 10.1103/PhysRevE.86.011112


    A general approach for investigating transport phenomena in porous media is presented. This approach has the capacity to represent various kinds of irregularity in porous media without the need for excessive detail or computational effort. The overall method combines a generalized Effective Medium Approximation (EMA) with a macroscopic continuum model in order to derive a transport equation with explicit analytical expressions for the transport coefficients. The proposed form of the EMA is an anisotropic and heterogeneous extension of Kirkpatrick's EMA [Rev. Mod. Phys. 45, 574 (1973)] which allows the overall model to account for microscopic alterations in connectivity (with the locations of the pores and the orientation and length of the throat) as well as macroscopic variations in transport properties. A comparison to numerical results for randomly generated networks with different properties is given, indicating the potential for this methodology to handle cases that would pose significant difficulties to ...

  9. Identification of a Long Rice Spikelet Mutant

    Institute of Scientific and Technical Information of China (English)

    WU Xian-jun; WANG Bin; HAN Zan-ping; XIE Zhao-hui; MOU Chun-hong; WANG Xu-dong


    A spontaneously occurring rice (Oryza sativa L. ) mutant, characterized by homeotic conversion in glumes and stamens, was found in the progeny of a cross. The mutant showed long glumes and glumaceous lodicules and morphological transformation of stamens into pistils. Mutant florets consisted of 1 to 3 completely developed pistils, some pistilloid stamens with filaments, but tipped by bulged tissue and 0 to 3 stigmas. It seens that the mutant phenotype of the homeotic conversions in glumes and stamens is similar to that of the B loss-of-function mutants in Arabidopsis and Antirrhinum. The mutant is controlled by a single recessive gene as a segregation ratio of 3:1 (wild type to mutant plants) was observed in the F2 generation.

  10. Factors Determining the Pore Shape in Polycarbonate Track Membranes

    CERN Document Server

    Apel, P Yu; Orelovich, O L; Akimenko, S N; Sartowska, B; Dmitriev, S N


    The process of pore formation in ion-irradiated polycarbonate films on treatment with alkali solutions in the presence of a surfactant is studied. It is found that the pore shape depends on both the structure of the initial films and the peculiarities of the interaction of the surfactant with the polymer surface and the transport of the surfactant into tracks. Due to heterogeneity of the films the cross-section of a track pore channel changes along its length. The presence of the surfactant results in a further effect. Surfactant molecules adsorb on the polymer surface at the pore entries and reduce the etch rate which leads to formation of cigar-like pore channels. The use of surfactant as a component of chemical etchant enables one to control the pore shape in track membranes thus optimizing their retention and permeation characteristics.

  11. Experimental study on pore water pressure dissipation of mucky soil

    Institute of Scientific and Technical Information of China (English)

    Xianwei ZHANG; Changming WANG; Junxia LI; Bin WANG


    Pore water pressure has an important influence on mechanical properties of soil. The authors studied the characteristics of pore water pressure dissipating of mucky soil under consolidated-drained condition by using refitted triaxial instrument and analyzed the variation of pore pressure coefficient with consolidation pressure. The results show that the dissipating of pore water pressure behaves in different ways depends on different styles of loading. What is more, the pore water pressure coefficient of mucky soil is less than 1. As the compactness of soil increases and moisture content reduces, the value of B reduces. There is a staggered dissipating in the process of consolidation, in which it is a mutate point when U/P is 80%. It is helpful to establish the pore water pressure model and study the strength-deformation of soil in process of consolidation.

  12. Vapor intrusion in soils with multimodal pore-size distribution


    Alfaro Soto Miguel; Hung Kiang Chang


    The Johnson and Ettinger [1] model and its extensions are at this time the most widely used algorithms for estimating subsurface vapor intrusion into buildings (API [2]). The functions which describe capillary pressure curves are utilized in quantitative analyses, although these are applicable for porous media with a unimodal or lognormal pore-size distribution. However, unaltered soils may have a heterogeneous pore distribution and consequently a multimodal pore-size distribution [3], which ...

  13. Fusion Pore Diameter Regulation by Cations Modulating Local Membrane Anisotropy

    Directory of Open Access Journals (Sweden)

    Doron Kabaso


    Full Text Available The fusion pore is an aqueous channel that is formed upon the fusion of the vesicle membrane with the plasma membrane. Once the pore is open, it may close again (transient fusion or widen completely (full fusion to permit vesicle cargo discharge. While repetitive transient fusion pore openings of the vesicle with the plasma membrane have been observed in the absence of stimulation, their frequency can be further increased using a cAMP-increasing agent that drives the opening of nonspecific cation channels. Our model hypothesis is that the openings and closings of the fusion pore are driven by changes in the local concentration of cations in the connected vesicle. The proposed mechanism of fusion pore dynamics is considered as follows: when the fusion pore is closed or is extremely narrow, the accumulation of cations in the vesicle (increased cation concentration likely leads to lipid demixing at the fusion pore. This process may affect local membrane anisotropy, which reduces the spontaneous curvature and thus leads to the opening of the fusion pore. Based on the theory of membrane elasticity, we used a continuum model to explain the rhythmic opening and closing of the fusion pore.

  14. Enzyme screening with synthetic multifunctional pores: Focus on biopolymers (United States)

    Sordé, Nathalie; Das, Gopal; Matile, Stefan


    This report demonstrates that a single set of identical synthetic multifunctional pores can detect the activity of many different enzymes. Enzymes catalyzing either synthesis or degradation of DNA (exonuclease III or polymerase I), RNA (RNase A), polysaccharides (heparinase I, hyaluronidase, and galactosyltransferase), and proteins (papain, ficin, elastase, subtilisin, and pronase) are selected to exemplify this key characteristic of synthetic multifunctional pore sensors. Because anionic, cationic, and neutral substrates can gain access to the interior of complementarily functionalized pores, such pores can be the basis for very user-friendly screening of a broad range of enzymes. PMID:14530413

  15. Pore-size-distribution of cationic polyacrylamide hydrogels. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    Kremer, M.; Prausnitz, J.M.


    The pore size distribution of a AAm/MAPTAC (acrylamide copolymerized with (3-methacrylamidopropyl)trimethylammonium chloride) hydrogel was investigated using Kuga`s mixed-solute-exclusion method, taking into account the wall effect. A Brownian-motion model is also used. Results show the feasibility of determining pore-size distribution of porous materials using the mixed-solute-exclusion method in conjunction with solution of the Fredholm equation; good agreement was obtained with experiment, even for bimodal pore structures. However, different pore size distributions were calculated for the two different probe-solutes (Dextran and poly(ethylene glycol/oxide)). Future work is outlined. 32 figs, 25 refs.

  16. Pore-size-distribution of cationic polyacrylamide hydrogels

    Energy Technology Data Exchange (ETDEWEB)

    Kremer, M.; Prausnitz, J.M.


    The pore size distribution of a AAm/MAPTAC (acrylamide copolymerized with (3-methacrylamidopropyl)trimethylammonium chloride) hydrogel was investigated using Kuga's mixed-solute-exclusion method, taking into account the wall effect. A Brownian-motion model is also used. Results show the feasibility of determining pore-size distribution of porous materials using the mixed-solute-exclusion method in conjunction with solution of the Fredholm equation; good agreement was obtained with experiment, even for bimodal pore structures. However, different pore size distributions were calculated for the two different probe-solutes (Dextran and poly(ethylene glycol/oxide)). Future work is outlined. 32 figs, 25 refs.

  17. Ultrafast laser fabrication of submicrometer pores in borosilicate glass. (United States)

    An, Ran; Uram, Jeffrey D; Yusko, Erik C; Ke, Kevin; Mayer, Michael; Hunt, Alan J


    We demonstrate rapid fabrication of submicrometer-diameter pores in borosilicate glass using femtosecond laser machining and subsequent wet-etch techniques. This approach allows direct and repeatable fabrication of high-quality pores with diameters of 400-800 nm. Such small pores coupled with the desirable electrical and chemical properties of glass enable sensitive resistive-pulse analysis to determine the size and concentration of macromolecules and nanoparticles. Plasma-enhanced chemical vapor deposition allows further reduction of pore diameters to below 300 nm.

  18. Relationship between elastic moduli and pore radius in clay aggregates

    DEFF Research Database (Denmark)

    Fabricius, Ida Lykke


    Available experimental data on elastic velocities of clay-air mixtures and clay-brine mixtures as a function of porosity are re-interpreted. Pore radius as calculated from porosity and specific surface measured by BET seems to be the factor controlling stiffness of these un-cemented sediments....... For each of the two pore fluids: air or brine smectitic clay and kaolinitic clay seem to have similar power law relationships between a given elastic modulus and pore radius. These results indicate that pore radius and thus permeability of shale in the depth interval of mechanical compaction may...

  19. Tension-induced vesicle fusion: pathways and pore dynamics

    DEFF Research Database (Denmark)

    Shillcock, Julian C.


    and eventually opens a pore to complete the fusion process. In pathway II, at higher tension, a stalk is formed during the fusion process that is then transformed by transmembrane pore formation into a fusion pore. Whereas the latter pathway II resembles stalk pathways as observed in other simulation studies...... fusion time on membrane tension implies that the fusion process is completed by overcoming two energy barriers with scales of 13kBT and 11kBT. The fusion pore radius as a function of time has also been extracted from the simulations, and provides a quantitative measure of the fusion dynamics which...

  20. Charges in the cytoplasmic pore control intrinsic inward rectification and single-channel properties in Kir1.1 and Kir2.1 channels. (United States)

    Chang, Hsueh-Kai; Yeh, Shih-Hao; Shieh, Ru-Chi


    An E224G mutation of the Kir2.1 channel generates intrinsic inward rectification and single-channel fluctuations in the absence of intracellular blockers. In this study, we showed that positively charged residues H226, R228 and R260, near site 224, regulated the intrinsic inward rectification and single-channel properties of the E224G mutant. By carrying out systematic mutations, we found that the charge effect on the intrinsic inward rectification and single-channel conductance is consistent with a long-range electrostatic mechanism. A Kir1.1 channel where the site equivalent to E224 in the Kir2.1 channel is a glycine residue does not show inward rectification or single-channel fluctuations. The G223K and N259R mutations of the Kir1.1 channel induced intrinsic inward rectification and reduced the single-channel conductance but did not generate large open-channel fluctuations. Substituting the cytoplasmic pore of the E224G mutant into the Kir1.1 channel induced open-channel fluctuations and intrinsic inward rectification. The single-channel conductance of the E224G mutant showed inward rectification. Also, a voltage-dependent gating mechanism decreased open probability during depolarization and contributed to the intrinsic inward rectification in the E224G mutant. In addition to an electrostatic effect, a close interaction of K(+) with channel pore may be required for generating open-channel fluctuations in the E224G mutant.

  1. Dominant negative phenotype of Bacillus thuringiensis Cry1Ab, Cry11Aa and Cry4Ba mutants suggest hetero-oligomer formation among different Cry toxins.

    NARCIS (Netherlands)

    Carmona, D.; Rodriguez-Almazan, C.; Munoz-Garay, C.; Portugal, L.; Perez, C.; Maagd, de R.A.; Bakker, P.; Soberon, M.; Bravo, A.


    Background - Bacillus thuringiensis Cry toxins are used worldwide in the control of different insect pests important in agriculture or in human health. The Cry proteins are pore-forming toxins that affect the midgut cell of target insects. It was shown that non-toxic Cry1Ab helix a-4 mutants had a d

  2. Rock Physics of Reservoir Rocks with Varying Pore Water Saturation and Pore Water Salinity

    DEFF Research Database (Denmark)

    Katika, Konstantina

    be performed on specific geological structures and why it is sometimes successful; has yet to be established. The presence of both oil and water in the pore space, several different ions present in the injected water that contact the pore walls, possible changes in the fluid wetting the surface of the grains......Advanced waterflooding (injection of water with selective ions in reservoirs) is a method of enhanced oil recovery (EOR) that has attracted the interest of oil and gas companies that exploit the Danish oil and gas reservoirs. This method has been applied successfully in oil reservoirs...... and in the Smart Water project performed in a laboratory scale in order to evaluate the EOR processes in selected core plugs. A major step towards this evaluation is to identify the composition of the injected water that leads to increased oil recovery in reservoirs and to define changes in the petrophysical...

  3. Entropy of Shortest Distance (ESD as Pore Detector and Pore-Shape Classifier

    Directory of Open Access Journals (Sweden)

    Klaudia Oleschko


    Full Text Available The entropy of shortest distance (ESD between geographic elements (“elliptical intrusions”, “lineaments”, “points” on a map, or between "vugs", "fractures" and "pores" in the macro- or microscopic images of triple porosity naturally fractured vuggy carbonates provides a powerful new tool for the digital processing, analysis, classification and space/time distribution prognostic of mineral resources as well as the void space in carbonates, and in other rocks. The procedure is applicable at all scales, from outcrop photos, FMI, UBI, USI (geophysical imaging techniques to micrographs, as we shall illustrate through some examples. Out of the possible applications of the ESD concept, we discuss in details the sliding window entropy filtering for nonlinear pore boundary enhancement, and propose this procedure as unbiased thresholding technique.

  4. Pore morphologies of root induced biopores from single pore to network scale investigated by XRCT (United States)

    Peth, Stephan; Wittig, Marlen C.; Uteau Puschmann, Daniel; Pagenkemper, Sebastian; Haas, Christoph; Holthusen, Dörthe; Horn, Rainer


    Biopores are assumed to be an important factor for nutrient acquisition by providing biologically highly active soil-root interfaces to re-colonizing roots and controlling oxygen and water flows at the pedon scale and within the rhizosphere through the formation of branching channel networks which potentially enhance microbial turnover processes. Characteristic differences in pore morphologies are to be expected depending on the genesis of biopores which, for example, can be earthworm-induced or root-induced or subsequently modified by one of the two. Our understanding of biophysical interactions between plants and soil can be significantly improved by quantifying 3D biopore architectures across scales ranging from single biopores to pedon scale pore networks and linking pore morphologies to microscale measurements of transport processes (e.g. oxygen diffusion). While a few studies in the past have investigated biopore networks on a larger scale yet little is known on the micro-morphology of root-induces biopores and their associated rhizosphere. Also little data is available on lateral transport of oxygen through the rhizosphere which will strongly influence microbial turnover processes and consequently control the release and uptake of nutrients. This paper highlights results gathered within a research unit on nutrient acquisition from the subsoil. Here we focus on X-ray microtomography (XRCT) studies ranging from large soil columns (70 cm length and 20 cm diameter) to individual biopores and its surrounding rhizosphere. Samples were collected from sites with different preceding crops (fescue, chicory, alfalfa) and various cropping durations (1-3 years). We will present an approach for quantitative image analysis combined with micro-sensor measurements of oxygen diffusion and spatial gradients of O2 partial pressures to relate pore structure with transport functions. Implications of various biopore architectures for the accessibility of nutrient resources in

  5. Effects of supersaturation on pore shape in solid (United States)

    Wei, P. S.; Hsiao, S. Y.


    The shape of a pore resulting from a bubble entrapped by a solidification front with different supersaturation ratios is predicted in this work. Supersaturation ratio, representing the ratio between solute concentration and saturation solute concentration, determines nucleation of a bubble and development of the pore shape in the early stage. Pore formation and its shape in solid influence contemporary issues of biology, engineering, foods, geophysics and climate change, etc. This work extends and combines previous models accounting for realistic mass and momentum transport, and physico-chemical equilibrium of solute gas across the bubble cap to self-consistently determine shape of the bubble cap beyond the solidification front and the pore shape in solid. The study also deal with that pore formation can be resulted from three different mechanisms, depending on the directions and magnitude of solute gas transport across the bubble cap. Case 1 is subject to solute transport from the pore across the cap into the surrounding liquid in the early stage. Cases 2a and 2b indicate opposite direction of solute transport. In contrast to Case 2b, the effect of solute transport on solute gas pressure in the pore in Case 2a is stronger than that of pore volume expansionin the last stage. The results find that an increase in supersaturation ratio decreases pore radius and time for bubble entrapment in Case 1. The bubble cannot be entrapped in Case 2. The predicted pore shape in solid agrees with experimental data. Understanding, prediction and control of the growth of the pore shape have therefore been obtained.

  6. Reversible Self-Actuated Thermo-Responsive Pore Membrane (United States)

    Park, Younggeun; Gutierrez, Maria Paz; Lee, Luke P.


    Smart membranes, which can selectively control the transfer of light, air, humidity and temperature, are important to achieve indoor climate regulation. Even though reversible self-actuation of smart membranes is desirable in large-scale, reversible self-regulation remains challenging. Specifically, reversible 100% opening/closing of pore actuation showing accurate responsiveness, reproducibility and structural flexibility, including uniform structure assembly, is currently very difficult. Here, we report a reversible, thermo-responsive self-activated pore membrane that achieves opening and closing of pores. The reversible, self-actuated thermo-responsive pore membrane was fabricated with hybrid materials of poly (N-isopropylacrylamide), (PNIPAM) within polytetrafluoroethylene (PTFE) to form a multi-dimensional pore array. Using Multiphysics simulation of heat transfer and structural mechanics based on finite element analysis, we demonstrated that pore opening and closing dynamics can be self-activated at environmentally relevant temperatures. Temperature cycle characterizations of the pore structure revealed 100% opening ratio at T = 40 °C and 0% opening ratio at T = 20 °C. The flexibility of the membrane showed an accurate temperature-responsive function at a maximum bending angle of 45°. Addressing the importance of self-regulation, this reversible self-actuated thermo-responsive pore membrane will advance the development of future large-scale smart membranes needed for sustainable indoor climate control.

  7. Hydrodemetallization of residue (Part 3). Influence of catalyst pore size

    Energy Technology Data Exchange (ETDEWEB)

    Hinuma, Yutaka; Kushiyama, Akira; Kobayashi, Satoru; Aizawa, Reiji; Inoue, Keiichi; Shimizu, Yoshikazu (National Research Inst. for Pollution and Resources, Tsukuba (Japan))


    Influence of catalyst pore size on hydrodemetallization of residues was studied, Starting oils were Khafji atmospheric residue and Morichal crude and two catalysts of alumina which have 93A and 160A mean pore diameters respectively and Mo was supported, were used. The rate at which asphaltene(A) was decomposed to maltene(M), related large to the demetallization of whole heavy oil and larger the pore diameter of catalyst, higher the demetallizing activity. But in the demetallization from A and M, the influence of pore size was different and vanadium(V) removal from A was preferable for larger pore size but was reverse from M. These were caused by the difference of diffusion in their pores. Ni was more difficult to remove than V. Catalyst of small pore size was preferable for simultaneous desulfurization, because sulfur compounds were composed of smaller molecular weight distribution. The decomposition of A started preferentially from part having large number of peripheral carbon in condensed aromatic ring or f{sub a} (aromaticity) and the catalyst of large pore size affected large on the structural change of A. 12 ref., 7 figs., 5 tabs.

  8. The Pore Structure of Direct Methanol Fuel Cell Electrodes

    DEFF Research Database (Denmark)

    Lund, Peter Brilner


    The pore structure and morphology of direct methanol fuel cell electrodes are characterized using mercury intrusion porosimetry and scanning electron microscopy. It is found that the pore size distributions of printed primer and catalyst layers are largely dictated by the powders used to make the...

  9. Reversible Self-Actuated Thermo-Responsive Pore Membrane. (United States)

    Park, Younggeun; Gutierrez, Maria Paz; Lee, Luke P


    Smart membranes, which can selectively control the transfer of light, air, humidity and temperature, are important to achieve indoor climate regulation. Even though reversible self-actuation of smart membranes is desirable in large-scale, reversible self-regulation remains challenging. Specifically, reversible 100% opening/closing of pore actuation showing accurate responsiveness, reproducibility and structural flexibility, including uniform structure assembly, is currently very difficult. Here, we report a reversible, thermo-responsive self-activated pore membrane that achieves opening and closing of pores. The reversible, self-actuated thermo-responsive pore membrane was fabricated with hybrid materials of poly (N-isopropylacrylamide), (PNIPAM) within polytetrafluoroethylene (PTFE) to form a multi-dimensional pore array. Using Multiphysics simulation of heat transfer and structural mechanics based on finite element analysis, we demonstrated that pore opening and closing dynamics can be self-activated at environmentally relevant temperatures. Temperature cycle characterizations of the pore structure revealed 100% opening ratio at T = 40 °C and 0% opening ratio at T = 20 °C. The flexibility of the membrane showed an accurate temperature-responsive function at a maximum bending angle of 45°. Addressing the importance of self-regulation, this reversible self-actuated thermo-responsive pore membrane will advance the development of future large-scale smart membranes needed for sustainable indoor climate control.


    DEFF Research Database (Denmark)


    The present invention offers a method for separating dry matter from a medium. A separation chamber is at least partly defined by a plurality of rollers (2,7) and is capable of being pressure regulated. At least one of the rollers is a pore roller (7) having a surface with pores allowing permeabi...

  11. Bacteriocins : mechanism of membrane insertion and pore formation

    NARCIS (Netherlands)

    Moll, Gert N.; Konings, Wil N.; Driessen, Arnold J.M.


    Lactic acid bacteria produce several types of pore forming peptides. Class I bacteriocins are lantibiotics that contain (methyl)lanthionine residues that may form intramolecular thioether rings. These peptides generally have a broad spectrum of activity and form unstable pores. Class II bacteriocins

  12. Pore size distribution in tablets measured with a morphological sieve

    NARCIS (Netherlands)

    Wu, Yu San; van Vliet, Lucas J.; Frijlink, Henderik W.; van der Voort Maarschalk, Kees


    Porosity and pore structure are important characteristics of tablets, since they influence mechanical strength and many other proper-ties. This paper proposes an alternative method for the characterization of pore structure based on image analysis of SEM micrographs. SEM images were made of sodium c

  13. Pore structure and growth kinetics in carbon materials

    Energy Technology Data Exchange (ETDEWEB)

    Bose, S.


    Pore structure of glassy carbon (GC) and pyrolytic graphite (PG) have been investigated. GC is one of the most impervious of solids finding applications in prosthetic devices and fuel cells while PG is used extensively in the aerospace industry. One third of the microstructure of GC consists of closed pores inaccessible to fluids. The microstructure of this material has been characterized using x-ray diffraction (XRD) and high resolution electron microscopy. Small angle x-ray scattering (SAXS) has been used to measure the angstrom sized pores and to follow the evolution of pore surface area as a function of heat treatment temperature (HTT) and heat treatment time (HTt) at constant temperature. From these measurements an analysis of the surface area kinetics was made to find out if rate processes are involved and to locate graphitization occurring at pore surfaces. PG on the other hand has been found to have larger sized pores that comprise five percent of its volume. In addition to being closed these pores are oriented. Some pore models are proposed for PG and the existing scattering theory from oriented ellipsoids is modified to include the proposed shapes.

  14. Bacteriocins : mechanism of membrane insertion and pore formation

    NARCIS (Netherlands)

    Moll, G.N.; Konings, W.N; Driessen, A.J.M.


    Lactic acid bacteria produce several types of pore forming peptides. Class I bacteriocins are lantibiotics that contain (methyl)lanthionine residues that may form intramolecular thioether rings. These peptides generally have a broad spectrum of activity and form unstable pores. Class II bacteriocins

  15. Letter to the editor: Diffusion under pore saturation conditions

    NARCIS (Netherlands)

    Krishna, R.; van Baten, J.M.


    In the article "Diffusion Under Pore Saturaton Conditions", Lettat et al. (1) presents a model to describe mixture diffusion in MFI zeolite under conditions of pore saturation. As a motivation for developing their model they remark ‘‘the classical adsorbed-phase diffusion models based on the Maxwell

  16. Porous Boron Nitride with Tunable Pore Size. (United States)

    Dai, Jun; Wu, Xiaojun; Yang, Jinlong; Zeng, Xiao Cheng


    On the basis of a global structural search and first-principles calculations, we predict two types of porous boron-nitride (BN) networks that can be built up with zigzag BN nanoribbons (BNNRs). The BNNRs are either directly connected with puckered B (N) atoms at the edge (type I) or connected with sp(3)-bonded BN chains (type II). Besides mechanical stability, these materials are predicted to be thermally stable at 1000 K. The porous BN materials entail large surface areas, ranging from 2800 to 4800 m(2)/g. In particular, type-II BN material with relatively large pores is highly favorable for hydrogen storage because the computed hydrogen adsorption energy (-0.18 eV) is very close to the optimal adsorption energy (-0.15 eV) suggested for reversible hydrogen storage at room temperature. Moreover, the type-II materials are semiconductors with width-dependent direct bandgaps, rendering the type-II BN materials promising not only for hydrogen storage but also for optoelectronic and photonic applications.

  17. Freezing of charged colloids in slit pores. (United States)

    Grandner, Stefan; Klapp, Sabine H L


    Using Monte Carlo simulations in the grand canonical and isobaric ensembles we investigate freezing phenomena in a charged colloidal suspension confined to narrow slit pores. Our model involves only the macroions which interact via a Derjaguin-Landau-Verwey-Overbeek (DLVO) potential supplemented by a soft-sphere potential. We focus on DLVO parameters typical for moderately charged silica particles (with charges Z approximately 35) in solvents of low ionic strengths. The corresponding DLVO interactions are too weak to drive a (bulk) freezing transition. Nevertheless, for sufficiently small surface separations L(z) the confined systems display not only layering but also significant in-plane crystalline order at chemical potentials where the bulk system is a globally stable fluid (capillary freezing). At confinement conditions related to two-layer systems the observed in-plane structures are consistent with those detected in ground state calculations for perfect Yukawa bilayers [R. Messina and H. Lowen, Phys. Rev. Lett. 91, 146101 (2003)]. Here we additionally observe (at fixed L(z)) a compression-induced first-order phase transition from a two-layer to a three-layer system with different in-plane structure, in agreement with previous findings for pure hard spheres.

  18. Silicon pore optics developments and status (United States)

    Bavdaz, Marcos; Wille, Eric; Wallace, Kotska; Shortt, Brian; Collon, Maximilien; Ackermann, Marcelo; Olde Riekerink, Mark; Haneveld, Jeroen; van Baren, Coen; Erhard, Markus; Christensen, Finn; Krumrey, Michael; Burwitz, Vadim


    Silicon Pore Optics (SPO) is a lightweight high performance X-ray optics technology being developed in Europe, driven by applications in observatory class high energy astrophysics missions. An example of such application is the former ESA science mission candidate ATHENA (Advanced Telescope for High Energy Astrophysics), which uses the SPO technology for its two telescopes, in order to provide an effective area exceeding 1 m2 at 1 keV, and 0.5 m2 at 6 keV, featuring an angular resolution of 10” or better [1 to 24]. This paper reports on the development activities led by ESA, and the status of the SPO technology. The technology development programme has succeeded in maturing the SPO further and achieving important milestones, in each of the main activity streams: environmental compatibility, industrial production and optical performance. In order to accurately characterise the increasing performance of this innovative optical technology, the associated X-ray test facilities and beam-lines have been refined and upgraded.

  19. Evaluating transport in irregular pore networks (United States)

    Klimenko, Dimitri A.; Hooman, Kamel; Klimenko, Alexander Y.


    A general approach for investigating transport phenomena in porous media is presented. This approach has the capacity to represent various kinds of irregularity in porous media without the need for excessive detail or computational effort. The overall method combines a generalized effective medium approximation (EMA) with a macroscopic continuum model in order to derive a transport equation with explicit analytical expressions for the transport coefficients. The proposed form of the EMA is an anisotropic and heterogeneous extension of Kirkpatrick's EMA [Rev. Mod. Phys.RMPHAT0034-686110.1103/RevModPhys.45.574 45, 574 (1973)] which allows the overall model to account for microscopic alterations in connectivity (with the locations of the pores and the orientation and length of the throat) as well as macroscopic variations in transport properties. A comparison to numerical results for randomly generated networks with different properties is given, indicating the potential for this methodology to handle cases that would pose significant difficulties to many other analytical models.

  20. Biochemical and histological characterization of tomato mutants

    Directory of Open Access Journals (Sweden)

    Carolina C. Monteiro


    Full Text Available Biochemical responses inherent to antioxidant systems as well morphological and anatomical properties of photomorphogenic, hormonal and developmental tomato mutants were investigated. Compared to the non-mutant Micro-Tom (MT, we observed that the malondialdehyde (MDA content was enhanced in the diageotropica (dgt and lutescent (l mutants, whilst the highest levels of hydrogen peroxide (H2O2 were observed in high pigment 1 (hp1 and aurea (au mutants. The analyses of antioxidant enzymes revealed that all mutants exhibited reduced catalase (CAT activity when compared to MT. Guaiacol peroxidase (GPOX was enhanced in both sitiens (sit and notabilis (not mutants, whereas in not mutant there was an increase in ascorbate peroxidase (APX. Based on PAGE analysis, the activities of glutathione reductase (GR isoforms III, IV, V and VI were increased in l leaves, while the activity of superoxide dismutase (SOD isoform III was reduced in leaves of sit, epi, Never ripe (Nr and green flesh (gf mutants. Microscopic analyses revealed that hp1 and au showed an increase in leaf intercellular spaces, whereas sit exhibited a decrease. The au and hp1 mutants also exhibited a decreased in the number of leaf trichomes. The characterization of these mutants is essential for their future use in plant development and ecophysiology studies, such as abiotic and biotic stresses on the oxidative metabolism.Neste trabalho, analisamos as respostas bioquímicas inerentes ao sistema antioxidante, assim como propriedades morfológicas e anatômicas de mutantes fotomorfogenéticos e hormonais de tomateiro. Comparados ao não mutante Micro-Tom (MT, observamos que o conteúdo de malondialdeído (MDA aumentou nos mutantes diageotropica (dgt e lutescent (l, enquanto os maiores níveis de H2O2 foram encontrados nos mutantes high pigment 1 (hp1 e aurea (au. Análises de enzimas antioxidantes mostraram que todos os mutantes reduziram a atividade de catalase (CAT quando comparado a MT. A

  1. Influence of pore structure on compressive strength of cement mortar. (United States)

    Zhao, Haitao; Xiao, Qi; Huang, Donghui; Zhang, Shiping


    This paper describes an experimental investigation into the pore structure of cement mortar using mercury porosimeter. Ordinary Portland cement, manufactured sand, and natural sand were used. The porosity of the manufactured sand mortar is higher than that of natural sand at the same mix proportion; on the contrary, the probable pore size and threshold radius of manufactured sand mortar are finer. Besides, the probable pore size and threshold radius increased with increasing water to cement ratio and sand to cement ratio. In addition, the existing models of pore size distribution of cement-based materials have been reviewed and compared with test results in this paper. Finally, the extended Bhattacharjee model was built to examine the relationship between compressive strength and pore structure.

  2. Vapor intrusion in soils with multimodal pore-size distribution

    Directory of Open Access Journals (Sweden)

    Alfaro Soto Miguel


    Full Text Available The Johnson and Ettinger [1] model and its extensions are at this time the most widely used algorithms for estimating subsurface vapor intrusion into buildings (API [2]. The functions which describe capillary pressure curves are utilized in quantitative analyses, although these are applicable for porous media with a unimodal or lognormal pore-size distribution. However, unaltered soils may have a heterogeneous pore distribution and consequently a multimodal pore-size distribution [3], which may be the result of specific granulometry or the formation of secondary porosity related to genetic processes. The present paper was designed to present the application of the Vapor Intrusion Model (SVI_Model to unsaturated soils with multimodal pore-size distribution. Simulations with data from the literature show that the use of a multimodal model in soils with such pore distribution characteristics could provide more reliable results for indoor air concentration, rather than conventional models.

  3. Role of pore-forming toxins in neonatal sepsis. (United States)

    Sonnen, Andreas F-P; Henneke, Philipp


    Protein toxins are important virulence factors contributing to neonatal sepsis. The major pathogens of neonatal sepsis, group B Streptococci, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus, secrete toxins of different molecular nature, which are key for defining the disease. Amongst these toxins are pore-forming exotoxins that are expressed as soluble monomers prior to engagement of the target cell membrane with subsequent formation of an aqueous membrane pore. Membrane pore formation is not only a means for immediate lysis of the targeted cell but also a general mechanism that contributes to penetration of epithelial barriers and evasion of the immune system, thus creating survival niches for the pathogens. Pore-forming toxins, however, can also contribute to the induction of inflammation and hence to the manifestation of sepsis. Clearly, pore-forming toxins are not the sole factors that drive sepsis progression, but they often act in concert with other bacterial effectors, especially in the initial stages of neonatal sepsis manifestation.

  4. Surface pore tension and adsorption characteristics of polluted sediment

    Institute of Scientific and Technical Information of China (English)


    Most natural sediment particles have numerous pores and a complex surface texture which facilitates their adsorption of contaminants. Particle surface structure,therefore,is an important instrumental factor in the transport of contaminants,especially in water environments. This paper reports on the results of adsorption-desorption experiments to analyze polluted sediment surface pore tension characteristics performed on samples from the bottom of Guanting Reservoir. In our analysis,the Frenkel-Halsey-Hill(FHH) equation is applied to calculate the fractal dimensions of particles to quantify the surface roughness and pore tension characteristics. The results show that the surface fractal dimensions of sediment particle surfaces normally measure from 2.6 to 2.85. The volume of pores smaller than 10 nm changes significantly after being contaminated with pollutants and the fractal dimension decreases because the pores adsorb the contaminants.

  5. Pore capillary pressure and saturation of methane hydrate bearing sediments

    Institute of Scientific and Technical Information of China (English)

    SUN Shicai; LIU Changling; YE Yuguang; LIU Yufeng


    To better understand the relationship between the pore capillary pressure and hydrate saturation in sedi-ments, a new method was proposed. First, the phase equilibria of methane hydrate in fine-grained silica sands were measured. As to the equilibrium data, the pore capillary pressure and saturation of methane hydrate were calculated. The results showed that the phase equilibria of methane hydrates in fine-grained silica sands changed due to the depressed activity of pore water caused by the surface group and negatively charged characteristic of silica particles as well as the capillary pressure in small pores together. The capil-lary pressure increased with the increase of methane hydrate saturation due to the decrease of the available pore space. However, the capillary-saturation relationship could not yet be described quantitatively because of the stochastic habit of hydrate growth.

  6. Measurements of pore-scale flow through apertures.

    Energy Technology Data Exchange (ETDEWEB)

    Chojnicki, Kirsten


    Pore-scale aperture effects on flow in pore networks was studied in the laboratory to provide a parameterization for use in transport models. Four cases were considered: regular and irregular pillar/pore alignment with and without an aperture. The velocity field of each case was measured and simulated, providing quantitatively comparable results. Two aperture effect parameterizations were considered: permeability and transmission. Permeability values varied by an order of magnitude between the cases with and without apertures. However, transmission did not correlate with permeability. Despite having much greater permeability the regular aperture case permitted less transmission than the regular case. Moreover, both irregular cases had greater transmission than the regular cases, a difference not supported by the permeabilities. Overall, these findings suggest that pore-scale aperture effects on flow though a pore-network may not be adequately captured by properties such as permeability for applications that are interested in determining particle transport volume and timing.

  7. Biophysics, Pathophysiology and Pharmacology of Ion Channel Gating Pores

    Directory of Open Access Journals (Sweden)

    Adrien eMoreau


    Full Text Available Voltage sensor domain (VSDs are a feature of voltage gated ion channel (VGICs and voltage sensitive proteins. They are composed of four transmembrane (TM segments (S1 to S4. Currents leaking through VSDs are called omega or gating pore currents.Gating pores are caused by mutations of the highly conserved positively charged amino acids in the S4 segment that disrupt interactions between the S4 segment and the gating charge transfer center (GCTC. The GCTC separates the intracellular and extracellular water crevices. The disruption of S4–GCTC interactions allows these crevices to communicate and create a fast activating and non-inactivating alternative cation-selective permeation pathway of low conductance, or a gating pore.Gating pore currents have recently been shown to cause periodic paralysis phenotypes. There is also increasing evidence that gating pores are linked to several other familial diseases. For example, gating pores in Nav1.5 and Kv7.2 channels may underlie mixed arrhythmias associated with dilated cardiomyopathy (DCM phenotypes and peripheral nerve hyperexcitability (PNH respectively. There is little evidence for the existence of gating pore blockers. Moreover, it is known that a number of toxins bind to the VSD of a specific domain of Na+ channels. These toxins may thus modulate gating pore currents. This focus on the VSD motif opens up a new area of research centered on developing molecules to treat a number of cell excitability disorders such as epilepsy, cardiac arrhythmias, and pain.The purpose of the present review is to summarize existing knowledge of the pathophysiology, biophysics, and pharmacology of gating pore currents and to serve as a guide for future studies aimed at improving our understanding of gating pores and their pathophysiological roles.

  8. X-ray CT analysis of pore structure in sand (United States)

    Mukunoki, Toshifumi; Miyata, Yoshihisa; Mikami, Kazuaki; Shiota, Erika


    The development of microfocused X-ray computed tomography (CT) devices enables digital imaging analysis at the pore scale. The applications of these devices are diverse in soil mechanics, geotechnical and geoenvironmental engineering, petroleum engineering, and agricultural engineering. In particular, the imaging of the pore space in porous media has contributed to numerical simulations for single-phase and multiphase flows or contaminant transport through the pore structure as three-dimensional image data. These obtained results are affected by the pore diameter; therefore, it is necessary to verify the image preprocessing for the image analysis and to validate the pore diameters obtained from the CT image data. Moreover, it is meaningful to produce the physical parameters in a representative element volume (REV) and significant to define the dimension of the REV. This paper describes the underlying method of image processing and analysis and discusses the physical properties of Toyoura sand for the verification of the image analysis based on the definition of the REV. On the basis of the obtained verification results, a pore-diameter analysis can be conducted and validated by a comparison with the experimental work and image analysis. The pore diameter is deduced from Young-Laplace's law and a water retention test for the drainage process. The results from previous study and perforated-pore diameter originally proposed in this study, called the voxel-percolation method (VPM), are compared in this paper. In addition, the limitations of the REV, the definition of the pore diameter, and the effectiveness of the VPM for an assessment of the pore diameter are discussed.

  9. An undulation theory for condensation in open end slit pores: critical hysteresis temperature & critical hysteresis pore size. (United States)

    Fan, Chunyan; Zeng, Yonghong; Do, D D; Nicholson, D


    A new theory of condensation in an open end slit pore, based on the concept of temperature dependent undulation, at the interface separating the adsorbed phase and the gas-like region, is presented. The theory, describes, for the first time, the microscopic origin of the critical hysteresis temperature and the critical hysteresis pore size, properties which are not accessible to any classical theories.

  10. Investigating Hydrophilic Pores in Model Lipid Bilayers Using Molecular Simulations: Correlating Bilayer Properties with Pore-Formation Thermodynamics. (United States)

    Hu, Yuan; Sinha, Sudipta Kumar; Patel, Sandeep


    Cell-penetrating and antimicrobial peptides show a remarkable ability to translocate across physiological membranes. Along with factors such as electric-potential-induced perturbations of membrane structure and surface tension effects, experiments invoke porelike membrane configurations during the solute transfer process into vesicles and cells. The initiation and formation of pores are associated with a nontrivial free-energy cost, thus necessitating a consideration of the factors associated with pore formation and the attendant free energies. Because of experimental and modeling challenges related to the long time scales of the translocation process, we use umbrella sampling molecular dynamics simulations with a lipid-density-based order parameter to investigate membrane-pore-formation free energy employing Martini coarse-grained models. We investigate structure and thermodynamic features of the pore in 18 lipids spanning a range of headgroups, charge states, acyl chain lengths, and saturation. We probe the dependence of pore-formation barriers on the area per lipid, lipid bilayer thickness, and membrane bending rigidities in three different lipid classes. The pore-formation free energy in pure bilayers and peptide translocating scenarios are significantly coupled with bilayer thickness. Thicker bilayers require more reversible work to create pores. The pore-formation free energy is higher in peptide-lipid systems than in peptide-free lipid systems due to penalties to maintain the solvation of charged hydrophilic solutes within the membrane environment.

  11. Asymmetric pore occupancy in crystal structure of OmpF porin from Salmonella typhi. (United States)

    Balasubramaniam, D; Arockiasamy, Arulandu; Kumar, P D; Sharma, Amit; Krishnaswamy, S


    OmpF is a major general diffusion porin of Salmonella typhi, a Gram-negative bacterium, which is an obligatory human pathogen causing typhoid. The structure of S. typhi Ty21a OmpF (PDB Id: 3NSG) determined at 2.8 Å resolution by X-ray crystallography shows a 16-stranded β-barrel with three β-barrel monomers associated to form a trimer. The packing observed in S. typhi Ty21a rfOmpF crystals has not been observed earlier in other porin structures. The variations seen in the loop regions provide a starting point for using the S. typhi OmpF for structure-based multi-valent vaccine design. Along one side of the S. typhi Ty21a OmpF pore there exists a staircase arrangement of basic residues (20R, 60R, 62K, 65R, 77R, 130R and 16K), which also contribute, to the electrostatic potential in the pore. This structure suggests the presence of asymmetric electrostatics in the porin oligomer. Moreover, antibiotic translocation, permeability and reduced uptake in the case of mutants can be understood based on the structure paving the way for designing new antibiotics.

  12. Enhanced Retention of Chemotactic Bacteria in a Pore Network with Residual NAPL Contamination. (United States)

    Wang, Xiaopu; Lanning, Larry M; Ford, Roseanne M


    Nonaqueous-phase liquid (NAPL) contaminants are difficult to eliminate from natural aquifers due, in part, to the heterogeneous structure of the soil. Chemotaxis enhances the mixing of bacteria with contaminant sources in low-permeability regions, which may not be readily accessible by advection and dispersion alone. A microfluidic device was designed to mimic heterogeneous features of a contaminated groundwater aquifer. NAPL droplets (toluene) were trapped within a fine pore network, and bacteria were injected through a highly conductive adjacent macrochannel. Chemotactic bacteria (Pseudomonas putida F1) exhibited greater accumulation near the pore network at 0.5 m/day than both the nonchemotactic control and the chemotactic bacteria at a higher groundwater velocity of 5 m/day. Chemotactic bacteria accumulated in the vicinity of NAPL droplets, and the accumulation was 15% greater than a nonchemotactic mutant. Indirect evidence showed that chemotactic bacteria were retained within the contaminated low-permeability region longer than nonchemotactic bacteria at 0.25 m/day. This retention was diminished at 5 m/day. Numerical solutions of the bacterial-transport equations were consistent with the experimental results. Because toluene is degraded by P. putida F1, the accumulation of chemotactic bacteria around NAPL sources is expected to increase contaminant consumption and improve the efficiency of bioremediation.

  13. Muscle channelopathies: does the predicted channel gating pore offer new treatment insights for hypokalaemic periodic paralysis? (United States)

    Matthews, E; Hanna, M G


    Hypokalaemic periodic paralysis (hypoPP) is the archetypal skeletal muscle channelopathy caused by dysfunction of one of two sarcolemmal ion channels, either the sodium channel Nav1.4 or the calcium channel Cav1.1. Clinically, hypoPP is characterised by episodes of often severe flaccid muscle paralysis, in which the muscle fibre membrane becomes electrically inexcitable, and which may be precipitated by low serum potassium levels. Initial functional characterisation of hypoPP mutations failed to adequately explain the pathomechanism of the disease. Recently, as more pathogenic mutations involving loss of positive charge have been identified in the S4 segments of either channel, the hypothesis that an abnormal gating pore current may be important has emerged. Such an aberrant gating pore current has been identified in mutant Nav1.4 channels and has prompted potentially significant advances in this area. The carbonic anhydrase inhibitor acetazolamide has been used as a treatment for hypokalaemic periodic paralysis for over 40 years but its precise therapeutic mechanism of action is unclear. In this review we summarise the recent advances in the understanding of the molecular pathophysiology of hypoPP and consider how these may relate to the reported beneficial effects of acetazolamide. We also consider potential areas for future therapeutic development. PMID:20123788

  14. Pore Scale Dynamics of Microemulsion Formation. (United States)

    Unsal, Evren; Broens, Marc; Armstrong, Ryan T


    Experiments in various porous media have shown that multiple parameters come into play when an oleic phase is displaced by an aqueous solution of surfactant. In general, the displacement efficiency is improved when the fluids become quasi-miscible. Understanding the phase behavior oil/water/surfactant systems is important because microemulsion has the ability to generate ultralow interfacial tension (formation and the resulting properties under equilibrium conditions. However, the majority of applications where microemulsion is present also involve flow, which has received relatively less attention. It is commonly assumed that the characteristics of an oil/water/surfactant system under flowing conditions are identical to the one under equilibrium conditions. Here, we show that this is not necessarily the case. We studied the equilibrium phase behavior of a model system consisting of n-decane and an aqueous solution of olefin sulfonate surfactant, which has practical applications for enhanced oil recovery. The salt content of the aqueous solution was varied to provide a range of different microemulsion compositions and oil-water interfacial tensions. We then performed microfluidic flow experiments to study the dynamic in situ formation of microemulsion by coinjecting bulk fluids of n-decane and surfactant solution into a T-junction capillary geometry. A solvatochromatic fluorescent dye was used to obtain spatially resolved compositional information. In this way, we visualized the microemulsion formation and the flow of it along with the excess phases. A complex interaction between the flow patterns and the microemulsion properties was observed. The formation of microemulsion influenced the flow regimes, and the flow regimes affected the characteristics of the microemulsion formation. In particular, at low flow rates, slug flow was observed, which had profound consequences on the pore scale mixing behavior and resulting microemulsion properties.

  15. Study on culturing Trichodema mutants

    Institute of Scientific and Technical Information of China (English)

    CHEN Jian-ai; WANG Wei-ming


    @@ Trichodema mutants strains T5, T0803, T1010, T1003were cultured in different conditions and media, also in the presence of fungicides at 40 mg/kg (CK or procymidone + chlorothalonil, or maneb or phosethyl-Al) . The pH values of media were 5, 6, 7 and 8 and hyphae were grown at temperatures of 15, 20, 25 and 30 ℃. After being cultured for 3, 4, 5, or 6 days, the strains were transferred at a lower temperature to sporulate (20℃) Obtained data were analyzed statistically, with the orthogonal array and ranges (R) differing dependes on the treatments (R = 40.0,42.4, 48.0, 62.8,107.0). The results indicated that the most important factor was the nature of the strain (R =107.0), while the change in temperature and time of cultivation produced the lowest effect (R =40.0). Each factor variance was significant and A3B4C2D1E3 was the optimum combined condition, in which strain T1010 grew more quickly and sporulated most.

  16. CMPD: cancer mutant proteome database. (United States)

    Huang, Po-Jung; Lee, Chi-Ching; Tan, Bertrand Chin-Ming; Yeh, Yuan-Ming; Julie Chu, Lichieh; Chen, Ting-Wen; Chang, Kai-Ping; Lee, Cheng-Yang; Gan, Ruei-Chi; Liu, Hsuan; Tang, Petrus


    Whole-exome sequencing, which centres on the protein coding regions of disease/cancer associated genes, represents the most cost-effective method to-date for deciphering the association between genetic alterations and diseases. Large-scale whole exome/genome sequencing projects have been launched by various institutions, such as NCI, Broad Institute and TCGA, to provide a comprehensive catalogue of coding variants in diverse tissue samples and cell lines. Further functional and clinical interrogation of these sequence variations must rely on extensive cross-platforms integration of sequencing information and a proteome database that explicitly and comprehensively archives the corresponding mutated peptide sequences. While such data resource is a critical for the mass spectrometry-based proteomic analysis of exomic variants, no database is currently available for the collection of mutant protein sequences that correspond to recent large-scale genomic data. To address this issue and serve as bridge to integrate genomic and proteomics datasets, CMPD ( collected over 2 millions genetic alterations, which not only facilitates the confirmation and examination of potential cancer biomarkers but also provides an invaluable resource for translational medicine research and opportunities to identify mutated proteins encoded by mutated genes.

  17. Pore geometry as a control on rock strength (United States)

    Bubeck, A.; Walker, R. J.; Healy, D.; Dobbs, M.; Holwell, D. A.


    The strength of rocks in the subsurface is critically important across the geosciences, with implications for fluid flow, mineralisation, seismicity, and the deep biosphere. Most studies of porous rock strength consider the scalar quantity of porosity, in which strength shows a broadly inverse relationship with total porosity, but pore shape is not explicitly defined. Here we use a combination of uniaxial compressive strength measurements of isotropic and anisotropic porous lava samples, and numerical modelling to consider the influence of pore shape on rock strength. Micro computed tomography (CT) shows that pores range from sub-spherical to elongate and flat ellipsoids. Samples that contain flat pores are weaker if compression is applied parallel to the short axis (i.e. across the minimum curvature), compared to compression applied parallel to the long axis (i.e. across the maximum curvature). Numerical models for elliptical pores show that compression applied across the minimum curvature results in relatively broad amplification of stress, compared to compression applied across the maximum curvature. Certain pore shapes may be relatively stable and remain open in the upper crust under a given remote stress field, while others are inherently weak. Quantifying the shape, orientations, and statistical distributions of pores is therefore a critical step in strength testing of rocks.

  18. Impedance nanopore biosensor: influence of pore dimensions on biosensing performance. (United States)

    Kant, Krishna; Yu, Jingxian; Priest, Craig; Shapter, Joe G; Losic, Dusan


    Knowledge about electrochemical and electrical properties of nanopore structures and the influence of pore dimensions on these properties is important for the development of nanopore biosensing devices. The aim of this study was to explore the influence of nanopore dimensions (diameter and length) on biosensing performance using non-faradic electrochemical impedance spectroscopy (EIS). Nanoporous alumina membranes (NPAMs) prepared by self-ordered electrochemical anodization of aluminium were used as model nanopore sensing platforms. NPAMs with different pore diameters (25-65 nm) and lengths (4-18 μm) were prepared and the internal pore surface chemistry was modified by covalently attaching streptavidin and biotin. The performance of this antibody nanopore biosensing platform was evaluated using various concentrations of biotin as a model analyte. EIS measurements of pore resistivity and conductivity were carried out for pores with different diameters and lengths. The results showed that smaller pore dimensions of 25 nm and pore lengths up to 10 μm provide better biosensing performance.

  19. Superplastically foaming method to make closed pores inclusive porous ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Kishimoto, Akira; Hayashi, Hidetaka, E-mail: [Division of Molecular and Material Science, Graduate School of Natural Science and Technology, Okayama University Okayama (Japan)


    Porous ceramics incorporates pores to improve several properties including thermal insulation maintaining inherenet ceramic properties such as corrosion resistance and large mechanical strength. Conventional porous ceramics is usually fabricated through an insufficient sintering. Since the sintering accompanies the exclusion of pores, it must be terminated at the early stage to maintain the high porosity, leading to degraded strength and durability. Contrary to this, we have innovated superplastically foaming method to make ceramic foams only in the solid state. In this method, the previously inserted foam agent evaporates after the full densification of matrix at around the sintering temperature. Closed pores expand utilizing the superplastic deformation driven by the evolved gas pressure. The typical features of this superplastically foaming method are listed as follows, 1. The pores are introduced after sintering the solid polycrystal. 2. Only closed pores are introduced, improving the insulation of gas and sound in addition to heat. 3. The pore walls are fully densified expecting a large mechanical strength. 4. Compared with the melt foaming method, this method is practical because the fabrication temperature is far below the melting point and it does not need molds. 5. The size and the location pores can be controlled by the amount and position of the foam agent.

  20. Clostridial pore-forming toxins: powerful virulence factors. (United States)

    Popoff, Michel R


    Pore formation is a common mechanism of action for many bacterial toxins. More than one third of clostridial toxins are pore-forming toxins (PFTs) belonging to the β-PFT class. They are secreted as soluble monomers rich in β-strands, which recognize a specific receptor on target cells and assemble in oligomers. Then, they undergo a conformational change leading to the formation of a β-barrel, which inserts into the lipid bilayer forming functional pore. According to their structure, clostridial β-PFTs are divided into several families. Clostridial cholesterol-dependent cytolysins form large pores, which disrupt the plasma membrane integrity. They are potent virulence factors mainly involved in myonecrosis. Clostridial heptameric β-PFTs (aerolysin family and staphylococcal α-hemolysin family) induce small pores which trigger signaling cascades leading to different cell responses according to the cell types and toxins. They are mainly responsible for intestinal diseases, like necrotic enteritis, or systemic diseases/toxic shock from intestinal origin. Clostridial intracellularly active toxins exploit pore formation through the endosomal membrane to translocate the enzymatic component or domain into the cytosol. Single chain protein toxins, like botulinum and tetanus neurotoxins, use hydrophobic α-helices to form pores, whereas clostridial binary toxins encompass binding components, which are structurally and functionally related to β-PFTs, but which have acquired the specific activity to internalize their corresponding enzymatic components. Structural analysis suggests that β-PFTs and binding components share a common evolutionary origin.

  1. Nuclear Pore-Like Structures in a Compartmentalized Bacterium (United States)

    Sagulenko, Evgeny; Green, Kathryn; Yee, Benjamin; Morgan, Garry; Leis, Andrew; Lee, Kuo-Chang; Butler, Margaret K.; Chia, Nicholas; Pham, Uyen Thi Phuong; Lindgreen, Stinus; Catchpole, Ryan; Poole, Anthony M.; Fuerst, John A.


    Planctomycetes are distinguished from other Bacteria by compartmentalization of cells via internal membranes, interpretation of which has been subject to recent debate regarding potential relations to Gram-negative cell structure. In our interpretation of the available data, the planctomycete Gemmata obscuriglobus contains a nuclear body compartment, and thus possesses a type of cell organization with parallels to the eukaryote nucleus. Here we show that pore-like structures occur in internal membranes of G.obscuriglobus and that they have elements structurally similar to eukaryote nuclear pores, including a basket, ring-spoke structure, and eight-fold rotational symmetry. Bioinformatic analysis of proteomic data reveals that some of the G. obscuriglobus proteins associated with pore-containing membranes possess structural domains found in eukaryote nuclear pore complexes. Moreover, immunogold labelling demonstrates localization of one such protein, containing a β-propeller domain, specifically to the G. obscuriglobus pore-like structures. Finding bacterial pores within internal cell membranes and with structural similarities to eukaryote nuclear pore complexes raises the dual possibilities of either hitherto undetected homology or stunning evolutionary convergence. PMID:28146565

  2. Morphological Characterization of the Polyflux 210H Hemodialysis Filter Pores

    Directory of Open Access Journals (Sweden)

    A. Hedayat


    Full Text Available Background. Morphological characterization of hemodialysis membranes is necessary to improve pore design. Aim. To delineate membrane pore structure of a high flux filter, Polyflux 210H. Methods. We used a Joel JSM-6010LV scanning electron microscope (SEM and a SU6600 Hitachi field emission scanning electron microscope (FESEM to characterize the pore and fiber morphology. The maximal diameters of selected uremic toxins were calculated using the macromolecular modeling Crystallographic Object-Oriented Toolkit (COOT software. Results. The mean pore densities on the outermost and innermost surfaces of the membrane were 36.81% and 5.45%, respectively. The membrane exhibited a tortuous structure with poor connection between the inner and outer pores. The aperture’s width in the inner surface ranged between 34 and 45 nm, which is 8.76–11.60 times larger than the estimated maximum diameter of β2-microglobulin (3.88 nm. Conclusion. The results suggest that the diameter size of inner pore apertures is not a limiting factor to middle molecules clearance, the extremely diminished density is. Increasing inner pore density and improving channel structure are strategies to improve clearance of middle molecules.

  3. Size dependent pore size distribution of shales by gas physisorption (United States)

    Roshan, Hamid; Andersen, Martin S.; Yu, Lu; Masoumi, Hossein; Arandian, Hamid


    Gas physisorption, in particular nitrogen adsorption-desorption, is a traditional technique for characterization of geomaterials including the organic rich shales. The low pressure nitrogen is used together with adsorption-desorption physical models to study the pore size distribution (PSD) and porosity of the porous samples. The samples are usually crushed to a certain fragment size to measure these properties however there is not yet a consistent standard size proposed for sample crushing. Crushing significantly increases the surface area of the fragments e.g. the created surface area is differentiated from that of pores using BET technique. In this study, we show that the smaller fragment sizes lead to higher cumulative pore volume and smaller pore diameters. It is also shown that some of the micro-pores are left unaccounted because of the correction of the external surface area. In order to illustrate this, the nitrogen physisorption is first conducted on the identical organic rich shale samples with different sizes: 20-25, 45-50 and 63-71 µm. We then show that such effects are not only a function of pore structure changes induced by crushing, but is linked to the inability of the physical models in differentiating between the external surface area (BET) and micro-pores for different crushing sizes at relatively low nitrogen pressure. We also discuss models currently used in nano-technology such as t-method to address this issue and their advantages and shortcoming for shale rock characterization.

  4. Novel Techniques to Characterize Pore Size of Porous Materials

    KAUST Repository

    Alabdulghani, Ali J.


    Porous materials are implemented in several industrial applications such as water desalination, gas separation and pharmaceutical care which they are mainly governed by the pore size and the PSD. Analyzing shale reservoirs are not excluded from these applications and numerous advantages can be gained by evaluating the PSD of a given shale reservoir. Because of the limitations of the conventional characterization techniques, novel methods for characterizing the PSD have to be proposed in order to obtain better characterization results for the porous materials, in general, and shale rocks in particular. Thus, permporosimetry and evapoporometry (EP) technologies were introduced, designed and utilized for evaluating the two key parameters, pore size and pore size distribution. The pore size and PSD profiles of different shale samples from Norway and Argentina were analyzed using these technologies and then confirmed by mercury intrusion porosimeter (MIP). Norway samples showed an average pore diameter of 12.94 nm and 19.22 nm with an average diameter of 13.77 nm and 23.23 nm for Argentina samples using permporosimetry and EP respectively. Both techniques are therefore indicative of the heterogeneity of the shales. The results from permporosimetry are in good agreement with those obtained from MIP technique, but EP for most part over-estimates the average pore size. The divergence of EP results compared to permporosimetry results is referred to the fact that the latter technique measures only the active pores which is not the case with the former technique. Overall, both techniques are complementary to each other which the results from both techniques seem reasonable and reliable and provide two simple techniques to estimate the pore size and pore size distributions for shale rocks.

  5. Simple model of capillary condensation in cylindrical pores (United States)

    Szybisz, Leszek; Urrutia, Ignacio


    A simple model based on an approximation of the dropletlike model is formulated for studying adsorption of fluids into cylindrical pores. This model yields a nearly universal description of capillary condensation transitions for noble gases confined by alkali metals. The system's thermodynamical behavior is predicted from the values of two dimensionless parameters: D* (the reduced asymptotic strength of the fluid-adsorber interaction, a function of temperature) and R* (the reduced radius of the pore). The phenomenon of hysteresis inherently related to capillary condensation is discussed. The connection to a previously proposed universality for cylindrical pores is also established.

  6. Pore structure of SWNTs with high hydrogen storage capacity

    Institute of Scientific and Technical Information of China (English)

    杨全红; 刘畅; 刘敏; 樊彦贞; 成会明; 王茂章


    This paper reveals, by analyses of nitrogen cryo-adsorption isotherm, the energetic and structural heterogeneity of single-walled carbon nanotubes (SWNTs) which has a high hydrogen storage capacity. It was found that SWNTs had manifold pore structures and distributed surface energy. By comparison of the pore structures and energy distributions of SWNTs before and after hydrogen adsorption, it is preliminarily indicated that hydrogen adsorption occurred in micropores and mesopores with smaller diameter, and that the pores of different diameters determined different hydrogen adsorption processes and underwent different structure changes during hydrogen adsorption.

  7. A pore-forming toxin enables Serratia a nonlytic egress from host cells. (United States)

    Di Venanzio, Gisela; Lazzaro, Martina; Morales, Enrique S; Krapf, Darío; García Véscovi, Eleonora


    Several pathogens co-opt host intracellular compartments to survive and replicate, and they thereafter disperse progeny to prosper in a new niche. Little is known about strategies displayed by Serratia marcescens to defeat immune responses and disseminate afterwards. Upon invasion of nonphagocytic cells, Serratia multiplies within autophagosome-like vacuoles. These Serratia-containing vacuoles (SeCV) circumvent progression into acidic/degradative compartments, avoiding elimination. In this work, we show that ShlA pore-forming toxin (PFT) commands Serratia escape from invaded cells. While ShlA-dependent, Ca(2)(+) local increase was shown in SeCVs tight proximity, intracellular Ca(2)(+) sequestration prevented Serratia exit. Accordingly, a Ca(2)(+) surge rescued a ShlA-deficient strain exit capacity, demonstrating that Ca(2)(+) mobilization is essential for egress. As opposed to wild-type-SeCV, the mutant strain-vacuole was wrapped by actin filaments, showing that ShlA expression rearranges host actin. Moreover, alteration of actin polymerization hindered wild-type Serratia escape, while increased intracellular Ca(2)(+) reorganized the mutant strain-SeCV actin distribution, restoring wild-type-SeCV phenotype. Our results demonstrate that, by ShlA expression, Serratia triggers a Ca(2)(+) signal that reshapes cytoskeleton dynamics and ends up pushing the SeCV load out of the cell, in an exocytic-like process. These results disclose that PFTs can be engaged in allowing bacteria to exit without compromising host cell integrity.

  8. Microfiltration of distillery stillage: Influence of membrane pore size

    Directory of Open Access Journals (Sweden)

    Vasić Vesna M.


    Full Text Available Stillage is one of the most polluted waste products of the food industry. Beside large volume, the stillage contains high amount of suspended solids, high values of chemical oxygen demand and biological oxygen demand, so it should not be discharged in the nature before previous purification. In this work, three ceramic membranes for microfiltration with different pore sizes were tested for stillage purification in order to find the most suitable membrane for the filtration process. Ceramic membranes with a nominal pore size of 200 nm, 450 nm and 800 nm were used for filtration. The influence of pore size on permeate flux and removal efficiency was investigated. A membrane with the pore size of 200 nm showed the best filtration performance so it was chosen for the microfiltration process.

  9. Nano pores evolution in hydroxyapatite microsphere during spark plasma sintering

    Directory of Open Access Journals (Sweden)

    Lin C.


    Full Text Available Micron-spherical granules of hydroxyapatite (HAp nanoparticles were prepared by powder granulation methods. Through subsequent sintering, porous HAp microspheres with tailored pore and grain framework structures were obtained. Detailed microstructure investigation by SEM and TEM revealed the correlation of the pore structure and the necking strength with the sintering profiles that determine the coalescence features of the nanoparticles. The partially sintered porous HAp microspheres containing more than 50% porosity consisting of pores and grains both in nano-scale are active in inducing the precipitation of HAp in simulated body fluid. The nano-porous HAp microspheres with an extensive surface and interconnecting pores thus demonstrate the potential of stimulating the formation of collagen and bone and the integration with the newly formed bones during physiological bone remodeling.

  10. Bilayer Deformation, Pores, and Micellation Induced by Oxidized Lipids. (United States)

    Boonnoy, Phansiri; Jarerattanachat, Viwan; Karttunen, Mikko; Wong-Ekkabut, Jirasak


    The influence of different oxidized lipids on lipid bilayers was investigated with 16 individual 1 μs atomistic molecular dynamics (MD) simulations. Binary mixtures of lipid bilayers of 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidylcholine (PLPC) and its peroxide and aldehyde products were performed at different concentrations. In addition, an asymmetrical short chain lipid, 1-palmitoyl-2-decanoyl-sn-glycero-3-phosphatidylcholine (PDPC), was used to compare the effects of polar/apolar groups in the lipid tail on lipid bilayer. Although water defects occurred with both aldehyde and peroxide lipids, full pore formation was observed only for aldehyde lipids. At medium concentrations the pores were stable. At higher concentrations, however, the pores became unstable and micellation occurred. Data analysis shows that aldehyde lipids' propensity for pore formation is due to their shorter and highly mobile tail. The highly polar peroxide lipids are stabilized by strong hydrogen bonds with interfacial water.

  11. The Pore Structure and Hydration Performance of Sulphoaluminate MDF Cement

    Institute of Scientific and Technical Information of China (English)

    HUANG Cong-yun; YUAN Run-zhang; LONG Shi-zong


    The hydration and pore structure of sulphoaluminate MDF cement were studied by X-ray diffractometer ( XRD ), scanning electron microscope (SEM) and mercury intrusion porosimeter ( MIP ) etc. The ex-perimental results indicate that hydration products of the materials are entringites ( Aft ), aluminium hydroxide andCSH (Ⅰ) gel etc. Due to its very low water-cement ratio, hydration function is only confined to the surfaces of ce-ment grains, and there is a lot of sulphoaluminate cement in the hardenite which is unhydrated yet. Hydration re-action was rapidly carried under the condition of the heat-pressing. Therefore cement hydrates Aft, CSH (Ⅰ) andaluminium hydroxide gel fill in pores. The expansibility of Aft makes the porosity of MDF cement lower ( less than1 percent ) and the size of pore smaller (80 percent pore was less than 250A), and enhances its strength.

  12. Diffusion Pore Imaging by Hyperpolarized Xenon-129 Nuclear Magnetic Resonance

    CERN Document Server

    Kuder, Tristan Anselm; Windschuh, Johannes; Laun, Frederik Bernd


    Nuclear magnetic resonance (NMR) diffusion measurements are widely used to derive parameters indirectly related to the microstructure of biological tissues and porous media. However, a direct imaging of cell or pore shapes and sizes would be of high interest. For a long time, determining pore shapes by NMR diffusion acquisitions seemed impossible, because the necessary phase information could not be preserved. Here we demonstrate experimentally using the measurement technique which we have recently proposed theoretically that the shape of arbitrary closed pores can be imaged by diffusion acquisitions, which yield the phase information. For this purpose, we use hyperpolarized xenon gas in well-defined geometries. The signal can be collected from the whole sample which mainly eliminates the problem of vanishing signal at increasing resolution of conventional NMR imaging. This could be used to non-invasively gain structural information inaccessible so far such as pore or cell shapes, cell density or axon integri...

  13. Hypoxia and the hypoxic response pathway protect against pore-forming toxins in C. elegans.

    Directory of Open Access Journals (Sweden)

    Audrey Bellier


    Full Text Available Pore-forming toxins (PFTs are by far the most abundant bacterial protein toxins and are important for the virulence of many important pathogens. As such, cellular responses to PFTs critically modulate host-pathogen interactions. Although many cellular responses to PFTs have been recorded, little is understood about their relevance to pathological or defensive outcomes. To shed light on this important question, we have turned to the only genetic system for studying PFT-host interactions-Caenorhabditis elegans intoxication by Crystal (Cry protein PFTs. We mutagenized and screened for C. elegans mutants resistant to a Cry PFT and recovered one mutant. Complementation, sequencing, transgenic rescue, and RNA interference data demonstrate that this mutant eliminates a gene normally involved in repression of the hypoxia (low oxygen response pathway. We find that up-regulation of the C. elegans hypoxia pathway via the inactivation of three different genes that normally repress the pathway results in animals resistant to Cry PFTs. Conversely, mutation in the central activator of the hypoxia response, HIF-1, suppresses this resistance and can result in animals defective in PFT defenses. These results extend to a PFT that attacks mammals since up-regulation of the hypoxia pathway confers resistance to Vibrio cholerae cytolysin (VCC, whereas down-regulation confers hypersusceptibility. The hypoxia PFT defense pathway acts cell autonomously to protect the cells directly under attack and is different from other hypoxia pathway stress responses. Two of the downstream effectors of this pathway include the nuclear receptor nhr-57 and the unfolded protein response. In addition, the hypoxia pathway itself is induced by PFT, and low oxygen is protective against PFT intoxication. These results demonstrate that hypoxia and induction of the hypoxia response protect cells against PFTs, and that the cellular environment can be modulated via the hypoxia pathway to

  14. A structural model of the pore-forming region of the skeletal muscle ryanodine receptor (RyR1.

    Directory of Open Access Journals (Sweden)

    Srinivas Ramachandran


    Full Text Available Ryanodine receptors (RyRs are ion channels that regulate muscle contraction by releasing calcium ions from intracellular stores into the cytoplasm. Mutations in skeletal muscle RyR (RyR1 give rise to congenital diseases such as central core disease. The absence of high-resolution structures of RyR1 has limited our understanding of channel function and disease mechanisms at the molecular level. Here, we report a structural model of the pore-forming region of RyR1. Molecular dynamics simulations show high ion binding to putative pore residues D4899, E4900, D4938, and D4945, which are experimentally known to be critical for channel conductance and selectivity. We also observe preferential localization of Ca(2+ over K(+ in the selectivity filter of RyR1. Simulations of RyR1-D4899Q mutant show a loss of preference to Ca(2+ in the selectivity filter as seen experimentally. Electrophysiological experiments on a central core disease mutant, RyR1-G4898R, show constitutively open channels that conduct K(+ but not Ca(2+. Our simulations with G4898R likewise show a decrease in the preference of Ca(2+ over K(+ in the selectivity filter. Together, the computational and experimental results shed light on ion conductance and selectivity of RyR1 at an atomistic level.

  15. Diffusion in the pore water of compacted crushed salt

    Energy Technology Data Exchange (ETDEWEB)

    Fluegge, Judith; Herr, Sebastian; Lauke, Thomas; Meleshyn, Artur; Miehe, Ruediger; Ruebel, Andre


    Diffusion of dissolved radionuclides in the pore water of compacted crushed salt in the long-term is the most relevant process for the release of radionuclides from a dedicated repository for high-level waste in a salt formation as has been shown in latest safety assessments and research projects /BUH 16/. So far, diffusion coefficients for free water have been applied for the diffusion in pore water in models for long-term safety assessments. This conservative assumption was used, because data on the diffusion coefficient of dissolved substances in crushed salt have been missing. Furthermore, the diffusion coefficient in the pore water was assumed to be constant and independent from the degree of compaction of the crushed salt. The work presented in this report was intended to contribute to fill this gap of knowledge about how the diffusion of radionuclides takes place in the compacted backfill of a repository in salt. For the first time, the pore diffusion coefficient as well as its dependence on the porosity of the crushed salt was determined experimentally by means of through-diffusion experiments using caesium as tracer. The results achieved in this project suggest that the diffusion in compacted crushed salt is not fully comparable to that in a homogeneous, temporally stable porous medium like sand or clay. The results obtained from four diffusion experiments show a remarkably different behaviour and all yield unique concentration versus time plots which includes highly temporal variable tracer fluxes with even full interruptions of the flux for longer periods of time. This effect cannot be explained by assuming a tracer transport by diffusion in a temporarily invariant pore space and / or under temporally invariant experimental conditions. From our point of view, a restructuring of the pore space seems to lead to closed areas of pore water in the sample which may open up again after some time, leading to a variable pore space and hence variable diffusive

  16. Molecular biology and biophysical properties of ion channel gating pores. (United States)

    Moreau, Adrien; Gosselin-Badaroudine, Pascal; Chahine, Mohamed


    The voltage sensitive domain (VSD) is a pivotal structure of voltage-gated ion channels (VGICs) and plays an essential role in the generation of electrochemical signals by neurons, striated muscle cells, and endocrine cells. The VSD is not unique to VGICs. Recent studies have shown that a VSD regulates a phosphatase. Similarly, Hv1, a voltage-sensitive protein that lacks an apparent pore domain, is a self-contained voltage sensor that operates as an H⁺ channel. VSDs are formed by four transmembrane helices (S1-S4). The S4 helix is positively charged due to the presence of arginine and lysine residues. It is surrounded by two water crevices that extend into the membrane from both the extracellular and intracellular milieus. A hydrophobic septum disrupts communication between these water crevices thus preventing the permeation of ions. The septum is maintained by interactions between the charged residues of the S4 segment and the gating charge transfer center. Mutating the charged residue of the S4 segment allows the water crevices to communicate and generate gating pore or omega pore. Gating pore currents have been reported to underlie several neuronal and striated muscle channelopathies. Depending on which charged residue on the S4 segment is mutated, gating pores are permeant either at depolarized or hyperpolarized voltages. Gating pores are cation selective and seem to converge toward Eisenmann's first or second selectivity sequences. Most gating pores are blocked by guanidine derivatives as well as trivalent and quadrivalent cations. Gating pores can be used to study the movement of the voltage sensor and could serve as targets for novel small therapeutic molecules.

  17. Relationship between pore structure and compressive strength of concrete: Experiments and statistical modeling

    Indian Academy of Sciences (India)

    J BU; Z TIAN


    Properties of concrete are strongly dependent on its pore structure features, porosity being an important one among them. This study deals with developing an understanding of the pore structure-compressive strength relationship in concrete. Several concrete mixtures with different pore structures are proportioned and subjected to static compressive tests. The pore structure features such as porosity, pore size distribution are extracted using mercury intrusion porosimetry technique. A statistical model is developed to relate thecompressive strength to relevant pore structure features.

  18. Distributed pore model for bio-molecule chromatography. (United States)

    Coquebert de Neuville, Bertrand; Tarafder, Abhijit; Morbidelli, Massimo


    One of the main peculiarities in protein chromatography is that the adsorbing proteins and the adsorbent pores have comparable sizes. This has the consequence that the pore accessibility depends not only on the solute size but also on the loading conditions of the adsorbent because protein adsorption significantly reduces the size of the pores. A model that accounts for the pore size distribution of the stationary phase and for the pore shrinkage due to protein adsorption has been developed to describe mass transport and adsorption in the porous particles. This model has been shown to be equivalent to the general rate model (GRM) in the case of processes under highly diluted conditions with little adsorption. This implies that the model parameters determination follows the same procedure as for the classical GRM. The new pore model has been applied and compared to the GRM for the simulation of lysozyme breakthrough experiments and for the prediction of 5% dynamic binding capacity values solely based on static capacity measurements.

  19. Pore opening dynamics in the exocytosis of serotonin (United States)

    Ramirez-Santiago, Guillermo; Cercos, Montserrat G.; Martinez-Valencia, Alejandro; Salinas Hernandez, Israel; Rodríguez-Sosa, Leonardo; de-Miguel, Francisco F.


    The current view of the exocytosis of transmitter molecules is that it starts with the formation of a fusion pore that connects the intravesicular and the extracellular spaces, and is completed by the release of the rest of the transmitter contained in the vesicle upon the full fusion and collapse of the vesicle with the plasma membrane. However, under certain circumstances, a rapid closure of the pore before the full vesicle fusion produces only a partial release of the transmitter. Here we show that whole release of the transmitter occurs through fusion pores that remain opened for tens of milliseconds without vesicle collapse. This was demonstrated through amperometric measurements of serotonin release from electrodense vesicles in the axon of leech Retzius neurons and mathematical modelling. By modeling transmitter release with a diffusion equation subjected to boundary conditions that are defined by the experiment, we showed that those pores with a fast half rise time constant remained opened and allowed the full quantum release without vesicle collapse, whereas pores with a slow rise time constant closed rapidly, thus producing partial release. We conclude that a full transmitter release may occur through the fusion pore in the absence of vesicle collapse. This work was founded by a DGAPA-UNAM grants IN200914 and IN118410 CONACYT GRANT 130031, and CONACyT doctoral fellowships.

  20. Mangrove pore water exchange across a latitudinal gradient (United States)

    Tait, Douglas R.; Maher, Damien T.; Macklin, Paul A.; Santos, Isaac R.


    We combined observations of the natural tracer radon (222Rn) with hydrodynamic models across a broad latitudinal gradient covering several climate zones to estimate pore water exchange rates in mangroves. Pore water exchange ranged from 2.1 to 35.5 cm d-1 from temperate to tropical regions and averaged 16.3 ± 5.1 cm d-1. If upscaled to the global weighted mangrove area, pore water exchange in mangroves would recirculate the entire volume of water overlying the continental shelf in less than 153 years. Although pore water exchange (recirculated seawater) and river discharge represent different pathways for water entering the coastal ocean, the estimated global mangrove pore water exchange would be equal to approximately one third of annual global river discharge to the ocean (3.84 × 1013 m3 yr-1). Because biogeochemical processes in mangroves are largely dependent on pore water exchange, these large exchange rates have major implications for coastal nutrient, carbon, and greenhouse gas cycling in tropical marine systems.

  1. Patch-clamp detection of macromolecular translocation along nuclear pores

    Directory of Open Access Journals (Sweden)

    Bustamante J.O.


    Full Text Available The present paper reviews the application of patch-clamp principles to the detection and measurement of macromolecular translocation along the nuclear pores. We demonstrate that the tight-seal 'gigaseal' between the pipette tip and the nuclear membrane is possible in the presence of fully operational nuclear pores. We show that the ability to form a gigaseal in nucleus-attached configurations does not mean that only the activity of channels from the outer membrane of the nuclear envelope can be detected. Instead, we show that, in the presence of fully operational nuclear pores, it is likely that the large-conductance ion channel activity recorded derives from the nuclear pores. We conclude the technical section with the suggestion that the best way to demonstrate that the nuclear pores are responsible for ion channel activity is by showing with fluorescence microscopy the nuclear translocation of ions and small molecules and the exclusion of the same from the cisterna enclosed by the two membranes of the envelope. Since transcription factors and mRNAs, two major groups of nuclear macromolecules, use nuclear pores to enter and exit the nucleus and play essential roles in the control of gene activity and expression, this review should be useful to cell and molecular biologists interested in understanding how patch-clamp can be used to quantitate the translocation of such macromolecules into and out of the nucleus

  2. Pore growth in U-Mo/Al dispersion fuel

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Yeon Soo, E-mail: [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439 (United States); Jeong, G.Y.; Sohn, D.-S. [Ulsan National Institute of Science and Technology, 50 UNIST-gil, Eonyang-eup, Ulju-gun, Ulsan, 689-798 (Korea, Republic of); Jamison, L.M. [Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439 (United States)


    U-Mo/Al dispersion fuel is currently under development in the DOE’s Material Management and Minimization program to convert HEU-fueled research reactors to LEU-fueled reactors. In some demanding conditions in high-power and high-performance reactors, large pores form in the interaction layers between the U-Mo fuel particles and the Al matrix, which pose a potential to cause fuel failure. In this study, comprehension of the formation and growth of these pores was explored. As a product, a model to predict pore growth and porosity increase was developed. The model includes three major topics: fission gas release from the U-Mo and the IL to the pores, stress evolution in the fuel meat, and the effect of amorphous IL growth. Well-characterized in-pile data from reduced-size plates were used to fit the model parameters. A data set from full-sized plates, independent and distinctively different from those used to fit the model parameters, was used to examine the accuracy of the model. The model showed fair agreement with the measured data. The model suggested that the growth of the IL has a critical effect on pore growth, as both its material properties and energetics are favorable to pore formation. Therefore, one area of the current effort, focused on suppressing IL growth, appears to be on the right track to improve the performance of this fuel.

  3. Nonlinear electrokinetic transport in networks of microscale and nanoscale pores (United States)

    Alizadeh, Shima; Andersen, Mathias B.; Mani, Ali


    The objective of this study is to develop the understanding of nonlinear electrohydrodynamic effects in a wide range of systems including lab-on-a-chip systems, electroosmotic pumps, and, in general, porous media with random or fabricated pore morphology. We present a continuum model in which these systems are described as massive networks of long and thin pores. The thickness of the pores can vary from nanoscale to microscale, corresponding to the highly overlapped electric double layers (EDL) to the thin double layer limit. Within each pore the transport in the wall-normal direction is assumed to be in equilibrium leading to a reduced order model for the axial transport of species in the form of a transient one-dimensional partial differential equation (PDE). PDEs from different pores are coupled through boundary conditions at the pore intersections by proper implementation of the conservation laws. We show that this model can capture important nonlinear dynamics, which are typically ignored in homogenized models. Specifically, our model captures concentration polarization shocks and flow recirculation zones respectively formed when micropores and nanopores are connected in series and in parallel. We present a comparison between our model and recent experiments in microfluidics, and will discuss applications in porous media modeling for energy storage and water purification systems.

  4. Accumulation of formamide in hydrothermal pores to form prebiotic nucleobases (United States)

    Niether, Doreen; Afanasenkau, Dzmitry; Dhont, Jan K. G.


    Formamide is one of the important compounds from which prebiotic molecules can be synthesized, provided that its concentration is sufficiently high. For nucleotides and short DNA strands, it has been shown that a high degree of accumulation in hydrothermal pores occurs, so that temperature gradients might play a role in the origin of life [Baaske P, et al. (2007) Proc Natl Acad Sci USA 104(22):9346-9351]. We show that the same combination of thermophoresis and convection in hydrothermal pores leads to accumulation of formamide up to concentrations where nucleobases are formed. The thermophoretic properties of aqueous formamide solutions are studied by means of Infrared Thermal Diffusion Forced Rayleigh Scattering. These data are used in numerical finite element calculations in hydrothermal pores for various initial concentrations, ambient temperatures, and pore sizes. The high degree of formamide accumulation is due to an unusual temperature and concentration dependence of the thermophoretic behavior of formamide. The accumulation fold in part of the pores increases strongly with increasing aspect ratio of the pores, and saturates to highly concentrated aqueous formamide solutions of ˜85 wt% at large aspect ratios. Time-dependent studies show that these high concentrations are reached after 45-90 d, starting with an initial formamide weight fraction of 10-310-3 wt % that is typical for concentrations in shallow lakes on early Earth.

  5. Atomic structure of anthrax protective antigen pore elucidates toxin translocation. (United States)

    Jiang, Jiansen; Pentelute, Bradley L; Collier, R John; Zhou, Z Hong


    Anthrax toxin, comprising protective antigen, lethal factor, and oedema factor, is the major virulence factor of Bacillus anthracis, an agent that causes high mortality in humans and animals. Protective antigen forms oligomeric prepores that undergo conversion to membrane-spanning pores by endosomal acidification, and these pores translocate the enzymes lethal factor and oedema factor into the cytosol of target cells. Protective antigen is not only a vaccine component and therapeutic target for anthrax infections but also an excellent model system for understanding the mechanism of protein translocation. On the basis of biochemical and electrophysiological results, researchers have proposed that a phi (Φ)-clamp composed of phenylalanine (Phe)427 residues of protective antigen catalyses protein translocation via a charge-state-dependent Brownian ratchet. Although atomic structures of protective antigen prepores are available, how protective antigen senses low pH, converts to active pore, and translocates lethal factor and oedema factor are not well defined without an atomic model of its pore. Here, by cryo-electron microscopy with direct electron counting, we determine the protective antigen pore structure at 2.9-Å resolution. The structure reveals the long-sought-after catalytic Φ-clamp and the membrane-spanning translocation channel, and supports the Brownian ratchet model for protein translocation. Comparisons of four structures reveal conformational changes in prepore to pore conversion that support a multi-step mechanism by which low pH is sensed and the membrane-spanning channel is formed.

  6. Pore Scale Analysis of Oil Shale/Sands Pyrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chen-Luh [Univ. of Utah, Salt Lake City, UT (United States); Miller, Jan [Univ. of Utah, Salt Lake City, UT (United States)


    There are important questions concerning the quality and volume of pore space that is created when oil shale is pyrolyzed for the purpose of producing shale oil. In this report, 1.9 cm diameter cores of Mahogany oil shale were pyrolyzed at different temperatures and heating rates. Detailed 3D imaging of core samples was done using multiscale X-ray computed tomography (CT) before and after pyrolysis to establish the pore structure. The pore structure of the unreacted material was not clear. Selected images of a core pyrolyzed at 400oC were obtained at voxel resolutions from 39 microns (Οm) to 60 nanometers (nm). Some of the pore space created during pyrolysis was clearly visible at these resolutions and it was possible to distinguish between the reaction products and the host shale rock. The pore structure deduced from the images was used in Lattice Boltzmann simulations to calculate the permeability in the pore space. The permeabilities of the pyrolyzed samples of the silicate-rich zone were on the order of millidarcies, while the permeabilities of the kerogen-rich zone after pyrolysis were very anisotropic and about four orders of magnitude higher.

  7. Pore growth in U-Mo/Al dispersion fuel (United States)

    Kim, Yeon Soo; Jeong, G. Y.; Sohn, D.-S.; Jamison, L. M.


    U-Mo/Al dispersion fuel is currently under development in the DOE's Material Management and Minimization program to convert HEU-fueled research reactors to LEU-fueled reactors. In some demanding conditions in high-power and high-performance reactors, large pores form in the interaction layers between the U-Mo fuel particles and the Al matrix, which pose a potential to cause fuel failure. In this study, comprehension of the formation and growth of these pores was explored. As a product, a model to predict pore growth and porosity increase was developed. The model includes three major topics: fission gas release from the U-Mo and the IL to the pores, stress evolution in the fuel meat, and the effect of amorphous IL growth. Well-characterized in-pile data from reduced-size plates were used to fit the model parameters. A data set from full-sized plates, independent and distinctively different from those used to fit the model parameters, was used to examine the accuracy of the model. The model showed fair agreement with the measured data. The model suggested that the growth of the IL has a critical effect on pore growth, as both its material properties and energetics are favorable to pore formation. Therefore, one area of the current effort, focused on suppressing IL growth, appears to be on the right track to improve the performance of this fuel.

  8. Software Image J to study soil pore distribution

    Directory of Open Access Journals (Sweden)

    Sabrina Passoni


    Full Text Available In the soil science, a direct method that allows the study of soil pore distribution is the bi-dimensional (2D digital image analysis. Such technique provides quantitative results of soil pore shape, number and size. The use of specific softwares for the treatment and processing of images allows a fast and efficient method to quantify the soil porous system. However, due to the high cost of commercial softwares, public ones can be an interesting alternative for soil structure analysis. The objective of this work was to evaluate the quality of data provided by the Image J software (public domain used to characterize the voids of two soils, characterized as Geric Ferralsol and Rhodic Ferralsol, from the southeast region of Brazil. The pore distribution analysis technique from impregnated soil blocks was utilized for this purpose. The 2D image acquisition was carried out by using a CCD camera coupled to a conventional optical microscope. After acquisition and treatment of images, they were processed and analyzed by the software Noesis Visilog 5.4® (chosen as the reference program and ImageJ. The parameters chosen to characterize the soil voids were: shape, number and pore size distribution. For both soils, the results obtained for the image total porosity (%, the total number of pores and the pore size distribution showed that the Image J is a suitable software to be applied in the characterization of the soil sample voids impregnated with resin.

  9. Proline Scan of the hERG Channel S6 Helix Reveals the Location of the Intracellular Pore Gate (United States)

    Thouta, Samrat; Sokolov, Stanislav; Abe, Yuki; Clark, Sheldon J.; Cheng, Yen M.; Claydon, Tom W.


    In Shaker-like channels, the activation gate is formed at the bundle crossing by the convergence of the inner S6 helices near a conserved proline-valine-proline motif, which introduces a kink that allows for electromechanical coupling with voltage sensor motions via the S4-S5 linker. Human ether-a-go-go-related gene (hERG) channels lack the proline-valine-proline motif and the location of the intracellular pore gate and how it is coupled to S4 movement is less clear. Here, we show that proline substitutions within the S6 of hERG perturbed pore gate closure, trapping channels in the open state. Performing a proline scan of the inner S6 helix, from Ile655 to Tyr667 revealed that gate perturbation occurred with proximal (I655P-Q664P), but not distal (R665P-Y667P) substitutions, suggesting that Gln664 marks the position of the intracellular gate in hERG channels. Using voltage-clamp fluorimetry and gating current analysis, we demonstrate that proline substitutions trap the activation gate open by disrupting the coupling between the voltage-sensing unit and the pore of the channel. We characterize voltage sensor movement in one such trapped-open mutant channel and demonstrate the kinetics of what we interpret to be intrinsic hERG voltage sensor movement. PMID:24606930

  10. Determining pore length scales and pore surface relaxivity of rock cores by internal magnetic fields modulation at 2MHz NMR. (United States)

    Liu, Huabing; Nogueira d'Eurydice, Marcel; Obruchkov, Sergei; Galvosas, Petrik


    Pore length scales and pore surface relaxivities of rock cores with different lithologies were studied on a 2MHz Rock Core Analyzer. To determine the pore length scales of the rock cores, the high eigenmodes of spin bearing molecules satisfying the diffusion equation were detected with optimized encoding periods in the presence of internal magnetic fields Bin. The results were confirmed using a 64MHz NMR system, which supports the feasibility of high eigenmode detection at fields as low as 2MHz. Furthermore, this methodology was combined with relaxometry measurements to a two-dimensional experiment, which provides correlation between pore length and relaxation time. This techniques also yields information on the surface relaxivity of the rock cores. The estimated surface relaxivities were then compared to the results using an independent NMR method.

  11. Accurate relations between pore size and the pressure of capillary condensation and the evaporation of nitrogen in cylindrical pores. (United States)

    Morishige, Kunimitsu; Tateishi, Masayoshi


    To examine the theoretical and semiempirical relations between pore size and the pressure of capillary condensation or evaporation proposed so far, we constructed an accurate relation between the pore radius and the capillary condensation and evaporation pressure of nitrogen at 77 K for the cylindrical pores of the ordered mesoporous MCM-41 and SBA-15 silicas. Here, the pore size was determined from a comparison between the experimental and calculated X-ray diffraction patterns due to X-ray structural modeling recently developed. Among the many theoretical relations that differ from each other in the degree of theoretical improvements, a macroscopic thermodynamic approach based on Broekhoff-de Boer equations was found to be in fair agreement with the experimental relation obtained in the present study.

  12. Muscle development in mdx mutant mice. (United States)

    Dangain, J; Vrbova, G


    Mechanical and contractile properties of tibialis anterior (TA) muscles from X-linked muscular dystrophic (mdx) mutant mice at different stages of development are compared to those of muscles from normal control animals. There is no difference between the tension output, speeds of contraction and relaxation, and weight of TA muscles from mutant adults and normal control animals. However, it is found that in 3-4-week-old mutant animals, tension output and muscle weight are very much reduced, and half relaxation time is prolonged. Thus, during this stage of development, muscles from mdx mice do not function properly. Histological examination of these muscles provides further evidence that, in these animals, rapid muscle destruction occurs at a particular time of development and that it is followed by complete recovery. This new mutant therefore presents an interesting case of muscle destruction and rapid regeneration. However, it is not an adequate model for Duchenne muscular dystrophy.

  13. Targeting ESR1-Mutant Breast Cancer (United States)


    AWARD NUMBER: W81XWH-14-1-0359 TITLE: Targeting ESR1-Mutant Breast Cancer PRINCIPAL INVESTIGATOR: Dr. Sarat Chandarlapaty CONTRACTING...31 Aug 2015 4. TITLE AND SUBTITLE Targeting ESR1-Mutant Breast Cancer 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH-14-1-0359 5c. PROGRAM ELEMENT...mutations found in breast cancer using both structural and cell based assays. We have now have evidence for the effects of the most recurrent

  14. Targeting ESR1-Mutant Breast Cancer (United States)


    Introduction Approximately 70% of ER+ breast cancers harbor expression of the estrogen receptor and are dependent upon its activity for various aspects of the...resistance to current FDA approved ER antagonists, but that more potent and selective estrogen receptor antagonists will be sufficiently active to...antagonists and their potency against ER mutants both in vitro and in vivo . Targeting ESR1-Mutant Breast Cancer W81XWH-14-1-0359 9 4. Impact A) Impact

  15. Unified method for the total pore volume and pore size distribution of hierarchical zeolites from argon adsorption and mercury intrusion. (United States)

    Kenvin, Jeffrey; Jagiello, Jacek; Mitchell, Sharon; Pérez-Ramírez, Javier


    A generalized approach to determine the complete distribution of macropores, mesopores, and micropores from argon adsorption and mercury porosimetry is developed and validated for advanced zeolite catalysts with hierarchically structured pore systems in powder and shaped forms. Rather than using a fragmented approach of simple overlays from individual techniques, a unified approach that utilizes a kernel constructed from model isotherms and model intrusion curves is used to calculate the complete pore size distribution and the total pore volume of the material. An added benefit of a single full-range pore size distribution is that the cumulative pore area and the area distribution are also obtained without the need for additional modeling. The resulting complete pore size distribution and the kernel accurately model both the adsorption isotherm and the mercury porosimetry. By bridging the data analysis of two primary characterization tools, this methodology fills an existing gap in the library of familiar methods for porosity assessment in the design of materials with multilevel porosity for novel technological applications.

  16. Pore-forming pyocin S5 utilizes the FptA ferripyochelin receptor to kill Pseudomonas aeruginosa. (United States)

    Elfarash, Ameer; Dingemans, Jozef; Ye, Lumeng; Hassan, Ahmed Amir; Craggs, Michael; Reimmann, Cornelia; Thomas, Mark S; Cornelis, Pierre


    Pyocins are toxic proteins produced by some strains of Pseudomonas aeruginosa that are lethal for related strains of the same species. Some soluble pyocins (S2, S3 and S4) were previously shown to use the pyoverdine siderophore receptors to enter the cell. The P. aeruginosa PAO1 pore-forming pyocin S5 encoding gene (PAO985) was cloned into the expression vector pET15b, and the affinity-purified protein product tested for its killing activity against different P. aeruginosa strains. The results, however, did not show any correlation with a specific ferripyoverdine receptor. To further identify the S5 receptor, transposon mutants were generated. Pooled mutants were exposed to pyocin S5 and the resistant colonies growing in the killing zone were selected. The majority of S5-resistant mutants had an insertion in the fptA gene encoding the receptor for the siderophore pyochelin. Complementation of an fptA transposon mutant with the P. aeruginosa fptA gene in trans restored the sensitivity to S5. In order to define the receptor-binding domain of pyocin S5, two hybrid pyocins were constructed containing different regions from pyocin S5 fused to the C-terminal translocation and DNase killing domains of pyocin S2. Only the protein containing amino acid residues 151 to 300 from S5 showed toxicity, indicating that the pyocin S5 receptor-binding domain is not at the N-terminus of the protein as in other S-type pyocins. Pyocin S5 was, however, unable to kill Burkholderia cenocepacia strains producing a ferripyochelin FptA receptor, nor was the B. cenocepacia fptA gene able to restore the sensitivity of the resistant fptA mutant P. aeruginosa strain.

  17. Fractal classification and natural classification of coal pore structure based on migration of coal bed methane

    Institute of Scientific and Technical Information of China (English)

    FU Xuehai; QIN Yong; ZHANG Wanhong; WEI Chongtao; ZHOU Rongfu


    According to the data of 146 coal samples measured by mercury penetration, coal pores are classified into two levels of <65 nm diffusion pore and >65 nm seeping pore by fractal method based on the characteristics of diffusion, seepage of coal bed methane(CBM) and on the research results of specific pore volume and pore structure. The diffusion pores are further divided into three categories: <8 nm micropore, 8-20 nm transitional pore, and 20-65 nm minipore based on the relationship between increment of specific surface area and diameter of pores, while seepage pores are further divided into three categories: 65-325 nm mesopore,325-1000 nm transitional pore, and >1000 nm macropore based on the abrupt change in the increment of specific pore volume.

  18. Analysis of microscopic pore structures of rocks before and after water absorption

    Institute of Scientific and Technical Information of China (English)

    Li Dejian; Wang Guilian; Han Liqiang; Liu Peiyu; He Manchao; Yang Guoxing; Tai Qimin; Chen Cheng


    Hydrophilic characteristics of rocks are affected by their microscopic pore structures, which clearly change after water absorption. Water absorption tests and scanning electron microscopic (SEM) experiments on rock samples, located at a site in Tibet, China, were carried out. Changes of rock pore structures before and after water absorption were studied with the distribution of pore sizes and fractal characteristics of pores. The results show that surface porosities, fractal dimensions of pores and the complexity of pore structures increased because the number of new small pores produced increased or the original macropore flow channels were expanded after rocks absorbed water. There were points of inflection on their water absorption curves. After water absorption of other rocks, surface porosities and fractal dimensions of pores and complexity of pore structures decreased as the original pore flow channels became filled. Water absorption curves did not change. Surface porosity and the pore fractal dimensions of rocks have good linear relationships before and after water absorption.

  19. Heuristic Approach to Understanding the Accumulation Process in Hydrothermal Pores

    Directory of Open Access Journals (Sweden)

    Doreen Niether


    Full Text Available One of the central questions of humankind is: which chemical and physical conditions are necessary to make life possible? In this “origin-of-life” context, formamide plays an important role, because it has been demonstrated that prebiotic molecules can be synthesized from concentrated formamide solutions. Recently, it could be shown, using finite-element calculations combining thermophoresis and convection processes in hydrothermal pores, that sufficiently high formamide concentrations could be accumulated to form prebiotic molecules (Niether et al. (2016. Depending on the initial formamide concentration, the aspect ratio of the pores, and the ambient temperature, formamide concentrations up to 85 wt % could be reached. The stationary calculations show an effective accumulation, only if the aspect ratio is above a certain threshold, and the corresponding transient studies display a sudden increase of the accumulation after a certain time. Neither of the observations were explained. In this work, we derive a simple heuristic model, which explains both phenomena. The physical idea of the approach is a comparison of the time to reach the top of the pore with the time to cross from the convective upstream towards the convective downstream. If the time to reach the top of the pore is shorter than the crossing time, the formamide molecules are flushed out of the pore. If the time is long enough, the formamide molecules can reach the downstream and accumulate at the bottom of the pore. Analysing the optimal aspect ratio as function of concentration, we find that, at a weight fraction of w = 0 . 5 , a minimal pore height is required for effective accumulation. At the same concentration, the transient calculations show a maximum of the accumulation rate.

  20. Final Report for Subcontract B541028, Pore-Scale Modeling to Support "Pore Connectivity" Research Work

    Energy Technology Data Exchange (ETDEWEB)

    Ewing, R P


    This report covers modeling aspects of a combined experimental and modeling task in support of the DOE Science and Technology Program (formerly OSTI) within the Office of Civilian Radioactive Waste Management (OCRWM). Research Objectives The research for this project dealt with diffusive retardation: solute moving through a fracture diffuses into and out of the rock matrix. This diffusive exchange retards overall solute movement, and retardation both dilutes waste being released, and allows additional decay. Diffusive retardation involves not only fracture conductivity and matrix diffusion, but also other issues and processes: contaminants may sorb to the rock matrix, fracture flow may be episodic, a given fracture may or may not flow depending on the volume of flow and the fracture's connection to the overall fracture network, the matrix imbibes water during flow episodes and dries between episodes, and so on. The objective of the project was to improve understanding of diffusive retardation of radionuclides due to fracture / matrix interactions. Results from combined experimental/modeling work were to (1) determine whether the current understanding and model representation of matrix diffusion is valid, (2) provide insights into the upscaling of laboratory-scale diffusion experiments, and (3) help in evaluating the impact on diffusive retardation of episodic fracture flow and pore connectivity in Yucca Mountain tuffs. Questions explored included the following: (1) What is the relationship between the diffusion coefficient measured at one scale, to that measured or observed at a different scale? In classical materials this relationship is trivial; in low-connectivity materials it is not. (2) Is the measured diffusivity insensitive to the shape of the sample? Again, in classical materials there should be no sample shape effect. (3) Does sorption affect diffusive exchange in low-connectivity media differently than in classical media? (4) What is the effect of

  1. Idealized Shale Sorption Isotherm Measurements to Determine Pore Volume, Pore Size Distribution, and Surface Area (United States)

    Holmes, R.; Wang, B.; Aljama, H.; Rupp, E.; Wilcox, J.


    One method for mitigating the impacts of anthropogenic CO2-related climate change is the sequestration of CO2 in depleted gas and oil reservoirs, including shale. The accurate characterization of the heterogeneous material properties of shale, including pore volume, surface area, pore size distributions (PSDs) and composition is needed to understand the interaction of CO2 with shale. Idealized powdered shale sorption isotherms were created by varying incremental amounts of four essential components by weight. The first two components, organic carbon and clay, have been shown to be the most important components for CO2 uptake in shales. Organic carbon was represented by kerogen isolated from a Silurian shale, and clay groups were represented by illite from the Green River shale formation. The rest of the idealized shale was composed of equal parts by weight of SiO2 to represent quartz and CaCO3 to represent carbonate components. Baltic, Eagle Ford, and Barnett shale sorption measurements were used to validate the idealized samples. The idealized and validation shale sorption isotherms were measured volumetrically using low pressure N2 (77K) and CO2 (273K) adsorbates on a Quantachrome Autosorb IQ2. Gravimetric isotherms were also produced for a subset of these samples using CO2 and CH4adsorbates under subsurface temperature and pressure conditions using a Rubotherm magnetic suspension balance. Preliminary analyses were inconclusive in validating the idealized samples. This could be a result of conflicting reports of total organic carbon (TOC) content in each sample, a problem stemming from the heterogeneity of the samples and different techniques used for measuring TOC content. The TOC content of the validation samples (Eagle Ford and Barnett) was measured by Rock-Eval pyrolysis at Weatherford Laboratories, while the TOC content in the Baltic validation samples was determined by LECO TOC. Development of a uniform process for measuring TOC in the validation samples is

  2. Three-Dimensional Quantification of Pore Space in Flocculated Sediments (United States)

    Lawrence, Tom; Spencer, Kate; Bushby, Andy; Manning, Andrew


    Flocculated sediment structure plays a vital role in determining sediment dynamics within the water column in fresh and saline water bodies. The porosity of flocs contributes to their specific density and therefore their settling characteristics, and can also affect settling characteristics via through-flow. The process of settling and resuspension of flocculated material causes the formation of larger and more complex individual flocs, about which little is known quantitatively of the internal micro-structure and therefore porosity. Hydrological and sedimentological modelling software currently uses estimations of porosity, because it is difficult to capture and analyse flocs. To combat this, we use a novel microscopy method usually performed on biological material to scan the flocs, the output of which can be used to quantify the dimensions and arrangement of pores. This involves capturing flocculated sediment, staining the sample with heavy metal elements to highlight organic content in the Scanning Electron Microscope later, and finally setting the sample in resin. The overall research aim is to quantitatively characterise the dimensions and distribution of pore space in flocs in three dimensions. In order to gather data, Scanning Electron Microscopy and micro-Computed Tomography have been utilised to produce the necessary images to identify and quantify the pore space. The first objective is to determine the dimensional limits of pores in the structure (i.e. what area do they encapsulate? Are they interconnected or discreet?). This requires a repeatable definition to be established, so that all floc pore spaces can be quantified using the same parameters. The LabSFLOC settling column and dyes will be used as one possible method of determining the outer limits of the discreet pore space. LabSFLOC is a sediment settling column that uses a camera to record the flocs, enabling analysis of settling characteristics. The second objective is to develop a reliable

  3. Breaking the hydrophobicity of the MscL pore: insights into a charge-induced gating mechanism.

    Directory of Open Access Journals (Sweden)

    Balasubramanian Chandramouli

    Full Text Available The mechanosensitive channel of large conductance (MscL is a protein that responds to membrane tension by opening a transient pore during osmotic downshock. Due to its large pore size and functional reconstitution into lipid membranes, MscL has been proposed as a promising artificial nanovalve suitable for biotechnological applications. For example, site-specific mutations and tailored chemical modifications have shown how MscL channel gating can be triggered in the absence of tension by introducing charged residues at the hydrophobic pore level. Recently, engineered MscL proteins responsive to stimuli like pH or light have been reported. Inspired by experiments, we present a thorough computational study aiming at describing, with atomistic detail, the artificial gating mechanism and the molecular transport properties of a light-actuated bacterial MscL channel, in which a charge-induced gating mechanism has been enabled through the selective cleavage of photo-sensitive alkylating agents. Properties such as structural transitions, pore dimension, ion flux and selectivity have been carefully analyzed. Besides, the effects of charge on alternative sites of the channel with respect to those already reported have been addressed. Overall, our results provide useful molecular insights into the structural events accompanying the engineered MscL channel gating and the interplay of electrostatic effects, channel opening and permeation properties. In addition, we describe how the experimentally observed ionic current in a single-subunit charged MscL mutant is obtained through a hydrophobicity breaking mechanism involving an asymmetric inter-subunit motion.

  4. An investigation of pore collapse in asymmetric polysulfone membranes (United States)

    Subrahmanyan, Sumitra


    Porous polysulfone membranes prepared by phase inversion can be tailored to suit filtration requirements by the choice of solvent and coagulant. In the current research polysulfone membranes were prepared by inverting a solution in N-methyl pyrrolidinone (NMP) in isopropanol to form uniform sized pores. Phase inversion resulted in the formation of an asymmetric membrane. The membranes have a characteristic "skin" which is supported by a highly porous substructure. Water-wet membranes experience capillary force during water evaporation. Since the modulus of the membranes is lower than the capillary force, the membrane walls shrink and thicken giving rise to a condensed structure. The "skin" regulates permeation through the membranes which is essential for filtration. A change in the pore structure of the skin alters the permeability. The current research investigates the influence of amine plasma treatments on the surface pore structure of polysulfone membranes. The permeation of a rhodamine dye through the plasma treated membranes and through non-plasma treated membranes is used to examine the influence of the plasma treatment. Furthermore, the influence of plasma treatment on the loss of water from the membranes leading to pore collapse is also explored. The study revealed that a plasma ablates the skin, increasing the permeation. An ammonia plasma treatment produced more etching, and hence increased permeation compared to permeation for an aniline plasma-treated membrane. A one-minute aniline plasma treatment only caused a moderate increase in permeation. Plasma treatments introduced significant surface modification by the introduction of new functionalities. However, permeation was not influenced by the surface modification. Water trapped in the pores is essential to maintain the pore structure of the membrane. The surface treatment dictates the pore size and therefore, the convection allowing water evaporation, leading to pore collapse. Heat treating also


    Directory of Open Access Journals (Sweden)

    Piet Stroeven


    Full Text Available This paper concentrates on discrete element computer-simulation of concrete. It is argued on the basis of stochastic heterogeneity theory that modern concurrent-algorithm-based systems should be employed for the assessment of pore characteristics underlying durability performance of cementitious materials. The SPACE system was developed at Delft University of Technology for producing realistic schematizations of realcrete for a wide range of other particle packing problems, involving aggregate and fresh cement, and for the purpose of exploring characteristics in the hardened state of concrete, including of the pore network structure because of obvious durability problems. Since structure-sensitive properties are involved, schematization of reality should explicitly deal with the configuration of the cement particles in the fresh state. The paper concentrates on the stereological and mathematical morphology operations executed to acquire information on particle size, global porosity, and on distribution of porosity and of the connected pore fraction as a result of the near neighbourhood of aggregate grains. Goal is to provide information obtained along different exploration routes of concrete's pore space for setting up a pore network modelling approach. This type of methodological papers is scarce in concrete technology, if not missing at all. Technical publications that report on obtained results in our investigations are systematically referred to.

  6. Role of Pore-Forming Toxins in Neonatal Sepsis

    Directory of Open Access Journals (Sweden)

    Andreas F.-P. Sonnen


    Full Text Available Protein toxins are important virulence factors contributing to neonatal sepsis. The major pathogens of neonatal sepsis, group B Streptococci, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus, secrete toxins of different molecular nature, which are key for defining the disease. Amongst these toxins are pore-forming exotoxins that are expressed as soluble monomers prior to engagement of the target cell membrane with subsequent formation of an aqueous membrane pore. Membrane pore formation is not only a means for immediate lysis of the targeted cell but also a general mechanism that contributes to penetration of epithelial barriers and evasion of the immune system, thus creating survival niches for the pathogens. Pore-forming toxins, however, can also contribute to the induction of inflammation and hence to the manifestation of sepsis. Clearly, pore-forming toxins are not the sole factors that drive sepsis progression, but they often act in concert with other bacterial effectors, especially in the initial stages of neonatal sepsis manifestation.

  7. Fabrication of polymeric scaffolds with a controlled distribution of pores. (United States)

    Capes, J S; Ando, H Y; Cameron, R E


    The design of tissue engineering scaffolds must take into account many factors including successful vascularisation and the growth of cells. Research has looked at refining scaffold architecture to promote more directed growth of tissues through well-defined anisotropy in the pore structure. In many cases it is also desirable to incorporate therapeutic ingredients, such as growth factors, into the scaffold so that their release occurs as the scaffold degrades. Therefore, scaffold fabrication techniques must be found to precisely control, not only the overall porosity of scaffolds, but also the pore size, shape and spatial distribution. This work describes the use of a regularly shaped porogen, sugar spheres, to manufacture polymeric scaffolds. Results show that pre-assembling the spheres created scaffolds with a constant porosity of 60%, but with varying pores sizes from 200-800 microm, leading to a variation in the surface area and likely degradation rate of the scaffolds. Employing different polymer impregnation techniques tailored the number of pores present with a diameter of less than 100 microm to suit different functions, and altering the packing structure of the sugar spheres created scaffolds with novel layered porosity. Replacing sugar spheres with sugar strands formed scaffolds with pores aligned in one direction.

  8. The Bicomponent Pore-Forming Leucocidins of Staphylococcus aureus (United States)

    Alonzo, Francis


    SUMMARY The ability to produce water-soluble proteins with the capacity to oligomerize and form pores within cellular lipid bilayers is a trait conserved among nearly all forms of life, including humans, single-celled eukaryotes, and numerous bacterial species. In bacteria, some of the most notable pore-forming molecules are protein toxins that interact with mammalian cell membranes to promote lysis, deliver effectors, and modulate cellular homeostasis. Of the bacterial species capable of producing pore-forming toxic molecules, the Gram-positive pathogen Staphylococcus aureus is one of the most notorious. S. aureus can produce seven different pore-forming protein toxins, all of which are believed to play a unique role in promoting the ability of the organism to cause disease in humans and other mammals. The most diverse of these pore-forming toxins, in terms of both functional activity and global representation within S. aureus clinical isolates, are the bicomponent leucocidins. From the first description of their activity on host immune cells over 100 years ago to the detailed investigations of their biochemical function today, the leucocidins remain at the forefront of S. aureus pathogenesis research initiatives. Study of their mode of action is of immediate interest in the realm of therapeutic agent design as well as for studies of bacterial pathogenesis. This review provides an updated perspective on our understanding of the S. aureus leucocidins and their function, specificity, and potential as therapeutic targets. PMID:24847020

  9. Pore-forming activity of clostridial binary toxins. (United States)

    Knapp, O; Benz, R; Popoff, M R


    Clostridial binary toxins (Clostridium perfringens Iota toxin, Clostridium difficile transferase, Clostridium spiroforme toxin, Clostridium botulinum C2 toxin) as Bacillus binary toxins, including Bacillus anthracis toxins consist of two independent proteins, one being the binding component which mediates the internalization into cell of the intracellularly active component. Clostridial binary toxins induce actin cytoskeleton disorganization through mono-ADP-ribosylation of globular actin and are responsible for enteric diseases. Clostridial and Bacillus binary toxins share structurally and functionally related binding components which recognize specific cell receptors, oligomerize, form pores in endocytic vesicle membrane, and mediate the transport of the enzymatic component into the cytosol. Binding components retain the global structure of pore-forming toxins (PFTs) from the cholesterol-dependent cytotoxin family such as perfringolysin. However, their pore-forming activity notably that of clostridial binding components is more related to that of heptameric PFT family including aerolysin and C. perfringens epsilon toxin. This review focuses upon pore-forming activity of clostridial binary toxins compared to other related PFTs. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale.

  10. Regulation of soil organic C mineralisation at the pore scale. (United States)

    Ruamps, Léo S; Nunan, Naoise; Pouteau, Valérie; Leloup, Julie; Raynaud, Xavier; Roy, Virginie; Chenu, Claire


    Little is known about the factors that regulate C mineralisation at the soil pore scale or how these factors vary throughout the pore network. This study sought to understand how the decomposition of organic carbon varies within the soil pore network and to determine the relative importance of local environmental properties relative to biological properties as controlling factors. This was achieved by sterilising samples of soil and reinoculating them with axenic bacterial suspensions using the matric potential to target different locations in the pore network. Carbon mineralisation curves were described with two-compartment first-order models to distinguish CO2 derived from the labile organic carbon released during sterilisation from CO2 derived from organic C unaffected by sterilisation. The data indicated that the size of the labile pool of organic C, possibly of microbial origin, varied as a function of location in the pore network but that the organic carbon unaffected by sterilisation did not. The mineralisation rate of the labile C varied with the bacterial type inoculated, but the mineralisation rate of the organic C unaffected by sterilisation was insensitive to bacterial type. Taken together, the results suggest that microbial metabolism is a less significant regulator of soil organic carbon decomposition than are microbial habitat properties.

  11. Tunable ultrathin membranes with nonvolatile pore shape memory. (United States)

    Kuroki, Hidenori; Islam, Crescent; Tokarev, Igor; Hu, Heng; Liu, Guojun; Minko, Sergiy


    The concept of a responsive nanoporous thin-film gel membranes whose pores could be tuned to a desired size by a specific "molecular signal" and whose pore geometry becomes "memorized" by the gel is reported. The ∼100 nm thick membranes were prepared by dip-coating from a solution mixture of a random copolymer comprising responsive and photo-cross-linkable units and monodisperse latex nanoparticles used as a sacrificial colloidal template. After stabilization of the films by photo-cross-linking the latex template was removed, yielding nanoporous structures with a narrow pore size distribution and a high porosity. The thin-film membranes could be transferred onto porous supports to serve as tunable size-selective barriers in various colloids separation applications. The pore dimensions and hence the membrane's colloidal-particle-size cutoff were reversibly regulated by swelling-shrinking of the polymer network with a specially selected low-molar-mass compound. The attained pore shape was "memorized" in aqueous media and "erased" by treatment in special solvents reverting the membrane to the original state.

  12. To the Pore and Through the Pore: A Story of mRNA Export Kinetics (United States)

    Oeffinger, Marlene; Zenklusen, Daniel


    Summary The evolutionary ‘decision’ to store genetic information away from the place of protein synthesis, in a separate compartment, has forced eukaryotic cells to establish a system to transports mRNAs from the nucleus to the cytoplasm for translation. To ensure export to be fast and efficient, cells have evolved a complex molecular interplay that is tightly regulated. Over the last few decades, many of the individual players in this process have been described, starting with the composition of the nuclear pore complex to proteins that modulate co-transcriptional events required to prepare an mRNP for export to the cytoplasm. How the interplay between all the factors and processes results in the efficient and selective export of mRNAs from the nucleus and how the export process itself is executed within cells, however, is still not fully understood. Recent advances in using proteomic and single molecule microscopy approaches have provided important insights into the process and its kinetics. This review summarizes these recent advances and how they led to the current view on how cells orchestrate the export of mRNAs. PMID:22387213

  13. Why liquid displacement methods are sometimes wrong in estimating the pore-size distribution

    NARCIS (Netherlands)

    Gijsbertsen-Abrahamse, A.J.; Boom, R.M.; Padt, van der A.


    The liquid displacement method is a commonly used method to determine the pore size distribution of micro- and ultrafiltration membranes. One of the assumptions for the calculation of the pore sizes is that the pores are parallel and thus are not interconnected. To show that the estimated pore size

  14. What controls open-pore and residual currents in the first sensing zone of alpha-hemolysin nanopore? Combined experimental and theoretical study (United States)

    de Biase, Pablo M.; Ervin, Eric N.; Pal, Prithwish; Samoylova, Olga; Markosyan, Suren; Keehan, Michael G.; Barrall, Geoffrey A.; Noskov, Sergei Yu.


    The electrophoretic transport of single-stranded DNA through biological nanopores such as alpha-hemolysin (αHL) is a promising and cost-effective technology with the potential to revolutionize genomics. The rational design of pores with the controlled polymer translocation rates and high contrast between different nucleotides could improve significantly nanopore sequencing applications. Here, we apply a combination of theoretical and experimental methods in an attempt to elucidate several selective modifications in the pore which were proposed to be central for the effective discrimination between purines and pyrimidines. Our nanopore test set includes the wild type αHL and six mutants (E111N/M113X/K147N) in which the cross-section and chemical functionality of the first constriction zone of the pore are modified. Electrophysiological recordings were combined with all-atom Molecular Dynamics simulations (MD) and a recently developed Brownian Dynamics (BROMOC) protocol to investigate residual ion currents and pore-DNA interactions for two homo-polymers e.g. poly(dA)40 or poly(dC)40 blocking the pore. The calculated residual currents and contrast in the poly(dA)40/poly(dC)40 blocked pore are in qualitative agreement with the experimental recordings. We showed that a simple structural metric allows rationalization of key elements in the emergent contrast between purines and pyrimidines in the modified αHL mutants. The shape of the pore and its capacity for hydrogen bonding to a translocated polynucleotide are two essential parameters for contrast optimization. To further probe the impact of these two factors in the ssDNA sensing, we eliminated the effect of the primary constriction using serine substitutions (i.e. E111S/M113S/T145S/K147S) and increased the hydrophobic volume of the central residue in the secondary constriction (L135I). This pore modification sharply increased the contrast between Adenine (A) and Cytosine (C).The electrophoretic transport of single

  15. Nanometer to Centimeter Scale Analysis and Modeling of Pore Structures (United States)

    Wesolowski, D. J.; Anovitz, L.; Vlcek, L.; Rother, G.; Cole, D. R.


    The microstructure and evolution of pore space in rocks is a critically important factor controlling fluid flow. The size, distribution and connectivity of these confined geometries dictate how fluids including H2O and CO2, migrate into and through these micro- and nano-environments, wet and react with the solid. (Ultra)small-angle neutron scattering and autocorrelations derived from BSE imaging provide a method of quantifying pore structures in a statistically significant manner from the nanometer to the centimeter scale. Multifractal analysis provides additional constraints. These methods were used to characterize the pore features of a variety of potential CO2 geological storage formations and geothermal systems such as the shallow buried quartz arenites from the St. Peter Sandstone and the deeper Mt. Simon quartz arenite in Ohio as well as the Eau Claire shale and mudrocks from the Cranfield MS CO2 injection test and the normal temperature and high-temperature vapor-dominated parts of the Geysers geothermal system in California. For example, analyses of samples of St. Peter sandstone show total porosity correlates with changes in pores structure including pore size ratios, surface fractal dimensions, and lacunarity. These samples contain significant large-scale porosity, modified by quartz overgrowths, and neutron scattering results show significant sub-micron porosity, which may make up fifty percent or more of the total pore volume. While previous scattering data from sandstones suggest scattering is dominated by surface fractal behavior, our data are both fractal and pseudo-fractal. The scattering curves are composed of steps, modeled as polydispersed assemblages of pores with log-normal distributions. In some samples a surface-fractal overprint is present. There are also significant changes in the mono and multifractal dimensions of the pore structure as the pore fraction decreases. There are strong positive correlations between D(0) and image and total

  16. Fines Classification Based on Sensitivity to Pore-Fluid Chemistry

    KAUST Repository

    Jang, Junbong


    The 75-μm particle size is used to discriminate between fine and coarse grains. Further analysis of fine grains is typically based on the plasticity chart. Whereas pore-fluid-chemistry-dependent soil response is a salient and distinguishing characteristic of fine grains, pore-fluid chemistry is not addressed in current classification systems. Liquid limits obtained with electrically contrasting pore fluids (deionized water, 2-M NaCl brine, and kerosene) are combined to define the soil "electrical sensitivity." Liquid limit and electrical sensitivity can be effectively used to classify fine grains according to their fluid-soil response into no-, low-, intermediate-, or high-plasticity fine grains of low, intermediate, or high electrical sensitivity. The proposed methodology benefits from the accumulated experience with liquid limit in the field and addresses the needs of a broader range of geotechnical engineering problems. © ASCE.

  17. Highly Aminated Mesoporous Silica Nanoparticles with Cubic Pore Structure

    KAUST Repository

    Suteewong, Teeraporn


    Mesoporous silica with cubic symmetry has attracted interest from researchers for some time. Here, we present the room temperature synthesis of mesoporous silica nanoparticles possessing cubic Pm3n symmetry with very high molar ratios (>50%) of 3-aminopropyl triethoxysilane. The synthesis is robust allowing, for example, co-condensation of organic dyes without loss of structure. By means of pore expander molecules, the pore size can be enlarged from 2.7 to 5 nm, while particle size decreases. Adding pore expander and co-condensing fluorescent dyes in the same synthesis reduces average particle size further down to 100 nm. After PEGylation, such fluorescent aminated mesoporous silica nanoparticles are spontaneously taken up by cells as demonstrated by fluorescence microscopy.

  18. Formation of protein induced micro-pores in Chitosan membranes (United States)

    Begum, S. N. Suraiya; Aswal, V. K.; Ramasamy, Radha Perumal


    Polymer based nanocomposites are important class of materials and have wide applications. Blending two biopolymers can lead to the development of new materials with tailored properties. Chitosan is a naturally occurring polysaccharide with useful properties such as biodegradability and excellent film forming capacity. Bovine serum albumin (BSA) is a abundantly available globular protein. In our research the interaction of chitosan with BSA and the effect of formation of Au nanoparticles on chitosan-BSA system were investigated. Scanning electron microscope (SEM) of the films showed formation of micron sized pores and these pores were hindered with formation of Au nanoparticles. Small angle neutron scattering (SANS) analysis showed that BSA interacts with chitosan chain and affects the Rg value of chitosan. The formation of micro pores decreases the conductivity values (σ'), while the formation of Au nanoparticles increases σ'.

  19. Sound absorption property of open-pore aluminum foams

    Institute of Scientific and Technical Information of China (English)

    WANG Lu-cai; WANG Fang; WU Jian-guo; YOU Xiao-hong


    The sound absorption property of aluminum foam was studied by testing its sound absorption coefficients using standing wave tube method. The open-pore aluminum foams were prepared by infiltration process, with pore size of 0.5 mm to 3.2 mm and porosity of 54.2% to 77%. The frequency of indicted sound wave was ranging from 125 Hz to 10 kHz. The results show that the average values of sound absorption coefficients are all over 0.4 and the aluminum foam has better sound absorption property, its coefficients is influenced by frequency and pore structure, and reaches the maximum at about 1 kHz, with increasing porosity and decreasing cell diameter the sound absorption coefficient values increase.

  20. Integration of pore features into the evaluation of fingerprint evidence. (United States)

    Anthonioz, Alexandre; Champod, Christophe


    Fingerprint practitioners rely on level 3 features to make decisions in relation to the source of an unknown friction ridge skin impression. This research proposes to assess the strength of evidence associated with pores when shown in (dis)agreement between a mark and a reference print. Based upon an algorithm designed to automatically detect pores, a metric is defined in order to compare different impressions. From this metric, the weight of the findings is quantified using a likelihood ratio. The results obtained on four configurations and 54 donors show the significant contribution of the pore features and translate into statistical terms what latent fingerprint examiners have developed holistically through experience. The system provides LRs that are indicative of the true state under both the prosecution and the defense propositions. Not only such a system brings transparency regarding the weight to assign to such features, but also forces a discussion in relation to the risks of such a model to mislead.

  1. Pore-scale simulation of calcium carbonate precipitation and dissolution under highly supersaturated conditions in a microfludic pore network (United States)

    Yoon, H.; Dewers, T. A.; Valocchi, A. J.; Werth, C. J.


    Dissolved CO2 during geological CO2 storage may react with minerals in fractured rocks or confined aquifers and cause mineral precipitation. The overall rate of reaction can be affected by coupled processes among hydrodynamics, transport, and reactions at pore-scale. Pore-scale models of coupled fluid flow, reactive transport, and CaCO3 precipitation and dissolution are applied to account for transient experimental results of CaCO3 precipitation and dissolution under highly supersaturated conditions in a microfluidic pore network (i.e., micromodel). Pore-scale experiments in the micromodel are used as a basis for understanding coupled physics of systems perturbed by geological CO2 injection. In the micromodel, precipitation is induced by transverse mixing along the centerline in pore bodies. Overall, the pore-scale model qualitatively captured the governing physics of reactions such as precipitate morphology, precipitation rate, and maximum precipitation area in first few pore spaces. In particular, we found that proper estimation of the effective diffusion coefficient and the reactive surface area is necessary to adequately simulate precipitation and dissolution rates. As the model domain increases, the effect of flow patterns affected by precipitation on the overall reaction rate also increases. The model is also applied to account for the effect of different reaction rate laws on mineral precipitation and dissolution at pore-scale. Reaction rate laws tested include the linear rate law, nonlinear power law, and newly-developed rate law based on in-situ measurements at nano scale in the literature. Progress on novel methods for upscaling pore-scale models for reactive transport are discussed, and are being applied to mineral precipitation patterns observed in natural analogues. H.Y. and T. D. were supported as part of the Center for Frontiers of Subsurface Energy Security, an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of

  2. Phanerochaete mutants with enhanced ligninolytic activity

    Energy Technology Data Exchange (ETDEWEB)

    Kakar, S.N.; Perez, A.; Gonzales, J.


    In addition to lignin, the white rot fungus Phanerochaete chrysosporium has the ability to degrade a wide spectrum of recalcitrant organopollutants in soils and aqueous media. Although some of the organic compounds are degraded under nonligninolytic conditions, most are degraded under ligninolytic conditions with the involvement of the extracellular enzymes, lignin peroxidases, and manganese-dependent peroxidases, which are produced as secondary metabolites triggered by conditions of nutrient starvation (e.g., nitrogen limitation). The fungus and its enzymes can thus provide alternative technologies for bioremediation, biopulping, biobleaching, and other industrial applications. The efficiency and effectiveness of the fungus can be enhanced by increasing production and secretion of the important enzymes in large quantities and as primary metabolites under enriched conditions. One way this can be achieved is through isolation of mutants that are deregulated or are hyperproducers or supersecretors of key enzymes under enriched conditions. Through ultraviolet-light and gamma-rays mutagenesis we have isolated a variety of mutants, some of which produce key enzymes of the ligninolytic system under high-nitrogen growth conditions. One of the mutants produced 272 units (U) of lignin peroxidases enzyme activity per liter after nine days under high nitrogen. The mutant and the parent strains produced up to 54 U/L and 62 U/L, respectively, of the enzyme activity under low-nitrogen growth conditions during this period. In some experiments the mutant showed 281 U/L of enzyme activity under high nitrogen after 17 days.

  3. Evaluation of Colloid Retention Site Dominance in Variably Saturated Porous Media: An All Pores Pore-Scale Analysis (United States)

    Morales, Veronica; Perez-Reche, Francisco; Holzner, Markus; Kinzelbach, Wolfgang


    It is well accepted that colloid and nanoparticle transport processes in porous media differ substantially between water saturated and unsaturated conditions. Differences are frequently ascribed to particle immobilization by association with interfaces with the gas, as well as to restrictions of the liquid medium through which colloids are transported. Yet, the current understanding of the importance of particle retention at gas interfaces is based on observations of single pores or two-dimensional pore network representations, leaving open the question of their statistical significance when all pores in the medium are considered. In order to address this question, column experiments were performed using a model porous medium of glass beads through which Silver particles were transported for conditions of varying water content and water chemistry. X-ray microtomography was subsequently employed as a non-destructive imaging technique to obtain pore-scale information of the entire column regarding: i) the presence and distribution of the main locations where colloids can become retained (interfaces with the water-solid, air-water, air-solid, and air-water-solid, grain-grain contacts, and the bulk liquid), ii) deposition profiles of colloids along the column classified by the available retention location, and iii) channel widths of 3-dimensional pore-water network representations. The results presented provide a direct statistical evaluation on the significance of colloid retention by attachment to interfaces or by strainig at contact points where multiple interfaces meet.

  4. Pore-Width-Dependent Preferential Interaction of sp2 Carbon Atoms in Cyclohexene with Graphitic Slit Pores by GCMC Simulation

    Directory of Open Access Journals (Sweden)

    Natsuko Kojima


    Full Text Available The adsorption of cyclohexene with two sp2 and four sp3 carbon atoms in graphitic slit pores was studied by performing grand canonical Monte Carlo simulation. The molecular arrangement of the cyclohexene on the graphitic carbon wall depends on the pore width. The distribution peak of the sp2 carbon is closer to the pore wall than that of the sp3 carbon except for the pore width of 0.7 nm, even though the Lennard-Jones size of the sp2 carbon is larger than that of the sp3 carbon. Thus, the difference in the interactions of the sp2 and sp3 carbon atoms of cyclohexene with the carbon pore walls is clearly observed in this study. The preferential interaction of sp2 carbon gives rise to a slight tilting of the cyclohexene molecule against the graphitic wall. This is suggestive of a π-π interaction between the sp2 carbon in the cyclohexene molecule and graphitic carbon.

  5. Pore invasion dynamics during fluid front displacement in porous media determine functional pore size distribution and phase entrapment (United States)

    Moebius, F.; Or, D.


    Dynamics of fluid fronts in porous media shape transport properties of the unsaturated zone and affect management of petroleum reservoirs and their storage properties. What appears macroscopically as smooth and continuous motion of a displacement fluid front may involve numerous rapid interfacial jumps often resembling avalanches of invasion events. Direct observations using high-speed camera and pressure sensors in sintered glass micro-models provide new insights on the influence of flow rates, pore size, and gravity on invasion events and on burst size distribution. Fundamental differences emerge between geometrically-defined pores and "functional" pores invaded during a single burst (invasion event). The waiting times distribution of individual invasion events and decay times of inertial oscillations (following a rapid interfacial jump) are characteristics of different displacement regimes. An invasion percolation model with gradients and including the role of inertia provide a framework for linking flow regimes with invasion sequences and phase entrapment. Model results were compared with measurements and with early studies on invasion burst sizes and waiting times distribution during slow drainage processes by Måløy et al. [1992]. The study provides new insights into the discrete invasion events and their weak links with geometrically-deduced pore geometry. Results highlight factors controlling pore invasion events that exert strong influence on macroscopic phenomena such as front morphology and residual phase entrapment shaping hydraulic properties after the passage of a fluid front.

  6. Pore-scale modeling of competitive adsorption in porous media. (United States)

    Ryan, Emily M; Tartakovsky, Alexandre M; Amon, Cristina


    In this paper we present a smoothed particle hydrodynamics (SPH) pore-scale multicomponent reactive transport model with competitive adsorption. SPH is a Lagrangian, particle based modeling method which uses the particles as interpolation points to discretize and solve flow and transport equations. The theory and details of the SPH pore-scale model are presented along with a novel method for handling surface reactions, the continuum surface reaction (CSR) model. The numerical accuracy of the CSR model is validated with analytical and finite difference solutions, and the effects of spatial and temporal resolution on the accuracy of the model are also discussed. The pore-scale model is used to study competitive adsorption for different Damköhler and Peclet numbers in a binary system where a plume of species B is introduced into a system which initially contains species A. The pore-scale model results are compared with a Darcy-scale model to investigate the accuracy of a Darcy-scale reactive transport model for a wide range of Damköhler and Peclet numbers. The comparison shows that the Darcy model over estimates the mass fraction of aqueous and adsorbed species B and underestimates the mass fractions of species A. The Darcy-scale model also predicts faster transport of species A and B through the system than the pore-scale model. The overestimation of the advective velocity and the extent of reactions by the Darcy-scale model are due to incomplete pore-scale mixing. As the degree of the solute mixing decreases with increasing Peclet and Damköhler numbers, so does the accuracy of the Darcy-scale model. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Microfluidic Experiments Studying Pore Scale Interactions of Microbes and Geochemistry (United States)

    Chen, M.; Kocar, B. D.


    Understanding how physical phenomena, chemical reactions, and microbial behavior interact at the pore-scale is crucial to understanding larger scale trends in groundwater chemistry. Recent studies illustrate the utility of microfluidic devices for illuminating pore-scale physical-biogeochemical processes and their control(s) on the cycling of iron, uranium, and other important elements 1-3. These experimental systems are ideal for examining geochemical reactions mediated by microbes, which include processes governed by complex biological phenomenon (e.g. biofilm formation, etc.)4. We present results of microfluidic experiments using a model metal reducing bacteria and varying pore geometries, exploring the limitations of the microorganisms' ability to access tight pore spaces, and examining coupled biogeochemical-physical controls on the cycling of redox sensitive metals. Experimental results will provide an enhanced understanding of coupled physical-biogeochemical processes transpiring at the pore-scale, and will constrain and compliment continuum models used to predict and describe the subsurface cycling of redox-sensitive elements5. 1. Vrionis, H. A. et al. Microbiological and geochemical heterogeneity in an in situ uranium bioremediation field site. Appl. Environ. Microbiol. 71, 6308-6318 (2005). 2. Pearce, C. I. et al. Pore-scale characterization of biogeochemical controls on iron and uranium speciation under flow conditions. Environ. Sci. Technol. 46, 7992-8000 (2012). 3. Zhang, C., Liu, C. & Shi, Z. Micromodel investigation of transport effect on the kinetics of reductive dissolution of hematite. Environ. Sci. Technol. 47, 4131-4139 (2013). 4. Ginn, T. R. et al. Processes in microbial transport in the natural subsurface. Adv. Water Resour. 25, 1017-1042 (2002). 5. Scheibe, T. D. et al. Coupling a genome-scale metabolic model with a reactive transport model to describe in situ uranium bioremediation. Microb. Biotechnol. 2, 274-286 (2009).

  8. Pseudomonas aeruginosa Pore-Forming Exolysin and Type IV Pili Cooperate To Induce Host Cell Lysis (United States)

    Basso, Pauline; Ragno, Michel; Elsen, Sylvie; Reboud, Emeline; Golovkine, Guillaume; Bouillot, Stephanie; Huber, Philippe; Lory, Stephen; Faudry, Eric


    ABSTRACT   Clinical strains of Pseudomonas aeruginosa lacking the type III secretion system genes employ a toxin, exolysin (ExlA), for host cell membrane disruption. Here, we demonstrated that ExlA export requires a predicted outer membrane protein, ExlB, showing that ExlA and ExlB define a new active two-partner secretion (TPS) system of P. aeruginosa. In addition to the TPS signals, ExlA harbors several distinct domains, which include one hemagglutinin domain, five arginine-glycine-aspartic acid (RGD) motifs, and a C-terminal region lacking any identifiable sequence motifs. However, this C-terminal region is important for the toxic activity, since its deletion abolishes host cell lysis. Using lipid vesicles and eukaryotic cells, including red blood cells, we demonstrated that ExlA has a pore-forming activity which precedes cell membrane disruption of nucleated cells. Finally, we developed a high-throughput cell-based live-dead assay and used it to screen a transposon mutant library of an ExlA-producing P. aeruginosa clinical strain for bacterial factors required for ExlA-mediated toxicity. The screen resulted in the identification of proteins involved in the formation of type IV pili as being required for ExlA to exert its cytotoxic activity by promoting close contact between bacteria and the host cell. These findings represent the first example of cooperation between a pore-forming toxin of the TPS family and surface appendages in host cell intoxication. PMID:28119472

  9. The sensing of membrane microdomains based on pore-forming toxins. (United States)

    Skočaj, M; Bakrač, B; Križaj, I; Maček, P; Anderluh, G; Sepčić, K


    Membrane rafts are transient and unstable membrane microdomains that are enriched in sphingolipids, cholesterol, and specific proteins. They are involved in intracellular trafficking, signal transduction, pathogen entry, and attachment of various ligands. Increasing experimental evidence on the crucial biological roles of membrane rafts under normal and pathological conditions require new techniques for their structural and functional characterization. In particular, fluorescence-labeled cytolytic proteins that interact specifically with molecules enriched in rafts are of increasing interest. Cholera toxin subunit B interacts specifically with raft-residing ganglioside G(M1), and it has long been the lipid probe of choice for membrane rafts. Recently, four new pore-forming toxins have been proposed as selective raft markers: (i) equinatoxin II, a cytolysin from the sea anemone Actinia equina, which specifically recognizes free and membrane-embedded sphingomyelin; (ii) a truncated non-toxic mutant of a cytolytic protein, lysenin, from the earthworm Eisenia foetida, which specifically recognizes sphingomyelin-enriched membrane domains; (iii) a non-toxic derivative of the cholesterol-dependent cytolysin perfringolysin O, from the bacterium Clostridium perfringens, which selectively binds to membrane domains enriched in cholesterol; and (iv) ostreolysin, from the mushroom Pleurotus ostreatus, which does not bind to a single raft-enriched lipid component, but requires a specific combination of two of the most important raft-residing lipids: sphingomyelin and cholesterol. Nontoxic, raft-binding derivatives of cytolytic proteins have already been successfully used to explore both the structure and function of membrane rafts, and of raft-associated molecules. Here, we review these four new derivatives of pore-forming toxins as new putative markers of these membrane microdomains.

  10. Dependence of CO2 Reactivity of Carbon Anodes on Pore Structure (United States)

    Chen, Tong; Xue, Jilai; Lang, Guanghui; Liu, Rui; Gao, Shoulei; Wang, Zengjie


    The correlation between the CO2 reactivity and pore structure of carbon anodes was experimentally investigated. The pore structures of the anodes before and after CO2 oxidation were characterized using image analysis. The porosity, mean pore diameter, and the number of micro-cracks decreased with increasing anode forming pressure, while they increased with over-compaction. With prolonged CO2 oxidation time, the porosity, pore density, mean pore diameter, pore aspect ratio, and the number of micro-cracks increased due to the merging of small pores, increased pore connectivity, and generation of new pores. The activation energy decreased with increasing porosity of the anodes' pitch phase due to easier CO2 penetration and reaction within the anodes. The results confirm that the fine pitch-coke phase of anodes is preferentially consumed, a cause of carbon dusting. Optimization of the pore structures to balance the pitch, coke, and butt phases may potentially further reduce carbon dusting.

  11. A user-friendly modified pore-solid fractal model


    Dian-yuan Ding; Ying Zhao; Hao Feng; Bing-cheng Si; Robert Lee Hill


    The primary objective of this study was to evaluate a range of calculation points on water retention curves (WRC) instead of the singularity point at air-entry suction in the pore-solid fractal (PSF) model, which additionally considered the hysteresis effect based on the PSF theory. The modified pore-solid fractal (M-PSF) model was tested using 26 soil samples from Yangling on the Loess Plateau in China and 54 soil samples from the Unsaturated Soil Hydraulic Database. The derivation results s...

  12. Probing single nanometer-scale pores with polymeric molecular rulers (United States)

    Henrickson, Sarah E.; DiMarzio, Edmund A.; Wang, Qian; Stanford, Vincent M.; Kasianowicz, John J.


    We previously demonstrated that individual molecules of single-stranded DNA can be driven electrophoretically through a single Staphylococcus aureus α-hemolysin ion channel. Polynucleotides thread through the channel as extended chains and the polymer-induced ionic current blockades exhibit stable modes during the interactions. We show here that polynucleotides can be used to probe structural features of the α-hemolysin channel itself. Specifically, both the pore length and channel aperture profile can be estimated. The results are consistent with the channel crystal structure and suggest that polymer-based "molecular rulers" may prove useful in deducing the structures of nanometer-scale pores in general.

  13. A Dynamic Pore-Scale Model of Imbibition

    DEFF Research Database (Denmark)

    Mogensen, Kristian; Stenby, Erling Halfdan


    could not incorporate long-range correlations among pore and throat sizes in our network, but were limited to small-range correlations. Consequently, the gradual suppression of snap-off occurs within one order of magnitude of the capillary number. At capillary numbers around l0- to l0-, snap-off has......We present a dynamic pore-scale network model of imbibition, capable of calculating residual oil saturation for any given capillary number, viscosity ratio, contact angle and aspect ratio. Our goal is not to predict the outcome of core floods, but rather to perform a sensitivity analysis...

  14. Hydrochromic conjugated polymers for human sweat pore mapping. (United States)

    Lee, Joosub; Pyo, Minkyeong; Lee, Sang-hwa; Kim, Jaeyong; Ra, Moonsoo; Kim, Whoi-Yul; Park, Bum Jun; Lee, Chan Woo; Kim, Jong-Man


    Hydrochromic materials have been actively investigated in the context of humidity sensing and measuring water contents in organic solvents. Here we report a sensor system that undergoes a brilliant blue-to-red colour transition as well as 'Turn-On' fluorescence upon exposure to water. Introduction of a hygroscopic element into a supramolecularly assembled polydiacetylene results in a hydrochromic conjugated polymer that is rapidly responsive (polymer. As a result, the sensor can be used to construct a precise map of active sweat pores on fingertips. The sensor technology, developed in this study, has the potential of serving as new method for fingerprint analysis and for the clinical diagnosis of malfunctioning sweat pores.

  15. Binary hard-sphere mixtures within spherical pores

    CERN Document Server

    Kim, S C; Lee, C H


    The free-energy model, which is based on the fundamental geometric measures of the particles, has been employed to investigate the structural properties of nonuniform hard-sphere mixtures within spherical pores. Monte Carlo simulation has been performed to calculate the density profiles of hard-sphere mixtures confined in spherical pores, and the simulation has been compared with the calculated results. Comparisons between the theoretical results and the simulation data have shown that the free-energy model demonstrates reliable accuracy and reproduces the simulation data accurately even for larger size ratios of hard spheres.

  16. Hard Sphere Diffusion Behaviour of Polymer Translocating through Interacting Pores

    Institute of Scientific and Technical Information of China (English)

    SUN Li-Zhen; LUO Meng-Bo


    The translocation of polymer chain through a small pore from a high concentration side (cis side) to a low concentration side (trans side) is simulated by using Monte Carlo technique. The effect of the polymer-pore interaction on the translocation is studied. We find a special interaction at which the decay of the number of polymer chain, N, at the cis side obeys Fick's law, i.e. N decreases exponentially with time. The behaviour is analogous to the diffusion of hard sphere.

  17. Enhanced Retention of Chemotactic Bacteria in a Pore Network with Residual NAPL Contamination (United States)

    Ford, R.; Wang, X.


    Nonaqueous phase liquid (NAPL) contaminants are difficult to eliminate from natural aquifers due, in part, to the heterogeneous structure of the soil matrix. Residual NAPL ganglia remain trapped in regions where the hydraulic conductivity is relatively low. Bioremediation processes depend on adequate mixing of microbial populations and the groundwater contaminants that they degrade. The ability of bacteria to sense a chemical gradient and swim preferentially toward locations of higher concentration, known as chemotaxis, can enhance the mixing of bacteria with contaminant sources that may not be readily accessible by advection and dispersion alone. The impact of chemotaxis on bacterial abundance within a low conductivity NAPL-contaminated region of a well-characterized porous matrix was investigated. A microfluidic device was designed to mimic heterogeneous features of a contaminated groundwater system. NAPL ganglia (toluene) were trapped within a fine pore network, and bacteria were injected into the system through a highly conductive adjacent channel. Chemotactic bacteria (P. putida F1) migrated preferentially towards and accumulated in the vicinity of NAPL contaminant sources. The accumulation of chemotactic bacteria was 15% greater in comparison to a nonchemotactic mutant (P. putida F1 CheA). Bacteria in the microfluidic device were subjected to different flow velocities from 0.25 to 5 m/d encompassing the range of typical groundwater flow rates. Chemotactic bacteria exhibited greater accumulation near the intersection between the macrochannel and the porous network at a flow velocity of 0.5 m/d than both the nonchemotactic mutant control and the chemotactic bacteria at a higher flow velocity of 5 m/d. Breakthrough curves observed at the outlet provided indirect evidence that chemotactic bacteria were retained within the contaminated low permeable region for a longer time than the nonchemotactic bacteria at a flow velocity of 0.25 m/d. This retention was

  18. Enhanced cellulase production in mutants of Thermomonospora

    Energy Technology Data Exchange (ETDEWEB)

    Fennington, G.; Lupo, D.; Stutzenberger, F.


    Thermomonospora curvata, a thermophilic actinomycete, secretes multiple forms of endo-beta, 1-4-glucanase (EG) when grown on cellulose-mineral salts liquid medium. The EG activity (measured as carboxymethyl cellulose hydrolysis) was separated by ion exchange chromatography into three distinct components which differed in their kinetic properties. Exposure of T. curvata to ultraviolet light, N-nitrosoguanidine, or ethane methyl sulfonate produced mutants with enhanced EG production. Selection of colonies which cleared cellulose agar plants containing 2-deoxyglucose or glycerol yielded mutants having 1.5 to 2.6 times the extracellular EG and saccharifying activity (measured by filter-paper and cotton-fiber hydrolysis). The secretion of extracellular protein was increased proportionally in mutant cultures. (Refs. 40).

  19. High Persister Mutants in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Heather L Torrey

    Full Text Available Mycobacterium tuberculosis forms drug-tolerant persister cells that are the probable cause of its recalcitrance to antibiotic therapy. While genetically identical to the rest of the population, persisters are dormant, which protects them from killing by bactericidal antibiotics. The mechanism of persister formation in M. tuberculosis is not well understood. In this study, we selected for high persister (hip mutants and characterized them by whole genome sequencing and transcriptome analysis. In parallel, we identified and characterized clinical isolates that naturally produce high levels of persisters. We compared the hip mutants obtained in vitro with clinical isolates to identify candidate persister genes. Genes involved in lipid biosynthesis, carbon metabolism, toxin-antitoxin systems, and transcriptional regulators were among those identified. We also found that clinical hip isolates exhibited greater ex vivo survival than the low persister isolates. Our data suggest that M. tuberculosis persister formation involves multiple pathways, and hip mutants may contribute to the recalcitrance of the infection.

  20. Escherichia coli mutants with a temperature-sensitive alcohol dehydrogenase.


    Lorowitz, W; Clark, D.


    Mutants of Escherichia coli resistant to allyl alcohol were selected. Such mutants were found to lack alcohol dehydrogenase. In addition, mutants with temperature-sensitive alcohol dehydrogenase activity were obtained. These mutations, designated adhE, are all located at the previously described adh regulatory locus. Most adhE mutants were also defective in acetaldehyde dehydrogenase activity.

  1. Native Mutant Huntingtin in Human Brain (United States)

    Sapp, Ellen; Valencia, Antonio; Li, Xueyi; Aronin, Neil; Kegel, Kimberly B.; Vonsattel, Jean-Paul; Young, Anne B.; Wexler, Nancy; DiFiglia, Marian


    Huntington disease (HD) is caused by polyglutamine expansion in the N terminus of huntingtin (htt). Analysis of human postmortem brain lysates by SDS-PAGE and Western blot reveals htt as full-length and fragmented. Here we used Blue Native PAGE (BNP) and Western blots to study native htt in human postmortem brain. Antisera against htt detected a single band broadly migrating at 575–850 kDa in control brain and at 650–885 kDa in heterozygous and Venezuelan homozygous HD brains. Anti-polyglutamine antisera detected full-length mutant htt in HD brain. There was little htt cleavage even if lysates were pretreated with trypsin, indicating a property of native htt to resist protease cleavage. A soluble mutant htt fragment of about 180 kDa was detected with anti-htt antibody Ab1 (htt-(1–17)) and increased when lysates were treated with denaturants (SDS, 8 m urea, DTT, or trypsin) before BNP. Wild-type htt was more resistant to denaturants. Based on migration of in vitro translated htt fragments, the 180-kDa segment terminated ≈htt 670–880 amino acids. If second dimension SDS-PAGE followed BNP, the 180-kDa mutant htt was absent, and 43–50 kDa htt fragments appeared. Brain lysates from two HD mouse models expressed native full-length htt; a mutant fragment formed if lysates were pretreated with 8 m urea + DTT. Native full-length mutant htt in embryonic HD140Q/140Q mouse primary neurons was intact during cell death and when cell lysates were exposed to denaturants before BNP. Thus, native mutant htt occurs in brain and primary neurons as a soluble full-length monomer. PMID:22375012

  2. Aging Kit mutant mice develop cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Lei Ye

    Full Text Available Both bone marrow (BM and myocardium contain progenitor cells expressing the c-Kit tyrosine kinase. The aims of this study were to determine the effects of c-Kit mutations on: i. myocardial c-Kit(+ cells counts and ii. the stability of left ventricular (LV contractile function and structure during aging. LV structure and contractile function were evaluated (echocardiography in two groups of Kit mutant (W/Wv and W41/W42 and in wild type (WT mice at 4 and 12 months of age and the effects of the mutations on LV mass, vascular density and the numbers of proliferating cells were also determined. In 4 month old Kit mutant and WT mice, LV ejection fractions (EF and LV fractional shortening rates (FS were comparable. At 12 months of age EF and FS were significantly decreased and LV mass was significantly increased only in W41/W42 mice. Myocardial vascular densities and c-Kit(+ cell numbers were significantly reduced in both mutant groups when compared to WT hearts. Replacement of mutant BM with WT BM at 4 months of age did not prevent these abnormalities in either mutant group although they were somewhat attenuated in the W/Wv group. Notably BM transplantation did not prevent the development of cardiomyopathy in 12 month W41/W42 mice. The data suggest that decreased numbers and functional capacities of c-Kit(+ cardiac resident progenitor cells may be the basis of the cardiomyopathy in W41/W42 mice and although defects in mutant BM progenitor cells may prove to be contributory, they are not causal.

  3. Behavioral characterization of system xc- mutant mice. (United States)

    McCullagh, Elizabeth A; Featherstone, David E


    The slc7a11 gene encodes xCT, an essential component of 'system xc-', a plasma membrane exchanger that imports cystine and exports glutamate. Slc7a11 is expressed primarily in the brain, but its role there is not clear. We performed behavioral tests on two different strains of homozygous slc7a11 mutant mice ('sut' and 'xCT'), as well as heteroallelic offspring of these two strains ('xCT/sut') and their associated genetic backgrounds. Homozygous sut mutant males showed reduced spontaneous alternation in spontaneous alternation tasks as well as reduced movement in an open field maze, but xCT and xCT/sut strains did not show significant changes in these tasks compared to appropriate controls. Neither xCT nor sut mutants showed differences from controls in rotarod tests. Female behavioral phenotypes were independent of estrus cycle stage. To ensure that homozygous xCT, sut, and xCT/sut strains all represent protein null alleles, we measured whole brain xCT protein levels using immunoblots. xCT, sut and xCT/sut strains showed no detectable xCT protein expression, confirming them as null alleles. Previously published microdialysis experiments showed reduced striatal glutamate in xCT mutants. Using the same methods, we measured reduced interstitial glutamate levels in the striatum but not cerebellum of sut mutants. However, we detected no glutamate change in the striatum or cerebellum of sut/xCT mice. We detected no changes in whole brain EAAT-1, -2, or -3 expression. We conclude that the behavioral and chemical differences exist between slc7a11 mutant strains, but we were unable to definitively attribute any of these differences to loss of system xc-.

  4. Targeting ESR1-Mutant Breast Cancer (United States)


    cell line, biochemical and structural biology techniques to uncover the best candidate drugs for the clinical targeting of these mutants. Targeting...ESR1-­‐Mutant  Breast  Cancer   W81XWH-­‐14-­‐1-­‐0360   4   2. Keywords Estrogen Receptor Acquired Drug Resistance Metastatic Breast...preparations for publication: 1) “ESR1 Somatic Mutations Y537S and D538G Confer Breast Cancer Endocrine Resistance by Stabilizing the Active AF-2 conformation

  5. Residues involved in the pore-forming activity of the Clostridium perfringens iota toxin. (United States)

    Knapp, Oliver; Maier, Elke; Waltenberger, Eva; Mazuet, Christelle; Benz, Roland; Popoff, Michel R


    Clostridium perfringens iota toxin is a binary toxin that is organized into enzyme (Ia) and binding (Ib) components. Ib forms channels in lipid bilayers and mediates the transport of Ia into the target cells. Here we show that Ib residues 334-359 contain a conserved pattern of alternating hydrophobic and hydrophilic residues forming two amphipathic β-strands involved in membrane insertion and channel formation. This stretch of amino acids shows remarkable structural and functional analogies with the β-pore-forming domain of C. perfringens epsilon toxin. Several mutations within the two amphipathic β-strands affected pore formation, single-channel conductance and ion selectivity (S339E-S341E, Q345H N346E) confirming their involvement in channel formation. F454 of Ib corresponds to the Φ-clamp F427 of anthrax protective antigen and F428 of C2II binary toxins. The mutation F454A resulted in a loss of cytotoxicity and strong increase in single-channel conductance (500 pS as compared with 85 pS in 1 M KCl) with a slight decrease in cation selectivity, indicating that the Φ-clamp is highly conserved and crucial for binary toxin activity. In contrast, the mutants Q367D, N430D, L443E had no or only minor effects on Ib properties, while T360I, T360A and T360W caused a dramatic effect on ion selectivity and single-channel conductance, indicating gross disturbance of the oligomer structure. This suggests that, at least in the iota toxin family, T360 has a structural role in the pore organization. Moreover, introduction of charged residues within the channel (S339E-S341E) or in the vestibule (Q367D, N430D and L443E) had virtually no effect on chloroquine or Ia binding, whereas F454A, T360I, T360A and T360W strongly decreased the chloroquine and Ia affinity to Ib. These results support that distinct residues within the vestibule interact with chloroquine and Ia or are responsible for channel structure, while the channel lining amino acids play a less important role.

  6. Modern routes ro explore concrete's complex pore space

    NARCIS (Netherlands)

    Stroeven, P; Guo, Z.


    This paper concentrates on discrete element computer-simulation of concrete. It is argued on the basis of stochastic heterogeneity theory that modern concurrent-algorithm-based systems should be employed for the assessment of pore characteristics underlying durability performance of cementitious mat

  7. Septal Pore Caps in Basidiomycetes, Composition and Ultrastructure

    NARCIS (Netherlands)

    Driel, K.G.A. van


    Filamentous fungi, including Ascomycota and Basidiomycota, form mycelia that consist of a network of apical growing hyphae. These hyphae are separated into cellular compartments by septa that have pores of about 70 to 500 nm in diameter. The cytoplasm within the mycelium is thus continuous (coenocyt

  8. Development of Pore Pressure and Material Damping during Cyclic Loading

    DEFF Research Database (Denmark)

    Ibsen, Lars Bo


    The behaviour of sand during cyclic loading can be characterized as "stabilization", "instant stabilization". "pore pressure buildup" and "liquefaction". The terminologies can be defined exactly by a simple mathematical formulation based on the existence of a cyclic stable state. By introducing a...

  9. Capillary condensation of 4He in cylindrical pores (United States)

    Urrutia, Ignacio; Szybisz, Leszek


    The adsorption of superfluid 4He confined into cylindrical pores of alkali metals is illustrated by looking at the case of Na. A density functional formalism is utilized for the theoretical description. The energetics and density profiles are determined as a function of the radius of cylinders and the filling fraction. These results are compared with those provided by a simple model recently proposed.

  10. Pore-scale uncertainty quantification with multilevel Monte Carlo

    KAUST Repository

    Icardi, Matteo


    Computational fluid dynamics (CFD) simulations of pore-scale transport processes in porous media have recently gained large popularity. However the geometrical details of the pore structures can be known only in a very low number of samples and the detailed flow computations can be carried out only on a limited number of cases. The explicit introduction of randomness in the geometry and in other setup parameters can be crucial for the optimization of pore-scale investigations for random homogenization. Since there are no generic ways to parametrize the randomness in the porescale structures, Monte Carlo techniques are the most accessible to compute statistics. We propose a multilevel Monte Carlo (MLMC) technique to reduce the computational cost of estimating quantities of interest within a prescribed accuracy constraint. Random samples of pore geometries with a hierarchy of geometrical complexities and grid refinements, are synthetically generated and used to propagate the uncertainties in the flow simulations and compute statistics of macro-scale effective parameters.

  11. Alumina ceramics prepared with new pore-forming agents

    Directory of Open Access Journals (Sweden)

    Zuzana Živcová


    Full Text Available Porous ceramics have a wide range of applications at all length scales, ranging from fi ltration membranes and catalyst supports to biomaterials (scaffolds for bone ingrowths and thermally or acoustically insulating bulk materials or coating layers. Organic pore-forming agents (PFAs of biological origin can be used to control porosity, pore size and pore shape. This work concerns the characterization and testing of several less common pore-forming agents (lycopodium, coffee, fl our and semolina, poppy seed, which are of potential interest from the viewpoint of size, shape or availability. The performance of these new PFAs is compared to that of starch, which has become a rather popular PFA for ceramics during the last decade. The PFAs investigated in this work are in the size range from 5 μm (rice starch to approximately 1 mm (poppy seed, all with more or less isometric shape. The burnout behavior of PFAs is studied by thermal analysis, i.e. thermogravimetry and differential thermal analysis. For the preparation of porous alumina ceramics from alumina suspensions containing PFAs traditional slip casting (into plaster molds and starch consolidation casting (using metal molds are used in this work. The resulting microstructures are investigated using optical microscopy, combined with image analysis, as well as other methods (Archimedes method of double-weighing in water, mercury intrusion porosimetry.

  12. Quantifying uncertainty and computational complexity for pore-scale simulations (United States)

    Chen, C.; Yuan, Z.; Wang, P.; Yang, X.; Zhenyan, L.


    Pore-scale simulation is an essential tool to understand the complex physical process in many environmental problems, from multi-phase flow in the subsurface to fuel cells. However, in practice, factors such as sample heterogeneity, data sparsity and in general, our insufficient knowledge of the underlying process, render many simulation parameters and hence the prediction results uncertain. Meanwhile, most pore-scale simulations (in particular, direct numerical simulation) incur high computational cost due to finely-resolved spatio-temporal scales, which further limits our data/samples collection. To address those challenges, we propose a novel framework based on the general polynomial chaos (gPC) and build a surrogate model representing the essential features of the underlying system. To be specific, we apply the novel framework to analyze the uncertainties of the system behavior based on a series of pore-scale numerical experiments, such as flow and reactive transport in 2D heterogeneous porous media and 3D packed beds. Comparing with recent pore-scale uncertainty quantification studies using Monte Carlo techniques, our new framework requires fewer number of realizations and hence considerably reduce the overall computational cost, while maintaining the desired accuracy.

  13. Nanoporous carbide-derived carbon with tunable pore size (United States)

    Gogotsi, Yury; Nikitin, Alexei; Ye, Haihui; Zhou, Wei; Fischer, John E.; Yi, Bo; Foley, Henry C.; Barsoum, Michel W.


    Porous solids are of great technological importance due to their ability to interact with gases and liquids not only at the surface, but throughout their bulk. Although large pores can be produced and well controlled in a variety of materials, nanopores in the range of 2 nm and below (micropores, according to IUPAC classification) are usually achieved only in carbons or zeolites. To date, major efforts in the field of porous materials have been directed towards control of the size, shape and uniformity of the pores. Here we demonstrate that porosity of carbide-derived carbons (CDCs) can be tuned with subångström accuracy in a wide range by controlling the chlorination temperature. CDC produced from Ti3SiC2 has a narrower pore-size distribution than single-wall carbon nanotubes or activated carbons; its pore-size distribution is comparable to that of zeolites. CDCs are produced at temperatures from 200-1,200 °C as a powder, a coating, a membrane or parts with near-final shapes, with or without mesopores. They can find applications in molecular sieves, gas storage, catalysts, adsorbents, battery electrodes, supercapacitors, water/air filters and medical devices.

  14. Improving pore interconnectivity in polymeric scaffolds for tissue engineering. (United States)

    Aydin, H M; El Haj, A J; Pişkin, E; Yang, Y


    A new scaffold fabrication technique aiming to enhance pore interconnectivity for tissue engineering has been developed. Medical grade poly(lactic acid) was utilized to generate scaffolds by a solvent-evaporating/particulate-leaching technique, using a new dual-porogen system. Water-soluble sodium chloride particles were used to control macro-pore size in the range 106-255 microm, while organic naphthalene was utilized as a porogen to increase pore interconnections. The three-dimensional (3D) morphology of the scaffolds manufactured with and without naphthalene was examined by optical coherence tomography and scanning electron microscopy. The mechanical properties of the scaffolds were characterized by compression tests. MG63 osteoblast cells were seeded in the scaffolds to study the cell attachment and viability evaluated by confocal microscopy. It was revealed that introducing naphthalene as the second porogen in the solvent-evaporating/particulate-leaching process resulted in improvement of the pore interconnectivity. Cells grew in both scaffolds fabricated with and without naphthalene. They exhibited strong green fluorescence when using a live/dead fluorescent dye kit, indicating that the naphthalene in the scaffold process did not affect cell viability.

  15. Host defenses against bacterial pore-forming toxins

    NARCIS (Netherlands)

    Los, F.C.O.


    Pore-forming toxins (PFTs), the most common bacterial toxins, contribute to infection by perforating host cell membranes. Excessive use and lack of new development of antibiotics are causing increasing numbers of drug-resistant bacteria, like methicillin-resistant Staphylococcus aureus (MRSA) and

  16. Wafer-scale nanostructure formation inside vertical nano-pores

    NARCIS (Netherlands)

    Berenschot, Johan W.; Sun, Xingwu; Le The, Hai; Tiggelaar, Roald M.; de Boer, Meint J.; Eijkel, Jan C.T.; Gardeniers, Johannes G.E.; Tas, Niels Roelof; Sarajlic, Edin

    We propose a wafer-scale technique for nanostructure formation inside vertically oriented, through-membrane nano-pores. It uses 50 nm monocrystalline silicon pillars as a mold, embedded in a silicon nitride membrane formed in an innovative step. The proposed technique paves the way towards advanced

  17. Understanding gene expression with a pore forming toxin


    clamer, Massimiliano


    This thesis aimed to explore eukaryotic cellular processes upon the virulent attack of low doses of a well-known pore forming toxin (staphylococcal α-hemolysin (αHL)) and to develop a new biotech application using the same protein.

  18. Pore-scale Modelling of Capillarity in Swelling Granular Materials (United States)

    Hassanizadeh, S. M.; Sweijen, T.; Nikooee, E.; Chareyre, B.


    Capillarity in granular porous media is a common and important phenomenon in earth materials and industrial products, and therefore has been studied extensively. To model capillarity in granular porous media, one needs to go beyond current models which simulate either two-phase flow in porous media or mechanical behaviour in granular media. Current pore-scale models for two-phase flow such as pore-network models are tailored for rigid pore-skeletons, even though in many applications, namely hydro-mechanical coupling in soils, printing, and hygienic products, the porous structure does change during two-phase flow. On the other hand, models such as Discrete Element Method (DEM), which simulate the deformable porous media, have mostly been employed for dry or saturated granular media. Here, the effects of porosity change and swelling on the retention properties was studied, for swelling granular materials. A pore-unit model that was capable to construct the capillary pressure - saturation curve was coupled to DEM. Such that the capillary pressure - saturation curve could be constructed for varying porosities and amounts of absorbed water. The study material was super absorbent polymer particles, which are capable to absorb water 10's to 200 times their initial weight. We have simulated quasi-static primary imbibition for different porosities and amounts of absorbed water. The results reveal a 3 dimensional surface between capillary pressure, saturation, and porosity, which can be normalized by means of the entry pressure and the effective water saturation to a unique curve.

  19. Pore Water Pumping by Upside-Down Jellyfish (United States)

    Gaddam, Manikantam; Santhanakrishnan, Arvind


    Patchy aggregations of Cassiopea medusae, commonly called upside-down jellyfish, are found in sheltered marine environments with low-speed ambient flows. These medusae exhibit a sessile, non-swimming lifestyle, and are oriented such that their bells are attached to the substrate and oral arms point towards sunlight. Pulsations of their bells are used to generate currents for suspension feeding. Their pulsations have also been proposed to generate forces that can release sediment locked nutrients into the surrounding water. The goal of this study is to examine pore water pumping by Cassiopea individuals in laboratory aquaria, as a model for understanding pore water pumping in unsteady flows. Planar laser-induced fluorescence (PLIF) measurements were conducted to visualize the release of pore water via bell motion, using fluorescent dye introduced underneath the substrate. 2D particle image velocimetry (PIV) measurements were conducted on the same individuals to correlate PLIF-based concentration profiles with the jets generated by pulsing of medusae. The effects of varying bell diameter on pore water release and pumping currents will be discussed.

  20. Influence of pore roughness on high-frequency permeability

    NARCIS (Netherlands)

    Cortis, A.; Smeulders, D.M.J.; Guermond, J.L.; Lafarge, D.


    The high-frequency behavior of the fluid velocity patterns for smooth and corrugated pore channels is studied. The classical approach of Johnson et al. [J. Fluid Mech. 176, 379 (1987)] for smooth geometries is obtained in different manners, thus clarifying differences with Sheng and Zhou [Phys. Rev.

  1. Tuning the Pore Size in Ionic Nanoparticle Networks

    Directory of Open Access Journals (Sweden)

    Marie-Alexandra Neouze Gauthey


    Full Text Available Highly promising hybrid materials consisting of silica, titania, or zirconia nanoparticles linked with ionic liquid-like imidazolium units have been developed. The nanoparticle networks are prepared by click-chemistry-like process through a nucleophilic substitution reaction. The type of metal oxide nanoparticles appears to play a key role regarding the pore size of the hybrid material.

  2. Host defenses against bacterial pore-forming toxins

    NARCIS (Netherlands)

    Los, F.C.O.


    Pore-forming toxins (PFTs), the most common bacterial toxins, contribute to infection by perforating host cell membranes. Excessive use and lack of new development of antibiotics are causing increasing numbers of drug-resistant bacteria, like methicillin-resistant Staphylococcus aureus (MRSA) and My

  3. Wafer-scale nanostructure formation inside vertical nano-pores

    NARCIS (Netherlands)

    Berenschot, Johan W.; Sun, Xingwu; Le The, Hai; Tiggelaar, Roald M.; de Boer, Meint J.; Eijkel, Jan C.T.; Gardeniers, Johannes G.E.; Tas, Niels Roelof; Sarajlic, Edin


    We propose a wafer-scale technique for nanostructure formation inside vertically oriented, through-membrane nano-pores. It uses 50 nm monocrystalline silicon pillars as a mold, embedded in a silicon nitride membrane formed in an innovative step. The proposed technique paves the way towards advanced

  4. A pore scale study on turbulent combustion in porous media (United States)

    Jouybari, N. F.; Maerefat, M.; Nimvari, M. E.


    This paper presents pore scale simulation of turbulent combustion of air/methane mixture in porous media to investigate the effects of multidimensionality and turbulence on the flame within the pores of porous media. In order to investigate combustion in the pores of porous medium, a simple but often used porous medium consisting of a staggered arrangement of square cylinders is considered in the present study. Results of turbulent kinetic energy, turbulent viscosity ratio, temperature, flame speed, convective heat transfer and thermal conductivity are presented and compared for laminar and turbulent simulations. It is shown that the turbulent kinetic energy increases from the inlet of burner, because of turbulence created by the solid matrix with a sudden jump or reduction at the flame front due to increase in temperature and velocity. Also, the pore scale simulation revealed that the laminarization of flow occurs after flame front in the combustion zone and turbulence effects are important mainly in the preheat zone. It is shown that turbulence enhances the diffusion processes in the preheat zone, but it is not enough to affect the maximum flame speed, temperature distribution and convective heat transfer in the porous burner. The dimensionless parameters associated with the Borghi-Peters diagram of turbulent combustion have been analyzed for the case of combustion in porous media and it is found that the combustion in the porous burner considered in the present study concerns the range of well stirred reactor very close to the laminar flame region.

  5. Pore Characteristics of Chitosan Scaffolds Studied by Electrochemical Impedance Spectroscopy (United States)

    Tully-Dartez, Stephanie; Cardenas, Henry E.


    In this study, a novel approach, electrochemical impedance spectroscopy (EIS), was used to examine the pore characteristics of chitosan scaffolds under aqueous conditions. The EIS was run with a constant current of 0.1 mA with the frequency sweep of 106 to 10−4 Hz. The resulting complex impedance measurement was then used to calculate porosity, which was determined to be 71%. Scanning electron microscopy (SEM) and mercury intrusion porosimetry (MIP), two commonly used methods for scaffold characterization, were used to independently evaluate the pore characteristics and compare with that of EIS. The SEM and MIP were performed and analyzed under standard conditions. The pore diameter values found by SEM and MIP are 107 μm and 82 μm, respectively, indicating that both the image-based (SEM) and pressure-based (MIP) analyses provide similar results. The porosity of 73% calculated by MIP is comparable to that of EIS. From these results, it can be suggested that EIS, a relatively nondestructive test, is able to obtain comparable data on pore characteristics, as compared to SEM and MIP. The advantage of the EIS as an nondestructive test is that it can be performed under physiologically relevant conditions, whereas SEM and MIP require dry samples and vacuum conditions for measurement. These benefits make EIS a viable option for the characterization and long-term observation of tissue-engineered scaffolds. PMID:19580421

  6. How Lipid Membranes Affect Pore Forming Toxin Activity. (United States)

    Rojko, Nejc; Anderluh, Gregor


    Pore forming toxins (PFTs) evolved to permeate the plasma membrane of target cells. This is achieved in a multistep mechanism that usually involves binding of soluble protein monomer to the lipid membrane, oligomerization at the plane of the membrane, and insertion of part of the polypeptide chain across the lipid membrane to form a conductive channel. Introduced pores allow uncontrolled transport of solutes across the membrane, inflicting damage to the target cell. PFTs are usually studied from the perspective of structure-function relationships, often neglecting the important role of the bulk membrane properties on the PFT mechanism of action. In this Account, we discuss how membrane lateral heterogeneity, thickness, and fluidity influence the pore forming process of PFTs. In general, lipid molecules are more accessible for binding in fluid membranes due to steric reasons. When PFT specifically binds ordered domains, it usually recognizes a specific lipid distribution pattern, like sphingomyelin (SM) clusters or SM/cholesterol complexes, and not individual lipid species. Lipid domains were also suggested to act as an additional concentration platform facilitating PFT oligomerization, but this is yet to be shown. The last stage in PFT action is the insertion of the transmembrane segment across the membranes to build the transmembrane pore walls. Conformational changes are a spontaneous process, and sufficient free energy has to be available for efficient membrane penetration. Therefore, fluid bilayers are permeabilized more readily in comparison to highly ordered and thicker liquid ordered lipid phase (Lo). Energetically more costly insertion into the Lo phase can be driven by the hydrophobic mismatch between the thinner liquid disordered phase (Ld) and large protein complexes, which are unable to tilt like single transmembrane segments. In the case of proteolipid pores, membrane properties can directly modulate pore size, stability, and even selectivity. Finally

  7. Effects of pore-scale precipitation on permeability and flow (United States)

    Noiriel, Catherine; Steefel, Carl I.; Yang, Li; Bernard, Dominique


    The effects of calcite precipitation on porous media permeability and flow were evaluated with a combined experimental and modeling approach. X-ray microtomography images of two columns packed with glass beads and calcite (spar crystals) or aragonite (Bahamas ooids) injected with a supersaturated solution (log Ω = 1.42) were processed in order to calculate rates of calcite precipitation with a spatial resolution of 4.46 μm. Identification and localization of the newly precipitated crystals on the 3D images was performed and results used to calculate the crystal growth rates and velocities. The effects of carbonate precipitation were also evaluated in terms of the integrated precipitation rate over the length of the column, crystal shape, surface area and pore roughness changes. While growth was epitaxial on calcite spar, calcite rhombohedra formed on glass beads and clusters of polyhedrons formed on aragonite ooids. Near the column inlet, calcite precipitation occurred preferentially on carbonate grains compared to glass beads, with almost 100% of calcite spar surface area covered by new crystals versus 92% in the case of aragonite and 11% in the case of glass beads. Although the experimental chemistry and flow boundary conditions in the two columns were similar, their porosity-permeability evolution was different because the nucleation and subsequent crystal growth on the two substrates (i.e., calcite spar and aragonite ooids) was very different. The impact of mineral precipitation on pore-scale flow and permeability was evaluated using a pore-scale Stokes solver that accounted for the changes in pore geometry. For similar magnitude reductions in porosity, the decrease in permeability was highest within the sample that experienced the greatest increase in pore roughness. Various porous media models were generated to show the impact of different crystal growth patterns and pore roughness changes on flow and permeability-porosity relationship. Under constant flow

  8. Rapid Antibiotic Resistance Evolution of GASP Mutants (United States)

    Zhang, Qiucen; Kim, Hyunsung; Pourmand, Nader; Austin, Robert


    The GASP phenotype in bacteria is due to a mutation which enables the bacteria to grow under high stress conditions where other bacteria stop growing. We probe using our Death Galaxy microenvironment how rapidly the GASP mutant can evolve resistance to mutagenic antibiotics compared to wild-type bacteria, and explore the genomic landscape changes due to the evolution of resistance.

  9. Generation and characterization of pigment mutants of ...

    African Journals Online (AJOL)


    deleterious effects on living organisms (Shigaeva et al.,. 1994); they are also ... It was cultured at 25±0.5°C with a fluorescent light intensity of approximately 6 ... mutants) resulted in three new colonies characterized by different green colors ...

  10. A dominant semi dwarf mutant in rice

    Institute of Scientific and Technical Information of China (English)


    @@ In the winter of 1997, a semi dwarf mutant was found in the F6 population of M9056/ R8018 xuan in Hainan Province. In the spring of 1998, the seeds were sown in Hefei, Anhui Province and the plant height of the population was measured at maturity.

  11. Nicotinamide ribosyl uptake mutants in Haemophilus influenzae. (United States)

    Herbert, Mark; Sauer, Elizabeta; Smethurst, Graeme; Kraiss, Anita; Hilpert, Anna-Karina; Reidl, Joachim


    The gene for the nicotinamide riboside (NR) transporter (pnuC) was identified in Haemophilus influenzae. A pnuC mutant had only residual NR uptake and could survive in vitro with high concentrations of NR, but could not survive in vivo. PnuC may represent a target for the development of inhibitors for preventing H. influenzae disease.

  12. Mutant PTEN in Cancer : Worse Than Nothing

    NARCIS (Netherlands)

    Leslie, Nick R; den Hertog, Jeroen


    Tumor suppressors block the development of cancer and are often lost during tumor development. Papa et al. show that partial loss of normal PTEN tumor suppressor function can be compounded by additional disruption caused by the expression of inactive mutant PTEN protein. This has significant

  13. Colored HOMFLY polynomials can distinguish mutant knots

    CERN Document Server

    Nawata, Satoshi; Singh, Vivek Kumar


    We illustrate from the viewpoint of braiding operations on WZNW conformal blocks how colored HOMFLY polynomials with multiplicity structure can detect mutations. As an example, we explicitly evaluate the (2,1)-colored HOMFLY polynomials that distinguish a famous mutant pair, Kinoshita-Terasaka and Conway knot.

  14. Structural Insights into Clostridium perfringens Delta Toxin Pore Formation. (United States)

    Huyet, Jessica; Naylor, Claire E; Savva, Christos G; Gibert, Maryse; Popoff, Michel R; Basak, Ajit K


    Clostridium perfringens Delta toxin is one of the three hemolysin-like proteins produced by C. perfringens type C and possibly type B strains. One of the others, NetB, has been shown to be the major cause of Avian Nectrotic Enteritis, which following the reduction in use of antibiotics as growth promoters, has become an emerging disease of industrial poultry. Delta toxin itself is cytotoxic to the wide range of human and animal macrophages and platelets that present GM2 ganglioside on their membranes. It has sequence similarity with Staphylococcus aureus β-pore forming toxins and is expected to heptamerize and form pores in the lipid bilayer of host cell membranes. Nevertheless, its exact mode of action remains undetermined. Here we report the 2.4 Å crystal structure of monomeric Delta toxin. The superposition of this structure with the structure of the phospholipid-bound F component of S. aureus leucocidin (LukF) revealed that the glycerol molecules bound to Delta toxin and the phospholipids in LukF are accommodated in the same hydrophobic clefts, corresponding to where the toxin is expected to latch onto the membrane, though the binding sites show significant differences. From structure-based sequence alignment with the known structure of staphylococcal α-hemolysin, a model of the Delta toxin pore form has been built. Using electron microscopy, we have validated our model and characterized the Delta toxin pore on liposomes. These results highlight both similarities and differences in the mechanism of Delta toxin (and by extension NetB) cytotoxicity from that of the staphylococcal pore-forming toxins.

  15. Formation and decay of rudimentary penumbra around a pore

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Hiroko [Unit of Synergetic Studies for Space, Kyoto University, Yamashina-ku, Kyoto 607-8417 (Japan); Kitai, Reizaburo [Kwasan and Hida Observatories, Kyoto University, Yamashina-ku, Kyoto 607-8417 (Japan); Otsuji, Kenichi, E-mail: [Solar Observatory, National Astronomical Observatory, Mitaka, Tokyo 181-8588 (Japan)


    We analyze the evolution of a pore in the active region NOAA 10940 using the data obtained by the Hinode satellite on 2007 February 3. The pore we analyzed showed the formation of a rudimentary penumbra structure, succeeded by an abrupt disappearance after about 5 hr. The pore had an approximate radius of 3.5 Mm and a total magnetic flux of 3.0 × 10{sup 19} Mx, which is a little smaller than the necessary magnetic flux for penumbral formation supposed by Rucklidge et al. (1-1.5 × 10{sup 20} Mx). Our observation describes a rare phenomenon which was in the unstable phase between a pore and a sunspot. The area of the dark umbra gradually decreased when the rudimentary penumbral filaments formed the penumbral structure, meaning that the penumbra develops at the expense of the umbral magnetic flux. This statement was confirmed by a rough estimation of the magnetic flux variation observed by the Hinode Fe I magnetogram. Five hours after the formation phase, the decay phase began. In this decaying phase, multiple opposite polarity patches are found to appear in the exterior of the pore (a different location from the penumbra formation site). We interpret these opposite polarities as signatures of the horizontal magnetic field, which preferably appears in the course of the unstable reconfiguration of the magnetic field structure. During the course of the disappearance of the penumbra, the horizontal penumbral field seems to become vertical because of the dark umbral area that recovered by about 10%.

  16. Crystal structure of an invertebrate cytolysin pore reveals unique properties and mechanism of assembly (United States)

    Podobnik, Marjetka; Savory, Peter; Rojko, Nejc; Kisovec, Matic; Wood, Neil; Hambley, Richard; Pugh, Jonathan; Wallace, E. Jayne; McNeill, Luke; Bruce, Mark; Liko, Idlir; Allison, Timothy M.; Mehmood, Shahid; Yilmaz, Neval; Kobayashi, Toshihide; Gilbert, Robert J. C.; Robinson, Carol V.; Jayasinghe, Lakmal; Anderluh, Gregor


    The invertebrate cytolysin lysenin is a member of the aerolysin family of pore-forming toxins that includes many representatives from pathogenic bacteria. Here we report the crystal structure of the lysenin pore and provide insights into its assembly mechanism. The lysenin pore is assembled from nine monomers via dramatic reorganization of almost half of the monomeric subunit structure leading to a β-barrel pore ~10 nm long and 1.6-2.5 nm wide. The lysenin pore is devoid of additional luminal compartments as commonly found in other toxin pores. Mutagenic analysis and atomic force microscopy imaging, together with these structural insights, suggest a mechanism for pore assembly for lysenin. These insights are relevant to the understanding of pore formation by other aerolysin-like pore-forming toxins, which often represent crucial virulence factors in bacteria.

  17. Pore networks in continental and marine mudstones: Characteristics and controls on sealing behavior (United States)

    Heath, J.E.; Dewers, T.A.; McPherson, B.J.O.L.; Petrusak, R.; Chidsey, T.C.; Rinehart, A.J.; Mozley, P.S.


    Mudstone pore networks are strong modifiers of sedimentary basin fluid dynamics and have a critical role in the distribution of hydrocarbons and containment of injected fluids. Using core samples from continental and marine mudstones, we investigate properties of pore types and networks from a variety of geologic environments, together with estimates of capillary beam- scanning electron microscopy, suggest seven dominant mudstone pore types distinguished by geometry and connectivity. A dominant planar pore type occurs in all investigated mudstones and generally has high coordination numbers (i.e., number of neighboring connected pores). Connected networks of pores of this type contribute to high mercury capillary pressures due to small pore throats at the junctions of connected pores and likely control most matrix transport in these mudstones. Other pore types are related to authigenic (e.g., replacement or pore-lining precipitation) clay minerals and pyrite nodules; pores in clay packets adjacent to larger, more competent clastic grains; pores in organic phases; and stylolitic and microfracture-related pores. Pores within regions of authigenic clay minerals often form small isolated networks (environments and greater maximum depth of burial. ?? 2011 Geological Society of America.

  18. GAMPMS: Genetic algorithm managed peptide mutant screening. (United States)

    Long, Thomas; McDougal, Owen M; Andersen, Tim


    The prominence of endogenous peptide ligands targeted to receptors makes peptides with the desired binding activity good molecular scaffolds for drug development. Minor modifications to a peptide's primary sequence can significantly alter its binding properties with a receptor, and screening collections of peptide mutants is a useful technique for probing the receptor-ligand binding domain. Unfortunately, the combinatorial growth of such collections can limit the number of mutations which can be explored using structure-based molecular docking techniques. Genetic algorithm managed peptide mutant screening (GAMPMS) uses a genetic algorithm to conduct a heuristic search of the peptide's mutation space for peptides with optimal binding activity, significantly reducing the computational requirements of the virtual screening. The GAMPMS procedure was implemented and used to explore the binding domain of the nicotinic acetylcholine receptor (nAChR) α3β2-isoform with a library of 64,000 α-conotoxin (α-CTx) MII peptide mutants. To assess GAMPMS's performance, it was compared with a virtual screening procedure that used AutoDock to predict the binding affinity of each of the α-CTx MII peptide mutants with the α3β2-nAChR. The GAMPMS implementation performed AutoDock simulations for as few as 1140 of the 64,000 α-CTx MII peptide mutants and could consistently identify a set of 10 peptides with an aggregated binding energy that was at least 98% of the aggregated binding energy of the 10 top peptides from the exhaustive AutoDock screening.

  19. Phanerochaete mutants with enhanced ligninolytic activity

    Energy Technology Data Exchange (ETDEWEB)

    Kakar, S.N.; Perez, A.; Gonzales, J. (Argonne National Lab., IL (United States))

    In addition to lignin, the white rot fungus Phanerochaete chrysosporium has the ability to degrade a wide spectrum of recalcitrant organopollutants in soils and aqueous media. Most of the organic compounds are degraded under ligninolytic conditions with the involvement of the extracellular enzymes, lignin peroxidases, and manganese-dependent peroxidases, which are produced as secondary metabolites triggered by conditions of nutrient starvation (e.g., nitrogen limitation). The fungus and its enzymes can thus provide alternative technologies for bioremediation, biopulping, biobleaching, and other industrial applications. The efficiency and effectiveness of the fungus can be enhanced by increasing production and secretion of the important enzymes in large quantities and as primary metabolites under enriched conditions. One way this can be achieved is through isolation of mutants that are deregulated, or are hyperproducers or supersectors of key enzymes under enriched conditions. Through UV-light and [gamma]-ray mutagenesis, the authors have isolated a variety of mutants, some of which produce key enzymes of the ligninolytic system under high-nitrogen growth conditions. One of the mutants, 76UV, produced 272 U of lignin peroxidases enzyme activity/L after 9 d under high nitrogen (although the parent strain does not produce this enzyme under these conditions). The mutant and the parent strains produced up to 54 and 62 U/L, respectively, of the enzyme activity under low-nitrogen growth conditions during this period. In some experiments, the mutant showed 281 U/L of enzyme activity under high nitrogen after 17 d. 17 refs., 1 fig., 3 tabs.

  20. A specific two-pore domain potassium channel blocker defines the structure of the TASK-1 open pore. (United States)

    Streit, Anne K; Netter, Michael F; Kempf, Franca; Walecki, Magdalena; Rinné, Susanne; Bollepalli, Murali K; Preisig-Müller, Regina; Renigunta, Vijay; Daut, Jürgen; Baukrowitz, Thomas; Sansom, Mark S P; Stansfeld, Phillip J; Decher, Niels


    Two-pore domain potassium (K(2P)) channels play a key role in setting the membrane potential of excitable cells. Despite their role as putative targets for drugs and general anesthetics, little is known about the structure and the drug binding site of K(2P) channels. We describe A1899 as a potent and highly selective blocker of the K(2P) channel TASK-1. As A1899 acts as an open-channel blocker and binds to residues forming the wall of the central cavity, the drug was used to further our understanding of the channel pore. Using alanine mutagenesis screens, we have identified residues in both pore loops, the M2 and M4 segments, and the halothane response element to form the drug binding site of TASK-1. Our experimental data were used to validate a K(2P) open-pore homology model of TASK-1, providing structural insights for future rational design of drugs targeting K(2P) channels.

  1. A Specific Two-pore Domain Potassium Channel Blocker Defines the Structure of the TASK-1 Open Pore* (United States)

    Streit, Anne K.; Netter, Michael F.; Kempf, Franca; Walecki, Magdalena; Rinné, Susanne; Bollepalli, Murali K.; Preisig-Müller, Regina; Renigunta, Vijay; Daut, Jürgen; Baukrowitz, Thomas; Sansom, Mark S. P.; Stansfeld, Phillip J.; Decher, Niels


    Two-pore domain potassium (K2P) channels play a key role in setting the membrane potential of excitable cells. Despite their role as putative targets for drugs and general anesthetics, little is known about the structure and the drug binding site of K2P channels. We describe A1899 as a potent and highly selective blocker of the K2P channel TASK-1. As A1899 acts as an open-channel blocker and binds to residues forming the wall of the central cavity, the drug was used to further our understanding of the channel pore. Using alanine mutagenesis screens, we have identified residues in both pore loops, the M2 and M4 segments, and the halothane response element to form the drug binding site of TASK-1. Our experimental data were used to validate a K2P open-pore homology model of TASK-1, providing structural insights for future rational design of drugs targeting K2P channels. PMID:21362619

  2. Neuronal Goα and CAPS regulate behavioral and immune responses to bacterial pore-forming toxins.

    Directory of Open Access Journals (Sweden)

    Ferdinand C O Los

    Full Text Available Pore-forming toxins (PFTs are abundant bacterial virulence factors that attack host cell plasma membranes. Host defense mechanisms against PFTs described to date all function in the host tissue that is directly attacked by the PFT. Here we characterize a rapid and fully penetrant cessation of feeding of Caenorhabditis elegans in response to PFT attack. We demonstrate via analyses of C. elegans mutants that inhibition of feeding by PFT requires the neuronal G protein Goα subunit goa-1, and that maintenance of this response requires neuronally expressed calcium activator for protein secretion (CAPS homolog unc-31. Independently from their role in feeding cessation, we find that goa-1 and unc-31 are additionally required for immune protection against PFTs. We thus demonstrate that the behavioral and immune responses to bacterial PFT attack involve the cross-talk between the nervous system and the cells directly under attack.

  3. Neuronal Goα and CAPS regulate behavioral and immune responses to bacterial pore-forming toxins. (United States)

    Los, Ferdinand C O; Ha, Christine; Aroian, Raffi V


    Pore-forming toxins (PFTs) are abundant bacterial virulence factors that attack host cell plasma membranes. Host defense mechanisms against PFTs described to date all function in the host tissue that is directly attacked by the PFT. Here we characterize a rapid and fully penetrant cessation of feeding of Caenorhabditis elegans in response to PFT attack. We demonstrate via analyses of C. elegans mutants that inhibition of feeding by PFT requires the neuronal G protein Goα subunit goa-1, and that maintenance of this response requires neuronally expressed calcium activator for protein secretion (CAPS) homolog unc-31. Independently from their role in feeding cessation, we find that goa-1 and unc-31 are additionally required for immune protection against PFTs. We thus demonstrate that the behavioral and immune responses to bacterial PFT attack involve the cross-talk between the nervous system and the cells directly under attack.

  4. Choreography of importin-α/CAS complex assembly and disassembly at nuclear pores. (United States)

    Sun, Changxia; Fu, Guo; Ciziene, Danguole; Stewart, Murray; Musser, Siegfried M


    Nuclear pore complexes (NPCs) mediate the exchange of macromolecules between the cytoplasm and the nucleoplasm. Soluble nuclear transport receptors bind signal-dependent cargos to form transport complexes that diffuse through the NPC and are then disassembled. Although transport receptors enable the NPC's permeability barrier to be overcome, directionality is established by complex assembly and disassembly. Here, we delineate the choreography of importin-α/CAS complex assembly and disassembly in permeabilized cells, using single-molecule fluorescence resonance energy transfer and particle tracking. Monitoring interaction sequences in intact NPCs ensures spatiotemporal preservation of structures and interactions critical for activity in vivo. We show that key interactions between components are reversible, multiple outcomes are often possible, and the assembly and disassembly of complexes are precisely controlled to occur at the appropriate place and time. Importin-α mutants that impair interactions during nuclear import were used together with cytoplasmic Ran GTPase-activating factors to demonstrate that importin-α/CAS complexes form in the nuclear basket region, at the termination of protein import, and disassembly of importin-α/CAS complexes after export occurs in the cytoplasmic filament region of the NPC. Mathematical models derived from our data emphasize the intimate connection between transport and the coordinated assembly and disassembly of importin-α/CAS complexes for generating productive transport cycles.

  5. Effect of pore formers on properties of tape cast porous sheets for electrochemical flue gas purification

    DEFF Research Database (Denmark)

    Schmidt, Cristine Grings; Kammer Hansen, Kent; Andersen, Kjeld Bøhm


    permeability measurements, mercury porosimetry and pore orientation measurements, to investigate the role of the different pore formers on the properties after sintering at a temperature of 1250°C. Those tapes prepared from different non-spherical pore formers with comparable porosity of about 43%, showed...... significant differences in gas permeability which could be correlated to an increase in mean pore size and pore connectivity. The degree of pore orientation in the tape casting direction was determined by best-fit ellipse method and a modified linear intercept method and the obtained data were correlated...... with the corresponding gas permeability....

  6. Water retention, gas transport, and pore network complexity during short-term regeneration of soil structure

    DEFF Research Database (Denmark)

    Arthur, Emmanuel; Møldrup, Per; Schjønning, Per


    was done using water retention (pore size distribution), soil gas diffusivity, air permeability, and derived pore network complexity parameters. Significant decreases in bulk density (increased total porosity) and increases in pores > 100 1m was observed for incubated samples compared with SR samples....... The proportion of pores > 100 1m increased in order: smectite gas diffusivity, air permeability, and derived pore network indices was greater for incubated samples than SR. For illitic soils...... for convective air transport when analyzing pore network complexity. Overall, our results showed that short-term regeneration...

  7. Unstable Pore-Water Flow in Intertidal Wetlands (United States)

    Barry, D. A.; Shen, C.; Li, L.


    Salt marshes are important intertidal wetlands strongly influenced by interactions between surface water and groundwater. Bordered by coastal water, the marsh system undergoes cycles of inundation and exposure driven by the tide. This leads to dynamic, complex pore-water flow and solute transport in the marsh soil. Pore-water circulations occur over vastly different spatial and temporal scales with strong link to the marsh topography. These circulations control solute transport between the marsh soil and the tidal creek, and ultimately affect the overall nutrient exchange between the marsh and coastal water. The pore-water flows also dictate the soil condition, particularly aeration, which influences the marsh plant growth. Numerous studies have been carried out to examine the pore-water flow process in the marsh soil driven by tides, focusing on stable flow with the assumption of homogeneity in soil and fluid properties. This assumption, however, is questionable given the actual inhomogeneous conditions in the field. For example, the salinity of surface water in the tidal creek varies temporally and spatially due to the influence of rainfall and evapotranspiration as well as the freshwater input from upland areas to the estuary, creating density gradients across the marsh surface and within the marsh soil. Many marshes possess soil stratigraphy with low-permeability mud typically overlying high-permeability sandy deposits. Macropores such as crab burrows are commonly distributed in salt marsh sediments. All these conditions are prone to the development of non-uniform, unstable preferential pore-water flow in the marsh soil, for example, funnelling and fingering. Here we present results from laboratory experiments and numerical simulations to explore such unstable flow. In particular, the analysis aims to address how the unstable flow modifies patterns of local pore-water movement and solute transport, as well as the overall exchange between the marsh soil and

  8. Amuvatinib has cytotoxic effects against NRAS-mutant melanoma but not BRAF-mutant melanoma. (United States)

    Fedorenko, Inna V; Fang, Bin; Koomen, John M; Gibney, Geoffrey T; Smalley, Keiran S M


    Effective targeted therapy strategies are still lacking for the 15-20% of melanoma patients whose melanomas are driven by oncogenic NRAS. Here, we report on the NRAS-specific behavior of amuvatinib, a kinase inhibitor with activity against c-KIT, Axl, PDGFRα, and Rad51. An analysis of BRAF-mutant and NRAS-mutant melanoma cell lines showed the NRAS-mutant cohort to be enriched for targets of amuvatinib, including Axl, c-KIT, and the Axl ligand Gas6. Increasing concentrations of amuvatinib selectively inhibited the growth of NRAS-mutant, but not BRAF-mutant melanoma cell lines, an effect associated with induction of S-phase and G2/M-phase cell cycle arrest and induction of apoptosis. Mechanistically, amuvatinib was noted to either inhibit Axl, AKT, and MAPK signaling or Axl and AKT signaling and to induce a DNA damage response. In three-dimensional cell culture experiments, amuvatinib was cytotoxic against NRAS-mutant melanoma cell lines. Thus, we show for the first time that amuvatinib has proapoptotic activity against melanoma cell lines, with selectivity observed for those harboring oncogenic NRAS.

  9. Nanometer-Scale Pore Characteristics of Lacustrine Shale, Songliao Basin, NE China.

    Directory of Open Access Journals (Sweden)

    Min Wang

    Full Text Available In shale, liquid hydrocarbons are accumulated mainly in nanometer-scale pores or fractures, so the pore types and PSDs (pore size distributions play a major role in the shale oil occurrence (free or absorbed state, amount of oil, and flow features. The pore types and PSDs of marine shale have been well studied; however, research on lacustrine shale is rare, especially for shale in the oil generation window, although lacustrine shale is deposited widely around the world. To investigate the relationship between nanometer-scale pores and oil occurrence in the lacustrine shale, 10 lacustrine shale core samples from Songliao Basin, NE China were analyzed. Analyses of these samples included geochemical measurements, SEM (scanning electron microscope observations, low pressure CO2 and N2 adsorption, and high-pressure mercury injection experiments. Analysis results indicate that: (1 Pore types in the lacustrine shale include inter-matrix pores, intergranular pores, organic matter pores, and dissolution pores, and these pores are dominated by mesopores and micropores; (2 There is no apparent correlation between pore volumes and clay content, however, a weak negative correlation is present between total pore volume and carbonate content; (3 Pores in lacustrine shale are well developed when the organic matter maturity (Ro is >1.0% and the pore volume is positively correlated with the TOC (total organic carbon content. The statistical results suggest that oil in lacustrine shale mainly occurs in pores with diameters larger than 40 nm. However, more research is needed to determine whether this minimum pore diameter for oil occurrence in lacustrine shale is widely applicable.

  10. Network modelling of wettability and pore geometry effects on electrical resistivity and capillary pressure

    Energy Technology Data Exchange (ETDEWEB)

    Man, H.N.; Jing, X.D. [Centre for Petroleum Studies, T.H. Huxley School, Imperial College of Science, Technology and Medicine, Prince Consort Road, London (United Kingdom)


    Recent research efforts have focused on using simple non-circular cross-sectional pore shapes to honour the physics observed at the pore scale. For example, there is evidence to suggest variations of wettability occur at this level. These pores can exhibit water-wet and oil-wet regions, depending on the physics of wetting films, and hence the porous medium maybe of mixed-wettability character. For low water saturations, electrical resistivity cannot be physically simulated at the pore scale using cylindrical tubes, even though wetting film thickness' and pore constrictions are taken into account. A three-dimensional network model that investigates the petrophysical characteristics, electrical resistivity and capillary pressure, is presented. The influence of saturation history is also modelled. Key pore geometrical attributes such as pore shape, aspect ratio, pore coordination number (pore connectivity) and pore size distribution are included in the model. In addition, pore constrictions are introduced which may result in phase trapping via snap-off within the tube itself. Analysis of our developing network model starting from representing the pore shape as circular is presented. Using a simple non-circular cross-sectional pore shape we show bulk water retained in the crevices give rise to predictions that are in close agreement with electrical resistivity and capillary pressure trends observed in experiments. Numerical results are presented and compared with experimental data.

  11. Functional differences in pore properties between wild-type and cysteine-less forms of the CFTR chloride channel. (United States)

    Holstead, Ryan G; Li, Man-Song; Linsdell, Paul


    Studies of the structure and function of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel have been advanced by the development of functional channel variants in which all 18 endogenous cysteine residues have been mutated ("cys-less" CFTR). However, cys-less CFTR has a slightly higher single-channel conductance than wild-type CFTR, raising questions as to the suitability of cys-less as a model of the wild-type CFTR pore. We used site-directed mutagenesis and patch-clamp recording to investigate the origin of this conductance difference and to determine the extent of functional differences between wild-type and cys-less CFTR channel permeation properties. Our results suggest that the conductance difference is the result of a single substitution, of C343: the point mutant C343S has a conductance similar to cys-less, whereas the reverse mutation, S343C in a cys-less background, restores wild-type conductance levels. Other cysteine substitutions (C128S, C225S, C376S, C866S) were without effect. Substitution of other residues for C343 suggested that conductance is dependent on amino acid side chain volume at this position. A range of other functional pore properties, including interactions with channel blockers (Au[CN] (2) (-) , 5-nitro-2-[3-phenylpropylamino]benzoic acid, suramin) and anion permeability, were not significantly different between wild-type and cys-less CFTR. Our results suggest that functional differences between these two CFTR constructs are of limited scale and scope and result from a small change in side chain volume at position 343. These results therefore support the use of cys-less as a model of the CFTR pore region.

  12. Pore structure of natural and regenerated soil aggregates

    DEFF Research Database (Denmark)

    Naveed, Muhammad; Arthur, Emmanuel; de Jonge, Lis Wollesen


    of the soil type and organic matter amendment, and was vastly different from the state of natural aggregates. Aggregate porosity (> 30 Hm) was observed to be a good predictor for the mechanical properties of aggregates. In general, natural aggregates were stronger than lysimeter aggregates.......Quantitative characterization of aggregate pore structure can reveal the evolution of aggregates under different land use and management practices and their effects on soil processes and functions. Advances in X-ray Computed Tomography (CT) provide powerful means to conduct such characterization....... This study examined aggregate pore structure of three differently managed same textured Danish soils (mixed forage cropping, MFC; mixed cash cropping, MCC; cereal cash cropping, CCC) for (i) natural aggregates, and (ii) aggregates regenerated after 20 months of incubation. In total, 27 aggregates (8-16 mm...


    Energy Technology Data Exchange (ETDEWEB)

    Narasimhan, T. N.; Houston, W. N.; Nur, A. M.


    A Penrose Conference entitled, "The Role of Pore Pressure in Deformation in Geologic Processes" was convened by the authors at San Diego, California between November 9 and 13, 1979. The conference was sponsored by the Geological Society of America. This report is a summary of the highlights of the issues discussed during the conference. In addition, this report also includes a topical reference list relating to the different subject areas relevant to pore pressure and deformation. The references were compiled from a list suggested by the participants and were available for consultation during the conference. Although the list is far from complete, it should prove to be a good starting point for one who is looking for key papers in the field.

  14. Pore structure of the activated coconut shell charcoal carbon (United States)

    Budi, E.; Nasbey, H.; Yuniarti, B. D. P.; Nurmayatri, Y.; Fahdiana, J.; Budi, A. S.


    The development of activated carbon from coconut shell charcoal has been investigated by using physical method to determine the influence of activation parameters in term of temperature, argon gas pressure and time period on the pore structure of the activated carbon. The coconut shell charcoal was produced by pyrolisis process at temperature of about 75 - 150 °C for 6 hours. The charcoal was activated at various temperature (532, 700 and 868 °C), argon gas pressure (6.59, 15 and 23.4 kgf/cm2) and time period of (10, 60 and 120 minutes). The results showed that the pores size were reduced and distributed uniformly as the activation parameters are increased.

  15. Manufacture of Fine-Pored Ceramics by the Gelcasting Method

    Directory of Open Access Journals (Sweden)

    Bronisław Psiuk


    Full Text Available The fine-pored materials represent a wide range of applications and searches are being continued to develop methods of their manufacturing. In the article, based on measurements on fine-grained powders of Al2O3, TiO2, and SiO2, it has been demonstrated that gelcasting can be relatively simple method of obtaining of nanoporous materials with high values of both specific surface area and open porosity. The powders were dispersed in silica sol, and the gelling initiator was NH4Cl. The usefulness of experiment design theory for developing of fine-pored materials with high porosity and specific surface area was also shown.

  16. Soil Pore Network Visualisation and Quantification using ImageJ

    DEFF Research Database (Denmark)

    Garbout, Amin; Pajor, Radoslaw; Otten, Wilfred

    Abstract Soil is one of the most complex materials on the earth, within which many biological, physical and chemical processes that support life and affect climate change take place. A much more detailed knowledge of the soil system is required to improve our ability to develop soil management...... strategies to preserve this limited resource. Many of those processes occur at micro scales. For long our ability to study soils non-destructively at microscopic scales has been limited, but recent developments in the use of X-ray Computed Tomography has offered great opportunities to quantify the 3-D...... geometry of soil pores. In this study we look at how networks that summarize the geometry of pores in soil are affected by soil structure. One of the objectives is to develop a robust and reproducible image analysis technique to produce quantitative knowledge on soil architecture from high resolution 3D...

  17. Methane adsorption behavior on coal having different pore structures

    Institute of Scientific and Technical Information of China (English)

    Zhao; Yi; Jiang; Chengfa; Chu; Wei


    The adsorption of methane onto five dry coal samples was measured at 298 K over the pressure range from 0 to 3.5 MPa using a volumetric method.The isotherm data were fitted to the Langmuir and the Freundlich equations.The kinetic data were fitted to a pseudo second order equation,the linear driving force equation(LDF),and an intra-particle diffusion model.These results showed that higher methane adsorption is correlated with larger micro-pore volumes and specific surface areas.The adsorption was related to the narrow micro-pore size distribution when the previous two parameters are large.The kinetics study showed that the kinetics of methane adsorption onto these five dry coal samples followed a pseudo second order model very well.Methane adsorption rates are controlled by intra-particle diffusion.The faster the intra-particle diffusion,the faster the methane adsorption rate will be.

  18. Pore Size Distribution of High Performance Metakaolin Concrete

    Institute of Scientific and Technical Information of China (English)


    The Compressive strength, porosity and pore size distribution of high performance metakaolin (MK) concrete were investigated. Concretes containing 0,5%,10% and 20% metakaolin were prepared at a water/cementitious material ratio (W/C) of 0.30. In parallel, concrete mixtures with the replacement of cement by 20% fly ash or 5 and 10% silica fume were prepared for comparison. The specimens were cured in water at 27℃ for 3 to 90 days. The results show that at the early age of curing (3 days and 7 days), metakaolin replacements increase the compressive strength, but silica fume replacement slightly reduces the compressive strength. At the age of and after 28 days, the compressive strength of the concrete with metakaolin and silica fume replacement increases.A strong reduction in the total porosity and average pore diameter were observed in the concrete with MK 20% and 10% in the first 7 days.

  19. Silicon pore optics mirror modules for inner and outer radii (United States)

    Wille, Eric; Bavdaz, Marcos; Oosterbroek, Tim; Collon, Maximilien; Ackermann, Marcelo; Günther, Ramses; Vacanti, Giuseppe; Vervest, Mark; Yanson, Alexei; van Baren, Coen; Haneveld, Jeroen; Koelewijn, Arenda; Leenstra, Anne; Wijnperle, Maurice; Pareschi, Giovanni; Civitani, Marta; Conconi, Paolo; Spiga, Daniele; Valsecchi, Giuseppe; Marioni, Fabio; Zuknik, Karl-Heinz; Schweitzer, Mario


    Athena (Advanced Telescope for High Energy Astrophysics) is an x-ray observatory using a Silicon Pore Optics telescope and was selected as ESA's second L-class science mission for a launch in 2028. The x-ray telescope consists of several hundreds of mirror modules distributed over about 15-20 radial rings. The radius of curvature and the module sizes vary among the different radial positions of the rings resulting in different technical challenges for mirror modules for inner and outer radii. We present first results of demonstrating Silicon Pore Optics for the extreme radial positions of the Athena telescope. For the inner most radii (0.25 m) a new mirror plate design is shown which overcomes the challenges of larger curvatures, higher stress values and bigger plates. Preliminary designs for the mounting system and its mechanical properties are discussed for mirror modules covering all other radial positions up to the most outer radius of the Athena telescope.

  20. A general route towards defect and pore engineering in graphene. (United States)

    Xie, Guibai; Yang, Rong; Chen, Peng; Zhang, Jing; Tian, Xuezeng; Wu, Shuang; Zhao, Jing; Cheng, Meng; Yang, Wei; Wang, Duoming; He, Congli; Bai, Xuedong; Shi, Dongxia; Zhang, Guangyu


    Defect engineering in graphene is important for tailoring graphene's properties thus applicable in various applications such as porous membranes and ultra-capacitors. In this paper, we report a general route towards defect- and pore- engineering in graphene through remote plasma treatments. Oxygen plasma irradiation was employed to create homogenous defects in graphene with controllable density from a few to ≈10(3) (μm(-2)). The created defects can be further enlarged into nanopores by hydrogen plasma anisotropic etching with well-defined pore size of a few nm or above. The achieved smallest nanopores are ≈2 nm in size, showing the potential for ultra-small graphene nanopores fabrication. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. X-ray and Cryo-electron Microscopy Structures of Monalysin Pore-forming Toxin Reveal Multimerization of the Pro-form. (United States)

    Leone, Philippe; Bebeacua, Cecilia; Opota, Onya; Kellenberger, Christine; Klaholz, Bruno; Orlov, Igor; Cambillau, Christian; Lemaitre, Bruno; Roussel, Alain


    β-Barrel pore-forming toxins (β-PFT), a large family of bacterial toxins, are generally secreted as water-soluble monomers and can form oligomeric pores in membranes following proteolytic cleavage and interaction with cell surface receptors. Monalysin has been recently identified as a β-PFT that contributes to the virulence of Pseudomonas entomophila against Drosophila. It is secreted as a pro-protein that becomes active upon cleavage. Here we report the crystal and cryo-electron microscopy structure of the pro-form of Monalysin as well as the crystal structures of the cleaved form and of an inactive mutant lacking the membrane-spanning region. The overall structure of Monalysin displays an elongated shape, which resembles those of β-pore-forming toxins, such as Aerolysin, but is devoid of a receptor-binding domain. X-ray crystallography, cryo-electron microscopy, and light-scattering studies show that pro-Monalysin forms a stable doughnut-like 18-mer complex composed of two disk-shaped nonamers held together by N-terminal swapping of the pro-peptides. This observation is in contrast with the monomeric pro-form of the other β-PFTs that are receptor-dependent for membrane interaction. The membrane-spanning region of pro-Monalysin is fully buried in the center of the doughnut, suggesting that upon cleavage of pro-peptides, the two disk-shaped nonamers can, and have to, dissociate to leave the transmembrane segments free to deploy and lead to pore formation. In contrast with other toxins, the delivery of 18 subunits at once, nearby the cell surface, may be used to bypass the requirement of receptor-dependent concentration to reach the threshold for oligomerization into the pore-forming complex.

  2. A mechanistic view of mitochondrial death decision pores

    Directory of Open Access Journals (Sweden)

    J.E. Belizário


    Full Text Available Mitochondria increase their outer and inner membrane permeability to solutes, protons and metabolites in response to a variety of extrinsic and intrinsic signaling events. The maintenance of cellular and intraorganelle ionic homeostasis, particularly for Ca2+, can determine cell survival or death. Mitochondrial death decision is centered on two processes: inner membrane permeabilization, such as that promoted by the mitochondrial permeability transition pore, formed across inner membranes when Ca2+ reaches a critical threshold, and mitochondrial outer membrane permeabilization, in which the pro-apoptotic proteins BID, BAX, and BAK play active roles. Membrane permeabilization leads to the release of apoptogenic proteins: cytochrome c, apoptosis-inducing factor, Smac/Diablo, HtrA2/Omi, and endonuclease G. Cytochrome c initiates the proteolytic activation of caspases, which in turn cleave hundreds of proteins to produce the morphological and biochemical changes of apoptosis. Voltage-dependent anion channel, cyclophilin D, adenine nucleotide translocase, and the pro-apoptotic proteins BID, BAX, and BAK may be part of the molecular composition of membrane pores leading to mitochondrial permeabilization, but this remains a central question to be resolved. Other transporting pores and channels, including the ceramide channel, the mitochondrial apoptosis-induced channel, as well as a non-specific outer membrane rupture may also be potential release pathways for these apoptogenic factors. In this review, we discuss the mechanistic models by which reactive oxygen species and caspases, via structural and conformational changes of membrane lipids and proteins, promote conditions for inner/outer membrane permeabilization, which may be followed by either opening of pores or a rupture of the outer mitochondrial membrane.

  3. Track-etched membrane: dynamics of pore formation (United States)

    Ferain, E.; Legras, R.


    The dynamics of pore formation during etching of heavy ion (Ar 9+ - 4.5 MeV/amu) irradiated bisphenol-A polycarbonate (PC) and polyethylene terephthalate (PET) films is determined by a conductivity cell. This work presents the theoretical basis of this method and describes the experimental procedure. The obtained results allow the determination of the track ( Vt) and bulk ( Vg) etch rates, and an estimate of the damage zone diameter in PC before etching.

  4. An Investigation of Pore Collapse in Asymmetric Polysulfone Membranes


    Subrahmanyan, Sumitra


    Abstract Porous polysulfone membranes prepared by phase inversion can be tailored to suit filtration requirements by the choice of solvent and coagulant. In the current research polysulfone membranes were prepared by inverting a solution in N-methyl pyrrolidinone (NMP) in isopropanol to form uniform sized pores. Phase inversion resulted in the formation of an asymmetric membrane. The membranes have a characteristic "skin" which is supported by a highly porous substructure. Water-wet membra...

  5. Using Advanced Tensiometers to Monitor Temporal Variations in Pore Pressure (United States)

    Nichols, R. L.; Young, M. H.; Dixon, K. L.; Rossabi, J.; Hyde, W. K.; Holmes-Burns, H.


    The Savannah River Site has installed a comprehensive vadose zone monitoring system (VZMS) at it's low level radioactive waste disposal facility to collect the necessary information to calculate contaminant flux. The VZMS includes water content reflectometers, suction lysimeters, advanced tensiometers (ATs), water flux meters, access ports for neutron probes, and a tipping bucket rain gauge. Forty one ATs were installed from 1999 to 2001 at depths ranging from 2 to 60 feet and have been operated continuously. The installation depths were based on a hydrostatigraphic model developed from core logs, cone penetrometer logs, moisture content profiles, water retention curves model that were obtained during the phased installation of the VZMS. An AT consists of a porous cup installed at a prescribed depth with casing back to the surface and a pressure transducer that is lowered into the casing and connects with the porous cup. The pressure transducer transmits it's signal to a datalogger where the data is stored for future retrieval using a cellular phone communications package. Results from the 2 year operating period show that the AT calibrations are stable and t ATs are capable of extended monitoring of pore pressures in the 0 to 300 cm H2 O range. The ATs had sufficient resolution to detect the naturally occurring fluctuations in pore pressure (1 to 100 cm H2 O over 1 to 72 hours) that resulted from infiltration events at the site. The stable performance of the ATs combined with their ability to detect naturally occurring fluctuations in pore pressure make the ATs a useful tool in measuring temporal pore pressure variations for use in calibrating numerical models of fluid flow in variably saturated porous media.

  6. Membranes with functionalized carbon nanotube pores for selective transport (United States)

    Bakajin, Olgica; Noy, Aleksandr; Fornasiero, Francesco; Park, Hyung Gyu; Holt, Jason K; Kim, Sangil


    Provided herein composition and methods for nanoporous membranes comprising single walled, double walled, or multi-walled carbon nanotubes embedded in a matrix material. Average pore size of the carbon nanotube can be 6 nm or less. These membranes are a robust platform for the study of confined molecular transport, with applications in liquid and gas separations and chemical sensing including desalination, dialysis, and fabric formation.

  7. Laboratory tidal triggering in the presence of pore fluid (United States)

    Bartlow, N. M.; Lockner, D. A.; Beeler, N. M.


    The physical mechanism by which the low-frequency earthquakes (LFEs) that make up tremor are created is poorly understood. In many areas of the world, it is consistently observed that LFEs appear to be strongly tidally modulated, whereas ordinary earthquakes are not (e.g. Thomas et al., Nature, 2009; Vidale et al., JGR, 1998). Here we build upon the work of Lockner and Beeler, JGR, 1999, and Beeler and Lockner, JGR, 2003, which investigated the response of laboratory stick-slip to oscillatory, tide-like loading. These previous experiments determined ranges of amplitude and frequency of the oscillatory loading that resulted in tidally correlated populations, and explained the results in a theoretical framework. Two modes were found: the threshold failure mode in which the necessary amplitude for correlated populations decreased with increasing frequency, and the delayed failure mode in which the amplitude stayed the same or increased with increasing frequency. The frequency of transition between the two modes, which scales with event nucleation time, is predicted to depend on effective stress. This dependence was never tested, since all previous experiments were carried out at one effective stress. The previous experiments were also carried out using room dry samples of Westerly granite. Here we update these results with new experiments on Westerly granite, with the addition of varying effective stress and pore fluid at two pressures. The addition of pore fluid is especially important as pore fluid pressure is thought to be high in LFE regions. We verify the effective stress dependence of the mode transition predicted in Beeler and Lockner, JGR, 2003, allowing extrapolation of the results to other effective stresses. We also find that pore fluid effects become important at high frequencies, when the period of oscillation is comparable to the diffusion time over the sample. These results help constrain the conditions at depth that give rise to tidally modulated LFEs

  8. Host Glycan Recognition by a Pore Forming Toxin


    Bouyain, Samuel; Geisbrecht, Brian V.


    An exposed F-type lectin domain fused to the N-terminus of a cholesterol-dependent cytolysin scaffold allows Streptococcus mitis Lectinolysin to cluster at fucose-rich sites on target cell membranes, thereby leading to increased pore-forming toxin activity. In this issue of Structure, Feil and coworkers define the structural basis for Lectinolysin glycan-binding specificity (Feil et al, 2012).

  9. Smart Gating Multi-Scale Pore/Channel-Based Membranes. (United States)

    Hou, Xu


    Smart gating membranes are important and promising in membrane science and technology. Rapid progress in developing smart membranes is transforming technology in many different fields, from energy and environmental to the life sciences. How a specific smart behavior for controllable gating of porous membranes can be obtained, especially for nano- and micrometer-sized multi-scale pore/channel-based membrane systems is addressed.

  10. Pore Water Pressure Contribution to Debris Flow Mobility

    Directory of Open Access Journals (Sweden)

    Chiara Deangeli


    Full Text Available Problem statement: Debris flows are very to extremely rapid flows of saturated granular soils. Two main types of debris flow are generally recognized: Open slope debris flows and channelized debris flows. The former is the results of some form of slope failures, the latter can develop along preexisting stream courses by the mobilization of previously deposited debris blanket. The problem to be addressed is the influence of the mode of initiation on the subsequent mechanism of propagation. In particular the role of pore water pressure on debris flow mobility in both types was debated. Approach: Laboratory flume experiments were set up in order to analyze the behavior of debris flows generated by model sand slope failures. Failures were induced in sand slopes by raising the water level by seepage from a drain located at the top end of the flume, and by rainfall supplied by a set of pierced plastic pipes placed above the flume. Video recordings of the tests were performed to analyze debris flow characteristics. Results: In all the tests the sand water mixture flows were unsteady and non uniform and sand deposition along the channel bed was a relevant phenomenon. The flows were characterized by a behavioral stratification of the sand water mixture along the flow depth. Back analyzed pore water pressure were just in excess to the hydrostatic condition. The reliability of the experimental results was checked by comparison with other flume experiment data. Conclusion: Debris flow behavior was influenced by the mode of initiation, the inclination of the channel and grain size of the soils. These factors affected the attained velocities and the pore water pressure values. The mobility of debris flows was not always enhanced by high excess pore water pressure values.

  11. Pore Space Connectivity and the Transport Properties of Rocks

    Directory of Open Access Journals (Sweden)

    Bernabé Yves


    Full Text Available Pore connectivity is likely one of the most important factors affecting the permeability of reservoir rocks. Furthermore, connectivity effects are not restricted to materials approaching a percolation transition but can continuously and gradually occur in rocks undergoing geological processes such as mechanical and chemical diagenesis. In this study, we compiled sets of published measurements of porosity, permeability and formation factor, performed in samples of unconsolidated granular aggregates, in which connectivity does not change, and in two other materials, sintered glass beads and Fontainebleau sandstone, in which connectivity does change. We compared these data to the predictions of a Kozeny-Carman model of permeability, which does not account for variations in connectivity, and to those of Bernabé et al. (2010, 2011 model, which does [Bernabé Y., Li M., Maineult A. (2010 Permeability and pore connectivity: a new model based on network simulations, J. Geophys. Res. 115, B10203; Bernabé Y., Zamora M., Li M., Maineult A., Tang Y.B. (2011 Pore connectivity, permeability and electrical formation factor: a new model and comparison to experimental data, J. Geophys. Res. 116, B11204]. Both models agreed equally well with experimental data obtained in unconsolidated granular media. But, in the other materials, especially in the low porosity samples that had undergone the greatest amount of sintering or diagenesis, only Bernabé et al. model matched the experimental data satisfactorily. In comparison, predictions of the Kozeny-Carman model differed by orders of magnitude. The advantage of the Bernabé et al. model was its ability to account for a continuous, gradual reduction in pore connectivity during sintering or diagenesis. Although we can only speculate at this juncture about the mechanisms responsible for the connectivity reduction, we propose two possible mechanisms, likely to be active at different stages of sintering and diagenesis

  12. The C Terminus of the Herpes Simplex Virus UL25 Protein Is Required for Release of Viral Genomes from Capsids Bound to Nuclear Pores. (United States)

    Huffman, Jamie B; Daniel, Gina R; Falck-Pedersen, Erik; Huet, Alexis; Smith, Greg A; Conway, James F; Homa, Fred L


    The herpes simplex virus (HSV) capsid is released into the cytoplasm after fusion of viral and host membranes, whereupon dynein-dependent trafficking along microtubules targets it to the nuclear envelope. Binding of the capsid to the nuclear pore complex (NPC) is mediated by the capsid protein pUL25 and the capsid-tethered tegument protein pUL36. Temperature-sensitive mutants in both pUL25 and pUL36 dock at the NPC but fail to release DNA. The uncoating reaction has been difficult to study due to the rapid release of the genome once the capsid interacts with the nuclear pore. In this study, we describe the isolation and characterization of a truncation mutant of pUL25. Live-cell imaging and immunofluorescence studies demonstrated that the mutant was not impaired in penetration of the host cell or in trafficking of the capsid to the nuclear membrane. However, expression of viral proteins was absent or significantly delayed in cells infected with the pUL25 mutant virus. Transmission electron microscopy revealed capsids accumulated at nuclear pores that retained the viral genome for at least 4 h postinfection. In addition, cryoelectron microscopy (cryo-EM) reconstructions of virion capsids did not detect any obvious differences in the location or structural organization for the pUL25 or pUL36 proteins on the pUL25 mutant capsids. Further, in contrast to wild-type virus, the antiviral response mediated by the viral DNA-sensing cyclic guanine adenine synthase (cGAS) was severely compromised for the pUL25 mutant. These results demonstrate that the pUL25 capsid protein has a critical role in releasing viral DNA from NPC-bound capsids.IMPORTANCE Herpes simplex virus 1 (HSV-1) is the causative agent of several pathologies ranging in severity from the common cold sore to life-threatening encephalitic infection. Early steps in infection include release of the capsid into the cytoplasm, docking of the capsid at a nuclear pore, and release of the viral genome into the nucleus

  13. Characterization of a Legionella micdadei mip mutant

    DEFF Research Database (Denmark)

    O'Connell, W A; Bangsborg, Jette Marie; Cianciotto, N P


    The pathogenesis of Legionella micdadei is dependent upon its ability to infect alveolar phagocytes. To better understand the basis of intracellular infection by this organism, we examined the importance of its Mip surface protein. In Legionella pneumophila, Mip promotes infection of both human m...... into the phagocyte. Similarly, the mutant was less able to parasitize Hartmannella amoebae. Taken together, these data argue that Mip specifically potentiates intracellular growth by L. micdadei....

  14. Some Mutant Forms of Quantum Mechanics

    CERN Document Server

    Takeuchi, Tatsu; Lewis, Zachary; Minic, Djordje


    We construct a `mutant' form of quantum mechanics on a vector space over the finite Galois field GF(q). We find that the correlations in our model do not violate the Clauser-Horne-Shimony-Holt (CHSH) version of Bell's inequality, despite the fact that the predictions of this discretized quantum mechanics cannot be reproduced with any hidden variable theory. An alternative `mutation' is also suggested.

  15. Spontaneous Nif- mutants of Rhodopseudomonas capsulata.


    Wall, J D; Love, J.; Quinn, S P


    Revertible, spontaneous Nif- mutants of Rhodopseudomonas capsulata have been shown to accumulate in cultures growing photosynthetically with an amino acid as the nitrogen source such that H2 is maximally produced. The majority of such strains carry mutations which are clustered in a short region of the chromosome, probably representing one or two genes. Because this cluster includes temperature-sensitive mutations, it is also likely that it identifies the structural gene of a polypeptide. The...

  16. A user-friendly modified pore-solid fractal model (United States)

    Ding, Dian-Yuan; Zhao, Ying; Feng, Hao; Si, Bing-Cheng; Hill, Robert Lee


    The primary objective of this study was to evaluate a range of calculation points on water retention curves (WRC) instead of the singularity point at air-entry suction in the pore-solid fractal (PSF) model, which additionally considered the hysteresis effect based on the PSF theory. The modified pore-solid fractal (M-PSF) model was tested using 26 soil samples from Yangling on the Loess Plateau in China and 54 soil samples from the Unsaturated Soil Hydraulic Database. The derivation results showed that the M-PSF model is user-friendly and flexible for a wide range of calculation point options. This model theoretically describes the primary differences between the soil moisture desorption and the adsorption processes by the fractal dimensions. The M-PSF model demonstrated good performance particularly at the calculation points corresponding to the suctions from 100 cm to 1000 cm. Furthermore, the M-PSF model, used the fractal dimension of the particle size distribution, exhibited an accepted performance of WRC predictions for different textured soils when the suction values were ≥100 cm. To fully understand the function of hysteresis in the PSF theory, the role of allowable and accessible pores must be examined.

  17. Evolution of pore space in sandstones in relation to diagenesis

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, G.


    Results are described from studying the collector properties of a bed of sandstones of Yan-10 at the oil field Malin of the Ordosskiy oil and gas basin. The bed is represented by ancient river sandstones of the Jurassic age occurring on eroded surface of the Triassic deposits (Yangan series) and covered with bed of coal deposits Yan-9. The following conclusions were drawn from the results of the studies. Evolution of the pore space actually is the process of gradual substitution of the primary pores by secondary; in this case decrease in porosity in the quartz sandstones occurs mainly because of depositing in the pores of authigenic minerals, and in the sandstones whose grains consist of minerals with lower mechanical strength, because of packing. Secondary porosity develops because of dissolving and kaolinization in the sandstones with high content of feldspars. This process is possibly associated with decarboxilation of organic matter of interlayers of coal and calcareous shales under the influence of increased temperature in the submersion process. Since it has been established that considerable influence on porosity comes from the mineralogical position of the sandstones controlled by the sources of formation, in order to reveal the zones for development of primary and secondary porosity, it is very important to reveal the sources of removal of detrital rocks. A study was also made of the influence of diagenesis on uniformity and texture of the sandstones (by the methods of curves of capillary pressure).

  18. Image-based relative permeability upscaling from the pore scale (United States)

    Norouzi Apourvari, Saeid; Arns, Christoph H.


    High resolution images acquired from X-ray μ-CT are able to map the internal structure of porous media on which multiphase flow properties can be computed. While the resolution of a few micrometers may be sufficient for capturing the pore space of many sandstones, most carbonates exhibit a large amount of microporosity; pores which are below the image resolution and are not resolved at specific resolution. Neglecting the effect of micropores on fluid flow and transport properties of these rocks can cause erroneous results in particular at partial saturations. Current image-based pore scale models typically only consider macropores for simulating fluid flow. In this paper, we quantify the effect of microporosity on the effective permeability of the wetting phase for heterogeneous model structures with varying amount of micro-to-macro porosity. A multi-scale numerical approach is proposed to couple an average effect of micropores with an explicit representation of macropores. The Brinkman equation is solved using a lattice Boltzmann formulation to facilitate the coupling of Darcy and Stokes equations in micropores and macropores, respectively. The results show good agreement between the fine scale solution and the results of the upscaled models in which microporous regions are homogenised. The paper analyses in particular the choice of the momentum sink parameter at low wetting phase saturations. It is shown that this parameter can be found using either a flux-based calculation of permeability of microporous regions or chosen purely on the basis of the effective permeability of these regions.

  19. MCU encodes the pore conducting mitochondrial calcium currents. (United States)

    Chaudhuri, Dipayan; Sancak, Yasemin; Mootha, Vamsi K; Clapham, David E


    Mitochondrial calcium (Ca(2+)) import is a well-described phenomenon regulating cell survival and ATP production. Of multiple pathways allowing such entry, the mitochondrial Ca(2+) uniporter is a highly Ca(2+)-selective channel complex encoded by several recently-discovered genes. However, the identity of the pore-forming subunit remains to be established, since knockdown of all the candidate uniporter genes inhibit Ca(2+) uptake in imaging assays, and reconstitution experiments have been equivocal. To definitively identify the channel, we use whole-mitoplast voltage-clamping, the technique that originally established the uniporter as a Ca(2+) channel. We show that RNAi-mediated knockdown of the mitochondrial calcium uniporter (MCU) gene reduces mitochondrial Ca(2+) current (I MiCa ), whereas overexpression increases it. Additionally, a classic feature of I MiCa , its sensitivity to ruthenium red inhibition, can be abolished by a point mutation in the putative pore domain without altering current magnitude. These analyses establish that MCU encodes the pore-forming subunit of the uniporter channel. DOI:

  20. Formation of Anodic Aluminum Oxide with Branched and Meshed Pores. (United States)

    Kim, Byeol; Lee, Jin Seok


    Anodic aluminum oxide (AAO), with a self-ordered hexagonal array, is important for various applications in nanofabrication including as the fabrication of nanotemplates and other nanostructures. With the consideration, there have been many efforts to control the characteristic parameters of porous anodic alumina by adjustment of the anodizing conditions such as the electrolyte, temperature, applied potential, and Al purity. In particular, impurities in Al are changing the morphology of an alumina film; however, the formation mechanism has not yet been explained. In this work, we anodized a high purity (99.999%, Al(high)) and low purity (99.8%, Al(low)) aluminum foil by a two-step anodization process in an oxalic acid solution or phosphoric acid. It was found that the purity of aluminum foil has influenced the morphology of the alumina film resulting in branched and meshed pores. Also, electrochemical analysis indicated that the branched and meshed pores in the low-purity Al foil formed by the presence of impurities. Impurities act as defects and change the general growth mechanism for pore formation by inducing an electric field imbalance during anodization. This work contributes to the research field of topographical chemistry and applied fields including nanofabrication.

  1. Cationic PAMAM dendrimers as pore-blocking binary toxin inhibitors. (United States)

    Förstner, Philip; Bayer, Fabienne; Kalu, Nnanya; Felsen, Susanne; Förtsch, Christina; Aloufi, Abrar; Ng, David Y W; Weil, Tanja; Nestorovich, Ekaterina M; Barth, Holger


    Dendrimers are unique highly branched macromolecules with numerous groundbreaking biomedical applications under development. Here we identified poly(amido amine) (PAMAM) dendrimers as novel blockers for the pore-forming B components of the binary anthrax toxin (PA63) and Clostridium botulinum C2 toxin (C2IIa). These pores are essential for delivery of the enzymatic A components of the internalized toxins from endosomes into the cytosol of target cells. We demonstrate that at low μM concentrations cationic PAMAM dendrimers block PA63 and C2IIa to inhibit channel-mediated transport of the A components, thereby protecting HeLa and Vero cells from intoxication. By channel reconstitution and high-resolution current recording, we show that the PAMAM dendrimers obstruct transmembrane PA63 and C2IIa pores in planar lipid bilayers at nM concentrations. These findings suggest a new potential role for the PAMAM dendrimers as effective polyvalent channel-blocking inhibitors, which can protect human target cells from intoxication with binary toxins from pathogenic bacteria.

  2. Structure of Staphylococcal α-Hemolysin, a Heptameric Transmembrane Pore (United States)

    Song, Langzhou; Hobaugh, Michael R.; Shustak, Christopher; Cheley, Stephen; Bayley, Hagan; Gouaux, J. Eric


    The structure of the Staphylococcus aureus α-hemolysin pore has been determined to 1.9 overset{circ}{mathrm A} resolution. Contained within the mushroom-shaped homo-oligomeric heptamer is a solvent-filled channel, 100 overset{circ}{mathrm A} in length, that runs along the sevenfold axis and ranges from 14 overset{circ}{mathrm A} to 46 overset{circ}{mathrm A} in diameter. The lytic, transmembrane domain comprises the lower half of a 14-strand antiparallel β barrel, to which each protomer contributes two β strands, each 65 overset{circ}{mathrm A} long. The interior of the β barrel is primarily hydrophilic, and the exterior has a hydrophobic belt 28 overset{circ}{mathrm A} wide. The structure proves the heptameric subunit stoichiometry of the α-hemolysin oligomer, shows that a glycine-rich and solvent-exposed region of a water-soluble protein can self-assemble to form a transmembrane pore of defined structure, and provides insight into the principles of membrane interaction and transport activity of β barrel pore-forming toxins.

  3. Pool boiling on rectangular fins with tunnel-pore structure

    Directory of Open Access Journals (Sweden)

    Pastuszko A.


    Full Text Available Complex experimental investigations were conducted in the area of pool boiling heat transfer on extended surfaces with internal tunnels limited by perforated foil. The experiments were carried out for water and R-123 at atmospheric pressure. The tunnel surfaces were fabricated from 0.05 – 0.1 mm thick perforated copper foil (pore diameters: 0.3, 0.4, 0.5 mm sintered with mini-fins formed by 5 and 10 mm high rectangular fins and horizontal inter-fin surface. The effect of the main fin height, pore diameters and tunnel pitch on nucleate pool boiling was examined. Substantial enhancement of heat transfer coefficient was observed for the investigated surfaces. The highest increase in the heat transfer coefficient was obtained for the 10 mm high fins – about 50kW/m2K for water and 15 kW/m2K for R-123. The investigated surfaces showed boiling heat transfer coefficients similar to those of existing tunnel-pore structures.

  4. Nonlinear transport of soft droplets in pore networks (United States)

    Vernerey, Franck; Benet Cerda, Eduard; Koo, Kanghyeon

    A large number of biological and technological processes depend on the transport of soft colloidal particles through porous media; this includes the transport and separation of cells, viruses or drugs through tissues, membranes and microfluidic devices. In these systems, the interactions between soft particles, background fluid and the surrounding pore space yield complex, nonlinear behaviors such as non-Darcy flows, localization and jamming. We devise a computational strategy to investigate the transport of non-wetting and deformable water droplets in a microfluidic device made of a random distribution of cylindrical obstacles. We first derive scaling laws for the entry of the droplet in a single pore and discuss the role of surface tension, contact angle and size in this process. This information is then used to study the transport of multiple droplets in an obstacle network. We find that when the droplet size is close to the pore size, fluid flow and droplet trafficking strongly interact, leading to local redistributions in pressure fields, intermittent clogging and jamming. Importantly, it is found that the overall droplet and fluid transport display three different scaling regimes depending on the forcing pressure, and that these regimes can be related to droplet properties.

  5. Membrane Pore Formation by Amyloid beta (25-35) Peptide (United States)

    Kandel, Nabin; Tatulian, Suren

    Amyloid (A β) peptide contributes to Alzheimer's disease by a yet unidentified mechanism. One of the possible mechanisms of A β toxicity is formation of pores in cellular membranes. We have characterized the formation of pores in phospholipid membranes by the Aβ25 - 35 peptide (GSNKGAIIGLM) using fluorescence, Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD) techniques. CD and FTIR identified formation of β-sheet structure upon incubation of the peptide in aqueous buffer for 2 hours. Unilamellar vesicles composed of a zwitterionic lipid, 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), and 70 % POPC plus 30 % of an acidic lipid, 1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG), are made in 30 mM CaCl2. Quin-2, a fluorophore that displays increased fluorescence upon Ca2+ binding, is added to the vesicles externally. Peptide addition results in increased Quin-2 fluorescence, which is interpreted by binding of the peptide to the vesicles, pore formation, and Ca2+ leakage. The positive and negative control measurements involve addition of a detergent, Triton X-100, which causes vesicle rupture and release of total calcium, and blank buffer, respectively.

  6. Electrically tunable pore morphology in nanoporous gold thin films

    Institute of Scientific and Technical Information of China (English)

    Tatiana S. Dorofeeva; Erkin Seker


    Nanoporous gold (np-Au) is an emerging nanostructured material that exhibits many desirable properties, including high electrical and thermal conductivity, high surface area-to-volume ratio, tunable pore morphology well-established surface-binding chemistry, and compatibility with microfabrication. These features make np-Au a popular material for use in fuel cells, optical and electrical biosensors, drug delivery vehicles, neural electrode coatings, and as a model system for nanoscale mechanics. In each of its many applications, np-Au morphology plays an essential role in the overall device operation. Therefore, precise morphological control is necessary to attain optimal device performance. Traditionally thermal treatment by furnaces and hot plates is used to obtain np-Au with self-similar but coarser morphologies. However, this approach lacks the ability to create different morphologies on a single substrate and requires high temperatures (〉 250 ℃) incompatible with most plastic substrates. Herein, we report electro-annealing as a novel method that permits control of the extent and location of pore coarsening on a single substrate in one fast treatment step. The electro-annealing entails much lower temperatures (〈 150 ℃) than traditional thermal treatment, putatively due to electrically assisted phenomena contributing to the thermally activated surface diffusion of gold atoms, responsible for coarsening. Overall, this approach is easily scaled to display multiple pore morphologies on a single chip, therefore enabling high-throughput screening of optimal nanostructures for specific applications.

  7. Graphene-Induced Pore Formation on Cell Membranes (United States)

    Duan, Guangxin; Zhang, Yuanzhao; Luan, Binquan; Weber, Jeffrey K.; Zhou, Royce W.; Yang, Zaixing; Zhao, Lin; Xu, Jiaying; Luo, Judong; Zhou, Ruhong


    Examining interactions between nanomaterials and cell membranes can expose underlying mechanisms of nanomaterial cytotoxicity and guide the design of safer nanomedical technologies. Recently, graphene has been shown to exhibit potential toxicity to cells; however, the molecular processes driving its lethal properties have yet to be fully characterized. We here demonstrate that graphene nanosheets (both pristine and oxidized) can produce holes (pores) in the membranes of A549 and Raw264.7 cells, substantially reducing cell viability. Electron micrographs offer clear evidence of pores created on cell membranes. Our molecular dynamics simulations reveal that multiple graphene nanosheets can cooperate to extract large numbers of phospholipids from the membrane bilayer. Strong dispersion interactions between graphene and lipid-tail carbons result in greatly depleted lipid density within confined regions of the membrane, ultimately leading to the formation of water-permeable pores. This cooperative lipid extraction mechanism for membrane perforation represents another distinct process that contributes to the molecular basis of graphene cytotoxicity. PMID:28218295

  8. Alpha-tocopherol inhibits pore formation in the oxidized bilayers

    CERN Document Server

    Boonnoy, Phansiri; Wong-ekkabut, Jirasak


    In biological membranes, alpha-tocopherols ({\\alpha}-toc; vitamin E) protect polyunsaturated lipids from free radicals. Although the interactions of {\\alpha}-toc with non-oxidized lipid bilayers have been studied, their on oxidized bilayers remain unknown. In this study, atomistic molecular dynamics (MD) simulations of oxidized lipid bilayers were performed with varying concentrations of {\\alpha}-toc. Bilayers with 1-palmitoyl-2-lauroyl-sn-glycero-3-phosphocholine (PLPC) lipids and its aldehyde derivatives at 1:1 ratio were studied. Our simulations show that oxidized lipids self-assemble into aggregates with a water pore rapidly developing across the lipid bilayer. The free energy of transporting an {\\alpha}-toc molecule in a lipid bilayer suggests that {\\alpha}-tocs can passively adsorb into the bilayer. When {\\alpha}-toc molecules were present at low concentrations in bilayers containing oxidized lipids, the formation of water pores was slowed down. At high {\\alpha}-toc concentra-tions, no pores were observ...

  9. Hydrogeology and hydrodynamics of coral reef pore waters

    Energy Technology Data Exchange (ETDEWEB)

    Buddemeier, R.W.; Oberdorfer, J.A.


    A wide variety of forces can produce head gradients that drive the flow and advective mixing of internal coral reef pore waters. Oscillatory gradients that produce mixing result from wave and tide action. Sustained gradients result from wave and tide-induced setup and ponding, from currents impinging on the reef structure, from groundwater heads, and from density differenced (temperature or salinity gradients). These gradients and the permeabilities and porosities of reef sediments are such that most macropore environments are dominated by advection rather than diffusion. The various driving forces must be analyzed to determine the individual and combined magnitudes of their effects on a specific reef pore-water system. Pore-water movement controls sediment diagenesis, the exchange of nutrients between sediments and benthos, and coastal/island groundwater resources. Because of the complexity of forcing functions, their interactions with specific local reef environments, experimental studies require careful incorporation of these considerations into their design and interpretation. 8 refs., 3 figs., 1 tab.

  10. Mutant chaperonin proteins: new tools for nanotechnology

    Energy Technology Data Exchange (ETDEWEB)

    Li, Y [SETI Institute, 515 N Whisman Road, Mountain View, CA 94043 (United States); Paavola, C D [NASA Ames Research Center, Bioengineering Branch, Mail Stop 239-15, Moffett Field, CA 94035 (United States); Kagawa, H [SETI Institute, 515 N Whisman Road, Mountain View, CA 94043 (United States); Chan, S L [SETI Institute, 515 N Whisman Road, Mountain View, CA 94043 (United States); Trent, J D [NASA Ames Research Center, Bioengineering Branch, Mail Stop 239-15, Moffett Field, CA 94035 (United States)


    Much effort has gone into finding peptides that bind potentially useful nanoparticles, but relatively little effort has focused on the scaffolds that organize these peptides into useful nanostructures. Chaperonins are protein complexes with 14-18 protein subunits that self-assemble into double-ring complexes and function as scaffolds for peptides or amino acids that bind metallic and semiconductor quantum dots. The utility of chaperonins as scaffolds depends on their structure and their ability to self-assemble into double-rings and higher-order structures, such as filaments and two-dimensional arrays. To better understand the structure of chaperonins, we constructed a model of a group II chaperonin and, based on this model, genetically constructed five mutant subunits with significant deletions. We expressed these mutants as recombinant proteins and observed by native polyacrylamide gel electrophoresis (PAGE) and transmission electron microscopy (TEM) that they all self-assembled into double rings. Our model predicted and TEM confirmed that these deletions did not significantly change the 17 nm diameter of the wild-type double rings, but decreased their height and opened their central cavities. Four of the five mutants formed higher-order structures: chains of rings, bundles of chains or filaments, and two-dimensional arrays, which we suggest can be useful nanostructures.

  11. Understanding the effect of mean pore size on cell activity in collagen-glycosaminoglycan scaffolds

    National Research Council Canada - National Science Library

    Murphy, Ciara M; O'Brien, Fergal J


    Mean pore size is an essential aspect of scaffolds for tissue-engineering. If pores are too small cells cannot migrate in towards the center of the construct limiting the diffusion of nutrients and removal of waste products...

  12. Evaluation of Optimal Pore Size of (3-Aminopropyltriethoxysilane Grafted MCM-41 for Improved CO2 Adsorption

    Directory of Open Access Journals (Sweden)

    Zhilin Liu


    Full Text Available An array of new MCM-41 with substantially larger average pore diameters was synthesized through adding 1,3,5-trimethylbenzene (TMB as the swelling agent to explore the effect of pore size on final adsorbent properties. The pore expanded MCM-41 was also grafted with (3-Aminopropyltriethoxysilane (APTES to determine the optimal pore size for CO2 adsorption. The pore-expanded mesoporous MCM-41s showed relatively less structural regularity but significant increments of pore diameter (4.64 to 7.50 nm; the fraction of mesopore volume also illustrated an increase. The adsorption heat values were correlated with the order of the adsorption capacities for pore expanded MCM-41s. After amine functionalization, the adsorption capacities and heat values showed a significant increase. APTES-grafted pore-expanded MCM-41s depicted a high potential for CO2 capture regardless of the major drawback of the high energy required for regeneration.

  13. Isolation of a novel mutant from Bacillus subtilis natto. (United States)

    Yoshida, Kazuo


    For the construction of strains with full probiotics function in intestines, deoxycholate resistant mutants were isolated from Bacillus subtilis natto. The partial characterization of the mutants was carried out and described.

  14. Studying Pore Structure of Nonwovens with 3D Imaging and Modeling Permeability (United States)

    Baradari, Mehdi Gholipour

    Nonwovens are classified as a porous material and pore structure is named as the most important and complex feature of them. Since pore structure is out of control during any nonwovens manufacturing processes, many attempts have been made to measure the major characteristics of a pore network including: pore size, pore volume, pore surface area and pore shape. Among all pore characteristics, pore size due to its significant influence on many nonwovens applications such as filtration is counted as the most significant one. Generally, experiment, theoretical modeling and image analysis are the most common methods to measure pore size of nonwovens. Normally, pores in nonwovens make many convergences and divergences along the length and for this reason, many pore diameters could be assigned for a media. Due to inefficiency of the aforementioned techniques to measure all these diameters, they are not precise enough to study pore structure. The initial objective of this research is obtaining information of the pore structure, especially pore sizes, by applying image analysis techniques to a 3D image of nonwovens obtained through 3D imaging techniques such as DVI and micro CT. This 3D structure of the nonwoven media will be transformed to a graph, employing skeletonization through AvizoRTM software. The obtained graph exhibits topology, shape and connectivity of the pore structure for the utilized nonwoven. In this graph, each node and link would be a representative for pores intersection and body of pore, respectively. Saving the information of this graph results to some matrices/vectors including nodes coordinated, connectivity and nodes thickness, which exhibits the pore size. Therefore, all the pore sizes available in the structure will be extracted through this method. As expected, the information obtained from pore network is very complex consisting many numbers, so analyse them would be very difficult. Therefore, it was tried to use the saved information to model

  15. Molecular dynamics simulation of benzene in graphite and amorphous carbon slit pores. (United States)

    Fomin, Yu D


    It is well known that confining a liquid into a pore strongly alters the liquid behavior. Investigations of the effect of confinement are of great importance for many scientific and technological applications. Here, we present a study of the behavior of benzene confined in carbon slit pores. Two types of pores are considered-graphite and amorphous carbon ones. We show that the effect of different pore structure is of crucial importance for the benzene behavior.

  16. Study on Fractal Characteristics of Cracks and Pore Structure of Concrete based on Digital Image Technology


    Xianyu Jin; Bei Li; Ye Tian; Nanguo Jin; An Duan


    Based on the fractal theory, this study presents a numerical analysis on the fractal characteristics of cracks and pore structure of concrete with the help of digital image technology. The results show that concrete cracks and the micro pore distribution of concrete are of fractal characteristics and the fractal dimension ranges from 1 to 2. The fractal characteristics of pores in cracked concrete and un-cracked concrete is similar and the former fractal dimension of the micro pore structure ...

  17. Pore pressure development in hybrid fibre-reinforced high strength concrete at elevated temperatures


    Bangi, Mugume Rodgers; HORIGUCHI, Takashi


    The present experimental work investigates the build-up of pore pressure at different depths of High Strength Concrete (HSC) and Hybrid-Fibre-Reinforced High Strength Concrete (HFRHSC) when exposed to different heating rates. First, the effect of the measurement technique on maximum pore pressures measured was evaluated. The pressure measurement technique which utilized a sintered metal and silicon oil was found to be the most effective technique for pore pressure measurement. Pore pressure m...

  18. Pore Capture in Shales Due to Pervasive Micro-fracturing (United States)

    Hayman, N. W.; Daigle, H.; Kelly, E. D.; Milliken, K. L.; Jiang, H.


    Unconventional production from mudrock reservoirs is associated with steep production declines in individual wells. The decline curves may partly stem from the induced fracture spacing which is on the order of meters, whereas most of the hydrocarbon-bearing pores are at the nanoscale. Previous analysis of production data, however, indicates some permeability enhancement between the more widely spaced fractures that may be due to a small-scale microfracture network. In order to test the microfracture hypothesis, we fractured samples of shale under shear deformation and observed (post-experimental) changes in microstructure through scanning electron microscopy, CT-scanning and focused ion beam (FIB) imaging, low-pressure nitrogen sorption, and nuclear magnetic resonance. Our results indicate that a system of microcracks with apertures ranging from tens of nanometers to tens of microns forms in response to shear failure, which in some cases propagate into organic matter and intersect the organic-hosted pores, a process we refer to as "pore capture". This micromechanical process presumably underlies the bulk porosity response (increase) which is directly affected by the abundance of organic matter (OM), the fabric of the rock, and the direction of loading relative to any fabric. The trajectory of the fractures is governed by the contrasts in mechanical strength (and, potentially, the brittle-vs.-ductile response) among the different components of the shale and within the OM. Specifically, the fractures tend to follow trajectories through weaker material, which favors their propagation along grain contacts and into OM. However, such grain-boundary fractures do not typically intersect intergranular pores, possibly because these are preferentially located in mechanically stronger areas that resist deformation. The fractures we observed do not appear to contribute to enhanced connectivity of the overall intergranular pore system, potentially explaining why fracking yields

  19. Teardrop shapes minimize bending energy of fusion pores connecting planar bilayers (United States)

    Ryham, Rolf J.; Ward, Mark A.; Cohen, Fredric S.


    A numerical gradient flow procedure was devised to characterize minimal energy shapes of fusion pores connecting two parallel planar bilayer membranes. Pore energy, composed of splay, tilt, and stretching, was obtained by modeling each bilayer as two monolayers and treating each monolayer of a bilayer membrane as a freely deformable surface described with a mean lipid orientation field. Voids between the two monolayers were prevented by a steric penalty formulation. Pore shapes were assumed to possess both axial and reflectional symmetry. For fixed pore radius and bilayer separation, the gradient flow procedure was applied to initially toroidal pore shapes. Using initially elliptical pore shapes yielded the same final shape. The resulting minimal pore shapes and energies were analyzed as a function of pore dimension and lipid composition. Previous studies either assumed or confined pore shapes, thereby tacitly supplying an unspecified amount of energy to maintain shape. The shapes derived in the present study were outputs of calculations and an externally provided energy was not supplied. Our procedure therefore yielded energy minima significantly lower than those reported in prior studies. The membrane of minimal energy pores bowed outward near the pore lumen, yielding a pore length that exceeded the distance between the two fusing membranes.

  20. Reactive/Adsorptive transport in (partially-) saturated porous media: from pore scale to core scale

    NARCIS (Netherlands)

    Raoof, A.


    Pore-scale modeling provides opportunities to study transport phenomena in fundamental ways because detailed information is available at the microscopic pore scale. This offers the best hope for bridging the traditional gap that exists between pore scale and macro (lab) scale description of the proc


    Institute of Scientific and Technical Information of China (English)

    Deying Wang; Ziqiu Shen


    Scanning electronic microscope was adopted to investigate the pore structure of deposits formed during polypropylene fiber bundle filtration. The effects of flocculants, cationic polyacrylamide and polyaluminumchloride, on the pore structure and filtration process were examined. It is found from experimental results that the filter deposit has a self-similarity pore structure, which can be described in fractal dimensions.

  2. Hard, charged spheres in spherical pores. Grand canonical ensemble Monte Carlo calculations

    DEFF Research Database (Denmark)

    Sloth, Peter; Sørensen, T. S.


    A model consisting of hard charged spheres inside hard spherical pores is investigated by grand canonical ensemble Monte Carlo calculations. It is found that the mean ionic density profiles in the pores are almost the same when the wall of the pore is moderately charged as when it is uncharged...

  3. Effects of sand compaction and mixing on pore structure and the unsaturated soil hydraulic properties

    NARCIS (Netherlands)

    Mahmoodlu, Mojtaba Ghareh|info:eu-repo/dai/nl/357287746; Raoof, A.|info:eu-repo/dai/nl/304842338; Sweijen, T.|info:eu-repo/dai/nl/369415191; van Genuchten, M. Th


    The hydraulic properties of unsaturated porous media very much depend on their pore structure as defined by the size, arrangement, and connectivity of pores. Several empirical and quasi-empirical approaches have been used over the years to derive pore structure information from the particle size dis

  4. Performance of slotted pores in particle manufacture using rotating membrane emulsification

    Institute of Scientific and Technical Information of China (English)

    Qingchun Yuan; Nita Aryanti; Ruozhou Hou; Richard A.Williams


    This paper addresses the use of different slotted pores in rotating membrane emulsification technology.Pores of square and rectangular shapes were studied to understand the effect of aspect ratio (1-3.5) and their orientation on oil droplet formation.Increasing the membrane rotation speed decreased the droplet size,and the oil droplets produced were more uniform using slotted pores as compared to circular geometry.At a given rotation speed,the droplet size was mainly determined by the pore size and the fluid velocity of oil through the pore (pore fluid velocity).The ratio of droplet diameter to the equivalent diameter of the slotted pore increased with the pore fluid velocity.At a given pore fluid velocity and rotation speed,pore orientation significantly influences the droplet formation rate: horizontally disposed pores (with their longer side perpendicular to the membrane axis) generate droplets at double the rate of vertically disposed pores.This work indicates practical benefits in the use of slotted membranes over conventional methods.

  5. Experimental Study and Numerical Modeling of Wave Induced Pore Pressure Attenuation Inside a Rubble Mound Breakwater

    DEFF Research Database (Denmark)

    Troch, Peter; Rouck, Julien De; Burcharth, Hans Falk


    The main objective of this paper is to study the attenuation of the wave induced pore pressures inside the core of a rubble mound breakwater. The knowledge of the distribution and the attenuation of the pore pressures is important for the design of a stable and safe breakwater. The pore pressure...

  6. Inducement and identification of an endosperm mutant in maize

    African Journals Online (AJOL)

    ajl yemi


    Nov 30, 2011 ... “super sweet” phenotype were derived from the mutated offspring. ... characteristics and distinguished molecular mechanism to the previous mutants of gene sh2, these three mutant lines are ...... Physical association of starch biosynthetic ... reduced seedling mutant in oilseed rape, Brassica napus, for.

  7. Fluorescence-tracking of activation gating in human ERG channels reveals rapid S4 movement and slow pore opening.

    Directory of Open Access Journals (Sweden)

    Zeineb Es-Salah-Lamoureux

    Full Text Available BACKGROUND: hERG channels are physiologically important ion channels which mediate cardiac repolarization as a result of their unusual gating properties. These are very slow activation compared with other mammalian voltage-gated potassium channels, and extremely rapid inactivation. The mechanism of slow activation is not well understood and is investigated here using fluorescence as a direct measure of S4 movement and pore opening. METHODS AND FINDINGS: Tetramethylrhodamine-5-maleimide (TMRM fluorescence at E519 has been used to track S4 voltage sensor movement, and channel opening and closing in hERG channels. Endogenous cysteines (C445 and C449 in the S1-S2 linker bound TMRM, which caused a 10 mV hyperpolarization of the V((1/2 of activation to -27.5+/-2.0 mV, and showed voltage-dependent fluorescence signals. Substitution of S1-S2 linker cysteines with valines allowed unobstructed recording of S3-S4 linker E519C and L520C emission signals. Depolarization of E519C channels caused rapid initial fluorescence quenching, fit with a double Boltzmann relationship, F-V(ON, with V((1/2 (,1 = -37.8+/-1.7 mV, and V((1/2 (,2 = 43.5+/-7.9 mV. The first phase, V((1/2 (,1, was approximately 20 mV negative to the conductance-voltage relationship measured from ionic tail currents (G-V((1/2 = -18.3+/-1.2 mV, and relatively unchanged in a non-inactivating E519C:S620T mutant (V((1/2 = -34.4+/-1.5 mV, suggesting the fast initial fluorescence quenching tracked S4 voltage sensor movement. The second phase of rapid quenching was absent in the S620T mutant. The E519C fluorescence upon repolarization (V((1/2 = -20.6+/-1.2, k = 11.4 mV and L520C quenching during depolarization (V((1/2 = -26.8+/-1.0, k = 13.3 mV matched the respective voltage dependencies of hERG ionic tails, and deactivation time constants from -40 to -110 mV, suggesting they detected pore-S4 rearrangements related to ionic current flow during pore opening and closing. CONCLUSION: THE DATA INDICATE: 1

  8. Mutant prevention concentration and mutant selection window for 10 antimicrobial agents against Rhodococcus equi. (United States)

    Berghaus, Londa J; Giguère, Steeve; Guldbech, Kristen


    The objectives of this study were to determine the mutant prevention concentration (MPC), time above the MPC and mutant selection window for 10 antimicrobial agents against Rhodococcus equi and to determine if the combination of a macrolide with rifampin would decrease emergence of resistant mutants. Antimicrobial agents investigated (erythromycin, clarithromycin, azithromycin, rifampin, amikacin, gentamicin, enrofloxacin, vancomycin, imipenem, and doxycycline) were selected based on in vitro activity and frequency of use in foals or people infected with R. equi. Each antimicrobial agent or combination of agents was evaluated against four virulent strains of R. equi. MPC were determined using an agar plate assay. Pharmacodynamic parameters were calculated using published plasma and pulmonary pharmacokinetic variables. There was a significant (Pequi. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Characterisation of cuticular mutants in Arabidopsis thaliana


    Faust, Andrea


    Plants are protected by the extracellular cuticle, which is made up of cutin, cutan and waxes. The cutin composition of a variety of plants has been known and models of the biosynthesis of cutin monomers exist but not many enzymes have been identified. It is generally accepted that a defect in the cuticle leads to an organ fusion phenotype. In the model plant A. thaliana many fusion mutants have been identified but the identification of genes involved have not lead to a complete picture of th...

  10. In-pore exchange and diffusion of carbonate solvent mixtures in nanoporous carbon (United States)

    Alam, Todd M.; Osborn Popp, Thomas M.


    High resolution magic angle spinning (HRMAS) 1H NMR spectroscopy has been used to resolve different surface and in-pore solvent environments of ethylene carbonate (EC) and dimethyl carbonate (DMC) mixtures absorbed within nanoporous carbon (NPC). Two dimensional (2D) 1H HRMAS NMR exchange measurements revealed that the inhomogeneous broadened in-pore resonances have pore-to-pore exchange rates on the millisecond timescale. Pulsed-field gradient (PFG) NMR diffusometry revealed the in-pore self-diffusion constants for both EC and DMC were reduced by up to a factor of five with respect to the diffusion in the non-absorbed solvent mixtures.

  11. Diode-like properties of single- and multi-pore asymmetric track membranes

    Energy Technology Data Exchange (ETDEWEB)

    Zielinska, K., E-mail: [Flerov Laboratory of Nuclear Reactions, Joint Institute for Nuclear Research, Joliot-Curie str. 6, 141980 Dubna (Russian Federation); Faculty of Chemistry, Nicolaus Copernicus University, Gagarina str. 7, 87-100 Torun (Poland); Gapeeva, A.R.; Orelovich, O.L. [Flerov Laboratory of Nuclear Reactions, Joint Institute for Nuclear Research, Joliot-Curie str. 6, 141980 Dubna (Russian Federation); Apel, P.Yu. [Flerov Laboratory of Nuclear Reactions, Joint Institute for Nuclear Research, Joliot-Curie str. 6, 141980 Dubna (Russian Federation); The International University “Dubna”, Universitetskaya str. 19, 141980 Dubna (Russian Federation)


    In this work, we investigated the ionic transport properties of asymmetric polyethylene terephthalate (PET) track membranes with the thickness of 5 μm. The samples containing single pores and arrays of many pores were fabricated by irradiation with accelerated ions and subsequent physicochemical treatment. The method of etching in the presence of a surface-active agent was used to prepare the pores with highly-tapered tip. The transport of monovalent inorganic ions through the nano-scale holes was studied in a conductivity cell. The effective pore radii, electrical conductance and rectification ratios of pores were measured. The geometric characteristics of nanopores were investigated using FESEM.

  12. Experimental study on pore pressure in rock-soil slope during reservoir water level fluctuation

    Institute of Scientific and Technical Information of China (English)

    LIU; Yuewu; CHEN; Huixin; LIU; Qingquan; GONG; Xin; ZHANG


    A test system was developed for measuring the pore pressure in porous media, and a new model was devised for the pore pressure testing in both saturated and unsaturated rock-soil. Laboratory experiments were carried out to determine the pore pressure during water level fluctuation. The variations of transient pore pressure vs. time at different locations of the simulated rock-soil system were acquired and processed, and meanwhile the deformation and failure of the model are observed. The experiment results show that whether the porous media are saturated or not, the transient pore pressure is mainly dependent on the water level fluctuation, and coupled with the variation of the stress field.

  13. Effect of Fiber Properties on Nonwovens' Pore Structures with Fractal Geometry Analysis

    Institute of Scientific and Technical Information of China (English)


    Nonwovens' pore structures are very important to their mechanical and physical properties. And the pore structures are influenced by the fiber properties and fibers arrangement in web. In this paper, the fractal geometry, in combination with computer image analysis, is used to express the irregularity of pore size distribution in nonwovens, and the effect of fiber properties on fractal dimension of pore size distribution isdiscussed by using simulated images which are composed of nonlinear staple fibers. The results show that the fiber properties,such as crimp, diameer, angular distribution, and especially the number of fibers prominently influence the pore structure.

  14. Pore former induced porosity in LSM/CGO cathodes for electrochemical cells for flue gas purification

    DEFF Research Database (Denmark)

    Skovgaard, M.; Andersen, Kjeld Bøhm; Kammer Hansen, Kent


    In this study the effect of the characteristics of polymethyl methacrylate (PMMA) pore formers on the porosity, pore size distribution and the air flow through the prepared lanthanum strontium manganate/gadolinium-doped cerium oxide (LSM/CGO) cathodes was investigated. Porous cathodes were obtained...... and the highest porosity measured was 46.4% with an average pore diameter of 0.98 μm. The air flow through this cathode was measured to 5.8 ml/(min mm2). Also the effect of exposure time to the solvent was tested for the most promising PMMA pore former and it was found that the average pore diameter decreases...

  15. Conical tungsten stamps for the replication of pore arrays in anodic aluminium oxide films. (United States)

    LeClere, D J; Thompson, G E; Derby, B


    A tungsten master stamp has been generated by applying a novel procedure that includes two-step anodizing, followed by sequential anodizing and pore widening to develop nominally funnelled pores. These conical-shaped pores were filled with tungsten by sputter coating to manufacture a master stamp. Under a pressure of 65 MPa, the master stamp successfully embossed the surface of annealed and electropolished aluminium. The embossed surface was then used to control the position of pores created by anodizing under the conditions used to produce the original pore array.

  16. Pore-linked filaments in anura spermatocyte nuclei

    Directory of Open Access Journals (Sweden)

    Maria Luiza Beçak


    Full Text Available Pore-linked filaments were visualized in spreads of anuran spermatocyte nuclei using transmission electron microscope. We used Odontophrynus diplo and tetraploid species having the tetraploid frogs reduced metabolic activities. The filaments with 20-40 nm width are connected to a ring component of the nuclear pore complex with 90-120 nm and extend up to 1µm (or more into the nucleus. The filaments are curved and connect single or neighboring pores. The intranuclear filaments are associated with chromatin fibers and related to RNP particles of 20-25 nm and spheroidal structures of 0.5µm, with variations. The aggregates of several neighboring pores with the filaments are more commonly observed in 4n nuclei. We concluded that the intranuclear filaments may correspond to the fibrillar network described in Xenopus oocyte nucleus being probably related to RNA transport. The molecular basis of this RNA remains elusive. Nevertheless, the morphological aspects of the spheroidal structures indicate they could correspond to nucleolar chromatin or to nucleolus-derived structures. We also speculate whether the complex aggregates of neighboring pores with intranuclear filaments may correspond to pore clustering previously described in these tetraploid animals using freeze-etching experiments.Filamentos ligados a poros foram visualizados em núcleos de espermatócitos de anuros através da técnica de espalhamento para microscopia eletrônica de transmissão. Os animais usados pertencem ao gênero Odontophrynus com espécies cripticas diplo e tetraplóides naturais, tendo os tetraplóides atividade metabólica reduzida. Os filamentos com 20-40 nm de largura são ligados a um anel componente do complexo poro nuclear de 90-120 nm e estendem-se até 1 µm (ou mais para dentro do núcleo. Os filamentos são curvos e ligam poros simples ou poros vizinhos. Os filamentos intranucleares são associados a fibras de cromatina e relacionados a partículas de RNP de 20

  17. Pre-pore oligomer formation by Vibrio cholerae cytolysin: insights from a truncated variant lacking the pore-forming pre-stem loop. (United States)

    Paul, Karan; Chattopadhyay, Kausik


    Vibrio cholerae cytolysin (VCC), a β-barrel pore-forming toxin (β-PFT), induces killing of the target eukaryotic cells by forming heptameric transmembrane β-barrel pores. Consistent with the β-PFT mode of action, binding of the VCC toxin monomers with the target cell membrane triggers formation of pre-pore oligomeric intermediates, followed by membrane insertion of the β-strands contributed by the pre-stem motif within the central cytolysin domain of each protomer. It has been shown previously that blocking of membrane insertion of the VCC pre-stem motif arrests conversion of the pre-pore state to the functional transmembrane pore. Consistent with the generalized β-PFT mechanism, it therefore appears that the VCC pre-stem motif plays a critical role toward forming the structural scaffold of the transmembrane β-barrel pore. It is, however, still not known whether the pre-stem motif plays any role in the membrane interaction process, and subsequent pre-pore structure formation by VCC. In this direction, we have constructed a recombinant variant of VCC deleting the pre-stem region, and have characterized the effect(s) of physical absence of the pre-stem motif on the distinct steps of the membrane pore-formation process. Our results show that the deletion of the pre-stem segment does not affect membrane binding and pre-pore oligomer formation by the toxin, but it critically abrogates the functional pore-forming activity of VCC. Present study extends our insights regarding the structure-function mechanism associated with the membrane pore formation by VCC, in the context of the β-PFT mode of action.

  18. Structure of a C. perfringens enterotoxin mutant in complex with a modified Claudin-2 extracellular loop 2. (United States)

    Yelland, Tamas S; Naylor, Claire E; Bagoban, Tannya; Savva, Christos G; Moss, David S; McClane, Bruce A; Blasig, Ingolf E; Popoff, M; Basak, Ajit K


    CPE (Clostridium perfringens enterotoxin) is the major virulence determinant for C. perfringens type-A food poisoning, the second most common bacterial food-borne illness in the UK and USA. After binding to its receptors, which include particular human claudins, the toxin forms pores in the cell membrane. The mature pore apparently contains a hexamer of CPE, claudin and, possibly, occludin. The combination of high binding specificity with cytotoxicity has resulted in CPE being investigated, with some success, as a targeted cytotoxic agent for oncotherapy. In this paper, we present the X-ray crystallographic structure of CPE in complex with a peptide derived from extracellular loop 2 of a modified, CPE-binding Claudin-2, together with high-resolution native and pore-formation mutant structures. Our structure provides the first atomic-resolution data on any part of a claudin molecule and reveals that claudin's CPE-binding fingerprint (NPLVP) is in a tight turn conformation and binds, as expected, in CPE's C-terminal claudin-binding groove. The leucine and valine residues insert into the binding groove while the first residue, asparagine, tethers the peptide via an interaction with CPE's aspartate 225 and the two prolines are required to maintain the tight turn conformation. Understanding the structural basis of the contribution these residues make to binding will aid in engineering CPE to target tumor cells.

  19. Bax and Bak function as the outer membrane component of the mitochondrial permeability pore in regulating necrotic cell death in mice (United States)

    Karch, Jason; Kwong, Jennifer Q; Burr, Adam R; Sargent, Michelle A; Elrod, John W; Peixoto, Pablo M; Martinez-Caballero, Sonia; Osinska, Hanna; Cheng, Emily H-Y; Robbins, Jeffrey; Kinnally, Kathleen W; Molkentin, Jeffery D


    A critical event in ischemia-based cell death is the opening of the mitochondrial permeability transition pore (MPTP). However, the molecular identity of the components of the MPTP remains unknown. Here, we determined that the Bcl-2 family members Bax and Bak, which are central regulators of apoptotic cell death, are also required for mitochondrial pore-dependent necrotic cell death by facilitating outer membrane permeability of the MPTP. Loss of Bax/Bak reduced outer mitochondrial membrane permeability and conductance without altering inner membrane MPTP function, resulting in resistance to mitochondrial calcium overload and necrotic cell death. Reconstitution with mutants of Bax that cannot oligomerize and form apoptotic pores, but still enhance outer membrane permeability, permitted MPTP-dependent mitochondrial swelling and restored necrotic cell death. Our data predict that the MPTP is an inner membrane regulated process, although in the absence of Bax/Bak the outer membrane resists swelling and prevents organelle rupture to prevent cell death. DOI: PMID:23991283

  20. Bax and Bak function as the outer membrane component of the mitochondrial permeability pore in regulating necrotic cell death in mice. (United States)

    Karch, Jason; Kwong, Jennifer Q; Burr, Adam R; Sargent, Michelle A; Elrod, John W; Peixoto, Pablo M; Martinez-Caballero, Sonia; Osinska, Hanna; Cheng, Emily H-Y; Robbins, Jeffrey; Kinnally, Kathleen W; Molkentin, Jeffery D


    A critical event in ischemia-based cell death is the opening of the mitochondrial permeability transition pore (MPTP). However, the molecular identity of the components of the MPTP remains unknown. Here, we determined that the Bcl-2 family members Bax and Bak, which are central regulators of apoptotic cell death, are also required for mitochondrial pore-dependent necrotic cell death by facilitating outer membrane permeability of the MPTP. Loss of Bax/Bak reduced outer mitochondrial membrane permeability and conductance without altering inner membrane MPTP function, resulting in resistance to mitochondrial calcium overload and necrotic cell death. Reconstitution with mutants of Bax that cannot oligomerize and form apoptotic pores, but still enhance outer membrane permeability, permitted MPTP-dependent mitochondrial swelling and restored necrotic cell death. Our data predict that the MPTP is an inner membrane regulated process, although in the absence of Bax/Bak the outer membrane resists swelling and prevents organelle rupture to prevent cell death. DOI:

  1. The evolution of pore connectivity in volcanic rocks (United States)

    Colombier, Mathieu; Wadsworth, Fabian B.; Gurioli, Lucia; Scheu, Bettina; Kueppers, Ulrich; Di Muro, Andrea; Dingwell, Donald B.


    Pore connectivity is a measure of the fraction of pore space (vesicles, voids or cracks) in a material that is interconnected on the system length scale. Pore connectivity is fundamentally related to permeability, which has been shown to control magma outgassing and the explosive potential of magma during ascent in the shallowest part of the crust. Here, we compile a database of connectivity and porosity from published sources and supplement this with additional measurements, using natural volcanic rocks produced in a broad range of eruptive styles and with a range of bulk composition. The database comprises 2715 pairs of connectivity C and porosity ϕ values for rocks from 35 volcanoes as well as 116 products of experimental work. For 535 volcanic rock samples, the permeability k was also measured. Data from experimental studies constrain the general features of the relationship between C and ϕ associated with both vesiculation and densification processes, which can then be used to interpret natural data. To a first order, we show that a suite of rocks originating from effusive eruptive behaviour can be distinguished from rocks originating from explosive eruptive behaviour using C and ϕ. We observe that on this basis, a particularly clear distinction can be made between scoria formed in fire-fountains and that formed in Strombolian activity. With increasing ϕ, the onset of connectivity occurs at the percolation threshold ϕc which in turn can be hugely variable. We demonstrate that C is an excellent metric for constraining ϕc in suites of porous rocks formed in a common process and discuss the range of ϕc values recorded in volcanic rocks. The percolation threshold is key to understanding the onset of permeability, outgassing and compaction in shallow magmas. We show that this threshold is dramatically different in rocks formed during densification processes than in rocks formed in vesiculating processes and propose that this value is the biggest factor in

  2. Ring cycle for dilating and constricting the nuclear pore. (United States)

    Solmaz, Sozanne R; Blobel, Günter; Melcák, Ivo


    We recently showed that the three "channel" nucleoporins, Nup54, Nup58, and Nup62, interact with each other through only four distinct sites and established the crystal structures of the two resulting "interactomes," Nup54•Nup58 and Nup54•Nup62. We also reported instability of the Nup54•Nup58 interactome and previously determined the atomic structure of the relevant Nup58 segment by itself, demonstrating that it forms a twofold symmetric tetramer. Here, we report the crystal structure of the relevant free Nup54 segment and show that it forms a tetrameric, helical bundle that is structurally "conditioned" for instability by a central patch of polar hydrogen-bonded residues. Integrating these data with our previously reported results, we propose a "ring cycle" for dilating and constricting the nuclear pore. In essence, three homooligomeric rings, one consisting of eight modules of Nup58 tetramers, and two, each consisting of eight modules of Nup54 tetramers, are stacked in midplane and characterize a constricted pore of 10- to 20-nm diameter. In going to the dilated state, segments of one Nup58 and two Nup54 tetrameric modules reassort into a dodecameric module, eight of which form a single, heterooligomeric midplane ring, which is flexible in a diameter range of 40-50 nm. The ring cycle would be regulated by phenylalanine-glycine regions ("FG repeats") of channel nups. Akin to ligand-gated channels, the dilated state of the midplane ring may be stabilized by binding of [cargo•transport-factor] complexes to FG repeats, thereby linking the ratio of constricted to dilated nuclear pores to cellular transport need.

  3. Helium measurements of pore-fluids obtained from SAFOD drillcore

    Energy Technology Data Exchange (ETDEWEB)

    Ali, S.; Stute, M.; Torgersen, T.; Winckler, G.; Kennedy, B.M.


    {sup 4}He accumulated in fluids is a well established geochemical tracer used to study crustal fluid dynamics. Direct fluid samples are not always collectable; therefore, a method to extract rare gases from matrix fluids of whole rocks by diffusion has been adapted. Helium was measured on matrix fluids extracted from sandstones and mudstones recovered during the San Andreas Fault Observatory at Depth (SAFOD) drilling in California, USA. Samples were typically collected as subcores or from drillcore fragments. Helium concentration and isotope ratios were measured 4-6 times on each sample, and indicate a bulk {sup 4}He diffusion coefficient of 3.5 {+-} 1.3 x 10{sup -8} cm{sup 2}s{sup -1} at 21 C, compared to previously published diffusion coefficients of 1.2 x 10{sup -18} cm{sup 2}s{sup -1} (21 C) to 3.0 x 10{sup -15} cm{sup 2}s{sup -1} (150 C) in the sands and clays. Correcting the diffusion coefficient of {sup 4}He{sub water} for matrix porosity ({approx}3%) and tortuosity ({approx}6-13) produces effective diffusion coefficients of 1 x 10{sup -8} cm{sup 2}s{sup -1} (21 C) and 1 x 10{sup -7} (120 C), effectively isolating pore fluid {sup 4}He from the {sup 4}He contained in the rock matrix. Model calculations indicate that <6% of helium initially dissolved in pore fluids was lost during the sampling process. Complete and quantitative extraction of the pore fluids provide minimum in situ porosity values for sandstones 2.8 {+-} 0.4% (SD, n=4) and mudstones 3.1 {+-} 0.8% (SD, n=4).

  4. Using of AFLP to evaluate gamma-irradiated amaranth mutants

    Directory of Open Access Journals (Sweden)

    Labajová Mária


    Full Text Available To determine which of several gamma-irradiated mutants of amaranth Ficha cultivar and K-433 hybrid are most genetically similar to their non-irradiated control genotypes, we performed amplified fragment length polymorphism (AFLP based analysis. A total of 40 selective primer combinations were used in reported analyses. First analyses of gamma-irradiated amaranth mutant lines were done used the AFLP. In the study, primers with the differentiation ability for all analysed mutant lines are reported. The very specific changes in the mutant lines´ non-coding regions based on AFLP length polymorphism were analysed. Mutant lines of the Ficha cultivar (C15, C26, C27, C82, C236 shared a genetic dissimilarity of 0,11 and their ISSR profiles are more similar to the Ficha than those of K-433 hybrid mutant lines. The K-433 mutant lines (D54, D279, D282 shared genetic dissimilarity of 0,534 but are more distinct to their control plant as a whole, as those of the Ficha mutant lines. Different AFLP fingerprints patters of the mutant lines when compared to the Ficha cultivar and K-433 hybrid AFLP profiles may be a consequence of the complex response of the intergenic space of mutant lines to the gamma-radiance. Although a genetic polymorphism was detected within accessions, the AFLP markers successfully identified all the accessions. The AFLP results are discussed by a combination of biochemical characteristics of mutant lines and their control genotypes.

  5. Distribution of soluble amino acids in maize endosperm mutants

    Directory of Open Access Journals (Sweden)

    Toro Alejandro Alberto


    Full Text Available For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperm of wild-type, opaque and floury maize (Zea mays L. mutants were determined by HPLC. The total absolute concentration of soluble amino acids among the mutants varied depending on the mutant. The o11 and o13 mutants exhibited the highest average content, whereas o10, fl3 and fl1 exhibited the lowest average content. In general, the mutants exhibited similar concentrations of total soluble amino acids when compared to the wild-type lines, with the clear exception of mutants o11 and fl1, with the o11 mutant exhibiting a higher concentration of total soluble amino acids when compared to its wild-type counterpart W22 and the fl1 mutant a lower concentration when compared to its wild-type counterpart Oh43. For methionine, the mutants o2 and o11 and wild-type Oh43 exhibited the highest concentrations of this amino acid. Significant differences were not observed between mutants for other amino acids such as lysine and threonine. The high lysine concentrations obtained originally for these mutants may be due to the amino acids incorporated into storage proteins, but not those present in the soluble form.

  6. Functional Contributions of Positive Charges in the Pore-Lining Helix 3 of the Bordetella pertussis CyaA-Hemolysin to Hemolytic Activity and Ion-Channel Opening

    Directory of Open Access Journals (Sweden)

    Chattip Kurehong


    Full Text Available The Bordetella pertussis CyaA-hemolysin (CyaA-Hly domain was previously demonstrated to be an important determinant for hemolysis against target erythrocytes and ion-channel formation in planar lipid bilayers (PLBs. Here, net-charge variations in the pore-lining helix of thirteen related RTX cytolysins including CyaA-Hly were revealed by amino acid sequence alignments, reflecting their different degrees of hemolytic activity. To analyze possible functional effects of net-charge alterations on hemolytic activity and channel formation of CyaA-Hly, specific mutations were made at Gln574 or Glu581 in its pore-lining α3 of which both residues are highly conserved Lys in the three highly active RTX cytolysins (i.e., Escherichia coli α-hemolysin, Actinobacillus pleuropneumoniae toxin, and Aggregatibacter actinomycetemcomitans leukotoxin. All six constructed CyaA-Hly mutants that were over-expressed in E. coli as 126 kDa His-tagged soluble proteins were successfully purified via immobilized Ni2+-affinity chromatography. Both positive-charge substitutions (Q574K, Q574R, E581K, E581R and negative-charge elimination (E581Q appeared to increase the kinetics of toxin-induced hemolysis while the substitution with a negatively-charged side-chain (Q574E completely abolished its hemolytic activity. When incorporated into PLBs under symmetrical conditions (1.0 M KCl, pH 7.4, all five mutant toxins with the increased hemolytic activity produced clearly-resolved single channels with higher open probability and longer lifetime than the wild-type toxin, albeit with a half decrease in their maximum conductance. Molecular dynamics simulations for 50 ns of a trimeric CyaA-Hly pore model comprising three α2-loop-α3 transmembrane hairpins revealed a significant role of the positive charge at both target positions in the structural stability and enlarged diameter of the simulated pore. Altogether, our present data have disclosed functional contributions of positively

  7. Phosphomimetic mutation of cysteine string protein-α increases the rate of regulated exocytosis by modulating fusion pore dynamics in PC12 cells.

    Directory of Open Access Journals (Sweden)

    Ning Chiang

    Full Text Available BACKGROUND: Cysteine string protein-α (CSPα is a chaperone to ensure protein folding. Loss of CSPα function associates with many neurological diseases. However, its function in modulating regulated exocytosis remains elusive. Although cspα-knockouts exhibit impaired synaptic transmission, overexpression of CSPα in neuroendocrine cells inhibits secretion. These seemingly conflicting results lead to a hypothesis that CSPα may undergo a modification that switches its function in regulating neurotransmitter and hormone secretion. Previous studies implied that CSPα undergoes phosphorylation at Ser10 that may influence exocytosis by altering fusion pore dynamics. However, direct evidence is missing up to date. METHODOLOGY/PRINCIPAL FINDINGS: Using amperometry, we investigated how phosphorylation at Ser10 of CSPα (CSPα-Ser10 modulates regulated exocytosis and if this modulation involves regulating a specific kinetic step of fusion pore dynamics. The real-time exocytosis of single vesicles was detected in PC12 cells overexpressing control vector, wild-type CSPα (WT, the CSPα phosphodeficient mutant (S10A, or the CSPα phosphomimetic mutants (S10D and S10E. The shapes of amperometric signals were used to distinguish the full-fusion events (i.e., prespike feet followed by spikes and the kiss-and-run events (i.e., square-shaped flickers. We found that the secretion rate was significantly increased in cells overexpressing S10D or S10E compared to WT or S10A. Further analysis showed that overexpression of S10D or S10E prolonged fusion pore lifetime compared to WT or S10A. The fraction of kiss-and-run events was significantly lower but the frequency of full-fusion events was higher in cells overexpressing S10D or S10E compared to WT or S10A. Advanced kinetic analysis suggests that overexpression of S10D or S10E may stabilize open fusion pores mainly by inhibiting them from closing. CONCLUSIONS/SIGNIFICANCE: CSPα may modulate fusion pore dynamics

  8. Long n-alkanes isomerization by medium pore zeolites with pore mouth and key lock mechanisms; Isomerisation des paraffines longues par des zeolithes a pores moyens selon les mecanismes ouverture de pore et cle serrure

    Energy Technology Data Exchange (ETDEWEB)

    Claude, M.


    Skeletal isomerization of long n-alkanes is practiced to improve cold flow properties of diesel and lubricant fractions. In this work, model long n-alkanes (n-C{sub 10} - n-C{sub 24}) were hydro-isomerized in a fixed bed down flow vapour phase reactor loaded with bifunctional Pt/H-ZSM-22 zeolite catalyst. The skeletal isomers were analysed and identified with GC/MS. High isomer yields were obtained. The distribution of positional mono-methyl-branched isomers obtained from n-C{sub 12} to n-C{sub 24} are typically bimodal. This is explained by adsorption and reaction of the alkanes in pore mouths and locks on the external surface of the zeolite crystals. The pore mouth mode favours branching at C{sub 2} and C{sub 3}. The 'key lock' type proceeds by penetration of the two ends of the hydrocarbon chain into a different pore opening and favours more central mono-branching of the chain. The contribution of the key lock mode increases with increasing chain length and with the reaction temperature. The preferentially formed dimethyl-branched isomers have a separation between branchings of three up to fourteen carbon atoms. The formation of the second methyl-branching occurs preferentially from a centrally branched mono-methyl-branched isomer, so that the second branching is generated always more toward the end of the chain. Owing to the differences in adsorption entropy among the locks, at higher temperatures the largest lock is preferred and the distance between the two branching along the carbon chain in the preferred isomers is biggest. Thus the work resulted in the formulation of structure-selectivity relationships. n-C{sub 18} was hydro-isomerized on other zeolites. The nature and distribution of the isomers obtained suggest that the tubular 10-ring zeolites ZSM-23, ZSM-35 and SAPO-11 also operate according to pore mouth and key lock concepts. Zeolites with 12-rings show typical product patterns for catalysis in absence of steric hindrance. (author)

  9. Perforating the nuclear boundary - how nuclear pore complexes assemble. (United States)

    Weberruss, Marion; Antonin, Wolfram


    The nucleus is enclosed by the nuclear envelope, a double membrane which creates a selective barrier between the cytoplasm and the nuclear interior. Its barrier and transport characteristics are determined by nuclear pore complexes (NPCs) that are embedded within the nuclear envelope, and control molecular exchange between the cytoplasm and nucleoplasm. In this Commentary, we discuss the biogenesis of these huge protein assemblies from approximately one thousand individual proteins. We will summarize current knowledge about distinct assembly modes in animal cells that are characteristic for different cell cycle phases and their regulation.

  10. Lipopeptide surfactants: Production, recovery and pore forming capacity. (United States)

    Inès, Mnif; Dhouha, Ghribi


    Lipopeptides are microbial surface active compounds produced by a wide variety of bacteria, fungi and yeast. They are characterized by highly structural diversity and have the ability to decrease the surface and interfacial tension at the surface and interface, respectively. Surfactin, iturin and fengycin of Bacillus subtilis are among the most studied lipopeptides. This review will present the main factors encountering lipopeptides production along with the techniques developed for their extraction and purification. Moreover, we will discuss their ability to form pores and destabilize biological membrane permitting their use as antimicrobial, hemolytic and antitumor agents. These open great potential applications in biomediacal, pharmaceutic and agriculture fields.

  11. Explicit Pore Pressure Material Model in Carbon-Cloth Phenolic (United States)

    Gutierrez-Lemini, Danton; Ehle, Curt


    An explicit material model that uses predicted pressure in the pores of a carbon-cloth phenolic (CCP) composite has been developed. This model is intended to be used within a finite-element model to predict phenomena specific to CCP components of solid-fuel-rocket nozzles subjected to high operating temperatures and to mechanical stresses that can be great enough to cause structural failures. Phenomena that can be predicted with the help of this model include failures of specimens in restrained-thermal-growth (RTG) tests, pocketing erosion, and ply lifting

  12. Sound absorption property of open-pore aluminum foams

    Institute of Scientific and Technical Information of China (English)

    WANG Fang; WANG Lu-cai; WU Jian-guo; YOU Xiao-hong


    This paper presents a study on sound absorption property of aluminum foam by evaluating its sound absorption coefficients using standing wave tube method. Experimental results showed that the average values of sound absorption coefficients (over the test frequency range) are all above 0.4, which indicate very good sound absorption property of the aluminum foams. The sound absorption coefficient is affected by frequency and pore structure, and reaches its maximum value at around 1 000 Hz. With the increase of porosity and decrease of cell diameter, the sound absorption coefficient values increase.

  13. Migraine: Role of the TRESK two-pore potassium channel. (United States)

    Lafrenière, Ronald G; Rouleau, Guy A


    Migraine is a severe episodic headache disorder affecting one in five people. Genetic studies have identified mutations in the CACNA1, ATP1A2 and SCN1A genes in the rare familial hemiplegic migraine. Recently, a mutation in the KCNK18 gene, encoding the TRESK two-pore domain potassium channel, was described in a large family with migraine with aura. This review will elaborate on the possible role of the TRESK channel in regulating neuronal excitability, its role in migraine pathogenesis, and on promising therapeutic opportunities targeting this channel. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  14. Dendrimer-like hybrid particles with tunable hierarchical pores (United States)

    Du, Xin; Li, Xiaoyu; Huang, Hongwei; He, Junhui; Zhang, Xueji


    Dendrimer-like silica particles with a center-radial dendritic framework and a synergistic hierarchical porosity have attracted much attention due to their unique open three-dimensional superstructures with high accessibility to the internal surface areas; however, the delicate regulation of the hierarchical porosity has been difficult to achieve up to now. Herein, a series of dendrimer-like amino-functionalized silica particles with tunable hierarchical pores (HPSNs-NH2) were successfully fabricated by carefully regulating and optimizing the various experimental parameters in the ethyl ether emulsion systems via a one-pot sol-gel reaction. Interestingly, the simple adjustment of the stirring rate or reaction temperature was found to be an easy and effective route to achieve the controllable regulation towards center-radial large pore sizes from ca. 37-267 (148 +/- 45) nm to ca. 8-119 (36 +/- 21) nm for HPSNs-NH2 with particle sizes of 300-700 nm and from ca. 9-157 (52 +/- 28) nm to ca. 8-105 (30 +/- 16) nm for HPSNs-NH2 with particle sizes of 100-320 nm. To the best of our knowledge, this is the first successful regulation towards center-radial large pore sizes in such large ranges. The formation of HPSNs-NH2 may be attributed to the complex cross-coupling of two processes: the dynamic diffusion of ethyl ether molecules and the self-assembly of partially hydrolyzed TEOS species and CTAB molecules at the dynamic ethyl ether-water interface of uniform small quasi-emulsion droplets. Thus, these results regarding the elaborate regulation of center-radial large pores and particle sizes not only help us better understand the complicated self-assembly at the dynamic oil-water interface, but also provide a unique and ideal platform as carriers or supports for adsorption, separation, catalysis, biomedicine, and sensor.Dendrimer-like silica particles with a center-radial dendritic framework and a synergistic hierarchical porosity have attracted much attention due to their

  15. Investigating the effects of stress on the pore structures of nuclear grade graphites (United States)

    Taylor, Joshua E. L.; Hall, Graham N.; Mummery, Paul M.


    Graphite is used as a moderating material and as a structural component in a number of current generation nuclear reactors. During reactor operation stresses develop in the graphite components, causing them to deform. It is important to understand how the microstructure of graphite affects the material's response to these stresses. A series of experiments were performed to investigate how the pore structures of Pile Grade A and Gilsocarbon graphites respond to loading stresses. A compression rig was used to simulate the build-up of operational stresses in graphite components, and a confocal laser microscope was used to study variation of a number of important pore properties. Values of elastic modulus and Poisson's ratio were calculated and compared to existing literature to confirm the validity of the experimental techniques. Mean pore areas were observed to decrease linearly with increasing applied load, mean pore eccentricity increased linearly, and a small amount of clockwise pore rotation was observed. The response to build-up of stresses was dependent on the orientation of the pores and basal planes and the shapes of the pores with respect to the loading axis. It was proposed that pore closure and pore reorientation were competing processes. Pore separation was quantified using 'nearest neighbour' and Voronoi techniques, and non-pore regions were found to shrink linearly with increasing applied load.

  16. Using X-Ray Computed Tomography in Pore Structure Characterization for a Berea Sandstone: Resolution Effect

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Sheng; Hu, Qinhong; Dultz, Stefan; Zhang, Ming


    X-raycomputedtomography (XCT) is a powerful tool for detecting the micro-scale porestructure and has been applied to many natural and synthetic porous media. However, due to the resolution limitations, either non-representative view of the sample or inaccurate results can be produced from the XCT image processing. In this paper, two XCT (micro-CT and CT with synchrotron radiation) with different resolutions of 12.7 μm and 0.35 μm, as well as mercury intrusion porosimetry (MIP) with a minimum detection limit of 3 nm, were used for Berea sandstone to investigate the effect of detecting resolution on the porestructure. Several key porestructure parameters, including porosity, pore size distribution, pore connectivity, surface area, hydraulic radius, and aspect ratio were analyzed in a manner of quantitative comparison between different resolutions of XCT and MIP. The low resolution XCT can capture the large-pore porosity, while overestimates the pore size and pore connectivity. The high resolution XCT is more accurate in describing the pore shape, porosity, pore size; however, it is not representative since narrower detecting pore size range and small volume represented. A representative element volume related to large-pore porosity and probably large-pore connectivity with diameter and height of 2.8 mm is obtained through scale effect analysis. Therefore, selecting an appropriate resolution should be a compromise between the pore size and the representative element volume for the specific property or process of interest.

  17. Detectability of Pore Defect in Wind Turbine Blade Composites Using Image Correlation Technique

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jong Il; Huh, Yong Hak; Lee, Gun Chang [Korea Research institute of Standard and Science, Daejeon (Korea, Republic of)


    Defects that occur during the manufacturing process or operation of a wind turbine blade have a great influence on its life and safety. Typically, defects such as delamination, pore, wrinkle and matrix crack are found in a blade. In this study, the detectability of the pores, a type of defect that frequently occur during manufacturing, was examined from the full field strain distribution determined with the image correlation technique. Pore defects were artificially introduced in four-ply laminated GFRP composites with 0 .deg/{+-}45 .deg fiber direction. The artificial pores were introduced in consideration of their size and location. Three different-sized pores with diameter of 1, 2 and 3 mm were located on the top and bottom surface and embedded. By applying static loads of 0-200 MPa, the strain distributions over the specimen with the pore defects were determined using image correlation technique. It was found the pores with diameter exceeding 2 mm can be detected in diameter.

  18. Pro-apoptotic Bax molecules densely populate the edges of membrane pores. (United States)

    Kuwana, Tomomi; Olson, Norman H; Kiosses, William B; Peters, Bjoern; Newmeyer, Donald D


    How the pro-apoptotic Bax protein permeabilizes the mitochondrial outer membrane is not fully understood. Previously, using cryo-electron microscopy (cryo-EM), we showed that activated Bax forms large, growing pores. Whether formed in liposomes or in mitochondrial outer membranes, Bax-induced pores exhibit the same morphology, with negative curvature flanking the edges and with no visible protein structure protruding from the membranes. Here we used cryo-EM to show that gold-labeled Bax molecules, after activation by Bid, became localized strictly at pore edges. This argues that Bax acts at short range to deform the membrane. Also, Bax molecules populated the walls of both small and large pores at the same density, implying that Bax is continuously recruited to the pores as they widen. Moreover, because all Bax molecules became oligomerized after membrane insertion, we infer that Bax oligomers are present at pore edges. We suggest that oligomerization may promote pore enlargement.

  19. Direct measurement of the critical pore size in a model membrane

    CERN Document Server

    Ilton, Mark; Dalnoki-Veress, Kari


    We study pore nucleation in a model membrane system, a freestanding polymer film. Nucleated pores smaller than a critical size close, while pores larger than the critical size grow. Holes of varying size were purposefully prepared in liquid polymer films, and their evolution in time was monitored using optical and atomic force microscopy to extract a critical radius. The critical radius scales linearly with film thickness for a homopolymer film. The results agree with a simple model which takes into account the energy cost due to surface area at the edge of the pore. The energy cost at the edge of the pore is experimentally varied by using a lamellar-forming diblock copolymer membrane. The underlying molecular architecture causes increased frustration at the pore edge resulting in an enhanced cost of pore formation.

  20. Formation of spherical stomatocyte of high-genus vesicle under pore-size constraint

    CERN Document Server

    Noguchi, Hiroshi


    Nuclear pores have an approximately uniform distribution in the nuclear envelope of most living cells. Hence, the morphology of the nuclear envelope is a spherical stomatocyte with a high genus. We have investigated the morphology of high-genus vesicles under pore-size constraint using dynamically triangulated membrane simulations. Bending-energy minimization without volume or other constraints produces a circular-cage stomatocyte, where the pores are aligned in a circular line on an oblate inner bud. As the pore radius is reduced, the circular-pore alignment is more stabilized than a random pore distribution on a spherical bud. However, we have clarified the conditions for the formation of a spherical stomatocyte: a small reduced volume, osmotic pressure within the inner bud, and repulsion between the pores. When area-difference elasticity is taken into account, the formation of cylindrical or budded tubules from the stomatocyte and discoidal stomatocyte is found.

  1. Pore Structure Control of Ordered Mesoporous Silica Film Using Mixed Surfactants

    Directory of Open Access Journals (Sweden)

    Tae-Jung Ha


    Full Text Available Materials with nanosized and well-arranged pores have been researched actively in order to be applied to new technology fields. Especially, mesoporous material containing various pore structures is expected to have different pore structure. To form a mixed pore structure, ordered mesoporous silica films were prepared with a mixture of surfactant; Brij-76 and P-123 block copolymer. In mixed surfactant system, mixed pore structure was observed in the region of P-123/(Brij-76 + P-123 with about 50.0 wt.% while a single pore structure was observed in regions which have large difference in ratio between Brij-76 and P-123 through the X-ray diffraction analysis. Regardless of surfactant ratio, porosity was retained almost the same. It is expected that ordered mesoporous silica film with mixed pore structure can be one of the new materials which has distinctive properties.

  2. Auxin physiology of the tomato mutant diageotropica (United States)

    Daniel, S. G.; Rayle, D. L.; Cleland, R. E.


    The tomato (Lycopersicon esculentum, Mill.) mutant diageotropica (dgt) exhibits biochemical, physiological, and morphological abnormalities that suggest the mutation may have affected a primary site of auxin perception or action. We have compared two aspects of the auxin physiology of dgt and wild-type (VFN8) seedlings: auxin transport and cellular growth parameters. The rates of basipetal indole-3-acetic acid (IAA) polar transport are identical in hypocotyl sections of the two genotypes, but dgt sections have a slightly greater capacity for IAA transport. 2,3,5-Triiodobenzoic acid and ethylene reduce transport in both mutant and wild-type sections. The kinetics of auxin uptake into VFN8 and dgt sections are nearly identical. These results make it unlikely that an altered IAA efflux carrier or IAA uptake symport are responsible for the pleiotropic effects resulting from the dgt mutation. The lack of auxin-induced cell elongation in dgt plants is not due to insufficient turgor, as the osmotic potential of dgt cell sap is less (more negative) than that of VFN8. An auxin-induced increase in wall extensibility, as measured by the Instron technique, only occurs in the VFN8 plants. These data suggest dgt hypocotyls suffer a defect in the sequence of events culminating in auxin-induced cell wall loosening.

  3. Indy mutants: live long and prosper

    Directory of Open Access Journals (Sweden)

    Stewart eFrankel


    Full Text Available Indy encodes the fly homologue of a mammalian transporter of di and tricarboxylatecomponents of the Krebs cycle. Reduced expression of fly Indy or two of the C. elegansIndy homologs leads to an increase in life span. Fly and worm tissues that play key roles inintermediary metabolism are also the places where Indy genes are expressed. One of themouse homologs of Indy (mIndy is mainly expressed in the liver. It has been hypothesizedthat decreased INDY activity creates a state similar to caloric restriction (CR. Thishypothesis is supported by the physiological similarities between Indy mutant flies on highcalorie food and control flies on CR, such as increased physical activity and decreases inweight, egg production, triglyceride levels, starvation resistance, and insulin signaling. Inaddition, Indy mutant flies undergo changes in mitochondrial biogenesis also observed inCR animals. Recent findings with mIndy knockout mice support and extend the findingsfrom flies. mIndy-/- mice display an increase in hepatic mitochondrial biogenesis, lipidoxidation and decreased hepatic lipogenesis. When mIndy-/- mice are fed high calorie foodthey are protected from adiposity and insulin resistance. These findings point to INDY as apotential drug target for the treatment of metabolic syndrome, type 2 diabetes and obesity.

  4. Secretion and activation of the Serratia marcescens hemolysin by structurally defined ShlB mutants. (United States)

    Pramanik, Avijit; Könninger, Ulrich; Selvam, Arun; Braun, Volkmar


    The ShlA hemolysin of Serratia marcescens is secreted across the outer membrane by the ShlB protein; ShlB belongs to the two-partner secretion system (type Vb), a subfamily of the Omp85 outer membrane protein assembly and secretion superfamily. During secretion, ShlA is converted from an inactive non-hemolytic form into an active hemolytic form. The structure of ShlB is predicted to consist of the N-terminal α-helix H1, followed by the two polypeptide-transport-associated domains POTRA P1 and P2, and the β-barrel of 16 β-strands. H1 is inserted into the pore of the β-barrel in the outer membrane; P1 and P2 are located in the periplasm. To obtain insights into the secretion and activation of ShlA by ShlB, we isolated ShlB mutants impaired in secretion and/or activation. The triple H1 P1 P2 mutant did not secrete ShlA. The P1 and P2 deletion derivatives secreted reduced amounts of ShlA, of which P1 showed some hemolysis, whereas P2 was inactive. Deletion of loop 6 (L6), which is conserved among exporters of the Omp85 family, compromised activation but retained low secretion. Secretion-negative mutants generated by random mutagenesis were located in loop 6. The inactive secreted ShlA derivatives were complemented in vitro to active ShlA by an N-terminal ShlA fragment (ShlA242) secreted by ShlB. Deletion of H1 did not impair secretion of hemolytic ShlA. The study defines domains of ShlB which are important for ShlA secretion and activation.

  5. Simulation of Pore Width and Pore Charge Effects on Selectivities of CO2 vs. H2 from a Syngas-like Mixture in Carbon Mesopores

    NARCIS (Netherlands)

    Trinh, T.T.; Vlugt, T.J.H.; Hägg, M.B.; Bedeaux, D.; Kjelstrup, S.


    Classical molecular dynamics simulations were performed to study the effect of pore width and surface charge in carbon mesoporous materials on adsorption and diffusion selectivities of CO2/H2 in a syngas-like mixture (mole fraction of CO2 = 0.30). The pore width of the graphite slit varied from 2.5

  6. Pore-scale supercritical CO2 dissolution and mass transfer under drainage conditions (United States)

    Chang, Chun; Zhou, Quanlin; Oostrom, Mart; Kneafsey, Timothy J.; Mehta, Hardeep


    Recently, both core- and pore-scale imbibition experiments have shown non-equilibrium dissolution of supercritical CO2 (scCO2) and a prolonged depletion of residual scCO2. In this study, pore-scale scCO2 dissolution and mass transfer under drainage conditions were investigated using a two-dimensional heterogeneous micromodel and a novel fluorescent water dye with a sensitive pH range between 3.7 and 6.5. Drainage experiments were conducted at 9 MPa and 40 °C by injecting scCO2 into the sandstone-analogue pore network initially saturated by water without dissolved CO2 (dsCO2). During the experiments, time-lapse images of dye intensity, reflecting water pH, were obtained. These images show non-uniform pH in individual pores and pore clusters, with average pH levels gradually decreasing with time. Further analysis on selected pores and pore clusters shows that (1) rate-limited mass transfer prevails with slowly decreasing pH over time when the scCO2-water interface area is low with respect to the volume of water-filled pores and pore clusters, (2) fast scCO2 dissolution and phase equilibrium occurs when scCO2 bubbles invade into water-filled pores, significantly enhancing the area-to-volume ratio, and (3) a transition from rate-limited to diffusion-limited mass transfer occurs in a single pore when a medium area-to-volume ratio is prevalent. The analysis also shows that two fundamental processes - scCO2 dissolution at phase interfaces and diffusion of dsCO2 at the pore scale (10-100 μm) observed after scCO2 bubble invasion into water-filled pores without pore throat constraints - are relatively fast. The overall slow dissolution of scCO2 in the millimeter-scale micromodel can be attributed to the small area-to-volume ratios that represent pore-throat configurations and characteristics of phase interfaces. This finding is applicable for the behavior of dissolution at pore, core, and field scales when water-filled pores and pore clusters of varying size are surrounded

  7. Pore-scale supercritical CO2 dissolution and mass transfer under drainage conditions

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Chun; Zhou, Quanlin; Oostrom, Mart; Kneafsey, Timothy J.; Mehta, Hardeep


    Abstract: Recently, both core- and pore-scale imbibition experiments have shown non-equilibrium dissolution of supercritical CO2 (scCO2) and a prolonged depletion of residual scCO2. In this study, pore-scale scCO2 dissolution and mass transfer under drainage conditions were investigated using a two-dimensional heterogeneous micromodel and a novel fluorescent water dye with a sensitive pH range between 3.7 and 6.5. Drainage experiments were conducted at 9 MPa and 40 °C by injecting scCO2 into the sandstone-analogue pore network initially saturated by water without dissolved CO2 (dsCO2). During the experiments, time-lapse images of dye intensity, reflecting water pH, were obtained. These images show non-uniform pH in individual pores and pore clusters, with average pH levels gradually decreasing with time. Further analysis on selected pores and pore clusters shows that (1) rate-limited mass transfer prevails with slowly decreasing pH over time when the scCO2-water interface area is low with respect to the volume of water-filled pores and pore clusters, (2) fast scCO2 dissolution and phase equilibrium occurs when scCO2 bubbles invade into water-filled pores, significantly enhancing the area-to-volume ratio, and (3) a transition from rate-limited to diffusion-limited mass transfer occurs in a single pore when a medium area-to-volume ratio is prevalent. The analysis also shows that two fundamental processes – scCO2 dissolution at phase interfaces and diffusion of dsCO2 at the pore scale (10-100 µm) observed after scCO2 bubble invasion into water-filled pores without pore throat constraints – are relatively fast. The overall slow dissolution of scCO2 in the millimeter-scale micromodel can be attributed to the small area-to-volume ratios that represent pore-throat configurations and characteristics of phase

  8. Dominant negative mutants of Bacillus thuringiensis Cry1Ab toxin function as anti-toxins: demonstration of the role of oligomerization in toxicity.

    Directory of Open Access Journals (Sweden)

    Claudia Rodríguez-Almazán

    Full Text Available BACKGROUND: Bacillus thuringiensis Cry toxins, that are used worldwide in insect control, kill insects by a mechanism that depends on their ability to form oligomeric pores that insert into the insect-midgut cells. These toxins are being used worldwide in transgenic plants or spray to control insect pests in agriculture. However, a major concern has been the possible effects of these insecticidal proteins on non-target organisms mainly in ecosystems adjacent to agricultural fields. METHODOLOGY/PRINCIPAL FINDINGS: We isolated and characterized 11 non-toxic mutants of Cry1Ab toxin affected in different steps of the mechanism of action namely binding to receptors, oligomerization and pore-formation. These mutant toxins were analyzed for their capacity to block wild type toxin activity, presenting a dominant negative phenotype. The dominant negative phenotype was analyzed at two levels, in vivo by toxicity bioassays against susceptible Manduca sexta larvae and in vitro by pore formation activity in black lipid bilayers. We demonstrate that some mutations located in helix alpha-4 completely block the wild type toxin activity at sub-stoichiometric level confirming a dominant negative phenotype, thereby functioning as potent antitoxins. CONCLUSIONS/SIGNIFICANCE: This is the first reported case of a Cry toxin dominant inhibitor. These data demonstrate that oligomerization is a fundamental step in Cry toxin action and represent a potential mechanism to protect special ecosystems from the possible effect of Cry toxins on non-target organisms.

  9. Mutants of Saccharomyces cerevisiae with defects in acetate metabolism: isolation and characterization of Acn- mutants. (United States)

    McCammon, M T


    The two carbon compounds, ethanol and acetate, can be oxidatively metabolized as well as assimilated into carbohydrate in the yeast Saccharomyces cerevisiae. The distribution of acetate metabolic enzymes among several cellular compartments, mitochondria, peroxisomes, and cytoplasm makes it an intriguing system to study complex metabolic interactions. To investigate the complex process of carbon catabolism and assimilation, mutants unable to grow on acetate were isolated. One hundred five Acn- ("ACetate Nonutilizing") mutants were sorted into 21 complementation groups with an additional 20 single mutants. Five of the groups have defects in TCA cycle enzymes: MDH1, CIT1, ACO1, IDH1, and IDH2. A defect in RTG2, involved in the retrograde communication between the mitochondrion and the nucleus, was also identified. Four genes encode enzymes of the glyoxylate cycle and gluconeogenesis: ICL1, MLS1, MDH2, and PCK1. Five other genes appear to be defective in regulating metabolic activity since elevated levels of enzymes in several metabolic pathways, including the glyoxylate cycle, gluconeogenesis, and acetyl-CoA metabolism, were detected in these mutants: ACN8, ACN9, ACN17, ACN18, and ACN42. In summary, this analysis has identified at least 22 and as many as 41 different genes involved in acetate metabolism.

  10. Energetics of Transport through the Nuclear Pore Complex.

    Directory of Open Access Journals (Sweden)

    Ali Ghavami

    Full Text Available Molecular transport across the nuclear envelope in eukaryotic cells is solely controlled by the nuclear pore complex (NPC. The NPC provides two types of nucleocytoplasmic transport: passive diffusion of small molecules and active chaperon-mediated translocation of large molecules. It has been shown that the interaction between intrinsically disordered proteins that line the central channel of the NPC and the transporting cargoes is the determining factor, but the exact mechanism of transport is yet unknown. Here, we use coarse-grained molecular dynamics simulations to quantify the energy barrier that has to be overcome for molecules to pass through the NPC. We focus on two aspects of transport. First, the passive transport of model cargo molecules with different sizes is studied and the size selectivity feature of the NPC is investigated. Our results show that the transport probability of cargoes is significantly reduced when they are larger than ∼5 nm in diameter. Secondly, we show that incorporating hydrophobic binding spots on the surface of the cargo effectively decreases the energy barrier of the pore. Finally, a simple transport model is proposed which characterizes the energy barrier of the NPC as a function of diameter and hydrophobicity of the transporting particles.

  11. A Fence-like Coat for the Nuclear Pore Membrane

    Energy Technology Data Exchange (ETDEWEB)

    Debler, E.; Ma, Y; Seo, H; Hsia, K; Noriega, T; Blobel, G; Hoelz, A


    We recently proposed a cylindrical coat for the nuclear pore membrane in the nuclear pore complex (NPC). This scaffold is generated by multiple copies of seven nucleoporins. Here, we report three crystal structures of the nucleoporin pair Seh1{center_dot}Nup85, which is part of the coat cylinder. The Seh1{center_dot}Nup85 assembly bears resemblance in its shape and dimensions to that of another nucleoporin pair, Sec13{center_dot}Nup145C. Furthermore, the Seh1{center_dot}Nup85 structures reveal a hinge motion that may facilitate conformational changes in the NPC during import of integral membrane proteins and/or during nucleocytoplasmic transport. We propose that Seh1{center_dot}Nup85 and Sec13{center_dot}Nup145C form 16 alternating, vertical rods that are horizontally linked by the three remaining nucleoporins of the coat cylinder. Shared architectural and mechanistic principles with the COPII coat indicate a common evolutionary origin and support the notion that the NPC coat represents another class of membrane coats.

  12. Toward highly stable electrocatalysts via nanoparticle pore confinement. (United States)

    Galeano, Carolina; Meier, Josef C; Peinecke, Volker; Bongard, Hans; Katsounaros, Ioannis; Topalov, Angel A; Lu, Anhui; Mayrhofer, Karl J J; Schüth, Ferdi


    The durability of electrode materials is a limiting parameter for many electrochemical energy conversion systems. In particular, electrocatalysts for the essential oxygen reduction reaction (ORR) present some of the most challenging instability issues shortening their practical lifetime. Here, we report a mesostructured graphitic carbon support, Hollow Graphitic Spheres (HGS) with a specific surface area exceeding 1000 m(2) g(-1) and precisely controlled pore structure, that was specifically developed to overcome the long-term catalyst degradation, while still sustaining high activity. The synthetic pathway leads to platinum nanoparticles of approximately 3 to 4 nm size encapsulated in the HGS pore structure that are stable at 850 °C and, more importantly, during simulated accelerated electrochemical aging. Moreover, the high stability of the cathode electrocatalyst is also retained in a fully assembled polymer electrolyte membrane fuel cell (PEMFC). Identical location scanning and scanning transmission electron microscopy (IL-SEM and IL-STEM) conclusively proved that during electrochemical cycling the encapsulation significantly suppresses detachment and agglomeration of Pt nanoparticles, two of the major degradation mechanisms in fuel cell catalysts of this particle size. Thus, beyond providing an improved electrocatalyst, this study describes the blueprint for targeted improvement of fuel cell catalysts by design of the carbon support.

  13. Wave-induced stresses and pore pressures near a mudline

    Directory of Open Access Journals (Sweden)

    Andrzej Sawicki


    Full Text Available Conventional methods for the determination of water-wave induced stresses inseabeds composed of granular soils are based on Biot-type models, in which the soilskeleton is treated as an elastic medium. Such methods predict effective stressesin the soil that are unacceptable from the physical point of view, as they permittensile stresses to occur near the upper surface of the seabed. Therefore, in thispaper the granular soil is assumed to behave as an elastic-ideally plastic material,with the Coulomb-Mohr yield criterion adopted to bound admissible stress states inthe seabed. The governing equations are solved numerically by a~finite differencemethod. The results of simulations, carried out for the case of time-harmonicwater waves, illustrate the depth distributions of the excess pore pressures and theeffective stresses in the seabed, and show the shapes of zones of soil in the plastic state.~In particular, the effects on the seabed behaviour of suchparameters as the degree of pore water saturation, the soil permeability, and theearth pressure coefficient, are illustrated.

  14. The effect of pore fluid on seismicity: a computer model

    Institute of Scientific and Technical Information of China (English)


    The influence of fluid on seismicity of a computerized system is analyzed in this paper. The diffusion equation of fluid in a crustal fault area is developed and used in the calculation of a spring-slide-damper model. With mirror imagin boundary condition and three initial conditions, the equation is solved for a dynamic model that consists of six seismic belts and eight seismogenous sources in each belt with both explicit algorithm and implicit algorithm. The analysis of the model with water sources shows that the implicit algorithm is better to be used to calculate the model. Taking a constant proportion of the pore pressure of a broken element to that of its neighboring elements, the seismicity of the model is calculated with mirror boundary condition and no-water-source initial condition. The results shows that the frequency and magnitude of shocks are both higher than those in the model with no water pore pressure, which provides more complexity to earthquake prediction.

  15. Estimating pore and cement volumes in thin section (United States)

    Halley, R.B.


    Point count estimates of pore, grain and cement volumes from thin sections are inaccurate, often by more than 100 percent, even though they may be surprisingly precise (reproducibility + or - 3 percent). Errors are produced by: 1) inclusion of submicroscopic pore space within solid volume and 2) edge effects caused by grain curvature within a 30-micron thick thin section. Submicroscopic porosity may be measured by various physical tests or may be visually estimated from scanning electron micrographs. Edge error takes the form of an envelope around grains and increases with decreasing grain size and sorting, increasing grain irregularity and tighter grain packing. Cements are greatly involved in edge error because of their position at grain peripheries and their generally small grain size. Edge error is minimized by methods which reduce the thickness of the sample viewed during point counting. Methods which effectively reduce thickness include use of ultra-thin thin sections or acetate peels, point counting in reflected light, or carefully focusing and counting on the upper surface of the thin section.

  16. In situ structural analysis of the human nuclear pore complex. (United States)

    von Appen, Alexander; Kosinski, Jan; Sparks, Lenore; Ori, Alessandro; DiGuilio, Amanda L; Vollmer, Benjamin; Mackmull, Marie-Therese; Banterle, Niccolo; Parca, Luca; Kastritis, Panagiotis; Buczak, Katarzyna; Mosalaganti, Shyamal; Hagen, Wim; Andres-Pons, Amparo; Lemke, Edward A; Bork, Peer; Antonin, Wolfram; Glavy, Joseph S; Bui, Khanh Huy; Beck, Martin


    Nuclear pore complexes are fundamental components of all eukaryotic cells that mediate nucleocytoplasmic exchange. Determining their 110-megadalton structure imposes a formidable challenge and requires in situ structural biology approaches. Of approximately 30 nucleoporins (Nups), 15 are structured and form the Y and inner-ring complexes. These two major scaffolding modules assemble in multiple copies into an eight-fold rotationally symmetric structure that fuses the inner and outer nuclear membranes to form a central channel of ~60 nm in diameter. The scaffold is decorated with transport-channel Nups that often contain phenylalanine-repeat sequences and mediate the interaction with cargo complexes. Although the architectural arrangement of parts of the Y complex has been elucidated, it is unclear how exactly it oligomerizes in situ. Here we combine cryo-electron tomography with mass spectrometry, biochemical analysis, perturbation experiments and structural modelling to generate, to our knowledge, the most comprehensive architectural model of the human nuclear pore complex to date. Our data suggest previously unknown protein interfaces across Y complexes and to inner-ring complex members. We show that the transport-channel Nup358 (also known as Ranbp2) has a previously unanticipated role in Y-complex oligomerization. Our findings blur the established boundaries between scaffold and transport-channel Nups. We conclude that, similar to coated vesicles, several copies of the same structural building block--although compositionally identical--engage in different local sets of interactions and conformations.

  17. Epsilon toxin: a fascinating pore-forming toxin. (United States)

    Popoff, Michel R


    Epsilon toxin (ETX) is produced by strains of Clostridium perfringens classified as type B or type D. ETX belongs to the heptameric β-pore-forming toxins including aerolysin and Clostridium septicum alpha toxin, which are characterized by the formation of a pore through the plasma membrane of eukaryotic cells consisting in a β-barrel of 14 amphipatic β strands. By contrast to aerolysin and C. septicum alpha toxin, ETX is a much more potent toxin and is responsible for enterotoxemia in animals, mainly sheep. ETX induces perivascular edema in various tissues and accumulates in particular in the kidneys and brain, where it causes edema and necrotic lesions. ETX is able to pass through the blood-brain barrier and stimulate the release of glutamate, which accounts for the symptoms of nervous excitation observed in animal enterotoxemia. At the cellular level, ETX causes rapid swelling followed by cell death involving necrosis. The precise mode of action of ETX remains to be determined. ETX is a powerful toxin, however, it also represents a unique tool with which to vehicle drugs into the central nervous system or target glutamatergic neurons.

  18. Making the ATHENA optics using silicon pore optics (United States)

    Collon, Maximilien J.; Ackermann, Marcelo; Günther, Ramses; Chatbi, Abdelhakim; Vacanti, Giuseppe; Vervest, Mark; Yanson, Alex; Beijersbergen, Marco W.; Bavdaz, Marcos; Wille, Eric; Haneveld, Jeroen; Olde Riekerink, Mark; Koelewijn, Arenda; van Baren, Coen; Müller, Peter; Krumrey, Michael; Burwitz, Vadim; Sironi, Giorgia; Ghigo, Mauro


    Silicon Pore Optics, after 10 years of development, forms now the basis for future large (L) class astrophysics Xray observatories, such as the ATHENA mission to study the hot and energetic universe, matching the L2 science theme recently selected by ESA for launch in 2028. The scientific requirements result in an optical design that demands high angular resolution (5") and large effective area (2 m2 at a few keV) of an X-ray lens with a focal length of 12 to14 m. Silicon Pore Optics was initially based on long (25 to 50 m) focal length telescope designs, which could achieve several arc second angular resolution by curving the silicon mirror in only one direction (conical approximation). With the advent of shorter focal length missions we started to develop mirrors having a secondary curvature, allowing the production of Wolter-I type optics, which are on axis aberration-free. In this paper we will present the new manufacturing process, discuss the impact of the ATHENA optics design on the technology development and present the results of the latest X-ray test campaigns.

  19. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

    Directory of Open Access Journals (Sweden)

    Vincent Sanchis


    Full Text Available The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic effects of a number of virulence products that promote intestinal cell destruction and/or resistance to the host immune system. This review focuses on the pore-forming haemolysins produced by B. cereus: haemolysin I (cereolysin O, haemolysin II, haemolysin III and haemolysin IV (CytK. Haemolysin I belongs to the cholesterol-dependent cytolysin (CDC family whose best known members are listeriolysin O and perfringolysin O, produced by L. monocytogenes and C. perfringens respectively. HlyII and CytK are oligomeric ß-barrel pore-forming toxins related to the α-toxin of S. aureus or the ß-toxin of C. perfringens. The structure of haemolysin III, the least characterized haemolytic toxin from the B. cereus, group has not yet been determined.

  20. The effect of pore structure on ebullition from peat (United States)

    Ramirez, Jorge A.; Baird, Andy J.; Coulthard, Tom J.


    The controls on methane (CH4) bubbling (ebullition) from peatlands are uncertain, but evidence suggests that physical factors related to gas transport and storage within the peat matrix are important. Variability in peat pore size and the permeability of layers within peat can produce ebullition that ranges from steady to erratic in time and can affect the degree to which CH4 bubbles bypass consumption by methanotrophic bacteria and enter the atmosphere. Here we investigate the role of peat structure on ebullition in structurally different peats using a physical model that replicates bubble production using air injection into peat. We find that the frequency distributions of number of ebullition events per time and the magnitude of bubble loss from the physical model were similar in shape to ebullition from peatlands and incubated peats. This indicates that the physical model could be a valid proxy for naturally occurring ebullition from peat. For the first time, data on bubble sizes from peat were collected to conceptualize ebullition, and we find that peat structure affects bubble sizes. Using a new method to measure peat macrostructure, we collected evidence that supports the hypothesis that structural differences in peat determine if bubble release is steady or erratic and extreme. Collected pore size data suggest that erratic ebullition occurs when large amounts of gas stored at depth easily move through shallower layers of open peat. In contrast, steady ebullition occurs when dense shallower layers of peat regulate the flow of gas emitted from peat.

  1. Defective kernel mutants of maize. I. Genetic and lethality studies. (United States)

    Neuffer, M G; Sheridan, W F


    A planting of 3,919 M(1) kernels from normal ears crossed by EMS-treated pollen produced 3,461 M(1) plants and 3,172 selfed ears. These plants yielded 2,477 (72%) total heritable changes; the selfed ears yielded 2,457 (78%) recessive mutants, including 855 (27%) recessive kernel mutants and 8 (0.23%) viable dominant mutants. The ratio of recessive to dominant mutants was 201:1. The average mutation frequency for four known loci was three per 3,172 genomes analyzed. The estimated total number of loci mutated was 535 and the estimated number of kernel mutant loci mutated was 285. Among the 855 kernel mutants, 432 had a nonviable embryo, and 59 germinated but had a lethal seedling. A sample of 194 of the latter two types was tested for heritability, lethality, chromosome arm location and endosperm-embryo interaction between mutant and nonmutant tissues in special hyper-hypoploid combinations produced by manipulation of B-A translocations. The selected 194 mutants were characterized and catalogued according to endosperm phenotype and investigated to determine their effects on the morphology and development of the associated embryo. The possibility of rescuing some of the lethal mutants by covering the mutant embryo with a normal endosperm was investigated. Ninety of these 194 mutants were located on 17 of the 18 chromosome arms tested. Nineteen of the located mutants were examined to determine the effect of having a normal embryo in the same kernel with a mutant endosperm, and vice versa, as compared to the expression observed in kernels with both embryo and endosperm in a mutant condition. In the first situation, for three of the 19 mutants, the mutant endosperm was less extreme (the embryo helped); for seven cases, the mutant endosperm was more extreme (the embryo hindered); and for nine cases, there was no change. In the reverse situation, for four cases the normal endosperm helped the mutant embryo; for 14 cases there was no change and one case was inconclusive.

  2. The Solvent-Exposed C-Terminus of the Cytolysin A Pore-Forming Toxin Directs Pore Formation and Channel Function in Membranes. (United States)

    Sathyanarayana, Pradeep; Desikan, Rajat; Ayappa, K Ganapathy; Visweswariah, Sandhya S


    Pore-forming toxins (PFTs) bind to cell membranes and form nanoscale pores that allow leakage of cellular components, resulting in cell death. The water-soluble, monomeric form of these toxins shows a dramatic conformational change during pore formation, as exemplified by crystal structures of the monomer and functional pore of cytolysin A (ClyA). The solvent-exposed, C-terminal residues of the protein are essential for activity, but the mechanism by which this region regulates pore formation remains unknown. We show here that deletion of the C-terminus of ClyA did not alter its ability to bind to the membrane or oligomerize in detergent. However, the truncated toxin lysed erythrocytes poorly, was more susceptible to proteolysis and thermal unfolding, and showed low calcein leakage from small unilamellar vesicles. Using fully atomistic molecular dynamics (MD) simulations, we find that deletion of C-terminal residues from the ClyA monomer significantly altered stability and unfolding trajectories in the transmembrane N-terminal helix, a region that is pivotal in maintaining the structural integrity of the helical bundle. MD simulations of pores with or without the C-terminus showed minor differences, implying that if oligomerization could be induced prior to the addition to vesicles, then an active pore could be generated. Via generation of oligomers in a detergent prior to the addition to vesicles, the truncated toxin could induce calcein leakage from vesicles, albeit to a lower extent. Therefore, regions of pore-forming toxins, not directly involved in the pore structure, are not passive players but have important roles in undergoing the transition through intermediary steps leading to successful pore formation in a membrane environment.

  3. Mutants of downy mildew resistance in Lactuca sativa (lettuce). (United States)

    Okubara, P A; Anderson, P A; Ochoa, O E; Michelmore, R W


    As part of our investigation of disease resistance in lettuce, we generated mutants that have lost resistance to Bremia lactucae, the casual fungus of downy mildew. Using a rapid and reliable screen, we identified 16 distinct mutants of Latuca sativa that have lost activity of one of four different downy mildew resistance genes (Dm). In all mutants, only a single Dm specificity was affected. Genetic analysis indicated that the lesions segregated as single, recessive mutations at the Dm loci. Dm3 was inactivated in nine of the mutants. One of five Dm 1 mutants was selected from a population of untreated seeds and therefore carried a spontaneous mutation. All other Dm1, Dm3, Dm5/8 and Dm7 mutants were derived from gamma- or fast neutron-irradiated seed. In two separate Dm 1 mutants and in each of the eight Dm3 mutants analyzed, at least one closely linked molecular marker was absent. Also, high molecular weight genomic DNA fragments that hybridized to a tightly linked molecular marker in wild type were either missing entirely or were truncated in two of the Dm3 mutants, providing additional evidence that deletions had occurred in these mutants. Absence of mutations at loci epistatic to the Dm genes suggested that such loci were either members of multigene families, were critical for plant survival, or encoded components of duplicated pathways for resistance; alternatively, the genes determining downy mildew resistance might be limited to the Dm loci.

  4. Forward genetic screen for auxin-deficient mutants by cytokinin. (United States)

    Wu, Lei; Luo, Pan; Di, Dong-Wei; Wang, Li; Wang, Ming; Lu, Cheng-Kai; Wei, Shao-Dong; Zhang, Li; Zhang, Tian-Zi; Amakorová, Petra; Strnad, Miroslav; Novák, Ondřej; Guo, Guang-Qin


    Identification of mutants with impairments in auxin biosynthesis and dynamics by forward genetic screening is hindered by the complexity, redundancy and necessity of the pathways involved. Furthermore, although a few auxin-deficient mutants have been recently identified by screening for altered responses to shade, ethylene, N-1-naphthylphthalamic acid (NPA) or cytokinin (CK), there is still a lack of robust markers for systematically isolating such mutants. We hypothesized that a potentially suitable phenotypic marker is root curling induced by CK, as observed in the auxin biosynthesis mutant CK-induced root curling 1 / tryptophan aminotransferase of Arabidopsis 1 (ckrc1/taa1). Phenotypic observations, genetic analyses and biochemical complementation tests of Arabidopsis seedlings displaying the trait in large-scale genetic screens showed that it can facilitate isolation of mutants with perturbations in auxin biosynthesis, transport and signaling. However, unlike transport/signaling mutants, the curled (or wavy) root phenotypes of auxin-deficient mutants were significantly induced by CKs and could be rescued by exogenous auxins. Mutants allelic to several known auxin biosynthesis mutants were re-isolated, but several new classes of auxin-deficient mutants were also isolated. The findings show that CK-induced root curling provides an effective marker for discovering genes involved in auxin biosynthesis or homeostasis.

  5. Neurobehavioral Mutants Identified in an ENU Mutagenesis Project

    Energy Technology Data Exchange (ETDEWEB)

    Cook, Melloni N. [University of Memphis; Dunning, Jonathan P [University of Memphis; Wiley, Ronald G [Vanderbilt University and Veterans Administration, Nashville, TN; Chesler, Elissa J [ORNL; Johnson, Dabney K [ORNL; Goldowitz, Daniel [University of Tennessee Health Science Center, Memphis


    We report on a behavioral screening test battery that successfully identified several neurobehavioral mutants among a large-scale ENU-mutagenized mouse population. Large numbers of ENU mutagenized mice were screened for abnormalities in central nervous system function based on abnormal performance in a series of behavior tasks. We developed and employed a high-throughput screen of behavioral tasks to detect behavioral outliers. Twelve mutant pedigrees, representing a broad range of behavioral phenotypes, have been identified. Specifically, we have identified two open field mutants (one displaying hyper-locomotion, the other hypo-locomotion), four tail suspension mutants (all displaying increased immobility), one nociception mutant (displaying abnormal responsiveness to thermal pain), two prepulse inhibition mutants (displaying poor inhibition of the startle response), one anxiety-related mutant (displaying decreased anxiety in the light/dark test), and one learning and memory mutant (displaying reduced response to the conditioned stimulus) These findings highlight the utility of a set of behavioral tasks used in a high throughput screen to identify neurobehavioral mutants. Further analysis (i.e., behavioral and genetic mapping studies) of mutants is in progress with the ultimate goal of identification of novel genes and mouse models relevant to human disorders as well as the identification of novel therapeutic targets.

  6. Changes in Astrocyte Shape Induced by Sublytic Concentrations of the Cholesterol-Dependent Cytolysin Pneumolysin Still Require Pore-Forming Capacity

    Directory of Open Access Journals (Sweden)

    Christina Förtsch


    Full Text Available Streptococcus pneumoniae is a common pathogen that causes various infections, such as sepsis and meningitis. A major pathogenic factor of S. pneumoniae is the cholesterol-dependent cytolysin, pneumolysin. It produces cell lysis at high concentrations and apoptosis at lower concentrations. We have shown that sublytic amounts of pneumolysin induce small GTPase-dependent actin cytoskeleton reorganization and microtubule stabilization in human neuroblastoma cells that are manifested by cell retraction and changes in cell shape. In this study, we utilized a live imaging approach to analyze the role of pneumolysin’s pore-forming capacity in the actin-dependent cell shape changes in primary astrocytes. After the initial challenge with the wild-type toxin, a permeabilized cell population was rapidly established within 20–40 minutes. After the initial rapid permeabilization, the size of the permeabilized population remained unchanged and reached a plateau. Thus, we analyzed the non-permeabilized (non-lytic population, which demonstrated retraction and shape changes that were inhibited by actin depolymerization. Despite the non-lytic nature of pneumolysin treatment, the toxin’s lytic capacity remained critical for the initiation of cell shape changes. The non-lytic pneumolysin mutants W433F-pneumolysin and delta6-pneumolysin, which bind the cell membrane with affinities similar to that of the wild-type toxin, were not able to induce shape changes. The initiation of cell shape changes and cell retraction by the wild-type toxin were independent of calcium and sodium influx and membrane depolarization, which are known to occur following cellular challenge and suggested to result from the ion channel-like properties of the pneumolysin pores. Excluding the major pore-related phenomena as the initiation mechanism of cell shape changes, the existence of a more complex relationship between the pore-forming capacity of pneumolysin and the actin cytoskeleton

  7. Preparation of Metakaolin Based Geopolymer and Its Three- dimensional Pore Structural Characterization

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yunsheng; ZHANG Wenhua; SUN Wei; LI Zongjin; LIU Zhiyong


    Three types of pure geopolymer pastes (poly-sialate PS, poly- sialate-siloxo PSS, and poly-sialate-siloxo PSDS) werefi rst prepared by alkali (NaOH and KOH) activated metakaolin. Then a void space network was employed to simulate the 3-D pore-throat distribution across the unit cell of the various hardened geopolymer pastes with reference to their experimental mercury intrusion curves. Based on the simulated 3-D pore-throat structure models, a wide range of pore-level properties such as porosity, connectivity, permeability and tortuosity of various geopolymer pastes were calculated. The 3-D structural model and calculated parameters showed that most of the pores in Na-PS geopolymer paste was very small size pores ranging from 0 to 100 nm. A few very large pores were spread amongst the small pores, resulting in a very high penetration pressure, permeability resistance. Unlike Na-PS geopolymer paste, pore size with medium size of Na-PSS, K-PS and K-PSS geopolymer pastes distributed uniformly across the unit cell, and the size changes of adjacent pores in the 3 geopolymer pastes were little, producing higher penetration pressure, lower permeability, smaller connectivity and larger tortuosity. In contrast, pores in Na-PSDS and K-PSDS geopolymer pastes were relatively large and distributed concentratively, which caused samples to be easily penetrated by mercury, methane and nitrogenetc under relatively low pressures.

  8. Simulations of Pore Formation in Lipid Membranes: Reaction Coordinates, Convergence, Hysteresis, and Finite-Size Effects. (United States)

    Awasthi, Neha; Hub, Jochen S


    Transmembrane pores play an important role in various biophysical processes such as membrane permeation, membrane fusion, and antimicrobial peptide activity. In principal, all-atom molecular dynamics (MD) simulations provide an accurate model of pore formation in lipid membranes. However, the free energy landscape of transmembrane pore formation remains poorly understood, partly because potential of mean force (PMF) calculations of pore formation strongly depend on the choice of the reaction coordinate. In this study, we used umbrella sampling to compute PMFs for pore formation using three different reaction coordinates, namely, (i) a coordinate that steers the lipids in the lateral direction away from the pore center, (ii) the distance of a single lipid phosphate group from the membrane center, and (iii) the average water density inside a membrane-spanning cylinder. Our results show that while the three reaction coordinates efficiently form pores in membranes, they suffer from strong hysteresis between pore-opening and pore-closing simulations, suggesting that they do not restrain the systems close to the transition state for pore formation. The two reaction coordinates that act via restraining the lipids lead to more pronounced hysteresis compared with the coordinate acting on the water molecules. By comparing PMFs computed from membranes with different numbers of lipids, we observed significant artifacts from the periodic boundary conditions in small simulation systems. Further analysis suggests that the formation and disruption of a continuous hydrogen-bonding network across the membrane corresponds to the transition state for pore formation. Our study provides molecular insights into the critical steps of transmembrane pore formation, and it may guide the development of efficient reaction coordinates for pore formation.

  9. The Description of Shale Reservoir Pore Structure Based on Method of Moments Estimation (United States)

    Li, Wenjie; Wang, Changcheng; Shi, Zejin; Wei, Yi; Zhou, Huailai; Deng, Kun


    Shale has been considered as good gas reservoir due to its abundant interior nanoscale pores. Thus, the study of the pore structure of shale is of great significance for the evaluation and development of shale oil and gas. To date, the most widely used approaches for studying the shale pore structure include image analysis, radiation and fluid invasion methods. The detailed pore structures can be studied intuitively by image analysis and radiation methods, but the results obtained are quite sensitive to sample preparation, equipment performance and experimental operation. In contrast, the fluid invasion method can be used to obtain information on pore size distribution and pore structure, but the relative simple parameters derived cannot be used to evaluate the pore structure of shale comprehensively and quantitatively. To characterize the nanoscale pore structure of shale reservoir more effectively and expand the current research techniques, we proposed a new method based on gas adsorption experimental data and the method of moments to describe the pore structure parameters of shale reservoir. Combined with the geological mixture empirical distribution and the method of moments estimation principle, the new method calculates the characteristic parameters of shale, including the mean pore size (x¯), standard deviation (σ), skewness (Sk) and variation coefficient (c). These values are found by reconstructing the grouping intervals of observation values and optimizing algorithms for eigenvalues. This approach assures a more effective description of the characteristics of nanoscale pore structures. Finally, the new method has been applied to analyze the Yanchang shale in the Ordos Basin (China) and Longmaxi shale from the Sichuan Basin (China). The results obtained well reveal the pore characteristics of shale, indicating the feasibility of this new method in the study of the pore structure of shale reservoir. PMID:26992168

  10. Plant genetics: increased outcrossing in hothead mutants. (United States)

    Peng, Peng; Chan, Simon W-L; Shah, Govind A; Jacobsen, Steve E


    Arising from: S. J. Lolle, J. L. Victor, J. M. Young & R. E. Pruitt 434, 505-509 (2005); Lolle et al. reply. Lolle et al. report that loss-of-function alleles of the HOTHEAD (HTH) gene in Arabidopsis thaliana are genetically unstable, giving rise to wild-type revertants. On the basis of the reversion of many other genetic markers in hth plants, they suggested a model in which a cache of extragenomic information could cause genes to revert to the genotype of previous generations. In our attempts to reproduce this phenomenon, we discovered that hth mutants show a marked tendency to outcross (unlike wild-type A. thaliana, which is almost exclusively self-fertilizing). Moreover, when hth plants are grown in isolation, their genetic inheritance is completely stable. These results may provide an alternative explanation for the genome wide non-mendelian inheritance reported by Lolle et al.

  11. Google: a narrativa de uma marca mutante

    Directory of Open Access Journals (Sweden)

    Elizete de Azevedo Kreutz


    Full Text Available As marcas mutantes já fazem parte de nossa realidade, embora ainda não totalmente percebidas e/ou aceitas como tal. O presente artigo busca refletir sobre a relevância dessas novas estratégias de comunicação e branding, identificando suas principais características. Para isso, utilizamos o método de estudo de caso, o Google, ancorado nos métodos de pesquisa bibliográfica e de internet. A escolha foi intencional, posto que a organização é referência em sua categoria, mecanismo de busca, e reflete essa estratégia comunicacional contemporânea. Como resultado, as informações obtidas nos possibilitam compreender essa tendência de comportamento de marca que busca a interação com seus públicos.

  12. A fast Laplace solver approach to pore scale permeability (United States)

    Arns, Christoph; Adler, Pierre


    The permeability of a porous medium can be derived by solving the Stokes equations in the pore space with no slip at the walls. The resulting velocity averaged over the pore volume yields the permeability KS by application of the Darcy law. The Stokes equations can be solved by a number of different techniques such as finite differences, finite volume, Lattice Boltzmann, but whatever the technique it remains a heavy task since there are four unknowns at each node (the three velocity components and the pressure) which necessitate the solution of four equations (the projection of Newton's law on each axis and mass conservation). By comparison, the Laplace equation is scalar with a single unknown at each node. The objective of this work is to replace the Stokes equations by an elliptical equation with a space dependent permeability. More precisely, the local permeability k is supposed to be proportional to (r-alpha)**2 where r is the distance of the voxel to the closest wall, and alpha a constant; k is zero in the solid phase. The elliptical equation is div(k gradp)=0. A macroscopic pressure gradient is assumed to be exerted on the medium and again the resulting velocity averaged over space yields a permeability K_L. In order to validate this method, systematic calculations have been performed. First, elementary shapes (plane channel, circular pipe, rectangular channels) were studied for which flow occurs along parallel lines in which case KL is the arithmetic average of the k's. KL was calculated for various discretizations of the pore space and various values of alpha. For alpha=0.5, the agreement with the exact analytical value of KS is excellent for the plane and rectangular channels while it is only approximate for circular pipes. Second, the permeability KL of channels with sinusoidal walls was calculated and compared with analytical results and numerical ones provided by a Lattice Boltzmann algorithm. Generally speaking, the discrepancy does not exceed 25% when

  13. Creating transient cell membrane pores using a standard inkjet printer. (United States)

    Owczarczak, Alexander B; Shuford, Stephen O; Wood, Scott T; Deitch, Sandra; Dean, Delphine


    Bioprinting has a wide range of applications and significance, including tissue engineering, direct cell application therapies, and biosensor microfabrication. Recently, thermal inkjet printing has also been used for gene transfection. The thermal inkjet printing process was shown to temporarily disrupt the cell membranes without affecting cell viability. The transient pores in the membrane can be used to introduce molecules, which would otherwise be too large to pass through the membrane, into the cell cytoplasm. The application being demonstrated here is the use of thermal inkjet printing for the incorporation of fluorescently labeled g-actin monomers into cells. The advantage of using thermal ink-jet printing to inject molecules into cells is that the technique is relatively benign to cells. Cell viability after printing has been shown to be similar to standard cell plating methods. In addition, inkjet printing can process thousands of cells in minutes, which is much faster than manual microinjection. The pores created by printing have been shown to close within about two hours. However, there is a limit to the size of the pore created (~10 nm) with this printing technique, which limits the technique to injecting cells with small proteins and/or particles. A standard HP DeskJet 500 printer was modified to allow for cell printing. The cover of the printer was removed and the paper feed mechanism was bypassed using a mechanical lever. A stage was created to allow for placement of microscope slides and coverslips directly under the print head. Ink cartridges were opened, the ink was removed and they were cleaned prior to use with cells. The printing pattern was created using standard drawing software, which then controlled the printer through a simple print command. 3T3 fibroblasts were grown to confluence, trypsinized, and then resuspended into phosphate buffered saline with soluble fluorescently labeled g-actin monomers. The cell suspension was pipetted into the

  14. Mutants of Cercospora kikuchii altered in cercosporin synthesis and pathogenicity

    Energy Technology Data Exchange (ETDEWEB)

    Upchurch, R.G.; Walker, D.C.; Rollins, J.A.; Ehrenshaft, M.; Daub, M.E. (North Carolina State Univ., Raleigh (United States))


    The authors have obtained spontaneous and UV-induced stable mutants, altered in the synthesis of cercosporin, of the fungal soybean pathogen Cercospora kikuchii. The mutants were isolated on the basis of colony color on minimal medium. The UV-induced mutants accumulated, at most, 2% of wild-type cercosporin levels on all media tested. In contrast, cercosporin accumulation by the spontaneous mutants was strongly medium regulated, occurring only on potato dextrose medium but at concentrations comparable to those produced by the wild-type strain. UV-induced mutants unable to synthesize cercosporin on any medium were unable to incite lesions when inoculated onto the soybean host. Cercosporin was reproducibly isolated from all inoculated leaves showing lesions. Although cercosporin involvement in disease has been indirectly suggested by many previous studies, this is the first report in which mutants blocked in cercosporin synthesis have been used to demonstrate that cercosporin is a crucial pathogenicity factor for this fungal genus.

  15. Mutants of Cercospora kikuchii Altered in Cercosporin Synthesis and Pathogenicity. (United States)

    Upchurch, R G; Walker, D C; Rollins, J A; Ehrenshaft, M; Daub, M E


    We have obtained spontaneous and UV-induced stable mutants, altered in the synthesis of cercosporin, of the fungal soybean pathogen Cercospora kikuchii. The mutants were isolated on the basis of colony color on minimal medium. The UV-induced mutants accumulated, at most, 2% of wild-type cercosporin levels on all media tested. In contrast, cercosporin accumulation by the spontaneous mutants was strongly medium regulated, occurring only on potato dextrose medium but at concentrations comparable to those produced by the wild-type strain. UV-induced mutants unable to synthesize cercosporin on any medium were unable to incite lesions when inoculated onto the soybean host. Cercosporin was reproducibly isolated from all inoculated leaves showing lesions. Although cercosporin involvement in disease has been indirectly suggested by many previous studies, this is the first report in which mutants blocked in cercosporin synthesis have been used to demonstrate that cercosporin is a crucial pathogenicity factor for this fungal genus.

  16. Colony mutants of compatible nocardiae displaying variations in recombining capacity. (United States)

    Brownell, G H; Walsh, R S


    Colonial morphology mutants of Nocardia erythropolis were isolated following ultraviolet (UV) irradiation. The alleles rou-1/smo-1 were located by recombinant analysis and found to be linked to previously mapped characters. On the basis of recombinant class type patterns obtained from various selective characters it was postulated that the rou-1 allele may span a region of unique nucleotides in the Mat-Ce genome. Recombination frequencies of rou-1 and smo-2 bearing mutants of the Mat-Ce mating type were found to differ by over 1000 fold. Attempts to demonstrate that low recombination frequencies produced by the Smo mutants were due to Rec(-) genes were unsuccessful. No increased sensitivity to either UV or X irradiation was observed by the Smo mutants. Acriflavine treatment of either Rou or Smo colony mutants failed to accelerate reversion or to alter the recombining potentials of the mutants.

  17. Screening and identification of mutants of Magnaporthe grisea by REMI

    Institute of Scientific and Technical Information of China (English)

    XIONG Ruyi; LIU Juan; ZHOU Yijun; FAN Yongjian; ZHENG Xiaobo


    The plasmid pUCATPH was used to establish a transformation system in wild-type isolate M131 of Magnaporthe grisea.Six hundred and thirty-nine transformants were obtained by restriction enzyme-mediated integration (REMI) with hygromycin B (hyg B) resistance as a tag.Morphological analysis of two of the REMI mutants confirmed that they produced little melanin under black light and continued for three generations.Pathogenicity identification of six mutants screened proved that they made pathogenicity changes on three sets of differential varieties with different resistance genes.Rep-PCR analyses showed that two morphological mutants and two pathogenicity mutants differed from wild-type isolate M131 at the molecular level.RFLP analyses were performed to study the four mutants at the molecular level and the integration sites of the plasmid DNA.The results showed that the plasmid was inserted into all four mutants and that the insertion sites were random.

  18. Mutant p53: multiple mechanisms define biologic activity in cancer

    Directory of Open Access Journals (Sweden)

    Michael Paul Kim


    Full Text Available The functional importance of p53 as a tumor suppressor gene is evident through its pervasiveness in cancer biology. The p53 gene is the most commonly altered gene in human cancer; however, not all genetic alterations are biologically equivalent. The majority of p53 alterations involve missense mutations that result in the production of mutant p53 proteins. Such mutant p53 proteins lack normal p53 function and may acquire novel functions, often with deleterious effects. Here, we review characterized mechanisms of mutant p53 gain of function in multiple model systems. In addition, we review mutant p53 addiction as emerging evidence suggests that tumors may depend on sustained mutant p53 activity for continued growth. We also discuss the role of p53 in stromal elements and their contribution to tumor initiation and progression. Lastly, current genetic mouse models of mutant p53 are reviewed and their limitations discussed.

  19. Nanopore analysis of wild-type and mutant prion protein (PrP(C: single molecule discrimination and PrP(C kinetics.

    Directory of Open Access Journals (Sweden)

    Nahid N Jetha

    Full Text Available Prion diseases are fatal neurodegenerative diseases associated with the conversion of cellular prion protein (PrP(C in the central nervous system into the infectious isoform (PrP(Sc. The mechanics of conversion are almost entirely unknown, with understanding stymied by the lack of an atomic-level structure for PrP(Sc. A number of pathogenic PrP(C mutants exist that are characterized by an increased propensity for conversion into PrP(Sc and that differ from wild-type by only a single amino-acid point mutation in their primary structure. These mutations are known to perturb the stability and conformational dynamics of the protein. Understanding of how this occurs may provide insight into the mechanism of PrP(C conversion. In this work we sought to explore wild-type and pathogenic mutant prion protein structure and dynamics by analysis of the current fluctuations through an organic α-hemolysin nanometer-scale pore (nanopore in which a single prion protein has been captured electrophoretically. In doing this, we find that wild-type and D178N mutant PrP(C, (a PrP(C mutant associated with both Fatal Familial Insomnia and Creutzfeldt-Jakob disease, exhibit easily distinguishable current signatures and kinetics inside the pore and we further demonstrate, with the use of Hidden Markov Model signal processing, accurate discrimination between these two proteins at the single molecule level based on the kinetics of a single PrP(C capture event. Moreover, we present a four-state model to describe wild-type PrP(C kinetics in the pore as a first step in our investigation on characterizing the differences in kinetics and conformational dynamics between wild-type and D178N mutant PrP(C. These results demonstrate the potential of nanopore analysis for highly sensitive, real-time protein and small molecule detection based on single molecule kinetics inside a nanopore, and show the utility of this technique as an assay to probe differences in stability between

  20. Preliminary study on a gravity-insensitive rice mutant

    Institute of Scientific and Technical Information of China (English)

    金静; 朱诚; 张红心; 孙宗修


    A gravity-insensitive mutant was isolated from rice (Oryza sativa L. Cv. Zhonghua 11) transformed by Agrobacterium tumefaciens. The mutant's shoot growth (prostrate growth) was insensitive to gravity; whereas root growth displayed a normal positive gravitropism.Histological observation of root caps and leaf sheaths indicated that there was no significant difference in the number and size of amyloplasts in cells of the mutant and cells of the wild type

  1. Preliminary study on a gravity-insensitive rice mutant

    Institute of Scientific and Technical Information of China (English)

    金静; 朱诚; 张红心; 孙宗修


    A gravity-insensitive mutant was isolated from rice (Oryza sativa L. cv. Zhonghua 11) transformed by Agrobacterium tumefaciens. The mutant's shoot growth (prostrate growth) was insensitive to gravity; whereas root growth displayed a normal positive gravitropism. Histological observation of root caps and leaf sheaths indicated that there was no significant difference in the number and size of amyloplasts in cells of the mutant and cells of the wild type.

  2. [Eremothecium ashbyii mutants resistant to 2,6-diaminopurine]. (United States)

    Stepanov, A I; Beburov, M Iu; Zhdanov, V G


    3 groups of Eremothecium ashbyii mutants resistant to 5-10(-3) M 2,6-diaminopurine (DAP) ahve been obtained. The mutants of the 1st group (Dap-r) are selected from the initial susceptible strain by the ability to grow in the presence of 5-10(-3) M DAP. The mutants of the 2nd group (Azg-Dap-r) are selected in the selective background of two analogues of 5-10(-3) M DAP and 10(-4) M 8-azaguanine (AG). The mutants of the 3rd group (Azg-r - DAP-r) are isolated from the mutant Azg-r 34 resistant to 10(-4) M AG. The results of studying cross-resistance of mutants to DAP, AG and 8-azaadenine (AA) show that Dap-r and Azg-Dap-r mutants in contrast to Azg-r - Dap-r, have common phenotypic properties and can grow only on the analogues of adenine. DAP, but not AA, eliminates the inhibitory effect of AG on the growth of these mutants. This effect is probably due to deaminating DAP to guanine. Mutants Azg-r - Dap-r retain the initial resistance to 10(-4) M AG, but are susceptible to higher concentrations of AG and in this case DAP does not eliminate the inhibitory effect of AG. In all mutants obtained the effectiveness of the incorporation of 14C-adenine (but not 14C-guanine) is sharply reduced, thus indicating the absence of adenosine-monophosphate pyrophosphorylase activity. The mutants do not excrete purine-like compounds into the medium. In the course of the continuous growth of mutants in the presence of DAP but not of guanine the red intracellular pigment is formed which seems to be a complex of riboflavin with DAP. A disturbance in the synthesis of adenosine monophosphate pyrophosphorylase does not influence practically the level of the synthesis of riboflavin in E. ashbyii.

  3. Fatty acid biosynthesis in novel ufa mutants of Neurospora crassa. (United States)

    Goodrich-Tanrikulu, M; Stafford, A E; Lin, J T; Makapugay, M I; Fuller, G; McKeon, T A


    New mutants of Neurospora crassa having the ufa phenotype have been isolated. Two of these mutants, like previously identified ufa mutants, require an unsaturated fatty acid for growth and are almost completely blocked in the de novo synthesis of unsaturated fatty acids. The new mutations map to a different chromosomal location than previously characterized ufa mutations. This implies that at least one additional genetic locus controls the synthesis of unsaturated fatty acids in Neurospora.

  4. Transport and Deposition of Nanoparticles in the Pore Network of a Reservoir Rock: Effects of Pore Surface Heterogeneity and Radial Diffusion (United States)

    Pham, Ngoc; Papavassiliou, Dimitrios


    In this study, transport behavior of nanoparticles under different pore surface conditions of consolidated Berea sandstone is numerically investigated. Micro-CT scanning technique is applied to obtain 3D grayscale images of the rock sample geometry. Quantitative characterization, which is based on image analysis is done to obtain physical properties of the pore network, such as the pore size distribution and the type of each pore (dead-end, isolated, and fully connected pore). Transport of water through the rock is simulated by employing a 3D lattice Boltzmann method. The trajectories of nanopaticles moving under convection in the simulated flow field and due to molecular diffusion are monitored in the Lagrangian framework. It is assumed in the model that the particle adsorption on the pore surface, which is modeled as a pseudo-first order adsorption, is the only factor hindering particle propagation. The effect of pore surface heterogeneity to the particle breakthrough is considered, and the role of particle radial diffusion is also addressed in details. The financial support of the Advanced Energy Consortium (AEC BEG08-022) and the computational support of XSEDE (CTS090017) are acknowledged.

  5. The Structure Inventory of the Nuclear Pore Complex. (United States)

    Schwartz, Thomas U


    The nuclear pore complex (NPC) is the principal gateway for molecular exchange between nucleus and cytoplasm across the nuclear envelope. Due to its sheer size of estimated 50-112MDa and its complex buildup from about 500-1000 individual proteins, it is a difficult object to study for structural biologists. Here, I review the extensive ensemble of high-resolution structures of the building blocks of the NPC. Concurrent with the increase in size and complexity, these latest, large structures and assemblies can now be used as the basis for hybrid approaches, primarily in combination with cryo-electron microscopic analysis, generating the first structure-based assembly models of the NPC. Going forward, the structures will be critically important for a detailed analysis of the NPC, including function, evolution, and assembly.

  6. Pathophysiological role of omega pore current in channelopathies

    Directory of Open Access Journals (Sweden)

    Karin eJurkat-Rott


    Full Text Available In voltage-gated cation channels, a recurrent pattern for mutations is the neutralization of positively charged residues in the voltage-sensing S4 transmembrane segments. These mutations cause dominant ion channelopathies affecting many tissues such as brain, heart, and skeletal muscle. Recent studies suggest that the pathogenesis of associated phenotypes is not limited to alterations in the gating of the ion-conducting alpha pore. Instead, aberrant so-called omega currents facilitated by the movement of the S4 segments during activation and during recovery are thought to cause symptoms. Surprisingly, these omega currents display uni- or bi-directionality and conduct cations with varying ion selectivity. Additionally, the voltage-sensitivity enables the channels to conduct different omega currents in the various voltage ranges. This review gives an overview of voltage sensor channelopathies in general and focuses on pathogenesis of skeletal muscle S4 disorders for which current knowledge is most advanced.

  7. Dynamics of polynucleotide transport through nanometre-scale pores

    CERN Document Server

    Meller, A


    The transport of biopolymers through large membrane channels is a ubiquitous process in biology. It is central to processes such as gene transfer by transduction and RNA transport through nuclear pore complexes. The transport of polymers through nanoscopic channels is also of interest to physicists and chemists studying the effects of steric, hydrodynamic, and electrostatic interactions between polymers and confining walls. Single-channel ion current measurements have been recently used to study the transport of biopolymers, and in particular single-stranded DNA and RNA molecules, through nanometre-size channels. Under the influence of an electric field, the negatively charged polynucleotides can be captured and drawn through the channel in a process termed 'translocation'. During translocation, the ion current flowing through the channel is mostly blocked, indicating the presence of the polymer inside the channel. The current blockades were found to be sensitive to the properties of the biopolymers such as t...

  8. Boiling visualization on vertical fins with tunnel-pore structures

    Directory of Open Access Journals (Sweden)

    Kaniowski Robert


    Full Text Available The paper presents experimental studies of nucleate boiling heat transfer from a system of connected horizontal and vertical subsurface tunnels. The experiments were carried out for water at atmospheric pressure. The tunnel external covers were manufactured out of perforated copper foil (holes diameter 0.3 mm, sintered with the mini-fins, formed on the vertical side of the 10 mm high rectangular fins and horizontal inter-fin surface. The image acquisition speed was 493 fps (at resolution 400 × 300 pixels with Photonfocus PHOT MV-D1024-160-CL camera. Visualization investigations aimed to identify nucleation sites and flow patterns and to determine the bubble departure diameter and frequency at various superheats for vertical tunnels. At low superheat vapor bubbles are generated nearly exclusively by the vertical tunnel. At medium values of superheat, pores of the horizontal tunnel activate.

  9. Bax and Bak Pores: Are We Closing the Circle? (United States)

    Cosentino, Katia; García-Sáez, Ana J


    Bax and its homolog Bak are key regulators of the mitochondrial pathway of apoptosis. On cell stress Bax and Bak accumulate at distinct foci on the mitochondrial surface where they undergo a conformational change, oligomerize, and mediate cytochrome c release, leading to cell death. The molecular mechanisms of Bax and Bak assembly and mitochondrial permeabilization have remained a longstanding question in the field. Recent structural and biophysical studies at several length scales have shed light on key aspects of Bax and Bak function that have shifted how we think this process occurs. These discoveries reveal an unexpected molecular mechanism in which Bax (and likely Bak) dimers assemble into oligomers with an even number of molecules that fully or partially delineate pores of different sizes to permeabilize the mitochondrial outer membrane (MOM) during apoptosis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. The mitochondrial permeability transition pore: a mystery solved? (United States)

    Bernardi, Paolo


    The permeability transition (PT) denotes an increase of the mitochondrial inner membrane permeability to solutes with molecular masses up to about 1500 Da. It is presumed to be mediated by opening of a channel, the permeability transition pore (PTP), whose molecular nature remains a mystery. Here I briefly review the history of the PTP, discuss existing models, and present our new results indicating that reconstituted dimers of the FOF1 ATP synthase form a channel with properties identical to those of the mitochondrial megachannel (MMC), the electrophysiological equivalent of the PTP. Open questions remain, but there is now promise that the PTP can be studied by genetic methods to solve the large number of outstanding problems. PMID:23675351

  11. Integrated landslide monitoring: rainfalls, pore water pressures and surface movements (United States)

    Berti, M.; Casula, G.; Elmi, C.; Fabris, M.; Ghirotti, M.; Loddo, F.; Mora, P.; Pesci, A.; Simoni, A.


    Rainfall-induced landslides involving clay-rich soils are widely represented in the Apennines. They cover up to 30% of the slopes forming the relief constituted by chaotic clayey units and are typically subject to repeated reactivations of the movement which are often triggered by a series of discrete failures located in the upper part (headscarp). Failures and movement can then propagate downslope and reactivate the whole landslide deposit which displays a typical elongated body, limited depth and a fan-shaped toe as a result of successive slow earth-flow like movements. An experimental monitoring programme was designed and is currently operating on the Rocca Pitigliana landslide whose characteristics well represent the above described type of movements. Its last parossistic movement date back to 1999 and, since then, remedial works were realized on behalf of local authorities. They basically consist of surficial and deep drainage works located on the landslide body. Experimental activities focus on the main headscarp whose morphology and sub-surface water circulation scheme were unaffected by the interventions. The monitoring approach includes measuring rainfalls and pore-pressure responses in both saturated and unsaturated soils. Surficial movements are continuously measured by means of GPS permanent stations and by wire extensometers which allow real time control of headscarp activity. Main aim of the monitoring activities is to provide experimental data, which can be used to test various existing hydrologic models and to identify triggering conditions. Since the ‘70s, many hydrologic models have been proposed to describe the pore water pressure distribution within the soil and its response to precipitation. The topic has recently drawn growing attention because of the recognized importance in landslide triggering but still experimental data are very much needed in order to obtain and validate capable predicting tools. This is mostly due to the multiple and

  12. A biomimetic nanosponge that absorbs pore-forming toxins (United States)

    Hu, Che-Ming J.; Fang, Ronnie H.; Copp, Jonathan; Luk, Brian T.; Zhang, Liangfang


    Detoxification treatments such as toxin-targeted anti-virulence therapy offer ways to cleanse the body of virulence factors that are caused by bacterial infections, venomous injuries and biological weaponry. Because existing detoxification platforms such as antisera, monoclonal antibodies, small-molecule inhibitors and molecularly imprinted polymers act by targeting the molecular structures of toxins, customized treatments are required for different diseases. Here, we show a biomimetic toxin nanosponge that functions as a toxin decoy in vivo. The nanosponge, which consists of a polymeric nanoparticle core surrounded by red blood cell membranes, absorbs membrane-damaging toxins and diverts them away from their cellular targets. In a mouse model, the nanosponges markedly reduce the toxicity of staphylococcal alpha-haemolysin (α-toxin) and thus improve the survival rate of toxin-challenged mice. This biologically inspired toxin nanosponge presents a detoxification treatment that can potentially treat a variety of injuries and diseases caused by pore-forming toxins.

  13. Density profile of nitrogen in cylindrical pores of MCM-41 (United States)

    Soper, Alan K.; Bowron, Daniel T.


    A straightforward approach using radiation scattering (X-ray or neutron) combined with atomistic modelling is used to accurately assess the pore dimensions in the porous silica, MCM-41. The method is used to calculate the density profile of nitrogen absorbed in this material at a variety of fractional pressures, p/p0, where p0 is the saturated vapour pressure, up to p/p0 = 0.36 at T = 87 K in the present instance. At this pressure two distinct layers of liquid nitrogen occur on the silica surface, with a relatively sharp gas-liquid interface. It is suggested surface tension effects at this interface strongly influence the growth of further layers.

  14. Measurement of Restricted Atmospheric Barrier Discharge in Nonwoven Fiber Pores (United States)

    Kawabe, Masaaki

    The restricted dielectric barrier discharge in nonwoven pores has been investigated by observation of the current pulse, the Lissajous figure and light emissions. The current pulse measurement revealed that homogeneity of the discharge was relatively high and the amount of individual pulse was quite small on the order of 0.01nC. Such a small current pulse demonstrates that nonwoven fiber is effective as a dielectric barrier. Analysis of the Lissajous figure, indicates the calculated value of the gap voltage for the discharge starting point of nonwoven fiber layers is close to what was predicted using the Paschen curve. On the other hand, the measured value of the gap voltage in the Lissajous figure is larger than its calculated value, so the surface charge on the dielectrics dissipated relatively fast. The observations of light emissions also showed a high homogeneity.

  15. Diffusion of Macromolecules across the Nuclear Pore Complex

    CERN Document Server

    Chakrabarti, Rajarshi; Sebastian, K L


    Nuclear pore complexes (NPCs) are very selective filters that monitor the transport between the cytoplasm and the nucleoplasm. Two models have been suggested for the plug of the NPC. They are (i) it is a reversible hydrogel or (ii) it is a polymer brush. We propose a mesoscopic model for the transport of a protein through the plug, that is general enough to cover both. The protein stretches the plug and creates a local deformation. The bubble so created (prtoein+deformation) executes random walk in the plug. We find that for faster relaxation of the gel, the diffusion of the bubble is greater. Further, on using parameters appropriate for the brush, we find that the diffusion coefficient is much lower. Hence the gel model seems to be more likely explanation for the workings of the plug.

  16. Accurate and efficient maximal ball algorithm for pore network extraction (United States)

    Arand, Frederick; Hesser, Jürgen


    The maximal ball (MB) algorithm is a well established method for the morphological analysis of porous media. It extracts a network of pores and throats from volumetric data. This paper describes structural modifications to the algorithm, while the basic concepts are preserved. Substantial improvements to accuracy and efficiency are achieved as follows: First, all calculations are performed on a subvoxel accurate distance field, and no approximations to discretize balls are made. Second, data structures are simplified to keep memory usage low and improve algorithmic speed. Third, small and reasonable adjustments increase speed significantly. In volumes with high porosity, memory usage is improved compared to classic MB algorithms. Furthermore, processing is accelerated more than three times. Finally, the modified MB algorithm is verified by extracting several network properties from reference as well as real data sets. Runtimes are measured and compared to literature.

  17. Characterization of host-range mutants of cyanophage N-1. (United States)

    Sarma, T A; Kaur, B


    Fifteen host-range (h) mutants of cyanophage N-1 were characterized with reference to their efficiency of plating, time of appearance, morphology and size of plaques on Nostoc muscorum and its three phage-resistant (Nm 1/N-1, Nm 2/N-1 and Nm 8/N-1) mutants. While phage N-1 did not adsorb to the three phage-resistant mutants, the h mutants differed one from the other in having lower or higher adsorption rate constants on N. muscorum or the phage-resistant mutants. The inability of majority of h mutants isolated on Nm 1/N-1 to grow in Nm 8/N-1 was shown to be due to a failure of adsorption. The h mutants also differed one from the other in their reversion (back mutation) frequencies. The lethal doses (LD37) required to kill 37% of free phage particles after UV-irradiation, heating and ethylenediamine tetraacetate (EDTA) treatment greatly varied. Most of the h mutants were found to be considerably more sensitive to UV and thermic inactivation than N-1 while they were resistant to EDTA. The h mutants except five of them were unable to multiply at 40 degrees C. The significance of these features is discussed.

  18. plenty, a novel hypernodulation mutant in Lotus japonicus. (United States)

    Yoshida, Chie; Funayama-Noguchi, Sachiko; Kawaguchi, Masayoshi


    Nitrogen fixation in nodules that contain symbiotic rhizobial bacteria enables legumes to thrive in nitrogen-poor soils. However, this symbiosis is energy consuming. Therefore, legumes strictly control nodulation at both local and systemic levels. Mutants deficient in such controls exhibit a range of phenotypes from non-nodulation to hypernodulation. Here, we isolated a novel hypernodulation mutant from the M(2) progeny derived from Lotus japonicus MG-20 seeds mutagenized by irradiation with a carbon ion beam. We named the mutant 'plenty' because it formed more nodules than the wild-type MG-20. The nodulation zone in the plenty mutant was wider than that in the wild type, but not as enhanced as those in other previously reported hypernodulation mutants such as har1, klv or tml of L. japonicus. Unlike these hypernodulation mutants, the plenty mutant developed nodules of the same size as MG-20. Overall, the plenty mutant exhibited a unique phenotype of moderate hypernodulation. However, a biomass assay indicated that this unique pattern of hypernodulation was a hindrance to host plant growth. The plenty mutant displayed some tolerance to external nitrates and a normal triple response to ethylene. Grafting experiments demonstrated that the root of plenty was responsible for its hypernodulation phenotype. Genetic mapping indicated that the PLENTY gene was located on chromosome 2.

  19. Induction and selection of citrus mutant by gamma-irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, In Jung; Oh, Seung Kyu; Lee, Hyo Yeon [Jeju National University, Jeju (Korea, Republic of)


    We have subjected to gamma-irradiation to citrus buds and then grafted onto mature citrus tree. Mutant citrus branch lines have been induced. As a result of first selection, we found the several mutant lines showing interesting phenotypes such as higher sugar content. We have selected several branches showing good qualities such as higher sweetness and/or lower acidity. Some branch lines showed over 13 .deg. Brix sugar content and below 0.9% acidity. Other mutant branch lines showed the changes of shape, size, peel thickness, and fiber contents or distribution of fruits. The results suggest that gamma-irradiation is an effective tool for induction of citrus mutant lines.

  20. JNK3 phosphorylates Bax protein and induces ability to form pore on bilayer lipid membrane

    Directory of Open Access Journals (Sweden)

    Rajeev Gupta


    Full Text Available Bax is a pro-apoptotic cytosolic protein. In this work native (unphosphorylated and JNK3 phosphorylated Bax proteins are studied on artificial bilayer membranes for pore formation. Phosphorylated Bax formed pore on the bilayer lipid membrane whereas native one does not. In cells undergoing apoptosis the pore formed by the phosphorylated Bax could be important in cytochrome c release from the mitochondrial intermembrane space to the cytosol. The low conductance (1.5 nS of the open state of the phosphorylated Bax pore corresponds to pore diameter of 0.9 nm which is small to release cytochrome c (∼3.4 nm. We hypothesized that JNK3 phosphorylated Bax protein can form bigger pores after forming complexes with other mitochondrial proteins like VDAC, t-Bid etc. to release cytochrome c.

  1. Analysis of Pore Structures and Their Relations with Strength of Hardened Cement Paste

    Institute of Scientific and Technical Information of China (English)

    ZHANG Wensheng; LI Beixing; WANG Hongxia; WEI Jiangxiong; CHEN Yimin


    Three cement samples were prepared, including OPC consisted of 100wt% portland cement, PFA consisted of 70wt% portland cement and 30wt% fly-ash, and CA consisted of 70wt% portland cement and 30wt% modified fly ash. The strength of hardened cement paste of these samples was tested and their pore structures were determined by a mercury intrusion porosimeter. Moreover,the data of the pore structures of three samples were comprehensively analyzed. The relations between the pore structures and the compressive strength of the three samples were studied. The experimental results show that the relations between the porosity determined by the mercury intrusion porosimeter and the compressive strength are not notable, and the total pore surface area, the average pore diameter and the median pore diameter could be used to explain the difference of the strength of the tested samples.

  2. Impact of pore size variability and network coupling on electrokinetic transport in porous media (United States)

    Alizadeh, Shima; Bazant, Martin Z.; Mani, Ali


    We have developed and validated an efficient and robust computational model to study the coupled fluid and ion transport through electrokinetic porous media, which are exposed to external gradients of pressure, electric potential, and concentration. In our approach a porous media is modeled as a network of many pores through which the transport is described by the coupled Poisson-Nernst-Planck-Stokes equations. When the pore sizes are random, the interactions between various modes of transport may provoke complexities such as concentration polarization shocks and internal flow circulations. These phenomena impact mixing and transport in various systems including deionization and filtration systems, supercapacitors, and lab-on-a-chip devices. In this work, we present simulations of massive networks of pores and we demonstrate the impact of pore size variation, and pore-pore coupling on the overall electrokinetic transport in porous media.

  3. Quantification of Soil Pore Network Complexity with X-ray Computed Tomography and Gas Transport Measurements

    DEFF Research Database (Denmark)

    Katuwal, Sheela; Arthur, Emmanuel; Tuller, Markus


    Flow and transport of gases through soils are largely controlled by pore structural attributes. The quantification of pore network characteristics is therefore essential for accurate prediction of air permeability and gas diffusivity. In this study, the pore network characteristics of seven...... different soils subjected to 22 mo of field regeneration were quantified with X-ray computed tomography (CT) and compared with functional pore characteristics estimated from measurements of air permeability and gas diffusivity. Furthermore, predictive models for air permeability and gas diffusivity were...... equivalent pore diameter in predictive gas diffusivity and air permeability models significantly improved their performance. The obtained results suggest that the application of X-ray CT-derived pore-structural parameters has great potential for predicting gas diffusivity and air permeability....

  4. A Microfluidic Pore Network Approach to Investigate Water Transport in Fuel Cell Porous Transport Layers

    CERN Document Server

    Bazylak, A; Markicevic, B; Sinton, D; Djilali, N


    Pore network modelling has traditionally been used to study displacement processes in idealized porous media related to geological flows, with applications ranging from groundwater hydrology to enhanced oil recovery. Very recently, pore network modelling has been applied to model the gas diffusion layer (GDL) of a polymer electrolyte membrane (PEM) fuel cell. Discrete pore network models have the potential to elucidate transport phenomena in the GDL with high computational efficiency, in contrast to continuum or molecular dynamics modelling that require extensive computational resources. However, the challenge in studying the GDL with pore network modelling lies in defining the network parameters that accurately describe the porous media as well as the conditions of fluid invasion that represent realistic transport processes. In this work, we discuss the first stage of developing and validating a GDL-representative pore network model. We begin with a two-dimensional pore network model with a single mobile pha...

  5. Determination of a Pore Structure Parameter of Porous Media by Analysis of Percolation Network Model

    Institute of Scientific and Technical Information of China (English)


    According to the simulation of nitrogen sorption process in porous media with three-dimensional network model, and the analysis for such a process with percolation theory, a new method is proposed to determine a pore structure parameter-mean coordination number of pore network, which represents the connectivity among a great number of pores. Here the “chamber-throat” model and the Weibull distribution are used to describe the pore geometry and the pore size distribution respectively. This method is based on the scaling law of percolation theory after both effects of sorption thermodynamics and pore size on the sorption hysteresis loops are considered. The results show that it is an effective procedure to calculate the mean coordination number for micro- and meso-porous media.

  6. A quantitative investigation of the effect of pore morphology on soil aggregate stability (United States)

    Papadopoulos, A.


    Soil structure determines the operating environment for all physical, chemical and biological processes within the soil. Soil aggregate stability is an important measure for assessing soil structure quality. Non-destructive tomography techniques such as X-ray Computed Tomography (CT) offer great opportunities to quantitatively investigate the soil porous architecture which can provide important information for understanding soil processes and function in a multi-scale manner. For instance, the intra-aggregate pore space is of great importance for microbial activity, the sequestration of organic carbon and water flow. This paper investigates the effect of pore morphology on soil aggregate stability. Apparent porosity, pore size distribution, average pore size and fractal perimeter dimension (pore roughness) were measured from the images of the reconstructed 2-D image stacks. A new theoretical concept of soil aggregate stability is proposed. A strong relationship was observed between soil aggregate stability and pore morphological complexity.


    Institute of Scientific and Technical Information of China (English)

    Haidi Liu; Shufeng Ye; Yunfa Chen


    Mesoporous silica was prepared using tetrathoxysilane (TEOS), cetadecyltrimethylammonium bromide (CTAB), aqueous ammonia, acetone and water as silica source, template agent, precipitator and solvent respectively.Stearic acid and polyvinylpyrrolidone (PVP) were employed as additional templates to tailor the pore size in the resultant porous silica. BET, SAXRD and SEM analyses were used to characterize the surface area, pore size, pore structure,pore regularity and morphology of the sample. BET measurement results showed that PVP could increase the surface area but diminish the pore size while stearic acid could decrease the surface area but enlarge the pore size. SAXRD analysis indicated that more additional template introduction gave rise to less order-structured products. All these various results could be attributed to the differently modified CTAB micelles involving stearic acid and PVP addition.

  8. pH controlled gating of toxic protein pores by dendrimers (United States)

    Mandal, Taraknath; Kanchi, Subbarao; Ayappa, K. G.; Maiti, Prabal K.


    Designing effective nanoscale blockers for membrane inserted pores formed by pore forming toxins, which are expressed by several virulent bacterial strains, on a target cell membrane is a challenging and active area of research. Here we demonstrate that PAMAM dendrimers can act as effective pH controlled gating devices once the pore has been formed. We have used fully atomistic molecular dynamics (MD) simulations to characterize the cytolysin A (ClyA) protein pores modified with fifth generation (G5) PAMAM dendrimers. Our results show that the PAMAM dendrimer, in either its protonated (P) or non-protonated (NP) states can spontaneously enter the protein lumen. Protonated dendrimers interact strongly with the negatively charged protein pore lumen. As a consequence, P dendrimers assume a more expanded configuration efficiently blocking the pore when compared with the more compact configuration adopted by the neutral NP dendrimers creating a greater void space for the passage of water and ions. To quantify the effective blockage of the protein pore, we have calculated the pore conductance as well as the residence times by applying a weak force on the ions/water. Ionic currents are reduced by 91% for the P dendrimers and 31% for the NP dendrimers. The preferential binding of Cl- counter ions to the P dendrimer creates a zone of high Cl- concentration in the vicinity of the internalized dendrimer and a high concentration of K+ ions in the transmembrane region of the pore lumen. In addition to steric effects, this induced charge segregation for the P dendrimer effectively blocks ionic transport through the pore. Our investigation shows that the bio-compatible PAMAM dendrimers can potentially be used to develop therapeutic protocols based on the pH sensitive gating of pores formed by pore forming toxins to mitigate bacterial infections.Designing effective nanoscale blockers for membrane inserted pores formed by pore forming toxins, which are expressed by several virulent

  9. Pore structure effect on reservoir electrical properties and well logging evaluation

    Institute of Scientific and Technical Information of China (English)

    Bian Huan-Lin; Guan Ju; Mao Zhi-Qiang; Ju Xiao-Dong; Han Gui-Qing


    The reservoir pore structure controls the reservoir quality and resistivity response of hydrocarbon-bearing zones and thus, critically affects logging interpretation. We use petrophysical data in three types of reservoir with different pore structure characteristics to show that the complexity of pore structure had a significant effect on the effective porosity and permeability regardless of geological factors responsible for the formation of pore structure. Moreover,, the distribution and content of conductive fluids in the reservoir varies dramatically owing to pore structure differences, which also induces resistivity variations in reservoir rocks. Hence, the origin of low-resistivity hydrocarbon-bearing zones, except for those with conductive matrix and mud filtrate invasion, is attributed to the complexity of the pore structures. Consequently, reservoir-specific evaluation models, parameters, and criteria should be chosen for resistivity log interpretation to make a reliable evaluation of reservoir quality and fluids.

  10. Defect evolution and pore collapse in crystalline energetic materials (United States)

    Barton, Nathan R.; Winter, Nicholas W.; Reaugh, John E.


    This work examines the use of crystal based continuum mechanics in the context of dynamic loading. In particular, we examine model forms and simulations which are relevant to pore collapse in crystalline energetic materials. Strain localization and the associated generation of heat are important for the initiation of chemical reactions in this context. The crystal mechanics based model serves as a convenient testbed for the interactions among wave motion, slip kinetics, defect generation kinetics and physical length scale. After calibration to available molecular dynamics and single crystal gas gun data for HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine), the model is used to predict behaviors for the collapse of pores under various conditions. Implications for experimental observations are discussed. This document was prepared as an account of work sponsored by an agency of the United States government. Neither the United States government nor Lawrence Livermore National Security, LLC, nor any of their employees makes any warranty, expressed or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States government or Lawrence Livermore National Security, LLC. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States government or Lawrence Livermore National Security, LLC, and shall not be used for advertising or product endorsement purposes.

  11. Putative members of the Arabidopsis Nup107-160 nuclear pore sub-complex contribute to pathogen defense. (United States)

    Wiermer, Marcel; Cheng, Yu Ti; Imkampe, Julia; Li, Meilan; Wang, Dongmei; Lipka, Volker; Li, Xin


    In eukaryotic cells, transduction of external stimuli into the nucleus to induce transcription and export of mRNAs for translation in the cytoplasm is mediated by nuclear pore complexes (NPCs) composed of nucleoporin proteins (Nups). We previously reported that Arabidopsis MOS3, encoding the homolog of vertebrate Nup96, is required for plant immunity and constitutive resistance mediated by the de-regulated Toll interleukin 1 receptor/nucleotide-binding/leucine-rich repeat (TNL)-type R gene snc1. In vertebrates, Nup96 is a component of the conserved Nup107-160 nuclear pore sub-complex, and implicated in immunity-related mRNA export. Here, we used a reverse genetics approach to examine the requirement for additional subunits of the predicted Arabidopsis Nup107-160 complex in plant immunity. We show that, among eight putative complex members, beside MOS3, only plants with defects in Nup160 or Seh1 are impaired in basal resistance. Constitutive resistance in the snc1 mutant and immunity mediated by TNL-type R genes also depend on functional Nup160 and have a partial requirement for Seh1. Conversely, resistance conferred by coiled coil-type immune receptors operates largely independently of both genes, demonstrating specific contributions to plant defense signaling. Our functional analysis further revealed that defects in nup160 and seh1 result in nuclear accumulation of poly(A) mRNA, and, in the case of nup160, considerable depletion of EDS1, a key positive regulator of basal and TNL-triggered resistance. These findings suggest that Nup160 is required for nuclear mRNA export and full expression of EDS1-conditioned resistance pathways in Arabidopsis.

  12. Finite Element and Experimental Analysis of Closure and Contact Bonding of Pores During Hot Rolling of Steel (United States)

    Joo, Soo-Hyun; Jung, Jaimyun; Chun, Myung Sik; Moon, Chang Ho; Lee, Sunghak; Kim, Hyoung Seop


    The closure and contact bonding behavior of internal pores in steel slabs during hot rolling was studied using experiments and the finite element method (FEM). Effects of pore size and shape were investigated, and three different cases of pore closure results were observed: no closure, partial closure, and full closure. The FEM results well reproduced various closure events. Bonding strengths of unsuccessfully closed pores, measured by tensile tests, showed critical effects. Also, there was a difference in bonding strengths of several fully closed pores. Fracture surfaces showed that welded regions could be divided into three (not, partially, and perfectly) welded regions. The pressure-time curves obtained from the FEM results indicate that pore surface contact time and deformed surface length are important parameters in pore welding. Pore size, pore shape, time of pressure contact, and deformed surface length should be considered to completely eliminate pores in final products.

  13. Pore-scale Direct Numerical Simulation of Flow and Transport in Porous Media


    Pulloor Kuttanikkad, Sreejith


    This dissertation presents research on the pore-scale simulation of flow and transport in porous media and describes the application of a new numerical approach based on the discontinuous Galerkin (DG) finite elements to pore-scale modelling. In this approach, the partial differential equations governing the flow at the pore-scale are solved directly where the main advantage is that it does not require a body fitted grid and works on a structured partition of the domain. Furthermore this appr...

  14. A Pore-Network Model of In-Situ Combustion in Porous Media

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Chuan; Yortsos, Y.C.


    This report the use of dual pore networks (pores and solid sites) for modeling the effect of the microstructure on combustion processes in porous media is considered. The model accounts for flow and transport of the gas phase in the porespace, where convection predominates, and for heat transfer by conduction in the solid phase. Gas phase flow in the pore and throats is governed by Darcy's law.

  15. Imaging pore space in tight gas sandstone reservoir: insights from broad ion beam cross-sectioning


    Konstanty J.; Kukla P.A.; Urai J.L.; Baerle C.; Enzmann F.; Desbois G.


    Monetization of tight gas reservoirs, which contain significant gas reserves world-wide, represents a challenge for the entire oil and gas industry. The development of new technologies to enhance tight gas reservoir productivity is strongly dependent on an improved understanding of the rock properties and especially the pore framework. Numerous methods are now available to characterize sandstone cores. However, the pore space characterization at pore scale remains difficult due to the f...

  16. Seasonal variations in pore water and sediment geochemistry of littoral lake sediments (Asylum Lake, MI, USA)


    Miller Douglas; Haas Johnson R; Koretsky Carla M; Ndenga Noah T


    Abstract Background Seasonal changes in pore water and sediment redox geochemistry have been observed in many near-surface sediments. Such changes have the potential to strongly influence trace metal distribution and thus create seasonal fluctuations in metal mobility and bioavailability. Results Seasonal trends in pore water and sediment geochemistry are assessed in the upper 50 cm of littoral kettle lake sediments. Pore waters are always redox stratified, with the least compressed redox str...

  17. Pore structure of ore granular media by computerized tomography image processing

    Institute of Scientific and Technical Information of China (English)

    WU Ai-xiang; YANG Bao-hua; XI Yong; JIANG Huai-chun


    The pore structure images of ore particles located at different heights of leaching column were scanned with X-ray computerized tomography (CT) scanner, the porosity and pore size distribution were calculated and the geometrical shape and connectivity of pores were analyzed based on image process method, and the three dimensional reconstruction of pore structure images was realized. The results show that the porosity of ore particles bed in leaching column is 42.92%, 41.72%, 39.34% at top,middle and bottom zone, respectively. Obviously it has spatial variability and decreases appreciably along the height of the column.The overall average porosity obtained by image processing is 41.33% while the porosity gotten from general measurement method in laboratory is 42.77% showing the results of both methods are consistent well. The pore structure of ore granular media is characterized as a dynamical space network composed of interconnected pore bodies and pore throats. The ratio of throats with equivalent diameter less than 1.91 mm to the total pores is 29.31%, and that of the large pores with equivalent diameter more than 5.73 mm is 2.90%.

  18. Preparation and Selectivity of Molecularly Imprinted Polymer Coating on the Micro Pore Membrane of Polytetrafluoroethylene

    Institute of Scientific and Technical Information of China (English)

    Hong Sheng GUO; Yu Mei JIA; Xi Wen HE


    In order to obtain mechanically stable membrane for practical application, the imprintedpolymer was synthesized in the pores of polyfluoromembrane, the binding and transport ability ofthe membrane were studied.

  19. Optimization of hybrid laser arc welding of 42CrMo steel to suppress pore formation (United States)

    Zhang, Yan; Chen, Genyu; Mao, Shuai; Zhou, Cong; Chen, Fei


    The hybrid laser arc welding (HLAW) of 42CrMo quenched and tempered steel was conducted. The effect of the processing parameters, such as the relative positions of the laser and the arc, the shielding gas flow rate, the defocusing distance, the laser power, the wire feed rate and the welding speed, on the pore formation was analyzed, the morphological characteristics of the pores were analyzed using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The results showed that the majority of the pores were invasive. The pores formed at the leading a laser (LA) welding process were fewer than those at the leading a arc (AL) welding process. Increasing the shielding gas flow rate could also facilitate the reduction of pores. The laser power and the welding speed were two key process parameters to reduce the pores. The flow of the molten pool, the weld cooling rate and the pore escaping rate as a result of different parameters could all affect pore formation. An ideal pore-free weld was obtained for the optimal welding process parameters.

  20. Detection and Influence of Shrinkage Pores and Nonmetallic Inclusions on Fatigue Life of Cast Aluminum Alloys (United States)

    Tijani, Yakub; Heinrietz, André; Stets, Wolfram; Voigt, Patrick


    In the current study, test bars of cast aluminum alloys EN AC-AlSi8Cu3 and EN AC-AlSi7Mg0.3 were produced with a defined amounts of shrinkage pores and oxides. For this purpose, a permanent mold with heating and cooling devices for the generation of pores was constructed. The oxides were produced by contaminating the melt. The specimens and their corresponding defect distributions were examined and quantified by X-ray computer tomography (CT) and quantitative metallography, respectively. A special test algorithm for the simultaneous image analyses of pores and oxides was developed. Fatigue tests were conducted on the defective samples. It was found that the presence of shrinkage pores lowers the fatigue strength, and only few oxide inclusions were found to initiate fatigue cracks when shrinkage pores are present. The results show that the pore volume is not sufficient to characterize the influence of shrinkage pores on fatigue life. A parametric model for the calculation of fatigue life based on the pore parameters obtained from CT scans was implemented. The model accounts for the combined impact of pore location, size, and shape on fatigue life reduction.

  1. Investigation of the pore geometrical structure of nanofibrous membranes using statistical modelling (United States)

    Khanmohammadi Khoshui, Sedigheh; Hosseini Ravandi, Seyed Abdolkarim; Bagherzadeh, Roohollah; Saberi, Zahra; Karimi, Mohammad


    The pore size and its distribution are the two main geometrical properties of nanofibrous membranes in various applications such as filtration and tissue engineering. In the current paper, a modified approach (model) is suggested to predict pore size and its distribution in nanofibrous membranes. In the present work, inter-fibre pores are considered as polygons arising from the fibre contacts. For the first time, these polygons are assumed to be three-, four- and five-gons, and the hydraulic radius of the pores was obtained instead of the equal radius. The pore size of multilayer mats was provided with a different insight. The pore mean size and its distribution were obtained by statistical methods. In order to validate the model, polycaprolactone (PCL) nanofibrous mats were electrospun, and the mean pore size and its distribution were measured using porosimetry. It was found that the probability distribution function of the pore size in both single and multi nanofibrous layers was the Gamma function with two parameters. The effect of the fibre width and porosity raise was increasing of mean pore diameter of multilayer networks. A comparison between the modified model and previous models revealed that the modified approach was more realistic.

  2. Study into the correlation of dominant pore throat size and SIP relaxation frequency (United States)

    Kruschwitz, Sabine; Prinz, Carsten; Zimathies, Annett


    There is currently a debate within the SIP community about the characteristic textural length scale controlling relaxation time of consolidated porous media. One idea is that the relaxation time is dominated by the pore throat size distribution or more specifically the modal pore throat size as determined in mercury intrusion capillary pressure tests. Recently new studies on inverting pore size distributions from SIP data were published implying that the relaxation mechanisms and controlling length scale are well understood. In contrast new analytical model studies based on the Marshall-Madden membrane polarization theory suggested that two relaxation processes might compete: the one along the short narrow pore (the throat) with one across the wider pore in case the narrow pores become relatively long. This paper presents a first systematically focused study into the relationship of pore throat sizes and SIP relaxation times. The generality of predicted trends is investigated across a wide range of materials differing considerably in chemical composition, specific surface and pore space characteristics. Three different groups of relaxation behaviors can be clearly distinguished. The different behaviors are related to clay content and type, carbonate content, size of the grains and the wide pores in the samples.

  3. The study of the relationship between pore structure and photocatalysis of mesoporous TiO2

    Indian Academy of Sciences (India)

    Bing Guo; Hangyan Shen; Kangying Shu; Yaowu Zeng; Wensheng Ning


    Mesoporous titania was synthesized by a sol-gel method using the surfactants Span85 and X114 as the template. The pore structure was determined by the N2 adsorption/desorption method below 73 K and calculated using the BJH model. TEM characterizations show that the pores are formed through particle accumulation. Two kinds of channels, straight channels made of cylindrical capillaries and curved channels made of slit-shaped pores, exist in the bulk materials. The influence of the pore structure of mesoporous TiO2 on its photocatalytic performance was studied. The sample with higher porosity, better textural properties and straight channels are good for photocatalytic performance.

  4. Antamanide, a derivative of Amanita phalloides, is a novel inhibitor of the mitochondrial permeability transition pore.

    Directory of Open Access Journals (Sweden)

    Luca Azzolin

    Full Text Available Antamanide is a cyclic decapeptide derived from the fungus Amanita phalloides. Here we show that antamanide inhibits the mitochondrial permeability transition pore, a central effector of cell death induction, by targeting the pore regulator cyclophilin D. Indeed, (i permeability transition pore inhibition by antamanide is not additive with the cyclophilin D-binding drug cyclosporin A, (ii the inhibitory action of antamanide on the pore requires phosphate, as previously shown for cyclosporin A; (iii antamanide is ineffective in mitochondria or cells derived from cyclophilin D null animals, and (iv abolishes CyP-D peptidyl-prolyl cis-trans isomerase activity. Permeability transition pore inhibition by antamanide needs two critical residues in the peptide ring, Phe6 and Phe9, and is additive with ubiquinone 0, which acts on the pore in a cyclophilin D-independent fashion. Antamanide also abrogates mitochondrial depolarization and the ensuing cell death caused by two well-characterized pore inducers, clotrimazole and a hexokinase II N-terminal peptide. Our findings have implications for the comprehension of cyclophilin D activity on the permeability transition pore and for the development of novel pore-targeting drugs exploitable as cell death inhibitors.

  5. Porous hydrogels with well-defined pore structure for biomaterials applications (United States)

    Marshall, Andrew J.

    When any medical device is implanted inside the body, the natural inflammatory response causes the device to be encapsulated with a thin layer of dense, relatively avascular fibrous tissue, effectively sealing off the device from the surrounding tissue and isolating it from the rest of the body. For medical devices such as electrodes and glucose sensors, where functionality depends on the ability of the device to interact with the surrounding biochemistry, the "foreign body response" poses a formidable obstacle. Previous studies have demonstrated that porous materials with pore dimensions on the order of cell dimensions can induce a modified foreign body response, resulting in more vascularized capsule tissue. However, the utility of these studies is limited because the materials used had broad pore size distributions and poorly defined pore geometries. This thesis is motivated by the unavailability of biomaterials with well-defined and controlled pore size, and by the lack of understanding of the relationships between pore dimensions and the foreign body response. Our sphere templating technology permits the fabrication of open-pore structures with precisely controlled pore dimensions. We can produce these sphere-templated pore structures out of a variety of polymeric materials, including poly(2-hydroxyethyl methacrylate) (polyHEMA), silicone rubber, and degradable copolymers of polyHEMA and poly(epsilon-caprolactone). We applied our precision-engineered pore structures in vivo to investigate the role of pore size in the foreign body response. We implanted porous polyHEMA with various pore geometries under the skin of mice and found that the level of intra-pore vascularization increases with decreasing pore size, with vascular density directly proportional to the specific surface area of the implant, and that the threshold pore throat diameter for rapid tissue in-growth is approximately 8 mum. Based on our empirical results coupled with first principles, we

  6. Hierarchically templated beads with tailored pore structure for phosphopeptide capture and phosphoproteomics

    DEFF Research Database (Denmark)

    Wierzbicka, Celina; Torsetnes, Silje B.; Jensen, Ole N.


    and crosslinkers inside the pores of macroporous silica beads with both free and immobilized template. In the final step the silica was removed by fluoride etching resulting in mesoporous polymer replicas with narrow pore size distributions, pore diameters ≈ 10 nm and surface area > 260 m2 g-1. The beads displayed......Two templating approaches to produce imprinted phosphotyrosine capture beads with a controllable pore structure are reported and compared with respect to their ability to enrich phosphopeptides from a tryptic peptide mixture. The beads were prepared by the polymerization of urea-based host monomers...

  7. An elliptical-pore model for late-stage planar viscous sintering (United States)

    Crowdy, Darren G.


    A simple ‘elliptical-pore model’ of the shrinkage of compressible pores in late-stage planar viscous sintering is proposed. The model is in the spirit of matched asymptotics and relies on splitting the flow into an ‘inner’ and ‘outer’ problem. The inner problem in the vicinity of any given pore involves solving for its free-surface evolution exactly using complex-variable methods. The outer flow due to all other pores is assumed to be given by an assembly of point sinks/sources. As a test of the model, the evolution of a singly infinite periodic row of compressible pores is considered in detail. The effectiveness of the simple model is tested by comparison with a full numerical simulation. A novel boundary integral method based on Cauchy potentials and conformal mapping is used. In the case of pores with constant pressure, it is found that pores shrink faster than if in isolation. Compressible pores obeying the ideal gas law are also studied and are found to tend to a quasi-steady non-circular state. A higher-order model is also presented and compared with numerical simulations of the viscous sintering of a doubly periodic array of pores in Stokes flow.

  8. Effect of support structure on CO2 adsorption properties of pore-expanded hyperbranched aminosilicas

    KAUST Repository

    Drese, Jeffrey H.


    Hyperbranched aminosilica (HAS) CO 2 adsorbents are prepared by the ring-opening polymerization of aziridine from SBA-15 mesoporous silica, as in the original synthesis of HAS materials, as well as over an array of new support materials with substantially larger average pore diameters to elucidate the effect of support porosity on final adsorbent properties. Pore-expanded hyperbranched aminosilica (PEHAS) CO 2 adsorbents are prepared from several different pore-expanded, ordered mesoporous silicas including pore-expanded SBA-15, mesocellular foam, and a large-pore commercial silica. The effect of the nature of the silica support is determined by examining the degree of aziridine polymerization and the CO 2 adsorption kinetics and capacities of the resulting organic/inorganic hybrid materials. Comparisons are made to non-pore-expanded SBA-15 based HAS adsorbents, reported previously, where pores become blocked at higher amine loadings. The PEHAS materials unexpectedly possess lower amine loadings than the previously reported HAS materials and do not exhibit pore blocking. The use of acetic acid as a catalyst during PEHAS synthesis only marginally increases amine loading. The adsorption kinetics of PEHAS adsorbents are similar to HAS adsorbents with low amine loadings and do not show the detrimental effects of pore-blocking. However, the inability to synthesize PEHAS adsorbents with high amine loadings via this approach limits the total amount of CO 2 captured per gram of material, compared to HAS adsorbents with high amine loadings. © 2011 Elsevier Inc. All rights reserved.

  9. Shrinkage of large isolated pores during hot isostatic pressing of presintered alumina ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Oh, K.S.; Kim, D.Y. [Seoul National Univ. (Korea, Republic of). Dept. of Inorganic Materials Engineering; Cho, S.J. [Korea Research Inst. of Standards and Science, Taejon (Korea, Republic of)


    The shrinkage process of large pores during hot isostatic pressing (HIP) of presintered Al{sub 2}O{sub 3} and Al{sub 2}O{sub 3}-ZrO{sub 2} ceramics has been investigated. Large pores were observed to collapse by grain-boundary sliding, and then small pores resulting from the misfit of flowed grains disappeared, mainly by diffusion. Due to the high resistance to grain-boundary sliding, the pores in the Al{sub 2}O{sub 3}-ZrO{sub 2} ceramics were hard to eliminate.

  10. Ultrastructural aspects in perithecia hyphae septal pores of Glomerella cingulata F. SP. Phaseoli

    Directory of Open Access Journals (Sweden)

    Roca M. María Gabriela


    Full Text Available Glomerella cingulata (Stonem. Spauld. & Schrenk f. sp. phaseoli, better known in its anamorphic state Colletotrichum lindemuthianum (Sacc. & Magn. Briosi & Cav., is a causal agent of anthracnose in beans (Phaseolus vulgaris L.. Ultrastructural aspects of the perithecial hyphae of this pathogen were studied. The perithecia hyphae septal pores were found either plugged by a vesicle or unplugged. Some perithecia hyphae septa presented no pore. The Woronin bodies, close to the septal pores, appeared as globose structures which were more electron dense than the occlusions plugging the septal pore.

  11. Analysis of the effect of pore geometry in the physical properties of rocks

    Directory of Open Access Journals (Sweden)

    Luiz Alberto Oliveira Lima Roque


    Full Text Available Pore geometry is one of the main factors influencing the flow of reservoir fluids under pressure. Pores with narrower formats are more easily compressed when subject to pressure. Pressure modifies pore geometry by opening or closing cracks, causing increase or decrease in the elastic modulus, porosity, permeability, and other parameters. Rock physical properties depend on the size and shape of pores. Thus, in order to analyze changes on the physical properties behavior according to the pores geometry, it is necessary to study and improve mathematical models of the porous media by taking into account the pore shape factor for estimating rock elastic properties. Differential effective medium model (DEM, Hertz-Mindlin theory and coherent potential approximation (CPA are some of the theoretical paradigms that take into account pore geometry in changes in elastic moduli. Given the importance of the pore structure effect on the behavior of physical parameters, this article proposes an analysis of some mathematical models that consider the influence of pore shapes in the physical properties of rocks.

  12. Effect of Pore Size on the Biodegradation Rate of Silk Fibroin Scaffolds

    Directory of Open Access Journals (Sweden)

    Zuwei Luo


    Full Text Available Controlling the degradation rate of silk fibroin-based biomaterial is an important capability for the fabrication of silk-based tissue engineering scaffolds. In this study, scaffolds with different pore sizes were prepared by controlling the freezing temperature and the silk fibroin concentration. In vitro degradation results showed that the internal pore walls of the scaffolds with a larger pore size collapsed upon exposure to collagenase IA for times ranging from 6 to 12 days, and the silk scaffolds exhibited a faster rate of weight loss. The morphological and structural features of the silk scaffolds with a smaller pore size maintained structural integrity after incubation in the protease solution for 18 days, and the rate of weight loss was relatively slow. Scaffolds with a smaller pore size or a higher pore density degraded more slowly than scaffolds with a larger pore size or lower pore density. These results demonstrate that the pore size of silk biomaterials is crucial in controlling the degradation rate of tissue engineering scaffolds.

  13. Abnormal Nuclear Pore Formation Triggers Apoptosis in the Intestinal Epithelium of elys-Deficient Zebrafish

    NARCIS (Netherlands)

    de Jong-Curtain, Tanya A.; Parslow, Adam C.; Trotter, Andrew J.; Hall, Nathan E.; Verkade, Heather; Tabone, Tania; Christie, Elizabeth L.; Crowhurst, Meredith O.; Layton, Judith E.; Shepherd, Iain T.; Nixon, Susan J.; Parton, Robert G.; Zon, Leonard I.; Stainier, Didier Y. R.; Lieschke, Graham J.; Heath, Joan K.

    Background & Aims: Zebrafish mutants generated by ethylnitrosourea-mutagenesis provide a powerful toot for dissecting the genetic regulation of developmental processes, including organogenesis. One zebrafish mutant, "flotte lotte" (flo), displays striking defects in intestinal, liver, pancreas, and

  14. Abnormal Nuclear Pore Formation Triggers Apoptosis in the Intestinal Epithelium of elys-Deficient Zebrafish

    NARCIS (Netherlands)

    de Jong-Curtain, Tanya A.; Parslow, Adam C.; Trotter, Andrew J.; Hall, Nathan E.; Verkade, Heather; Tabone, Tania; Christie, Elizabeth L.; Crowhurst, Meredith O.; Layton, Judith E.; Shepherd, Iain T.; Nixon, Susan J.; Parton, Robert G.; Zon, Leonard I.; Stainier, Didier Y. R.; Lieschke, Graham J.; Heath, Joan K.


    Background & Aims: Zebrafish mutants generated by ethylnitrosourea-mutagenesis provide a powerful toot for dissecting the genetic regulation of developmental processes, including organogenesis. One zebrafish mutant, "flotte lotte" (flo), displays striking defects in intestinal, liver, pancreas, and

  15. Bacterial Chemotaxis Toward A NAPL Source Within A Pore-Scale Model Subject to A Range of Groundwater Flow Velocities (United States)

    Wang, X.; Ford, R. M.


    Organic solvents such as toluene are the most widely distributed pollutants in groundwater. Biodegradation of these industrial pollutants requires that microorganisms in the aqueous phase are brought in contact with sources of contamination, which may be dispersed as pore-size organic-phase droplets within the saturated soil matrix. Chemotaxis toward chemical pollutants provides a mechanism for bacteria to migrate to locations of high contamination, which may not normally be accessible to bacteria carried along by groundwater flow, and thus it may improve the efficiency of bioremediation. A microfluidic device was designed to mimic the dissolution of an organic-phase contaminant from a single pore into a larger macropore representing a preferred pathway for microorganisms that are carried along by groundwater flow. The glass windows of the µ-chip allowed image analysis of bacterial distributions within the vicinity of the organic contaminant. Concentrations of chemotactic bacteria P. putida F1 near the organic/aqueous interface were 25% greater than those of a nonchemotactic mutant in the vicinity of toluene for a fluid velocity of 0.5 m/d. For E. coli responding to phenol, the bacterial concentrations were 60% greater than the controls, also at a velocity of 0.5 m/d. Velocities in the macropore were varied over a range that is typical of groundwater velocities from 0.5 to 10 m/d. The accumulation of chemotactic bacteria near the NAPL (nonaqueous phase liquid) chemoattractant source decreased as the fluid velocity increased. At the higher velocities, accumulation of chemotactic bacteria was comparable to the non-chemotactic control experiments. Computer-based simulation using finite element analysis software (COMSOL) was also performed to understand the effects of various model parameters on bacterial chemotaxis to NAPL. There was good agreement between the simulations (generated using reasonable values of the model parameters) and the experimental data for P

  16. Chemotyping of yeast mutants using robotics. (United States)

    Rieger, K J; El-Alama, M; Stein, G; Bradshaw, C; Slonimski, P P; Maundrell, K


    By now, the EUROFAN programme for the functional analysis of genes from the yeast genome has attained its cruising speed. Indeed, several hundreds of yeast mutants with no phenotype as tested by growth on standard media and no significant sequence similarity to proteins of known function are available through the efforts of various laboratories. Based on the methodology initiated during the pilot project on yeast chromosome III (Yeast 13, 1547-1562, 1997) we adapted it to High Throughput Screening (HTS), using robotics. The first 100 different gene deletions from EUROSCARF, constructed in an FY1679 strain background, were run against a collection of about 300 inhibitors. Many of these inhibitors have not been reported until now to interfere in vivo with growth of Saccharomyces cerevisiae. In the present paper we provide a list of novel growth conditions and a compilation of 49 yeast deletants (from chromosomes II, IV, VII, X, XIV, XV) corresponding to 58% of the analysed genes, with at least one clear and stringent phenotype. The majority of these deletants are sensitive to one or two compounds (monotropic phenotype) while a distinct subclass of deletants displays a hyper-pleiotropic phenotype with sensitivities to a dozen or more compounds. Therefore, chemotyping of unknown genes with a large spectrum of drugs opens new vistas for a more in-depth functional analysis and a more precise definition of molecular targets.

  17. Pharmacological correctors of mutant CFTR mistrafficking

    Directory of Open Access Journals (Sweden)

    Nicoletta ePedemonte


    Full Text Available The lack of phenylalanine 508 (∆F508 mutation in the CFTR Cl- channel represents the most frequent cause of cystic fibrosis (CF, a genetic disease affecting multiple organs such lung, pancreas, and liver. ∆F508 causes instability and misfolding of CFTR protein leading to early degradation in the endoplasmic reticulum and accelerated removal from the plasma membrane. Pharmacological correctors of mutant CFTR protein have been identified by high-throughput screening of large chemical libraries, by in silico docking of virtual compounds on CFTR structure models, or by using compounds that affect the whole proteome (e.g. histone deacetylase inhibitors or a single CFTR-interacting protein. The presence of multiple defects caused at the CFTR protein level by ∆F508 mutation and the redundancy of quality control mechanisms detecting ∆F508-CFTR as a defective protein impose a ceiling to the maximal effect that a single compound (corrector may obtain. Therefore, treatment of patients with the most frequent CF mutation may require the optimized combination of two drugs having additive or synergic effects.

  18. New types of Escherichia coli recombination-deficient mutants. (United States)

    Freifelder, D


    A set of Escherichia coli mutants deficient in intramolecular recombination and different from those previously found is described. All have temperature-sensitive lethal mutations. The mutants have been characterized with respect to the following properties: the Pap phenotype, deoxyribonucleic acid synthesis, sensitivity to ultraviolet light, ability to support the growth of phage lambda, filament formation, and mutation frequency.

  19. Characterization of Gibberellin Receptor Mutants of Barley (Hordeum vulgare L.)

    Institute of Scientific and Technical Information of China (English)

    Peter M.Chandler; Carol A.Harding; Anthony R.Ashton; Mark D.Mulcair; Nicholas E.Dixon; Lewis N.Mander


    The sequence of Gidl (a gene for a gibberellin (GA) receptor from rice) was used to identify a putative orthoIogue from barley.This was expressed in E.coil,and produced a protein that was able to bind GA in vitro with both structural specificity and saturability.Its potential role in GA responses was investigated using barley mutants with reduced GA sensitivity (gsel mutants).Sixteen different gsel mutants each carried a unique nucleotide substitution in this sequence.In all but one case,these changes resulted in single amino acid substitutions,and,for the remaining mutant,a substitution in the 5' untranslated region of the mRNA is proposed to interfere with translation initiation.There was perfect linkage in segregating populations between new mutant alleles and the gsel phenotype,leading to the conclusion that the putative GID1 GA receptor sequence in barley corresponds to the Gsel locus.Determination of endogenous GA contents in one of the mutants revealed enhanced accumulation of bioactive GA1,and a deficit of C20 GA precursors.All of the gsel mutants had reduced sensitivity to exogenous GA3,and to AC94377 (a GA analogue) at concentrations that are normally 'saturating',but,at much higher concentrations,there was often a considerable response.The comparison between barley and rice mutants reveals interesting differences between these two cereal species in GA hormonal physiology.

  20. Mutants of Pseudomonas putida affected in poly-3-hydroxyalkanoate synthesis

    NARCIS (Netherlands)

    Ren, Q; Kessler, B; van der Leij, F; Witholt, B.


    The generation and characterization of Pseudomonas putida KT2442 mutants affected in poly-3-hydroxyalkanoate (PHA) synthesis are reported. The mutants from P. putida KT2442 carrying several copies of the PHA-polymerase-encoding gene (phaC) were isolated via N-methyl-N'-nitro-N-nitrosoguanidine chemi