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Sample records for gramicidin channel cs

  1. Gramicidin Channels: Versatile Tools

    NASA Astrophysics Data System (ADS)

    Andersen, Olaf S.; Koeppe, Roger E., II; Roux, Benoît

    Gramicidin channels are miniproteins in which two tryptophan-rich subunits associate by means of transbilayer dimerization to form the conducting channels. That is, in contrast to other ion channels, gramicidin channels do not open and close; they appear and disappear. Each subunit in the bilayer-spanning channel is tied to the bilayer/solution interface through hydrogen bonds that involve the indole NH groups as donors andwater or the phospholipid backbone as acceptors. The channel's permeability characteristics are well-defined: gramicidin channels are selective for monovalent cations, with no measurable permeability to anions or polyvalent cations; ions and water move through a pore whose wall is formed by the peptide backbone; and the single-channel conductance and cation selectivity vary when the amino acid sequence is varied, even though the permeating ions make no contact with the amino acid side chains. Given the plethora of available experimental information—for not only the wild-type channels but also for channels formed by amino acid-substituted gramicidin analogues—gramicidin channels continue to provide important insights into the microphysics of ion permeation through bilayer-spanning channels. For similar reasons, gramicidin channels constitute a system of choice for evaluating computational strategies for obtaining mechanistic insights into ion permeation through the more complex channels formed by integral membrane proteins.

  2. Gramicidin channels are internally gated.

    PubMed

    Jones, Tyson L; Fu, Riqiang; Nielson, Frederick; Cross, Timothy A; Busath, David D

    2010-04-21

    Gramicidin channels are archetypal molecular subjects for solid-state NMR studies and investigations of single-channel or cation conductance. Until now, the transitions between on and off conductance states have been thought, based on multichannel studies, to represent monomer <--> dimer reactions. Here we use a single-molecule deposition method (vesicle fusion to a planar bilayer) to show that gramicidin dimer channels do not normally dissociate when conductance terminates. Furthermore, the observation of two 13C peaks in solid-state NMR indicates very stable dichotomous conformations for both the first and second peptide bonds in the monomers, and a two-dimensional chemical exchange spectrum with a 12-s mixing time demonstrates that the Val1 carbonyl conformations exchange slowly, with lifetimes of several seconds. It is proposed that gramicidin channels are gated by small conformational changes in the channel near the permeation pathway. These studies demonstrate how regulation of conformations governing closed <--> open transitions may be achieved and studied at the molecular level.

  3. Temperature dependence of gramicidin channel conductance

    NASA Astrophysics Data System (ADS)

    Song, Hyundeok; Beck, Thomas

    2010-03-01

    The gramicidin channel is the smallest known biological ion channel, and it exhibits cation selectivity. Recently, Dr. John Cuppoletti's group at the University of Cincinnati has shown that the gramicidin channel can function at high temperatures with significant currents. This finding may have implications for fuel cell technologies. In order to explore the effect of temperature on channel conductance, we examined the gramicidin system at 300K, 330K, and 360K by computer simulation. Two forms of gramicidin, the head-to-head helical dimer and the intertwined double helix, were examined. Both the decrease of the free energy barrier and the increase of the diffusion of potassium ions inside the gramicidin channel at high temperatures imply an increase of current. We found that higher temperatures also affect the lifetime of hydrogen bonds, the distribution of the bending angle, the distribution of the distance between dimers, and the size of the pore radius for the helical dimer structure. These finding may be related to the gating of the gramicidin channel.

  4. Gramicidin channels that have no tryptophan residues.

    PubMed

    Fonseca, V; Daumas, P; Ranjalahy-Rasoloarijao, L; Heitz, F; Lazaro, R; Trudelle, Y; Andersen, O S

    1992-06-16

    In order to understand how aromatic residues modulate the function of membrane-spanning proteins, we examined the role of the four tryptophans in gramicidin A (gA) in determining the average duration and permeability characteristics of membrane-spanning gramicidin channels; the tryptophan residues were replaced by tyrosine (gramicidin T, gT), tyrosine O-benzyl ether [gramicidin T(Bzl), gT(Bzl)], naphthylalanine (gramicidin N, gN), and phenylalanine (gramicidin M enantiomer, gM-). These analogues form channels with durations and conductances that differ some 10- and 16-fold, respectively. The single-channel conductance was invariably decreased by the Trp----Yyy replacement, and the relative conductance alterations were similar in phosphatidylcholine (DPhPC) and monoglyceride (GMO) bilayers. The duration variations exhibited a more complex pattern, which was quite different in the two membrane environments: in DPhPC bilayers, gN channels have an average duration that is approximately 2-fold longer than that of gA channels; in GMO bilayers, the average duration of gN channels is about one-tenth that of gA channels. The sequence-dependent alterations in channel function do not result from alterations in the channels' peptide backbone structure, because heterodimers can form between the different analogues and gramicidine A, and there is no energetic cost associated with heterodimer formation [cf. Durkin, J. T., Koeppe, R. E., II, & Andersen, O. S. (1990) J. Mol. Biol. 211, 221]. The alterations in permeability properties are consistent with the notion that Trp residues alter the energy profile for ion permeation through long-range electrostatic interactions.

  5. Computer Simulation Studies of Gramicidin Channel

    NASA Astrophysics Data System (ADS)

    Song, Hyundeok; Beck, Thomas

    2009-04-01

    Ion channels are large membrane proteins, and their function is to facilitate the passage of ions across biological membranes. Recently, Dr. John Cuppoletti's group at UC showed that the gramicidin channel could function at high temperatures (360 -- 390K) with significant currents. This finding may have large implications for fuel cell technology. In this project, we will examine the experimental system by computer simulation. We will investigate how the temperature affects the current and differences in magnitude of the currents between two forms of Gramicidin, A and D. This research will help to elucidate the underlying molecular mechanism in this promising new technology.

  6. Gramicidins A, B, and C form structurally equivalent ion channels.

    PubMed Central

    Sawyer, D B; Williams, L P; Whaley, W L; Koeppe, R E; Andersen, O S

    1990-01-01

    The membrane structure of the naturally occurring gramicidins A, B, and C was investigated using circular dichroism (CD) spectroscopy and single-channel recording techniques. All three gramicidins form channels with fairly similar properties (Bamberg, E., K. Noda, E. Gross, and P. Läuger. 1976. Biochim. Biophys. Acta. 419:223-228.). When incorporated into lysophosphatidylcholine micelles, however, the CD spectrum of gramicidin B is different from that of gramicidin A or C (cf. Prasad, K. U., T. L. Trapane, D. Busath, G. Szabo, and D. W. Urry. 1983. Int. J. Pept. Protein Res. 22:341-347.). The structural identity of the channels formed by gramicidin B has, therefore, been uncertain. We find that when gramicidins A and B are incorporated into dipalmitoylphosphatidylcholine vesicles, their CD spectra are fairly similar, suggesting that the two channel structures could be similar. In planar bilayers, gramicidins A, B, and C all form hybrid channels with each other. The properties of the hybrid channels are intermediate to those of the symmetric channels, and the appearance rates of the hybrid channels (relative to the symmetric channels) corresponds to what would be predicted if all three gramicidin molecules were to form structurally equivalent channels. These results allow us to interpret the different behavior of channels formed by the three gramicidins solely on the basis of the amino acid substitution at position 11. PMID:1705449

  7. Uniformly oriented gramicidin channels embedded in thick monodomain lecithin multilayers.

    PubMed Central

    Huang, H W; Olah, G A

    1987-01-01

    Phosphatidylcholine multilayers, containing 20% water by total sample weight and gramicidin/lipid molar ratios up to 1:40 were aligned by low temperature annealing (less than 60 degrees C) and mechanical stressing. We were able to obtain large (greater than 80 micron thick X 40 mm2 area) monodomain defect-free multilayers containing approximately 10(17) uniformly oriented gramicidin channels. The alignment of lipid multilayers was monitored by conoscopy and polarized microscopy. The smectic defects, which appeared during the alignment process, were identified and dissolved. The incorporation of gramicidin into the multilayers in the form of transmembrane channels was indicated by its circular dichroic (CD) spectrum. A well-defined CD spectrum of uniformly oriented gramicidin channels was obtained. The oriented samples will allow spectroscopic studies of the ion channel in its conducting state and diffraction studies of the channel-channel organization in the membrane. Images FIGURE 1 PMID:2440487

  8. Lorentzian noise in single gramicidin A channel formamidinium currents.

    PubMed

    Fairbanks, T G; Andrus, C L; Busath, D D

    1999-01-01

    Seoh & Busath (1995) showed that in the presence of formamidinium, single gramicidin A channels were lengthened, had uniformly noisy currents at low voltages and had superlinear current-voltage relationships, all three properties being absent in gramicidin M- channels in which the interfacial tryptophan residues in gramicidin A are all replaced by phenylalanine. We measured the single channel noise power spectra (PSDs) in small monoolein (GMO) bilayers with formamidinium chloride solutions to help identify the mechanism of noise process. PSDs were Lorentzian with characteristic frequencies of 0.1-1.0 kHz in 0.1 and 0.3 M formamidinium chloride solutions, and from. 1-6 kHz in 1 M solution. Si(0), where measurable, ranged from approximately 50-200 fA2/Hz. The time course of the noise process could not be detected in these experiments. The low fc suggests slow motions or rare states of the blocking 'gates' which, judging from the result with gramicidin M-, must be equal to or related to the Trp residues.

  9. Effects of volatile anesthetic on channel structure of gramicidin A.

    PubMed Central

    Tang, Pei; Mandal, Pravat K; Zegarra, Martha

    2002-01-01

    Volatile anesthetic agent, 1-chloro-1,2,2-trifluorocyclobutane (F3), was found to alter gramicidin A channel function by enhancing Na(+) transport (. Biophys. J. 77:739-746). Whether this functional change is associated with structural alternation is evaluated by circular dichroism and nuclear magnetic resonance spectroscopy. The circular dichroism and nuclear magnetic resonance results indicate that at low millimolar concentrations, 1-chloro-1,2,2-trifluorocyclobutane causes minimal changes in gramicidin A channel structure in sodium dodecyl sulfate micelles. All hydrogen bonds between channel backbones are well maintained in the presence of 1-chloro-1,2,2-trifluorocyclobutane, and the channel structure is stable. The finding supports the notion that low affinity drugs such as volatile anesthetics and alcohols can cause significant changes in protein function without necessarily producing associated changes in protein structure. To understand the molecular mechanism of general anesthesia, it is important to recognize that in addition to structural changes, other protein properties, including dynamic characteristics of channel motions, may also be of functional significance. PMID:12202367

  10. Membrane organization and dynamics of "inner pair" and "outer pair" tryptophan residues in gramicidin channels.

    PubMed

    Haldar, Sourav; Chaudhuri, Arunima; Gu, Hong; Koeppe, Roger E; Kombrabail, Mamata; Krishnamoorthy, G; Chattopadhyay, Amitabha

    2012-09-13

    The linear ion channel peptide gramicidin serves as an excellent prototype for monitoring the organization, dynamics, and function of membrane-spanning channels. The tryptophan residues in gramicidin channels are crucial for establishing and maintaining the structure and function of the channel in the membrane bilayer. In order to address the basis of differential importance of tryptophan residues in the gramicidin channel, we monitored the effects of pairwise substitution of two of the four gramicidin tryptophans, the inner pair (Trp-9 and -11) and the outer pair (Trp-13 and -15), using a combination of steady state and time-resolved fluorescence approaches and circular dichroism spectroscopy. We show here that these double tryptophan gramicidin analogues adopt different conformations in membranes, suggesting that the conformational preference of double tryptophan gramicidin analogues is dictated by the positions of the tryptophans in the sequence. These results assume significance in the context of recent observations that the inner pair of tryptophans (Trp-9 and -11) is more important for gramicidin channel formation and channel conductance. These results could be potentially useful in analyzing the effect of tryptophan substitution on the functioning of ion channels and membrane proteins.

  11. Modulation of gramicidin channel conformation and organization by hydrophobic mismatch in saturated phosphatidylcholine bilayers.

    PubMed

    Kelkar, Devaki A; Chattopadhyay, Amitabha

    2007-05-01

    The matching of hydrophobic lengths of integral membrane proteins and the surrounding lipid bilayer is an important factor that influences both structure and function of integral membrane proteins. The ion channel gramicidin is known to be uniquely sensitive to membrane properties such as bilayer thickness and membrane mechanical properties. The functionally important carboxy terminal tryptophan residues of gramicidin display conformation-dependent fluorescence which can be used to monitor gramicidin conformations in membranes [S.S. Rawat, D.A. Kelkar, A. Chattopadhyay, Monitoring gramicidin conformations in membranes: a fluorescence approach, Biophys. J. 87 (2004) 831-843]. We have examined the effect of hydrophobic mismatch on the conformation and organization of gramicidin in saturated phosphatidylcholine bilayers of varying thickness utilizing the intrinsic conformation-dependent tryptophan fluorescence. Our results utilizing steady state and time-resolved fluorescence spectroscopic approaches, in combination with circular dichroism spectroscopy, show that gramicidin remains predominantly in the channel conformation and gramicidin tryptophans are at the membrane interfacial region over a range of mismatch conditions. Interestingly, gramicidin conformation shifts toward non-channel conformations in extremely thick gel phase membranes although it is not excluded from the membrane. In addition, experiments utilizing self quenching of tryptophan fluorescence indicate peptide aggregation in thicker gel phase membranes.

  12. An artificial primitive mimic of the Gramicidin-A channel.

    PubMed

    Barboiu, Mihail; Le Duc, Yann; Gilles, Arnaud; Cazade, Pierre-André; Michau, Mathieu; Marie Legrand, Yves; van der Lee, Arie; Coasne, Benoît; Parvizi, Paria; Post, Joshua; Fyles, Thomas

    2014-06-26

    Gramicidin A (gA) is the simplest known natural channel, and important progress in improving conduction activity has previously been obtained with modified natural gAs. However, simple artificial systems mimicking the gA functions are unknown. Here we show that gA can be mimicked using a simple synthetic triazole or 'T-channel' forming compound (TCT), having similar constitutional functions as the natural gAs. As in gA channels, the carbonyl moieties of the TCT, which point toward the T-channel core and surround the transport direction, are solvated by water. The net-dipolar alignment of water molecules along the chiral pore surfaces influences the conduction of protons/ions, envisioned to diffuse along dipolar hydrophilic pathways. Theoretical simulations and experimental assays reveal that the conduction through the T-channel, similar to that in gA, presents proton/water conduction, cation/anion selectivity and large open channel-conductance states. T-channels--associating supramolecular chirality with dipolar water alignment--represent an artificial primitive mimic of gA.

  13. Importance of indole N-H hydrogen bonding in the organization and dynamics of gramicidin channels.

    PubMed

    Chaudhuri, Arunima; Haldar, Sourav; Sun, Haiyan; Koeppe, Roger E; Chattopadhyay, Amitabha

    2014-01-01

    The linear ion channel peptide gramicidin represents an excellent model for exploring the principles underlying membrane protein structure and function, especially with respect to tryptophan residues. The tryptophan residues in gramicidin channels are crucial for the structure and function of the channel. In order to test the importance of indole hydrogen bonding for the biophysical properties of gramicidin channels, we monitored the effect of N-methylation of gramicidin tryptophans, using a combination of steady state and time-resolved fluorescence approaches along with circular dichroism spectroscopy. We show here that in the absence of the hydrogen bonding ability of tryptophans, tetramethyltryptophan gramicidin (TM-gramicidin) is unable to maintain the single stranded, head-to-head dimeric channel conformation in membranes. Our results show that TM-gramicidin displays a red-shifted fluorescence emission maximum, lower red edge excitation shift (REES), and higher fluorescence intensity and lifetime, consistent with its nonchannel conformation. This is in agreement with the measured location (average depth) of the 1-methyltryptophans in TM-gramicidin using the parallax method. These results bring out the usefulness of 1-methyltryptophan as a fluorescent tool to examine the hydrogen bonding ability of tryptophans in proteins and peptides. We conclude that changes in the hydrogen bonding ability of tryptophans, along with coupled changes in peptide backbone structure induce the loss of single stranded β(6.3) helical dimer conformation. These results agree with earlier results from size-exclusion chromatography and single-channel measurements for TM-gramicidin, and confirm the importance of indole hydrogen bonding for the conformation and function of ion channels and membrane proteins.

  14. Probing conformational changes of gramicidin ion channels by single-molecule patch-clamp fluorescence microscopy

    SciTech Connect

    Harms, Gregory S.; Orr, Galya; Montal, Mauricio; Thrall, Brian D.; Colson, Steve D.; Lu, H Peter

    2003-09-01

    Stochastic and inhomogeneous conformational changes often regulate the dynamics of ion channels. Such inhomogeneity makes it difficult, if not impossible; to be characterized not only by ensemble-averaged experiments by also by single-channel patch recording that does not specifically probe the associated conformational changes. Here, we report on our work using a new approach combining single-molecule fluorescence spectroscopy and single-channel patch recording to investigate conformational changes of individual gramicidin ion channels. We observed fluorescence self-quenching and single-pair fluorescence resonance energy transfer (spFRET) from dye-labeled gramicidin dimmers within the channel was open. We also observed that the efficiency of self-quenching and spFRETS is widely distributed when the channel is closed. Our results strongly suggest a hitherto undetectable correlation of multiple conformational states of the gramicidin channel associated with closed and open states under physiologically-related conditions.

  15. Ion movement through gramicidin A channels. Studies on the diffusion-controlled association step.

    PubMed Central

    Andersen, O S

    1983-01-01

    The permeability characteristics of gramicidin A channels are generally considered to reflect accurately the intrinsic properties of the channels themselves; i.e., the aqueous convergence regions are assumed to be negligible barriers for ion movement through the channels. The validity of this assumption has been examined by an analysis of gramicidin A single-channel current-voltage characteristics up to very high potentials (500 mV). At low permeant ion concentrations the currents approach a voltage-independent limiting value, whose magnitude is proportional to the permeant ion concentration. The magnitude of this current is decreased by experimental maneuvers that decrease the aqueous diffusion coefficient of the ions. It is concluded that the magnitude of this limiting current is determined by the diffusive ion movement through the aqueous convergence regions up to the channel entrance. It is further shown that the small-signal (ohmic) permeability properties also reflect the existence of the aqueous diffusion limitation. These results have considerable consequences for the construction of kinetic models for ion movement through gramicidin A channels. It is shown that the simple two-site-three-barrier model commonly used to interpret gramicidin A permeability data may lead to erroneous conclusions, as biionic potentials will be concentration dependent even when the channel is occupied by at most one ion. The aqueous diffusion limitation must be considered explicitly in the analysis of gramicidin A permeability characteristics. Some implications for understanding the properties of ion-conducting channels in biological membranes will be considered. PMID:6188502

  16. Effect of pyrrolidinium based ionic liquid on the channel form of gramicidin in lipid vesicles.

    PubMed

    Singh, Upendra Kumar; Dohare, Neeraj; Mishra, Prabhash; Singh, Prashant; Bohidar, Himadri B; Patel, Rajan

    2015-08-01

    The present work is focused on the interaction between membrane bound gramicidin and 1-butyl-1-methyl-2-oxopyrrolidinium bromide (BMOP) ionic liquid. Ionic liquids (ILs) are solvents that are often liquid at room temperature and composed of organic cation and appropriate anion. The gramicidin peptide forms prototypical ion channels for cations, which have been extensively used to study the organization, dynamics, and function of membrane spanning channels. The interaction was studied by circular dichroism, steady state, time-resolved fluorescence spectroscopy in combination with dynamic surface tension and field emission scanning electron microscopic methods (FESEM). The results obtained from circular dichroism shows that the BMOP interacts with the channel form of gramicidin in lipid vesicle without any considerable effect on its conformation. The Red-edge excitation shift (REES) also supported the above findings. In addition, the fluorescence studies suggested that BMOP makes ground state complex with ion channel, which was further supported by time resolved measurements. Furthermore, dynamic surface tension analysis shows the faster adsorption of BMOP with membrane bound gramicidin at the air-water interface. Additionally, FESEM results indicated that BMOP forms a film around the membrane bound gramicidin at higher concentration. These results are potentially useful to analyze the effect of ionic liquids on the behaviour of membrane proteins.

  17. Channel and nonchannel forms of spin-labeled gramicidin in membranes and their equilibria.

    PubMed

    Dzikovski, Boris G; Borbat, Peter P; Freed, Jack H

    2011-01-13

    Channel and nonchannel forms of gramicidin A (GA) were studied by ESR in various lipid environments using new mono- and double-spin-labeled compounds. For GA channels, we demonstrate here how pulse dipolar ESR can be used to determine the orientation of the membrane-traversing molecule relative to the membrane normal and to study subtle effects of lipid environment on the interspin distance in the spin-labeled gramicidin channel. To study nonchannel forms of gramicidin, pulse dipolar ESR was used first to determine interspin distances corresponding to monomers and double-helical dimers of spin-labeled GA molecules in the organic solvents trifluoroethanol and octanol. The same distances were then observed in membranes. Since detection of nonchannel forms in the membrane is complicated by aggregation, we suppressed any dipolar spectra from intermolecular interspin distances arising from the aggregates by using double-labeled GA in a mixture with excess unlabeled GA. In hydrophobic mismatching lipids (L(β) phase of DPPC), gramicidin channels dissociate into free monomers. The backbone structure of the monomeric form is similar to a monomeric unit of the channel dimer. In addition to channels and monomers, the double-helical conformation of gramicidin is present in some membrane environments. In the gel phase of saturated phosphatidylcholines, the fraction of double helices increases in the following order: DLPC < DMPC < DSPC < DPPC. The equilibrium DHD/monomer ratio in DPPC was determined. In membranes, the double-helical form is present only in aggregates. In addition, we studied the effect of N-terminal substitution in the GA molecule upon channel formation. This work demonstrates how pulsed dipolar ESR may be utilized to study complex equilibria of peptides in membranes.

  18. Polar groups in membrane channels: consequences of replacing alanines with serines in membrane-spanning gramicidin channels.

    PubMed

    Daily, Anna E; Kim, Jung H; Greathouse, Denise V; Andersen, Olaf S; Koeppe, Roger E

    2010-08-17

    To explore the consequences of burying polar, hydrogen-bonding hydroxyl groups within the hydrocarbon core of lipid bilayer membranes, we examined the structural and functional effects of alanine-to-serine substitutions in bilayer-spanning gramicidin channels. A native Ala was replaced by Ser at position 3 or 5 in the gramicidin A (gA) sequence: formyl-VG(2)A(3)LA(5)VVVWLWLWLW-ethanolamide (d-residues underlined). In the head-to-head dimers that form the conducting, membrane-spanning gA channels, these sequence positions are located near the lipid bilayer center (and subunit interface). The sequence substitutions at positions 3 and 5 were tested within the context of having either Gly or d-Ala at position 2, because d-Ala(2) causes the channel lifetimes to increase 3-fold relative to Gly(2) [Mattice et al. (1995) Biochemistry 34, 6827]. Size-exclusion chromatograms and circular dichroism spectra show that the Ala --> Ser replacements are well tolerated and have little effect on channel structure. In planar bilayers, the Ser-substituted gramicidins form well-defined channels, with cation conductances that are approximately 60% of those of the reference channels. The Ser-substituted channels are structurally equivalent to native gramicidin channels, as demonstrated by the formation of heterodimeric channels between a Ser-containing subunit and a native gramicidin subunit. These hybrid channels exhibit rectification, attributable to asymmetric placement of the single Ser hydroxyl group with respect to the bilayer center. Compared to the corresponding Ala-containing reference channels, the polar Ser residues decrease the analogues' channel-forming potency by 3 orders of magnitude, indicating a substantial energetic penalty ( approximately 15 kJ/mol) for burying the polar Ser side chain in the bilayer hydrophobic core.

  19. Effect of membrane environment on proton permeation through gramicidin A channels.

    PubMed

    Qin, Zhen; Tepper, Harald L; Voth, Gregory A

    2007-08-23

    Multistate empirical valence bond simulations were employed to study proton transport through gramicidin A channels embedded in two different lipid bilayers, glycerol 1-monooleate (GMO) and diphytanolphosphatidylcholine (DiPhPC). Free energy barriers to proton permeation were derived using a new internal reaction coordinate describing the proton permeation process. The large quantitative and qualitative differences between the two systems are discussed in terms of local bilayer structures, ordering of interfacial water, and channel flexibility in the two environments.

  20. Ion channel stability of Gramicidin A in lipid bilayers: effect of hydrophobic mismatch.

    PubMed

    Basu, Ipsita; Chattopadhyay, Amitabha; Mukhopadhyay, Chaitali

    2014-01-01

    Hydrophobic mismatch which is defined as the difference between the lipid hydrophobic thickness and the peptide hydrophobic length is known to be responsible in altering the lipid/protein dynamics. Gramicidin A (gA), a 15 residue β helical peptide which is well recognized to form ion conducting channels in lipid bilayer, may change its structure and function in a hydrophobic mismatched condition. We have performed molecular dynamics simulations of gA dimer in phospholipid bilayers to investigate whether or not the conversion from channel to non-channel form of gA dimer would occur under extreme negative hydrophobic mismatch. By varying the length of lipid bilayers from DLPC (1, 2-Dilauroyl-sn-glycero-3-phosphocholine) to DAPC (1, 2-Diarachidoyl-sn-glycero-3-phosphocholine), a broad range of mismatch was considered from nearly matching to extremely negative. Our simulations revealed that though the ion-channel conformation is retained by gA under a lesser mismatched situation, in extremely negative mismatched situation, in addition to bilayer thinning, the conformation of gA is changed and converted to a non-channel one. Our results demonstrate that although the channel conformation of Gramicidin A is the most stable structure, it is possible for gA to change its conformation from channel to non-channel depending upon the local environment of host bilayers.

  1. Binding constants of Li+, K+, and Tl+ in the gramicidin channel determined from water permeability measurements.

    PubMed Central

    Dani, J A; Levitt, D G

    1981-01-01

    In an open circuit there can be no net cation flux through membranes containing only cation-selective channels, because electroneutrality must be maintained. If the channels are so narrow that water and cations cannot pass by each other, then the net water flux through those "single-file" channels that contain a cation is zero. It is therefore possible to determine the cation binding constants from the decrease in the average water permeability per channel as the cation concentration in the solution is increased. Three different methods were used to determine the osmotic water permeability of gramicidin channels in lipid bilayer membranes. The osmotic water permeability coefficient per gramicidin channel in the absence of cations was found to be 6 x 10(-14) cm3/s. As the cation concentration was raised, the water permeability decreased and a binding constant was determined from a quantitative fit to the data. When the data were fitted assuming a maximum of one ion per channel, the dissociation constant was 115 mM for Li+, 69 mM for K+, and 2 mM for Tl+. PMID:6168310

  2. The Gramicidin A Transmembrane Channel: A Proposed π(L,D) Helix

    PubMed Central

    Urry, D. W.

    1971-01-01

    A lipophilic, left-handed helical structure is proposed for gramicidin A in which the C-O bonds alternately point toward the amino and carboxyl ends; it is a hybrid of the 4.314 and 4.416 helices. The C-O groups pointing toward the carboxyl end form part of 16-membered hydrogen-bonded rings, whereas the C-O moieties pointing toward the amino end form 14-membered hydrogenbonded rings. The proposed structure is based on conformational analysis combined with requirements for the gramicidin A transmembrane channel. Two helices combine to form the channel. The alternating C-O directions allow hydrogen-bonded dimerization by the unique possibilities of head-to-head and tail-to-tail attachment. The formyl group at the amino end allows for a favorable head-to-head attachment with no loss of structural continuity. Unpublished studies. by M. C. Goodall on the lipid bilayer conductance of deformyl gramicidin A strongly argue for head-to-head attachment. Such hydrogen-bonded association is not possible with previously described helices, as the C-O groups all point in the same direction. In relation to possible π(L,D) helices in mammalian systems, it should be noted that glycines would fill the role of D residues. The conformation can undergo ion-induced relaxations, which provide approximate tetrahedral coordination for the ion, with facile shifting of coordinations. The ready exchange of coordinations provides the mechanism for movement of the ion along the channel. Conceivably, such transmembrane channels could have application as models for ion transport across biological membranes—an application which may be as great as, or greater than, that of carriers such as valinomycin and nonactin. Specifically, biogenic amines and drugs containing aromatic groups could control access to the channel by interactions with the two tryptophan residues at the ethanolamine end and with the negative region provided by the three oxygens. Images PMID:5276779

  3. The study of volt-ampere characteristics of ionic channels formed by gramicidin A.

    PubMed

    Passechnik, V I; Flerov, M N; Hianik, T

    1985-02-01

    A method of measurement of the non-linearity coefficient of volt-ampere characteristics of the type i(U) approximately = U(1 + beta U2) has been developed for ionic channels formed by gramicidin A, using the third harmonic of the membrane current. The shape of the volt-ampere characteristics (VA) of ionic channels formed by gramicidin A did not depend on the antibiotic concentration in the membrane. The coefficient beta of non-linearity of VA of membranes modified by gramicidin A depended on electrolyte concentration "c" and it increased proportionally with the lg c from -17 V-2 at 0.03 mol/l KC1 to 8 V-2 at 3.4 mol/l KCl, and it was zero at co = 0.3 - 1 mol/l KCl. Egg lecithin and glycerol monooleate (GMO) membranes differ in their co values. The substitution of K+ for Li+ of the membrane solvent (n-heptane for n-hexadecane) did not influence the value of beta; the same applied for GMO membranes without any solvent. In a number of membranes, spontaneous change of the non-linearity coefficient with time observed after the membrane formation, as well as jumps of the non-linearity coefficient at a practically unchanged membrane conductivity. An analysis of some theoretical models of the ion transport through the channel has shown that, at voltages above 200 mV, these models provide rather small values of beta, or extremely high VA non-linearity.

  4. Use of weak acids to determine the bulk diffusion limitation of H+ ion conductance through the gramicidin channel.

    PubMed Central

    Decker, E R; Levitt, D G

    1988-01-01

    The addition of 2 M formic acid at pH 3.75 increased the single channel H+ ion conductance of gramicidin channels 12-fold at 200 mV. Other weak acids (acetic, lactic, oxalic) produce a similar, but smaller increase. Formic acid (and other weak acids) also blocks the K+ conductance at pH 3.75, but not at pH 6.0 when the anion form predominates. This increased H+ conductance and K+ block can be explained by formic acid (HF) binding to the mouth of the gramicidin channel (Km = 1 M) and providing a source of H+ ions. A kinetic model is derived, based on the equilibrium binding of formic acid to the channel mouth, that quantitatively predicts the conductance for different mixtures of H+, K+, and formic acid. The binding of the neutral formic acid to the mouth of the gramicidin channel is directly supported by the observation that a neutral molecule with a similar structure, formamide (and malonamide and acrylamide), blocks the K+ conductance at pH 6.0. The H+ conductance in the presence of formic acid provides a lower bound for the intrinsic conductance of the gramicidin channel when there is no diffusion limitation at the channel mouth. The 12-fold increase in conductance produced by formic acid suggests that greater than 90% of the total resistance to H+ results from diffusion limitation in the bulk solution. PMID:2449253

  5. Gramicidin channel-induced lipid membrane deformation energy: influence of chain length and boundary conditions.

    PubMed

    Ring, A

    1996-01-31

    The influence of boundary conditions on the deformation energy of a lipid membrane containing a gramicidin A channel was evaluated numerically. A liquid crystal model was used to calculate the relative contributions of compression, splay and surface tension. It is proposed that the nearest neighbor lipid molecules are displaced from the channel end in a direction perpendicular to the bilayer and it is concluded that surface tension is the major component of the deformation free energy for monoolein (gmo)/n-alkane membranes. This unexpected result supports the validity of the liquid crystal models of membrane deformation since gramicidin lifetime has been shown to correlate with surface tension for gmo membranes. The theory accurately predicts the experimentally measured relative lifetimes without the use of adjustable parameters. For conditions where splay may be neglected surface tension is always the major component of the deformation energy, irrespective of the magnitude of the compression coefficient. The deformation may extend for hundreds of angstroms from the peptide. The results obtained here are expected to be important for the characterization of protein-membrane interactions in general.

  6. Energetics of double-ion occupancy in the gramicidin A channel.

    PubMed

    Li, Yuhui; Andersen, Olaf S; Roux, Benoît

    2010-11-01

    To understand the energetics of double-ion occupancy in gramicidin A (gA) channels, the 2D potential of mean force (PMF) is calculated for two ions at different positions along the channel axis. The cross sections of this 2D PMF are compared with available one-ion PMFs to highlight the effect of one ion on the permeation dynamics of the other. It is found that, if the first ion stays on one side in the channel, the second ion has to pass over an additional barrier to move into the outer binding site. At the same time, both outer and inner binding sites for the second ion become shallower than those in the one-ion PMF. The calculated ion-ion repulsion for a doubly occupied channel is about 2 kcal/mol, in good agreement with previous experimental estimates. The number of water molecules inside the channel and their dipole moment are calculated to interpret the energetics of double-ion occupancy. As the first ion moves into the outer binding site and then further into the channel, the oxygen atoms of the single-file water column in the channel are oriented to point toward the ion. The observed dipole moment distribution of a singly occupied channel has only one sharp peak, and the water alignment is essentially perfect once the ion is in the inner binding site. For this reason, there is an energy penalty to accommodate a second ion at the opposite end of the channel.

  7. On the origin of closing flickers in gramicidin channels: a new hypothesis.

    PubMed

    Armstrong, Kathryn M; Cukierman, Samuel

    2002-03-01

    The submillisecond closing events (flickers) and the single channel conductances to protons (g(H)) were studied in native gramicidin A (gA) and in the SS and RR diastereoisomers of dioxolane-linked gA channels in planar bilayers. Bilayers were formed from glycerylmonooleate (GMO) in various solvents. In GMO/decane (thick) bilayers, the largest flicker frequency occurred in the SS channel (39 s(-1)), followed by the RR (4 s(-1)) and native gA channels (3 s(-1)). These frequencies were attenuated in GMO/squalene (thin) bilayers by 100-, 30-, and 70-fold in the SS, RR, and native gA channels, respectively. In thin bilayers, the average burst duration of native gA channels was 30-fold longer than in thick bilayers. The RR dioxolane-linked gA dimer "inactivated" in GMO/decane but not in squalene-containing bilayers. The mean closed time of flickers (approximately 0.12 ms) was essentially the same in various gA channels. In thin bilayers, g(H) values were larger by approximately 10% (SS), 30% (RR), and 20% (native gA) in relation to thick bilayers. It is concluded that flickers are not related to pre-dissociation or dissociation states of gA monomers, and do not seem to be caused by intrinsic conformational changes of channel proteins. It is proposed that flickers are caused by undulations of the bilayer that obliterate the openings of gA channels. Differences between flicker frequencies in various gA channels are likely to result from variations in channel geometries at the bilayer/channel interface. The smaller g(H) in thick bilayers suggests that the deformation of these bilayers around the gA channel creates a diffusional pathway next to the mouths of the channel that is longer and more restrictive than in thin GMO bilayers. A possible molecular interpretation for these effects is attempted.

  8. Can gramicidin ion channel affect the dipole potential of neighboring phospholipid headgroups?

    PubMed

    Becucci, Lucia; Guidelli, Rolando

    2015-12-01

    The cyclic voltammetry behavior of a mercury-supported tethered bilayer lipid membrane (tBLM) incorporating gramicidin A was investigated in aqueous 0.1 M KCl at pH 6.8, 5.4 and 3. The distal leaflet of the lipid bilayer consisted of dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylserine (DOPS), dioleoylphosphatidic acid or a DOPC/cholesterol mixture. In passing from pH 6.8 to pH 3, the midpoint potential between the negative current peak, due to K(+) inflow into the spacer, and the positive current peak, due to K(+) ejection into the aqueous solution, shifts toward more positive potentials, while the separation between these two peaks decreases. This behavior is interpreted quantitatively on the basis of a model relying on tBLM structural features estimated independently in previous works. The only adjustable parameter is the rate constant for cation translocation across a potential energy barrier located in the hydrocarbon tail region. The behavior is ascribed to a dragging of the lipid headgroups adjacent to the gramicidin channel mouth toward the hydrocarbon tail region, with a resulting decrease in the negative charge of the DOPC phosphate group, or of the DOPS carboxyl group, with decreasing pH.

  9. Peroxyl radicals promoted changes in water permeability through gramicidin channels in DPPC and lecithin-PC vesicles.

    PubMed

    Soto, M A; Sotomayor, C P; Lissi, E A

    2003-03-01

    Gramicidin incorporation to DPPC or lecithin-PC large unilamellar vesicles (LUVs) leads to pore formation that, under hyper-osmotic conditions, produces a noticeable increase in the rate of trans-membrane water flow. This pore formation is more efficient in the more fluid lecithin-PC LUVs. Exposure of these vesicles to peroxyl radicals generated in the aerobic thermolysis of 2,2'-azo-bis(2-amidinopropane) (AAPH), changes the physical properties of the bilayer (as sensed employing fluorescent probes), modifies gramicidin molecules (as sensed by the decrease in Trp fluorescence) and notably reduces the transbilayer rate of water outflow. In order to evaluate if this reduced water-transport capacity is due to changes in the membrane due to lipid-peroxidation and/or direct damage to gramicidin channels, results obtained in the oxidable vesicles (lecithin-PC) were compared to those obtained in DPPC vesicles. The data obtained show that most of the water transport efficiency loss can be ascribed to a direct disruption of gramicidin channels by AAPH derived peroxyl radicals. PMID:12637166

  10. Peroxyl radicals promoted changes in water permeability through gramicidin channels in DPPC and lecithin-PC vesicles.

    PubMed

    Soto, M A; Sotomayor, C P; Lissi, E A

    2003-03-01

    Gramicidin incorporation to DPPC or lecithin-PC large unilamellar vesicles (LUVs) leads to pore formation that, under hyper-osmotic conditions, produces a noticeable increase in the rate of trans-membrane water flow. This pore formation is more efficient in the more fluid lecithin-PC LUVs. Exposure of these vesicles to peroxyl radicals generated in the aerobic thermolysis of 2,2'-azo-bis(2-amidinopropane) (AAPH), changes the physical properties of the bilayer (as sensed employing fluorescent probes), modifies gramicidin molecules (as sensed by the decrease in Trp fluorescence) and notably reduces the transbilayer rate of water outflow. In order to evaluate if this reduced water-transport capacity is due to changes in the membrane due to lipid-peroxidation and/or direct damage to gramicidin channels, results obtained in the oxidable vesicles (lecithin-PC) were compared to those obtained in DPPC vesicles. The data obtained show that most of the water transport efficiency loss can be ascribed to a direct disruption of gramicidin channels by AAPH derived peroxyl radicals.

  11. Noncontact dipole effects on channel permeation. III. Anomalous proton conductance effects in gramicidin

    PubMed Central

    Phillips, LR; Cole, CD; Hendershot, RJ; Cotten, M; Cross, TA; Busath, DD

    1999-01-01

    Proton transport on water wires, of interest for many problems in membrane biology, is analyzed in side-chain analogs of gramicidin A channels. In symmetrical 0.1 N HCl solutions, fluorination of channel Trp(11), Trp-(13), or Trp(15) side chains is found to inhibit proton transport, and replacement of one or more Trps with Phe enhances proton transport, the opposite of the effects on K(+) transport in lecithin bilayers. The current-voltage relations are superlinear, indicating that some membrane field-dependent process is rate limiting. The interfacial dipole effects are usually assumed to affect the rate of cation translocation across the channel. For proton conductance, however, water reorientation after proton translocation is anticipated to be rate limiting. We propose that the findings reported here are most readily interpreted as the result of dipole-dipole interactions between channel waters and polar side chains or lipid headgroups. In particular, if reorientation of the water column begins with the water nearest the channel exit, this hypothesis explains the negative impact of fluorination and the positive impact of headgroup dipole on proton conductance. PMID:20540928

  12. Thermodynamic view of activation energies of proton transfer in various gramicidin A channels.

    PubMed

    Chernyshev, Anatoly; Cukierman, Samuel

    2002-01-01

    The temperature dependencies (range: 5-45 degrees C) of single-channel proton conductances (g(H)) in native gramicidin A (gA) and in two diastereoisomers (SS and RR) of the dioxolane-linked gA channels were measured in glycerylmonooleate/decane (GMO) and diphytanoylphosphatidylcholine/decane (DiPhPC) bilayers. Linear Arrhenius plots (ln (g(H)) versus K(-1)) were obtained for the native gA and RR channels in both types of bilayers, and for the SS channel in GMO bilayers only. The Arrhenius plot for proton transfer in the SS channel in DiPhPC bilayers had a break in linearity around 20 degrees C. This break seems to occur only when protons are the permeating cations in the SS channel. The activation energies (E(a)) for proton transfer in various gA channels (approximately 15 kJ/mol) are consistent with the rate-limiting step being in the channel and/or at the membrane-channel/solution interface, and not in bulk solution. E(a) values for proton transfer in gA channels are considerably smaller than for the permeation of nonproton currents in gA as well as in various other ion channels. The E(a) values for proton transfer in native gA channels are nearly the same in both GMO and DiPhPC bilayers. In contrast, for the dioxolane linked gA dimers, E(a) values were strongly modulated by the lipid environment. The Gibbs activation free energies (Delta G(#)(o)) for protons in various gA channels are within the range of 27-29 kJ/mol in GMO bilayers and of 20-22 kJ/mol in DiPhPC bilayers. The largest difference between Delta G(#)(o) for proton currents occurs between native gA (or SS channels) and the RR channel. In general, the activation entropy (Delta S) is mostly responsible for the differences between g(H) values in various gA channels, and also in distinct bilayers. However, significant differences between the activation enthalpies (Delta H(#)(o)) for proton transfer in the SS and RR channels occur in distinct membranes.

  13. Density-functional theory study of gramicidin A ion channel geometry and electronic properties.

    PubMed

    Todorović, Milica; Bowler, David R; Gillan, Michael J; Miyazaki, Tsuyoshi

    2013-12-01

    Understanding the mechanisms underlying ion channel function from the atomic-scale requires accurate ab initio modelling as well as careful experiments. Here, we present a density functional theory (DFT) study of the ion channel gramicidin A (gA), whose inner pore conducts only monovalent cations and whose conductance has been shown to depend on the side chains of the amino acids in the channel. We investigate the ground state geometry and electronic properties of the channel in vacuum, focusing on their dependence on the side chains of the amino acids. We find that the side chains affect the ground state geometry, while the electrostatic potential of the pore is independent of the side chains. This study is also in preparation for a full, linear scaling DFT study of gA in a lipid bilayer with surrounding water. We demonstrate that linear scaling DFT methods can accurately model the system with reasonable computational cost. Linear scaling DFT allows ab initio calculations with 10,000-100,000 atoms and beyond, and will be an important new tool for biomolecular simulations.

  14. Molecular mechanism of H+ conduction in the single-file water chain of the gramicidin channel.

    PubMed

    Pomès, Régis; Roux, Benoît

    2002-05-01

    The conduction of protons in the hydrogen-bonded chain of water molecules (or "proton wire") embedded in the lumen of gramicidin A is studied with molecular dynamics free energy simulations. The process may be described as a "hop-and-turn" or Grotthuss mechanism involving the chemical exchange (hop) of hydrogen nuclei between hydrogen-bonded water molecules arranged in single file in the lumen of the pore, and the subsequent reorganization (turn) of the hydrogen-bonded network. Accordingly, the conduction cycle is modeled by two complementary steps corresponding respectively to the translocation 1) of an ionic defect (H+) and 2) of a bonding defect along the hydrogen-bonded chain of water molecules in the pore interior. The molecular mechanism and the potential of mean force are analyzed for each of these two translocation steps. It is found that the mobility of protons in gramicidin A is essentially determined by the fine structure and the dynamic fluctuations of the hydrogen-bonded network. The translocation of H+ is mediated by spontaneous (thermal) fluctuations in the relative positions of oxygen atoms in the wire. In this diffusive mechanism, a shallow free-energy well slightly favors the presence of the excess proton near the middle of the channel. In the absence of H+, the water chain adopts either one of two polarized configurations, each of which corresponds to an oriented donor-acceptor hydrogen-bond pattern along the channel axis. Interconversion between these two conformations is an activated process that occurs through the sequential and directional reorientation of water molecules of the wire. The effect of hydrogen-bonding interactions between channel and water on proton translocation is analyzed from a comparison to the results obtained previously in a study of model nonpolar channels, in which such interactions were missing. Hydrogen-bond donation from water to the backbone carbonyl oxygen atoms lining the pore interior has a dual effect: it

  15. Reactive derivatives of gramicidin enable light- and ion-modulated ion channels

    NASA Astrophysics Data System (ADS)

    Macrae, Michael X.; Blake, Steven; Mayer, Thomas; Mayer, Michael; Yang, Jerry

    2009-08-01

    Detection of chemical processes on a single molecule scale is the ultimate goal of sensitive analytical assays. We have explored methods to detect chemical analytes in solution using synthetic derivatives of gramicidin A (gA). We exploited the functional properties of an ion channel-forming peptideg--gA--to report changes in the local environment near the opening of these semi-synthetic nanopores upon exposure to specific external stimuli. These peptide-based nanosensors detect reaction-induced changes in the chemical or physical properties of functional groups presented at the opening of the pore. This paper discusses the development of gA-based sensors for detecting external factors such as metal ions in solution or for detecting specific wavelengths of light. We propose that gA-based ion channel sensors offer tremendous potential for ultra sensitive functional detection since a single chemical modification of each individual sensing element can lead to readily detectable changes in channel conductance.

  16. Effects of lysine methylation on gramicidin A channel folding in lipid membranes.

    PubMed

    Wang, Fang; Qin, Luoheng; Wong, Patrick; Gao, Jianmin

    2013-11-01

    Protein-membrane interactions underlie numerous biological processes including folding of ion channels and signal transduction across lipid membranes. A detailed understanding of protein-lipid interactions is critical for designing membrane-active peptides as potential antibiotics, as well. Using gramicidin A (gA) as a model system, we investigated the effects of lysine methylation on peptide folding into transmembrane channels. The results are discussed in terms of the peptides' binding affinity to, translocation across, and structure formation in lipid membranes. The results show that gA mutants with N(ɛ)-trimethylated D-lysines (dMe3 K) are capable of folding into wild type-like channels that are selective for monovalent cations. Surprisingly, N(ɛ)-trimethylation in general reduces the peptide's binding affinity to lipid membranes despite the increased hydrophobicity. Further investigation reveals the critical contribution of the hydrogen bonding potential of lysine side chains to peptide-membrane association, which has previously been underappreciated. Importantly, methylation does give improved therapeutic indices for certain combinations of gA variant and bacterium, indicating that methylation can be an effective strategy to fine tune the performance of peptide antibiotics.

  17. Structure of gramicidin D-RbCl complex at atomic resolution from low-temperature synchrotron data: interactions of double-stranded gramicidin channel contents and cations with channel wall

    SciTech Connect

    Glówka, M.L.; Olczak, A.; Bojarska, J.; Szczesio, M.; Duax, W.L.; Burkhart, B.M.; Pangborn, W.A.; Langs, D.A.; Wawrzak, Z.

    2010-03-05

    Gramicidin D (gD) is a naturally occurring ionophoric antibiotic that forms membrane channels specific for monovalent cations. The crystal structure of the RbCl complex of gD has been determined at 1.14 {angstrom} resolution from low-temperature (100 K) synchrotron-radiation data with a final R of 16%. The structure was refined with anisotropic temperature factors for all non-H atoms and with partial occupancies for many of them. The asymmetric unit in the crystal contains four crystallographically independent molecules that form two right-handed antiparallel double-stranded dimers. There are seven distinct rubidium-binding sites in each dimeric channel. The occupancy factors of Rb cations are between 0.11 and 0.35 and the total ion contents of the two crystallographically independent channels are 1.59 and 1.22 ions, respectively. Although each channel is 'chemically symmetrical', the side-chain conformations, the distributions of rubidium cations and their binding sites in the two independent channels are not. Cations are 'coordinated' by delocalized {pi}-electrons of three to five carbonyl groups that together with peptide backbone chains form the gramicidin channel walls. The water:cation ratio in the channel interior is four or five:one, and five or six waters separate Rb cations during their passage through the channel.

  18. Investigation of polarization effects in the gramicidin A channel from ab initio molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Timko, Jeff; Kuyucak, Serdar

    2012-11-01

    Polarization is an important component of molecular interactions and is expected to play a particularly significant role in inhomogeneous environments such as pores and interfaces. Here we investigate the effects of polarization in the gramicidin A ion channel by performing quantum mechanics/molecular mechanics molecular dynamics (MD) simulations and comparing the results with those obtained from classical MD simulations with non-polarizable force fields. We consider the dipole moments of backbone carbonyl groups and channel water molecules as well as a number of structural quantities of interest. The ab initio results show that the dipole moments of the carbonyl groups and water molecules are highly sensitive to the hydrogen bonds (H-bonds) they participate in. In the absence of a K+ ion, water molecules in the channel are quite mobile, making the H-bond network highly dynamic. A central K+ ion acts as an anchor for the channel waters, stabilizing the H-bond network and thereby increasing their average dipole moments. In contrast, the K+ ion has little effect on the dipole moments of the neighboring carbonyl groups. The weakness of the ion-peptide interactions helps to explain the near diffusion-rate conductance of K+ ions through the channel. We also address the sampling issue in relatively short ab initio MD simulations. Results obtained from a continuous 20 ps ab initio MD simulation are compared with those generated by sampling ten windows from a much longer classical MD simulation and running each window for 2 ps with ab initio MD. Both methods yield similar results for a number of quantities of interest, indicating that fluctuations are fast enough to justify the short ab initio MD simulations.

  19. Investigation of polarization effects in the gramicidin A channel from ab initio molecular dynamics simulations.

    PubMed

    Timko, Jeff; Kuyucak, Serdar

    2012-11-28

    Polarization is an important component of molecular interactions and is expected to play a particularly significant role in inhomogeneous environments such as pores and interfaces. Here we investigate the effects of polarization in the gramicidin A ion channel by performing quantum mechanics/molecular mechanics molecular dynamics (MD) simulations and comparing the results with those obtained from classical MD simulations with non-polarizable force fields. We consider the dipole moments of backbone carbonyl groups and channel water molecules as well as a number of structural quantities of interest. The ab initio results show that the dipole moments of the carbonyl groups and water molecules are highly sensitive to the hydrogen bonds (H-bonds) they participate in. In the absence of a K(+) ion, water molecules in the channel are quite mobile, making the H-bond network highly dynamic. A central K(+) ion acts as an anchor for the channel waters, stabilizing the H-bond network and thereby increasing their average dipole moments. In contrast, the K(+) ion has little effect on the dipole moments of the neighboring carbonyl groups. The weakness of the ion-peptide interactions helps to explain the near diffusion-rate conductance of K(+) ions through the channel. We also address the sampling issue in relatively short ab initio MD simulations. Results obtained from a continuous 20 ps ab initio MD simulation are compared with those generated by sampling ten windows from a much longer classical MD simulation and running each window for 2 ps with ab initio MD. Both methods yield similar results for a number of quantities of interest, indicating that fluctuations are fast enough to justify the short ab initio MD simulations.

  20. High-resolution polypeptide structure and dynamics in anisotropic environments: The gramicidin channel

    SciTech Connect

    Cross, T.A.; Lee, K.C.; Ketchem, R.R.; Hu, W.; Lazo, N.D.; Huo, S.

    1994-12-01

    To understand the details of macromolecular function, high-resolution structural and dynamic detail is essential. The polypeptide fold of the gramicidin channel has been effectively modeled for the past 20 years, yet the functional changes in conductance and channel lifetime associated with amino acid substitutions cannot be predicted. To accomplish this goal, high-resolution electrostatic modeling and the precise orientation of all dipoles are required. Furthermore, an enhanced knowledge of the complex molecular environment of this membrane-bound peptide is needed. An aqueous environment is relatively uniform and achiral. The membrane environment is very heterogenous and chiral. A knowledge of the interactions, specific and nonspecific, between peptide and lipid will aid in developing a better understanding of this environment. To accomplish this goal, it is necessary to study the peptide in an extended lipid bilayer, rather than in a vesicular or micellar form. These latter environments are likely to possess increased dynamics, increased water penetration, and distorted interactions between the polypeptide and membrane surface. To perform NMR studies on bilayer bound peptides, solid state NMR methods are required, and for specific site information, isotopic labels are incorporated using solid phase peptide synthesis.

  1. Double-stranded helical twisted beta-sheet channels in crystals of gramicidin S grown in the presence of trifluoroacetic and hydrochloric acids.

    PubMed

    Llamas-Saiz, Antonio L; Grotenbreg, Gijsbert M; Overhand, Mark; van Raaij, Mark J

    2007-03-01

    Gramicidin S is a nonribosomally synthesized cyclic decapeptide antibiotic with twofold symmetry (Val-Orn-Leu-D-Phe-Pro)(2); a natural source is Bacillus brevis. Gramicidin S is active against Gram-positive and some Gram-negative bacteria. However, its haemolytic toxicity in humans limits its use as an antibiotic to certain topical applications. Synthetically obtained gramicidin S was crystallized from a solution containing water, methanol, trifluoroacetic acid and hydrochloric acid. The structure was solved and refined at 0.95 A resolution. The asymmetric unit contains 1.5 molecules of gramicidin S, two trifluoroacetic acid molecules and ten water molecules located and refined in 14 positions. One gramicidin S molecule has an exact twofold-symmetrical conformation; the other deviates from the molecular twofold symmetry. The cyclic peptide adopts an antiparallel beta-sheet secondary structure with two type II' beta-turns. These turns have the residues D-Phe and Pro at positions i + 1 and i + 2, respectively. In the crystals, the gramicidin S molecules line up into double-stranded helical channels that differ from those observed previously. The implications of the supramolecular structure for several models of gramicidin S conformation and assembly in the membrane are discussed.

  2. The permeation properties of small organic cations in gramicidin A channels.

    PubMed Central

    Seoh, S A; Busath, D

    1993-01-01

    The conductance properties of organic cations in single gramicidin A channels were studied using planar lipid bilayers. From measurements at 10 mM and at 27 mV the overall selectivity sequence was found to be NH4+ > K+ > hydrazinium > formamidinium > Na+ > methylammonium, which corresponds to Eisenman polyatomic cation sequence X'. Methylammonium and formamidinium exhibit self block, suggesting multiple occupancy and single filing. Formamidinium has an apparent dissociation constant (which is similar to those of alkali metal cations) for the first ion being 22 mM from the Eadie-Hofstee plot (G0 vs. G0/C), 12 mM from the rate constants of a three-step kinetic model. The rate-limiting step for formamidinium is translocation judging from supralinear I-V relations at low concentrations. 1 M formamidinium solutions yields exceptionally long single channel lifetimes, 20-fold longer than methylammonium, which yields lifetimes similar to those found with alkali metal cations. The average lifetime in formamidinium solution significantly decreases with increasing voltage up to 100 mV but is relatively voltage independent between 100 and 200 mV. At lower voltages (< or = 100 mV), the temperature and concentration dependences of the average lifetime of formamidinium were steep. At very low salt concentrations (0.01 M, 100 mV), there was no significant difference in average lifetime from that formed with 0.01 M methylammonium or hydrazinium. We conclude that formamidinium very effectively stabilizes the dimeric channel while inside the channel and speculate that it does so by affecting tryptophan-reorientation or tryptophan-lipid interactions at binding sites. PMID:7684267

  3. Design of Peptide-Membrane Interactions to Modulate Single-File Water Transport through Modified Gramicidin Channels

    PubMed Central

    Portella, Guillem; Polupanow, Tanja; Zocher, Florian; Boytsov, Danila A.; Pohl, Peter; Diederichsen, Ulf; de Groot, Bert L.

    2012-01-01

    Water permeability through single-file channels is affected by intrinsic factors such as their size and polarity and by external determinants like their lipid environment in the membrane. Previous computational studies revealed that the obstruction of the channel by lipid headgroups can be long-lived, in the range of nanoseconds, and that pore-length-matching membrane mimetics could speed up water permeability. To test the hypothesis of lipid-channel interactions modulating channel permeability, we designed different gramicidin A derivatives with attached acyl chains. By combining extensive molecular-dynamics simulations and single-channel water permeation measurements, we show that by tuning lipid-channel interactions, these modifications reduce the presence of lipid headgroups in the pore, which leads to a clear and selective increase in their water permeability. PMID:23083713

  4. Design of peptide-membrane interactions to modulate single-file water transport through modified gramicidin channels.

    PubMed

    Portella, Guillem; Polupanow, Tanja; Zocher, Florian; Boytsov, Danila A; Pohl, Peter; Diederichsen, Ulf; de Groot, Bert L

    2012-10-17

    Water permeability through single-file channels is affected by intrinsic factors such as their size and polarity and by external determinants like their lipid environment in the membrane. Previous computational studies revealed that the obstruction of the channel by lipid headgroups can be long-lived, in the range of nanoseconds, and that pore-length-matching membrane mimetics could speed up water permeability. To test the hypothesis of lipid-channel interactions modulating channel permeability, we designed different gramicidin A derivatives with attached acyl chains. By combining extensive molecular-dynamics simulations and single-channel water permeation measurements, we show that by tuning lipid-channel interactions, these modifications reduce the presence of lipid headgroups in the pore, which leads to a clear and selective increase in their water permeability.

  5. The effect of microwave radiation on the stability and formation of gramicidin-A channels in lipid bilayer membranes

    SciTech Connect

    Sandblom, J.; Theander, S. )

    1991-01-01

    The effects of microwaves on the single-channel kinetics of gramicidin-A channels in lipid bilayer membranes were examined. Attempts were made to separate thermal and athermal effects by accurate measurements of temperature at the site of the membrane and by relating the measured parameters to their previously characterized temperature dependence. It was found that microwave radiation does not affect single-channel conductance or channel life time to a degree that is significantly different from that expected of a purely thermal effect. On the other hand, the rate of channel formation is decreased during exposure, which is opposite to that expected of a purely thermal effect. The mechanism of this effect is discussed in terms of the dimerization process of channel formation.

  6. Noncontact dipole effects on channel permeation. I. Experiments with (5F-indole)Trp13 gramicidin A channels.

    PubMed Central

    Busath, D D; Thulin, C D; Hendershot, R W; Phillips, L R; Maughan, P; Cole, C D; Bingham, N C; Morrison, S; Baird, L C; Hendershot, R J; Cotten, M; Cross, T A

    1998-01-01

    Gramicidin A (gA), with four Trp residues per monomer, has an increased conductance compared to its Phe replacement analogs. When the dipole moment of the Trp13 side chain is increased by fluorination at indole position 5 (FgA), the conductance is expected to increase further. gA and FgA conductances to Na+, K+, and H+ were measured in planar diphytanoylphosphatidylcholine (DPhPC) or glycerylmonoolein (GMO) bilayers. In DPhPC bilayers, Na+ and K+ conductances increased upon fluorination, whereas in GMO they decreased. The low ratio in the monoglyceride bilayer was not reversed in GMO-ether bilayers, solvent-inflated or -deflated bilayers, or variable fatty acid chain monoglyceride bilayers. In both GMO and DPhPC bilayers, fluorination decreased conductance to H+ but increased conductance in the mixed solution, 1 M KCl at pH 2.0, where K+ dominates conduction. Eadie-Hofstee plot slopes suggest similar destabilization of K+ binding in both lipids. Channel lifetimes were not affected by fluorination in either lipid. These observations indicate that fluorination does not change the rotameric conformation of the side chain. The expected difference in the rate-limiting step for transport through channels in the two bilayers qualitatively explains all of the above trends. PMID:9826605

  7. Noncontact dipole effects on channel permeation. I. Experiments with (5F-indole)Trp13 gramicidin A channels.

    PubMed

    Busath, D D; Thulin, C D; Hendershot, R W; Phillips, L R; Maughan, P; Cole, C D; Bingham, N C; Morrison, S; Baird, L C; Hendershot, R J; Cotten, M; Cross, T A

    1998-12-01

    Gramicidin A (gA), with four Trp residues per monomer, has an increased conductance compared to its Phe replacement analogs. When the dipole moment of the Trp13 side chain is increased by fluorination at indole position 5 (FgA), the conductance is expected to increase further. gA and FgA conductances to Na+, K+, and H+ were measured in planar diphytanoylphosphatidylcholine (DPhPC) or glycerylmonoolein (GMO) bilayers. In DPhPC bilayers, Na+ and K+ conductances increased upon fluorination, whereas in GMO they decreased. The low ratio in the monoglyceride bilayer was not reversed in GMO-ether bilayers, solvent-inflated or -deflated bilayers, or variable fatty acid chain monoglyceride bilayers. In both GMO and DPhPC bilayers, fluorination decreased conductance to H+ but increased conductance in the mixed solution, 1 M KCl at pH 2.0, where K+ dominates conduction. Eadie-Hofstee plot slopes suggest similar destabilization of K+ binding in both lipids. Channel lifetimes were not affected by fluorination in either lipid. These observations indicate that fluorination does not change the rotameric conformation of the side chain. The expected difference in the rate-limiting step for transport through channels in the two bilayers qualitatively explains all of the above trends.

  8. Single-channel studies on linear gramicidins with altered amino acid sequences. A comparison of phenylalanine, tryptophane, and tyrosine substitutions at positions 1 and 11.

    PubMed Central

    Mazet, J L; Andersen, O S; Koeppe, R E

    1984-01-01

    The relation between chemical structure and permeability characteristics of transmembrane channels has been investigated with the linear gramicidins (A, B, and C), where the amino acid at position 1 was chemically replaced by phenylalanine, tryptophane or tyrosine. The purity of most of the compounds was estimated to be greater than 99.99%. The modifications resulted in a wide range of conductance changes in NaCl solutions: sixfold from tryptophane gramicidin A to tyrosine gramicidin B. The conductance changes induced by a given amino acid substitution at position 1 are not the same as at position 11. The only important change in the Na+ affinity was observed when the first amino acid was tyrosine. No major conformational changes of the polypeptide backbone structure could be detected on the basis of experiments with mixtures of different analogues and valine gramicidin A (except possibly with tyrosine at position 1), as all the compounds investigated could form hybrid channels with valine gramicidin A. The side chains are not in direct contact with the permeating ions. The results were therefore interpreted in terms of modifications of the energy profile for ion movement through the channel, possibly due to an electrostatic interaction between the dipoles of the side chains and ions in the channel. Images FIGURE 1 FIGURE 2 FIGURE 3 PMID:6201199

  9. Effect of gating modifier toxins on membrane thickness: implications for toxin effect on gramicidin and mechanosensitive channels.

    PubMed

    Chen, Rong; Chung, Shin-Ho

    2013-02-22

    Various gating modifier toxins partition into membranes and interfere with the gating mechanisms of biological ion channels. For example, GsMTx4 potentiates gramicidin and several bacterial mechanosensitive channels whose gating kinetics are sensitive to mechanical properties of the membrane, whereas binding of HpTx2 shifts the voltage-activity curve of the voltage-gated potassium channel Kv4.2 to the right. The detailed process by which the toxin partitions into membranes has been difficult to probe using molecular dynamics due to the limited time scale accessible. Here we develop a protocol that allows the spontaneous assembly of a polypeptide toxin into membranes in atomistic molecular dynamics simulations of tens of nanoseconds. The protocol is applied to GsMTx4 and HpTx2. Both toxins, released in water at the start of the simulation, spontaneously bind into the lipid bilayer within 50 ns, with their hydrophobic patch penetrated into the bilayer beyond the phosphate groups of the lipids. It is found that the bilayer is about 2 Å thinner upon the binding of a GsMTx4 monomer. Such a thinning effect of GsMTx4 on membranes may explain its potentiation effect on gramicidin and mechanosensitive channels.

  10. Gramicidin cation channel: An experimental determination of the right-handed helix sense and verification of. beta. -type hydrogen bonding

    SciTech Connect

    Nicholson, L.K.; Cross, T.A. )

    1989-11-28

    Due to the difficulty of obtaining protein/lipid cocrystals for diffraction studies, structural research on intrinsic membrane proteins and polypeptides has been largely restricted to indirect experimental techniques. Hence, many fundamental questions associated with peptide/lipid systems remain unanswered. In particular, the handedness of the gramicidin A transmembrane ion channel incorporated into lipid bilayers has been an open question for nearly two decades. In this study, solid-state {sup 15}N NMR spectroscopy is employed to probe directly the secondary structure of the polypeptide backbone. Recent determinations of the {sup 15}N chemical shift anisotropy tensor with respect to the molecular frame enable the quantitative evaluation of the {sup 15}N chemical shift resonances obtained from oriented dimyristoylphosphatidylcholine (DMPC) bilayer samples containing specific site {sup 15}N labeled gramicidin. This direct structural approach verifies the {beta}-sheet hydrogen-bonding pattern proposed by Urry and determines that in our DMPC bilayer preparations the gramicidin channel is right-handed. Additional structural information is provided by the {sup 15}N chemical shift data in the form of orientational constraints on the C{sub {alpha}}-C{sub {alpha}} axis orientation of individual peptides relative to the helix axis. The significance of these solid-state NMR results lies in the direct determination of the helix sense and the verification of the {beta}-type hydrogen bonding, in the development of the solid-state NMR methods for obtaining such information, and in emphasizing the importance of having direct structural data at atomic resolution.

  11. Proton mobilities in water and in different stereoisomers of covalently linked gramicidin A channels.

    PubMed Central

    Cukierman, S

    2000-01-01

    Proton conductivities in bulk solution (lambda(H)) and single-channel proton conductances (g(H)) in two different stereoisomers of the dioxolane-linked gramicidin A channel (the SS and RR dimers) were measured in a wide range of bulk proton concentrations ([H], 0.1-8000 mM). Proton mobilities (micro(H)) in water as well as in the SS and RR dimers were calculated from the conductivity data. In the concentration range of 0.1-2000 mM, a straight line with a slope of 0.75 describes the log (g(H))-log ([H]) relationship in the SS dimer. At [H] > 2000 mM, saturation is followed by a decline in g(H). The g(H)-[H] relationship in the SS dimer is qualitatively similar to the [H] dependence of lambda(H). However, the slope of the straight line in the log(lambda(H))-log([H]) plot is 0.96, indicating that the rate-limiting step for proton conduction through the SS dimer is not the diffusion of protons in bulk solution. The significant difference between the slopes of those linear relationships accounts for the faster decline of micro(H) as a function of [H] in the SS dimer in relation to bulk solution. In the high range of [H], saturation and decline of g(H) in the SS dimer can be accounted for by the significant decrease of micro(H) in bulk solution. At any given [H], g(H) in the RR dimer is significantly smaller than in the SS. Moreover, the g(H)-[H] relationship in the RR stereoisomer is qualitatively different from that in the SS. Between 1 and 50 mM [H], g(H) can be fitted with an adsorption isotherm, suggesting the presence of a proton-binding site inside the pore (pK(a) approximately 2), which limits proton exit from the channel. At 100 mM < [H] < 3000 mM, g(H) increases linearly with [H]. The distinctive shape of the g(H)-[H] relationship in the RR dimer suggests that the channel can be occupied simultaneously by more than one proton. At higher [H], the saturation and decline of g(H) in the RR dimer reflect the properties of micro(H) in bulk solution. In the entire

  12. Accurate Evaluation of Ion Conductivity of the Gramicidin A Channel Using a Polarizable Force Field without Any Corrections.

    PubMed

    Peng, Xiangda; Zhang, Yuebin; Chu, Huiying; Li, Yan; Zhang, Dinglin; Cao, Liaoran; Li, Guohui

    2016-06-14

    Classical molecular dynamic (MD) simulation of membrane proteins faces significant challenges in accurately reproducing and predicting experimental observables such as ion conductance and permeability due to its incapability of precisely describing the electronic interactions in heterogeneous systems. In this work, the free energy profiles of K(+) and Na(+) permeating through the gramicidin A channel are characterized by using the AMOEBA polarizable force field with a total sampling time of 1 μs. Our results indicated that by explicitly introducing the multipole terms and polarization into the electrostatic potentials, the permeation free energy barrier of K(+) through the gA channel is considerably reduced compared to the overestimated results obtained from the fixed-charge model. Moreover, the estimated maximum conductance, without any corrections, for both K(+) and Na(+) passing through the gA channel are much closer to the experimental results than any classical MD simulations, demonstrating the power of AMOEBA in investigating the membrane proteins. PMID:27171823

  13. Noncontact dipole effects on channel permeation. IV. Kinetic model of 5F-Trp(13) gramicidin A currents.

    PubMed

    Thompson, N; Thompson, G; Cole, C D; Cotten, M; Cross, T A; Busath, D D

    2001-09-01

    Nonlinear least squares fitting was used to assign rate constants for the three-barrier, two-site, double-occupancy, single-filing kinetic model for previously reported current-voltage relations of (5F-Indole)Trp(13) gramicidin A and gramicidin A channels (, 75:2830-2844). By judicious coupling of parameters, it was possible to reduce the parameter space from 64 parameters to 24, and a reasonable fit consistent with other experimental data was obtained. The main features of the fit were that fluorination increased the rate constant for translocation by a factor of 2.33, consistent with a free energy change in the translocation barrier of -0.50 kcal/mol, and increased first-ion binding affinity by a factor of 1.13, primarily by decreasing the first-ion exit rate constant. The translocation rate constant was 5.62 times slower in diphytanoyl phosphatidylcholine (DPhPC) bilayers than in monoolein (GMO) bilayers (coupled for the four combinations of peptide and salt), suggesting a 44.2-mV difference in the projection of the interfacial dipole into the channel. Thus fluorination caused increased currents in DPhPC bilayers, where a high interfacial dipole potential makes translocation more rate limiting because the translocation barrier was reduced, and decreased currents in GMO bilayers, where ion exit or entry is rate limiting because these barriers were increased.

  14. Photodynamic inactivation of gramicidin channels in bilayer lipid membranes: protective efficacy of singlet oxygen quenchers depends on photosensitizer location.

    PubMed

    Rokitskaya, T I; Firsov, A M; Kotova, E A; Antonenko, Y N

    2015-06-01

    The impact of double bonds in fatty acyl tails of unsaturated lipids on the photodynamic inactivation of ion channels formed by the pentadecapeptide gramicidin A in a planar bilayer lipid membrane was studied. The presence of unsaturated acyl tails protected gramicidin A against photodynamic inactivation, with efficacy depending on the depth of a photosensitizer in the membrane. The protective effect of double bonds was maximal with membrane-embedded chlorin e6-monoethylenediamine monoamide dimethyl ester, and minimal - in the case of water-soluble tri-sulfonated aluminum phthalocyanine (AlPcS3) known to reside at the membrane surface. By contrast, the protective effect of the hydrophilic singlet oxygen scavenger ascorbate was maximal for AlPcS3 and minimal for amide of chlorin e6 dimethyl ester. The depth of photosensitizer position in the lipid bilayer was estimated from the quenching of photosensitizer fluorescence by iodide. Thus, the protective effect of a singlet oxygen scavenger against photodynamic inactivation of the membrane-inserted peptide is enhanced upon location of the photosensitizer and scavenger molecules in close vicinity to each other.

  15. Ab initio molecular dynamics study of proton transfer in a polyglycine analog of the ion channel gramicidin A.

    PubMed Central

    Sagnella, D E; Laasonen, K; Klein, M L

    1996-01-01

    Proton transfer in biological systems is thought to often proceed through hydrogen-bonded chains of water molecules. The ion channel, gramicidin A (gA), houses within its helical structure just such a chain. Using the density functional theory based ab initio molecular dynamics Car-Parrinello method, the structure and dynamics of proton diffusion through a polyglycine analog of the gA ion channel has been investigated. In the channel, a proton, which is initially present as hydronium (H3O+), rapidly forms a strong hydrogen bond with a nearest neighbor water, yielding a transient H5O2+ complex. As in bulk water, strong hydrogen bonding of this complex to a second neighbor solvation shell is required for proton transfer to occur. Within gA, this second neighbor shell included not only a channel water molecule but also a carbonyl of the channel backbone. The present calculations suggest a transport mechanism in which a priori carbonyl solvation is a requirement for proton transfer. Images FIGURE 1 FIGURE 3 PMID:8873991

  16. Investigation of Ion Channel Activities of Gramicidin A in the Presence of Ionic Liquids Using Model Cell Membranes

    PubMed Central

    Ryu, Hyunil; Lee, Hwankyu; Iwata, Seigo; Choi, Sangbaek; Ki Kim, Moon; Kim, Young-Rok; Maruta, Shinsaku; Min Kim, Sun; Jeon, Tae-Joon

    2015-01-01

    Ionic liquids (ILs) are considered to be green solvents because of their non-volatility. Although ILs are relatively safe in the atmospheric environment, they may be toxic in other environments. Our previous research showed that the cytotoxicity of ILs to biological organisms is attributable to interference with cell membranes by IL insertion. However, the effects of ILs on ion channels, which play important roles in cell homeostasis, have not been comprehensively studied to date. In this work, we studied the interactions between ILs and lipid bilayer membranes with gramicidin A ion channels. We used two methods, namely electrical and fluorescence measurements of ions that permeate the membrane. The lifetimes of channels were increased by all the ILs tested in this work via stabilizing the compressed structure of the lipid bilayer and the rate of ion flux through gA channels was decreased by changing the membrane surface charge. The former effect, which increased the rate of ion flux, was dominant at high salt concentrations, whereas the latter, which decreased the rate of ion flux, was dominant at low salt concentrations. The effects of ILs increased with increasing concentration and alkyl chain length. The experimental results were further studied using molecular dynamics simulations. PMID:26189604

  17. Investigation of Ion Channel Activities of Gramicidin A in the Presence of Ionic Liquids Using Model Cell Membranes.

    PubMed

    Ryu, Hyunil; Lee, Hwankyu; Iwata, Seigo; Choi, Sangbaek; Kim, Moon Ki; Kim, Young-Rok; Maruta, Shinsaku; Kim, Sun Min; Jeon, Tae-Joon

    2015-07-20

    Ionic liquids (ILs) are considered to be green solvents because of their non-volatility. Although ILs are relatively safe in the atmospheric environment, they may be toxic in other environments. Our previous research showed that the cytotoxicity of ILs to biological organisms is attributable to interference with cell membranes by IL insertion. However, the effects of ILs on ion channels, which play important roles in cell homeostasis, have not been comprehensively studied to date. In this work, we studied the interactions between ILs and lipid bilayer membranes with gramicidin A ion channels. We used two methods, namely electrical and fluorescence measurements of ions that permeate the membrane. The lifetimes of channels were increased by all the ILs tested in this work via stabilizing the compressed structure of the lipid bilayer and the rate of ion flux through gA channels was decreased by changing the membrane surface charge. The former effect, which increased the rate of ion flux, was dominant at high salt concentrations, whereas the latter, which decreased the rate of ion flux, was dominant at low salt concentrations. The effects of ILs increased with increasing concentration and alkyl chain length. The experimental results were further studied using molecular dynamics simulations.

  18. The membrane interface dictates different anchor roles for "inner pair" and "outer pair" tryptophan indole rings in gramicidin A channels.

    PubMed

    Gu, Hong; Lum, Kevin; Kim, Jung H; Greathouse, Denise V; Andersen, Olaf S; Koeppe, Roger E

    2011-06-01

    We investigated the effects of substituting two of the four tryptophans (the "inner pair" Trp(9) and Trp(11) or the "outer pair" Trp(13) and Trp(15)) in gramicidin A (gA) channels. The conformational preferences of the doubly substituted gA analogues were assessed using circular dichroism spectroscopy and size-exclusion chromatography, which show that the inner tryptophans 9 and 11 are critical for the gA's conformational preference in lipid bilayer membranes. [Phe(13,15)]gA largely retains the single-stranded helical channel structure, whereas [Phe(9,11)]gA exists primarily as double-stranded conformers. Within this context, the (2)H NMR spectra from labeled tryptophans were used to examine the changes in average indole ring orientations, induced by the Phe substitutions and by the shift in conformational preference. Using a method for deuterium labeling of already synthesized gAs, we introduced deuterium selectively onto positions C2 and C5 of the remaining tryptophan indole rings in the substituted gA analogues for solid-state (2)H NMR spectroscopy. The (least possible) changes in orientation and overall motion of each indole ring were estimated from the experimental spectra. Regardless of the mixture of backbone folds, the indole ring orientations observed in the analogues are similar to those found previously for gA channels. Both Phe-substituted analogues form single-stranded channels, as judged from the formation of heterodimeric channels with the native gA. [Phe(13,15)]gA channels have Na(+) currents that are ~50% and lifetimes that are ~80% of those of native gA channels. The double-stranded conformer(s) of [Phe(9,11)]gA do not form detectable channels. The minor single-stranded population of [Phe(9,11)]gA forms channels with Na(+) currents that are ~25% and single-channel lifetimes that are ~300% of those of native gA channels. Our results suggest that Trp(9) and Trp(11), when "reaching" for the interface, tend to drive both monomer folding (to "open" a

  19. Modulation of proton transfer in the water wire of dioxolane-linked gramicidin channels by lipid membranes.

    PubMed

    de Godoy, C M; Cukierman, S

    2001-09-01

    Proton conductance (g(H)) in single SS stereoisomers of dioxolane-linked gramicidin A (gA) channels were measured in different phospholipid bilayers at different HCl concentrations. In particular, measurements were obtained in bilayers made of 1,2-diphytanoyl 3-phosphocholine (DiPhPC) or its ethylated derivative 1,2-diphytanoyl 3-ethyl-phosphocholine (et-DiPhPC,). The difference between these phospholipids is that in et-DiPhPC one of the phosphate oxygens is covalently linked to an ethyl group and cannot be protonated. In relatively dilute acid solutions, g(H) in DiPhPC is significantly higher than in et-DiPhPC. At high acid concentrations, g(H) is the same in both diphytanoyl bilayers. Such differences in g(H) can be accounted for by surface charge effects at the membrane/solution interfaces. In the linear portion of the log g(H)-log [H] relationship, g(H) values in diphytanoyl bilayers were significantly larger (approximately 10-fold) than in neutral glyceryl monooleate (GMO) membranes. The slopes of the linear log-log relationships between g(H) and [H] in diphytanoyl and GMO bilayers are essentially the same (approximately 0.76). This slope is significantly lower than the slope of the log-log plot of proton conductivity versus proton concentration in aqueous solutions (approximately 1.00). Because the chemical composition of the membrane-channel/solution interface is strikingly different in GMO and diphytanoyl bilayers, the reduced slope in g(H)-[HCl] relationships may be a characteristic of proton transfer in the water wire inside the SS channel. Values of g(H) in diphytanoyl bilayers were also significantly larger than in membranes made of the more common biological phospholipids 1-palmitoyl 2-oleoyl phosphocholine (POPC) or 1-palmitoyl 2-oleoyl phosphoethanolamine (POPE). These differences, however, cannot be accounted for by different surface charge effects or by different internal dipole potentials. On the other hand, maximum g(H) measured in the SS channel

  20. Highly sensitive and rapid assay of substance P and streptolysin O in human serum using immuno-liposomes and gramicidin channels.

    PubMed

    Sakamoto, Misato; Shoji, Atsushi; Sugawara, Masao

    2013-01-01

    A highly sensitive and rapid method for the determination of substance P (SP) and streptolysin O (SLO) in human serum is described. The assay is based on enriching the analyte by agglutination/precipitation of immuno-liposomes and enhancing the fluorescence intensity by gramicidin A channels. A mixture of the immuno-liposomes encapsulating a pH-sensitive fluorescence dye BCECF, gramicidin A and a given concentration of SP (or SLO) is preincubated in a solution and captured on anti-SP (or anti-SLO)-modified cover slips, followed by measuring fluorescence images after removing excess liposomes. The method allowed quantifying SP and SLO in the range from sub-pg mL(-1) to pg mL(-1), with detection limits of 0.32 pg mL(-1) and 8 fg mL(-1), respectively. The present method could determine SP and SLO in 50-125 times diluted human serum without any extraction steps. The assay can be completed within 60 min.

  1. A semi-microscopic Monte Carlo study of permeation energetics in a gramicidin-like channel: the origin of cation selectivity.

    PubMed Central

    Dorman, V; Partenskii, M B; Jordan, P C

    1996-01-01

    The influence of a gramicidin-like channel former on ion free energy barriers is studied using Monte Carlo simulation. The model explicitly describes the ion, the water dipoles, and the peptide carbonyls; the remaining degrees of freedom, bulk electrolyte, non-polar lipid and peptide regions, and electronic (high frequency) permittivity, are treated in continuum terms. Contributions of the channel waters and peptide COs are studied both separately and collectively. We found that if constrained to their original orientations, the COs substantially increase the cationic permeation free energy; with or without water present, CO reorientation is crucial for ion-CO interaction to lower cation free energy barriers; the translocation free energy profiles for potassium-, rubidium-, and cesium-like cations exhibit no broad barriers; the lipid-bound peptide interacts more effectively with anions than cations; anionic translocation free energy profiles exhibit well defined maxima. Using experimental data to estimate transfer free energies of ions and water from bulk electrolyte to a non-polar dielectric (continuum lipid), we found reasonable ion permeation profiles; cations bind and permeate, whereas anions cannot enter the channel. Cation selectivity arises because, for ions of the same size and charge, anions bind hydration water more strongly. PMID:8770192

  2. Gramicidin A directly inhibits mammalian Na(+)/K (+)-ATPase.

    PubMed

    Takada, Yohei; Matsuo, Kentaro; Kataoka, Takao

    2008-12-01

    The linear pentadecapeptide gramicidin A forms an ion channel in the lipid bilayer to selectively transport monovalent cations. Nevertheless, we have surprisingly found that gramicidin A directly inhibits mammalian Na(+)/K(+)-ATPase. Gramicidin A inhibited ATP hydrolysis by Na(+)/K(+)-ATPase from porcine cerebral cortex at the IC(50) value of 8.1 microM, while gramicidin S was approximately fivefold less active. The synthetic gramicidin A analog lacking N-terminal formylation and C-terminal ethanolamine exhibited a weaker inhibitory effect on the ATP-hydrolyzing activity of Na(+)/K(+)-ATPase than gramicidin A, indicating that these end modifications are necessary for gramicidin A to inhibit Na(+)/K(+)-ATPase activity. Moreover, Lineweaver-Burk analysis showed that gramicidin A exhibits a mixed type of inhibition. In addition to the most well-studied ionophore activity, our present study has disclosed a novel biological function of gramicidin A as a direct inhibitor of mammalian Na(+)/K(+)-ATPase activity.

  3. Thallium-205 NMR determination of the thermodynamic parameters for the binding of monovalent and divalent cations to gramicidin A and C incorporated into model phospholipid membranes: An equilibrium study

    SciTech Connect

    Fernandez, J.Q.

    1987-01-01

    Thermodynamic parameters have been determined for the binding of monovalent and divalent cations to gramicidin A and C incorporated in lyso-{alpha}-phosphatidylcholine dispersions. The thermodynamic analyses used equilibrium constants derived from the analysis of Tl-205 NMR chemical shifts using a one-site binding model of the gramicidin channel. Initial experiments determined the thermodynamic parameters of the Tl{sup +} ion binding to gramicidin A. Next, the competitive binding-Tl-205 NMR technique was used to obtain the thermodynamic parameters for the binding of Li{sup +}, Na{sup +}, K{sup +}, Rb{sup +}, Cs{sup +}, NH{sub 4}{sup +} and Ag{sup +} cations to gramicidin A. A similar experiment for the binding of the Li{sup +}, Na{sup +}, K{sup +} and Rb{sup +} cations to gramicidin C was performed. Similar experiments determined the thermodynamic parameter for the Cd{sup ++}, Mg{sup ++}, Ca{sup ++}, Sr{sup ++} and Ba{sup ++} cations. Thermodynamic parameters have also been determined for the binding of the Tl{sup +} ion to gramicidin A incorporated into dimyristoylphosphatidylcholine vesicles using Tl-205 NMR spectroscopy.

  4. The gramicidin channel ion permeation free-energy profile: direct and indirect effects of CHARMM force field improvements.

    PubMed

    Mustafa, Morad; Busath, David D

    2009-06-01

    A revised CHARMM force field for tryptophan residues is studied as well as a new grid-based correction algorithm, called CMAP, using molecular dynamics simulations of gramicidin A (1JNO) embedded in a lipid bilayer (DMPC) with 1 mol/kg NaCl or KCl saline solution. The conformational stability of the interfacial side chains is studied, which shows good stability on the 10 ns time scale. The revised force field for the tryptophan side chain produces, in the decomposition, a Na(+) PMF(Trp) profile that is consonant with the prediction from the experimental results, analyzed with rate theory by Durrant et al. (2006), but in stark contrast to the prediction of the original CHARMM force field, version 22. However, the effect is diluted in the PMF profile due to indirect effects mediated by other components of the system (polypeptide, lipid molecules, ions, and water molecules). CMAP corrections to the L-amino acids help reduce the excessive translocation barrier. Decomposition demonstrates that this effect is due to effects on the K(+) PMF(H(2)O) profile rather than on the K(+) PMF(gA) profile. The results have been confirmed to be robust using an alternative umbrella-potential method. Further force field balancing efforts (direct and indirect) are required for future studies to evaluate whether these effects give rise to predictions that are consistent with those observables extracted from real experiments.

  5. Channel waveguides on RbTiOPO4 by Cs+ ion exchange.

    PubMed

    Cugat, J; Solé, R; Carvajal, J J; Mateos, X; Massons, J; Lifante, G; Díaz, F; Aguiló, M

    2013-02-01

    In this Letter we report Cs(+) ion exchange channel waveguides on RbTiOPO(4) (RTP) for what we believe is the first time. A Ti channel mask was fabricated on an RTP substrate by conventional photolithography. The ion exchange process was carried out in a CsNO(3) melt, and the channels produced ranged from 6 to 11 μm in width. The near-field pattern of the modes was recorded, and type II second harmonic generation in waveguide regime was obtained, producing 512.5 nm green light. The optical characterization shows optical losses of 3.8 dB/cm.

  6. Spectroscopic studies of the interactions of the pyrethroid insecticide fenvalerate with gramicidin.

    PubMed

    Ghosh, J K; Sarkar, S N; Sikdar, S K

    1998-05-01

    Fenvalerate is a pyrethroid insecticide which interacts with ionic channels. Using circular dichroism technique we have studied the interaction of fenvalerate with gramicidin, a model channel peptide which transports ions. In most organic solvents, gramicidin exists as a double helix except in trifluoroethanol where it exists as a channel forming single stranded beta6.3 helical monomer. In model lipid membranes, under certain experimental conditions, gramicidin exists as a channel forming single stranded beta6.3 helical dimer. Our results show that fenvalerate interacts more with the single stranded beta6.3 helical monomer or dimer than with the double helical form of gramicidin. This was further confirmed by an increase in the rate of gramicidin mediated proton transport in liposomes by fenvalerate, using the pH sensitive fluorophore, pyranine.

  7. Preparation and properties of O-dansyltyrosine gramicidin C.

    PubMed

    Veatch, W R; Blout, E R

    1976-07-13

    Gramicidins A, B, and C are a family of poly-peptide antibiotics which facilitate the passive diffusion of alkali cations and protons through lipid bilayer membranes. It is clear that gramicidin forms a multimeric transmembrane channel and it has been suggested that the channel is an io-conducting dimer in equilibrium on the membrane with non-conducting monomer. We describe the preparation and purification of a derivative of gramicidin C in which the phenolic hydroxyl of the tyrosine at position 11 has been esterified to 8-dimethylaminonaphthalene-1-sulfonate (dansyl). This derivative fluoresces strongly in the visible with an emission maximun in dioxane of 530 nm, an emission lifetime of 16 ns, and a quantum yield of 0.8. Veatch et al. ((1975),J. Mol. Biol. 99, 75) have shown this 0-dansyltyrosine gamicidin C to be a fully active analogue of gramicidin A in artificial lipid bilayer membranes. We here utilize this derivative to further characterize the state of aggregation and rotational mobility of the four interconvertible conformational species formed by gramicidin in nonpolar organic solvents (Veatch et al. (1974), Biochemsitry 13, 5249; Veatch and Blout (1974), Biochemistry 13, 5257). Fluorescence energy transfer from the tryptophans of gramicidin A to the 0-dansyltyrosine of this derivatives supports the conclusion that all of these gramicidin isolated species are aggregates. Decay of fluorescence polarization anisotropy measurements yield a rotational correlation time of 1 ns for the 0-dansyltyrosine chromophore in ethanol in good agreement with the more detailed information previously obtained by 13C-nuclear magnetic resonance for the monomer in dimethyl sulfoxide (Fossel et al. (1974), Biochemistry 13, 5264). However, it is likely that the chromophore has much more rotational mobility than the rest of the gramicidin molecule in the aggregated comformational states. PMID:60127

  8. Gramicidin Induce Local Non-Uniform Distribution of Lipids in Multi-Component Membrane Domains

    NASA Astrophysics Data System (ADS)

    Mao, Yu; Hussain, Fazle; Huang, Juyang

    2015-03-01

    In lipid membranes, gramicidin form trans-membrane channels that are specific for monovalent cations. We performed Molecular Dynamics simulations of gramicidin in coexisting liquid-ordered (Lo) and liquid disordered (Ld) domains using GROMACS. The lipid compositions of Lo and Ld domains are DOPC/DSPC/Cholesterol = 6.5/52.6/40.9 and 74.4/10.6/15, respectively. In the Ld domain, the membrane thickness matches the hydrophobic length of gramicidin quite well, and water molecules can diffuse through the gramicidin channels. However, in the Lo lipid domain, the bilayer thickness is far greater than the hydrophobic length of gramicidin and majority of gramicidin do not form conducting channel. The simulation result explained our experimental finding that gramicidin partition favorably into the Ld domains. The calculated radial distribution functions of lipids indicate that gramicidin recruit a layer of short DOPC surrounding each protein and keep cholesterol and taller DSPC away from the protein-bilayer interface. Our result indicates that membrane proteins are capable of inducing non-uniform distributions of lipids and creating a local bilayer environment, which favors protein function.

  9. CSI feedback-based CS for underwater acoustic adaptive modulation OFDM system with channel prediction

    NASA Astrophysics Data System (ADS)

    Kuai, Xiao-yan; Sun, Hai-xin; Qi, Jie; Cheng, En; Xu, Xiao-ka; Guo, Yu-hui; Chen, You-gan

    2014-06-01

    In this paper, we investigate the performance of adaptive modulation (AM) orthogonal frequency division multiplexing (OFDM) system in underwater acoustic (UWA) communications. The aim is to solve the problem of large feedback overhead for channel state information (CSI) in every subcarrier. A novel CSI feedback scheme is proposed based on the theory of compressed sensing (CS). We propose a feedback from the receiver that only feedback the sparse channel parameters. Additionally, prediction of the channel state is proposed every several symbols to realize the AM in practice. We describe a linear channel prediction algorithm which is used in adaptive transmission. This system has been tested in the real underwater acoustic channel. The linear channel prediction makes the AM transmission techniques more feasible for acoustic channel communications. The simulation and experiment show that significant improvements can be obtained both in bit error rate (BER) and throughput in the AM scheme compared with the fixed Quadrature Phase Shift Keying (QPSK) modulation scheme. Moreover, the performance with standard CS outperforms the Discrete Cosine Transform (DCT) method.

  10. Hydrogen-bonded helical hydrazide oligomers and polymer that mimic the ion transport of gramicidin A.

    PubMed

    Xin, Pengyang; Zhu, Pingping; Su, Pei; Hou, Jun-Li; Li, Zhan-Ting

    2014-09-24

    A new series of hydrogen-bonded helical aromatic hydrazide oligomers and polymer that bear phenylalanine tripeptide chains have been designed and synthesized. It was revealed that the helical structures could insert into lipid bilayers to form unimolecular channels. The longest oligomeric and polymeric helical channels exhibited an NH4(+)/K(+) selectivity that was higher than that of natural gramicidin A, whereas the transport of a short helical channel for Tl(+) could achieve an efficiency as high as that of gramicidin A.

  11. Enhanced Eryptosis Following Gramicidin Exposure

    PubMed Central

    Malik, Abaid; Bissinger, Rosi; Liu, Guoxing; Liu, Guilai; Lang, Florian

    2015-01-01

    The peptide antibiotic and ionophore gramicidin has previously been shown to trigger apoptosis of nucleated cells. In analogy to apoptosis, the suicidal death of erythrocytes or eryptosis involves cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress, increase of cytosolic Ca2+ activity ([Ca2+]i), and ceramide. The present study explored, whether gramicidin triggers eryptosis. To this end phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, red blood cell distribution width (RDW) from electronic particle counting, reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, [Ca2+]i from Fluo3- and Fluo4 fluorescence, and ceramide abundance from binding of specific antibodies. As a result, a 24 h exposure of human erythrocytes to gramicidin significantly increased the percentage of annexin-V-binding cells (≥1 µg/mL), forward scatter (≥0.5 µg/mL) and hemolysis. Gramicidin enhanced ROS activity, [Ca2+]i and ceramide abundance at the erythrocyte surface. The stimulation of annexin-V-binding by gramicidin was significantly blunted but not abolished by removal of extracellular Ca2+. In conclusion, gramicidin stimulates phospholipid scrambling of the erythrocyte cell membrane, an effect at least partially due to induction of oxidative stress, increase of [Ca2+]i and up-regulation of ceramide abundance. Despite increase of [Ca2+]i, gramicidin increases cell volume and slightly reduces RWD. PMID:25915718

  12. Enhanced eryptosis following gramicidin exposure.

    PubMed

    Malik, Abaid; Bissinger, Rosi; Liu, Guoxing; Liu, Guilai; Lang, Florian

    2015-04-23

    The peptide antibiotic and ionophore gramicidin has previously been shown to trigger apoptosis of nucleated cells. In analogy to apoptosis, the suicidal death of erythrocytes or eryptosis involves cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress, increase of cytosolic Ca2+ activity ([Ca2+]i), and ceramide. The present study explored, whether gramicidin triggers eryptosis. To this end phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, red blood cell distribution width (RDW) from electronic particle counting, reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, [Ca2+]i from Fluo3- and Fluo4 fluorescence, and ceramide abundance from binding of specific antibodies. As a result, a 24 h exposure of human erythrocytes to gramicidin significantly increased the percentage of annexin-V-binding cells (≥1 µg/mL), forward scatter (≥0.5 µg/mL) and hemolysis. Gramicidin enhanced ROS activity, [Ca2+]i and ceramide abundance at the erythrocyte surface. The stimulation of annexin-V-binding by gramicidin was significantly blunted but not abolished by removal of extracellular Ca2+. In conclusion, gramicidin stimulates phospholipid scrambling of the erythrocyte cell membrane, an effect at least partially due to induction of oxidative stress, increase of [Ca2+]i and up-regulation of ceramide abundance. Despite increase of [Ca2+]i, gramicidin increases cell volume and slightly reduces RWD.

  13. The first crystal structure of a gramicidin complex with sodium: high-resolution study of a nonstoichiometric gramicidin D-NaI complex.

    PubMed

    Olczak, A; Główka, M L; Szczesio, M; Bojarska, J; Wawrzak, Z; Duax, W L

    2010-08-01

    The crystal structure of the nonstoichiometric complex of gramicidin D with NaI has been studied using synchrotron radiation at 100 K. The limiting resolution was 1.25 A and the R factor was 16% for 19 883 observed reflections. The general architecture of the antiparallel two-stranded gramicidin dimers in the studied crystal was a right-handed antiparallel double-stranded form that closely resembles the structures of other right-handed species published to date. However, there were several surprising observations. In addition to the significantly different composition of linear gramicidins identified in the crystal structure, including the absence of the gramicidin C form, only two cationic sites were found in each of the two independent dimers (channels), which were partially occupied by sodium, compared with the seven sites found in the RbCl complex of gramicidin. The sum of the partial occupancies of Na(+) was only 1.26 per two dimers and was confirmed by the similar content of iodine ions (1.21 ions distributed over seven sites), which was easily visible from their anomalous signal. Another surprising observation was the significant asymmetry of the distributions and occupancies of cations in the gramicidin dimers, which was in contrast to those observed in the high-resolution structures of the complexes of heavier alkali metals with gramicidin D, especially that of rubidium.

  14. The first crystal structure of a gramicidin complex with sodium: high-resolution study of a nonstoichiometric gramicidin D-NaI complex

    SciTech Connect

    Olczak, A.; Glówka, M.L.; Szczesio, M.; Bojarska, J.; Wawrzak, Z.; Duax, W.L.

    2010-11-15

    The crystal structure of the nonstoichiometric complex of gramicidin D with NaI has been studied using synchrotron radiation at 100 K. The limiting resolution was 1.25 {angstrom} and the R factor was 16% for 19,883 observed reflections. The general architecture of the antiparallel two-stranded gramicidin dimers in the studied crystal was a right-handed antiparallel double-stranded form that closely resembles the structures of other right-handed species published to date. However, there were several surprising observations. In addition to the significantly different composition of linear gramicidins identified in the crystal structure, including the absence of the gramicidin C form, only two cationic sites were found in each of the two independent dimers (channels), which were partially occupied by sodium, compared with the seven sites found in the RbCl complex of gramicidin. The sum of the partial occupancies of Na{sup +} was only 1.26 per two dimers and was confirmed by the similar content of iodine ions (1.21 ions distributed over seven sites), which was easily visible from their anomalous signal. Another surprising observation was the significant asymmetry of the distributions and occupancies of cations in the gramicidin dimers, which was in contrast to those observed in the high-resolution structures of the complexes of heavier alkali metals with gramicidin D, especially that of rubidium.

  15. Monitoring Gramicidin Conformations in Membranes: A Fluorescence Approach

    PubMed Central

    Rawat, Satinder S.; Kelkar, Devaki A.; Chattopadhyay, Amitabha

    2004-01-01

    We have monitored the membrane-bound channel and nonchannel conformations of gramicidin utilizing red-edge excitation shift (REES), and related fluorescence parameters. In particular, we have used fluorescence lifetime, polarization, quenching, chemical modification, and membrane penetration depth analysis in addition to REES measurements to distinguish these two conformations. Our results show that REES of gramicidin tryptophans can be effectively used to distinguish conformations of membrane-bound gramicidin. The interfacially localized tryptophans in the channel conformation display REES of 7 nm whereas the tryptophans in the nonchannel conformation exhibit REES of 2 nm which highlights the difference in their average environments in terms of localization in the membrane. This is supported by tryptophan penetration depth measurements using the parallax method and fluorescence lifetime and polarization measurements. Further differences in the average tryptophan microenvironments in the two conformations are brought out by fluorescence quenching experiments using acrylamide and chemical modification of the tryptophans by N-bromosuccinimide. In summary, we report novel fluorescence-based approaches to monitor conformations of this important ion channel peptide. Our results offer vital information on the organization and dynamics of the functionally important tryptophan residues in gramicidin. PMID:15298892

  16. Unsaturated lipids protect the integral membrane peptide gramicidin A from singlet oxygen.

    PubMed

    Rokitskaya, Tatyana I; Kotova, Elena A; Agapov, Igor I; Moisenovich, Mikhail M; Antonenko, Yuri N

    2014-05-01

    In contrast to expectations that unsaturated fatty acids contribute to oxidative stress by providing a source of lipid peroxides, we demonstrated the protective effect of double bonds in lipids on oxidative damage to membrane proteins. Photodynamic inactivation of gramicidin channels was decreased in unsaturated lipid compared to saturated lipid bilayers. By estimating photosensitizer (boronated chlorine e6 amide) binding to the membrane with the current relaxation technique, the decrease in gramicidin photoinactivation was attributed to singlet oxygen scavenging by double bonds in lipids rather than to the reduction in photosensitizer binding. Gramicidin protection by unsaturated lipids was also observed upon induction of oxidative stress with tert-butyl hydroperoxide.

  17. Stimulation of cation transport in mitochondria by gramicidin and truncated derivatives

    SciTech Connect

    Rottenberg, H.; Koeppe, R.E. II )

    1989-05-16

    Gramicidin and the truncated derivatives desformylgramicidin (desfor) and des(formylvalyl)gramicidin (desval) stimulate monovalent cation transport in rat liver mitochondria. Cation fluxes were compared indirectly from the effect of cations on the membrane potential at steady state (state 4) or from the associated stimulation of electron transport. Rb{sup +} transport was measured directly from the uptake of {sup 86}Rb. The truncated gramicidins show enhanced selectivity for K{sup +} and Rb{sup +} when compared to gramicidin. Moreover, the pattern of selectivity within the alkali cation series is altered. The cation fluxes through the truncated derivatives are more strongly dependent on the cation concentration. The presence of high concentrations of permeating cation enhances the transport of other cations through the truncated derivative channels, suggesting that cations are required for stabilizing the channel structure. In high concentrations of KCl, desfor and desval are nearly as effective as gramicidin in collapsing the mitochondrial membrane potential, and consequently, in the uncoupling of oxidative phosphorylation and enhancement of ATP hydrolysis. Preliminary experiments with liposomes show that {sup 86}Rb exchange is stimulated by desfor and desval almost to the same extend at gramicidin. These results strongly suggest that the truncated gramicidins form a novel conducting channel which differs from the gramicidin head-to-head, single-stranded {beta}{sup 6.3}-helical dimer (channel) in its conductance characteristic and its structure. On the basis of the secondary structure of the truncated derivatives, the authors suggest that the antiparallel double-stranded helix dimer (pore) is a likely alternative structure for this novel channel.

  18. Gramicidin conformational studies with mixed-chain unsaturated phospholipid bilayer systems

    SciTech Connect

    Cox, K.J.; Ho, Cojen; Lombardi, J.V.; Stubbs, C.D. )

    1992-02-04

    The transition of gramicidin from a nonchannel to a channel form was investigated using mixed-chain phosphatidylcholine lipid bilayers. Gramicidin and phospholipids were codispersed, after removal of the solvents chloroform/methanol or trifluoroethanol which resulted in nonchannel and channel conformations, respectively, as confirmed using circular dichroism (CD). The fluorescence emission maxima of the nonchannel form were shifted toward shorter wavelengths by heating at 60C (for 0-12 h), which converted it to a channel form, again as confirmed by CD. The channel form did not respond to heat treatment. Heat treatment also increased the fluorescence anisotropy of the nonchannel gramicidin tryptophans. The rate of transition from the nonchannel to channel conformation was found to be faster is phosphatidylethanolamine was present in combination with phosphatidylcholine compared to phosphatidylcholine alone. Using the fluorescence anisotropy of the membrane lipid probe, 1,6-diphenyl-1,3,5-hexatriene, it was also shown that the motional properties of the surrounding lipid acyl chains differed for the channel and nonchannel gramicidin conformations. The possibility that lipids tending to favor the hexagonal phase (H{sub II}) would enhance the rate of the nonchannel to channel transition was supported by {sup 31}P NMR which revealed the presence of some H{sub II} lipids in the channel preparations. The results of this study suggest that gramicidin may serve as a useful model for similar conformational transitions in other more complex membrane proteins.

  19. Sedimentation rates measurements in former channels of the upper Rhône river using Chernobyl 137Cs and 134Cs as tracers.

    PubMed

    Rostan, J C; Juget, J; Brun, A M

    1997-01-30

    Former river channels are aquatic ecosystems with a different geomorphology generated by fluvial dynamics more or less linked to the main channel. They present different ecological successions to become terrestrial ecosystems and are thus supposed to have different sedimentation rates. The aim of this paper is to assess this sedimentation rate using radioactive tracer methodology commonly used in lake studies. Chernobyl impacts, expressed in 137Cs concentration and 137Cs/134Cs ratio, were determined in sediment cores. Sites (21) were sampled in the alluvial plain of the Upper Rhône River from 1989 to 1994. The contamination presented a high spatial heterogeneity. The maximum values encountered by site ranged between 34 and 541 Bq/kg of dry matter. The method generally gave good core profiles. Sedimentation rate ranged between 0.14 and 0.70 cm/year for the former meanders and between 0.14 and 2.86 cm/year for the braided channels. The sediment accumulation rates ranged from 0.03 to 0.25 g/cm2 per year and 0.03 to 2.26 g/cm2 per year, respectively. These values are similar to those found for Lake Geneva. The importance of the former channels in relation to the main channel is enhanced by the higher contamination and radionuclides retention. The sediment accumulation rate is related to the organic carbon content in the sediment. A comparison between two former channels with different productivity showed that the the allogenous driven system presents a high organic sediment accumulation rate with a low organic content in the sediment and inversely, a low organic sediment accumulation rate with a high organic carbon content was found for the autogenous drive system.

  20. Dielectric boundary force and its crucial role in gramicidin

    NASA Astrophysics Data System (ADS)

    Nadler, Boaz; Hollerbach, Uwe; Eisenberg, R. S.

    2003-08-01

    In an electrostatic problem with nonuniform geometry, a charge Q in one region induces surface charges [called dielectric boundary charges (DBC)] at boundaries between different dielectrics. These induced surface charges, in return, exert a force [called dielectric boundary force (DBF)] on the charge Q that induced them. The DBF is often overlooked. It is not present in standard continuum theories of (point) ions in or near membranes and proteins, such as Gouy-Chapman, Debye-Huckel, Poisson-Boltzmann or Poisson-Nernst- Planck. The DBF is important when a charge Q is near dielectric interfaces, for example, when ions permeate through protein channels embedded in biological membranes. In this paper, we define the DBF and calculate it explicitly for a planar dielectric wall and for a tunnel geometry resembling the ionic channel gramicidin. In general, we formulate the DBF in a form useful for continuum theories, namely, as a solution of a partial differential equation with boundary conditions. The DBF plays a crucial role in the permeation of ions through the gramicidin channel. A positive ion in the channel produces a DBF of opposite sign to that of the fixed charge force (FCF) produced by the permanent charge of the gramicidin polypeptide, and so the net force on the positive ion is reduced. A negative ion creates a DBF of the same sign as the FCF and so the net (repulsive) force on the negative ion is increased. Thus, a positive ion can permeate the channel, while a negative ion is excluded from it. In gramicidin, it is this balance between the FCF and DBF that allows only singly charged positive ions to move into and through the channel. The DBF is not directly responsible, however, for selectivity between the alkali metal ions (e.g., Li+, Na+, K+): we prove that the DBF on a mobile spherical ion is independent of the ion’s radius.

  1. Carbon nanotube as a gramicidin analogue

    NASA Astrophysics Data System (ADS)

    Hilder, Tamsyn A.; Chung, Shin-Ho

    2011-01-01

    We have designed a carbon nanotube that is selectively permeable to monovalent cations, binds divalent cations and rejects anions. The nanotubes, with an effective radius of 4.53 Å and length of 36 Å, are terminated with hydrogen atoms and are exohydrogenated in two regions near the entrance and exit. Using molecular and stochastic dynamics simulations we examine the free energy, current-voltage-concentration profiles and ion binding sites. The characteristics of this channel are comparable to the antibiotic gramicidin-A, but the potassium current is six times larger. At 40 mM calcium concentration the current is reduced from 26 pA to 4 pA due to a calcium ion binding at the channel entrance.

  2. Gramicidin A blocks tumor growth and angiogenesis through inhibition of hypoxia-inducible factor in renal cell carcinoma.

    PubMed

    David, Justin M; Owens, Tori A; Inge, Landon J; Bremner, Ross M; Rajasekaran, Ayyappan K

    2014-04-01

    Ionophores are hydrophobic organic molecules that disrupt cellular transmembrane potential by permeabilizing membranes to specific ions. Gramicidin A is a channel-forming ionophore that forms a hydrophilic membrane pore that permits the rapid passage of monovalent cations. Previously, we found that gramicidin A induces cellular energy stress and cell death in renal cell carcinoma (RCC) cell lines. RCC is a therapy-resistant cancer that is characterized by constitutive activation of the transcription factor hypoxia-inducible factor (HIF). Here, we demonstrate that gramicidin A inhibits HIF in RCC cells. We found that gramicidin A destabilized HIF-1α and HIF-2α proteins in both normoxic and hypoxic conditions, which in turn diminished HIF transcriptional activity and the expression of various hypoxia-response genes. Mechanistic examination revealed that gramicidin A accelerates O(2)-dependent downregulation of HIF by upregulating the expression of the von Hippel-Lindau (VHL) tumor suppressor protein, which targets hydroxylated HIF for proteasomal degradation. Furthermore, gramicidin A reduced the growth of human RCC xenograft tumors without causing significant toxicity in mice. Gramicidin A-treated tumors also displayed physiologic and molecular features consistent with the inhibition of HIF-dependent angiogenesis. Taken together, these results demonstrate a new role for gramicidin A as a potent inhibitor of HIF that reduces tumor growth and angiogenesis in VHL-expressing RCC.

  3. Gramicidin A induces metabolic dysfunction and energy depletion leading to cell death in renal cell carcinoma cells.

    PubMed

    David, Justin M; Owens, Tori A; Barwe, Sonali P; Rajasekaran, Ayyappan K

    2013-11-01

    Ionophores are lipid-soluble organic molecules that disrupt cellular transmembrane potential by rendering biologic membranes permeable to specific ions. They include mobile-carriers that complex with metal cations and channel-formers that insert into the membrane to form hydrophilic pores. Although mobile-carriers possess anticancer properties, investigations on channel-formers are limited. Here, we used the channel-forming ionophore gramicidin A to study its effects on the growth and survival of renal cell carcinoma (RCC) cells. RCC is a histologically heterogeneous malignancy that is highly resistant to conventional treatments. We found that gramicidin A reduced the in vitro viability of several RCC cell lines at submicromolar concentrations (all IC50 < 1.0 μmol/L). Gramicidin A exhibited similar toxicity in RCC cells regardless of histologic subtype or the expression of either the von Hippel-Lindau tumor suppressor gene or its downstream target, hypoxia-inducible factor-1α. Gramicidin A decreased cell viability equal to or greater than the mobile-carrier monensin depending on the cell line. Mechanistic examination revealed that gramicidin A blocks ATP generation by inhibiting oxidative phosphorylation and glycolysis, leading to cellular energy depletion and nonapoptotic cell death. Finally, gramicidin A effectively reduced the growth of RCC tumor xenografts in vivo. These results show a novel application of gramicidin A as a potential therapeutic agent for RCC therapy.

  4. Free energy calculations of gramicidin dimer dissociation.

    PubMed

    Wanasundara, Surajith N; Krishnamurthy, Vikram; Chung, Shin-Ho

    2011-11-24

    Molecular dynamics simulations, combined with umbrella sampling, is used to study how gramicidin A (gA) dimers dissociate in the lipid bilayer. The potential of mean force and intermolecular potential energy are computed as functions of the distance between center of masses of the two gA monomers in two directions of separation: parallel to the bilayer surface and parallel to the membrane normal. Results from this study show that the dissociation of gA dimers occurs via lateral displacement of gA monomers followed by tilting of dimers with respect to the lipid bilayer normal. It is found that the dissociation energy of gA dimers in the dimyristoylphosphatidylcholine bilayer is 14 kcal mol(-1) (~22 kT), which is approximately equal to the energy of breaking six intermolecular hydrogen bonds that stabilize the gA channel dimer.

  5. Effect of gramicidin A on the dipole potential of phospholipid membranes.

    PubMed Central

    Shapovalov, V L; Kotova, E A; Rokitskaya, T I; Antonenko, Y N

    1999-01-01

    The effect of channel-forming peptide gramicidin A on the dipole potential of phospholipid monolayers and bilayers has been studied. Surface pressure and surface potential isotherms of monolayers have been measured with a Langmuir trough equipped with a Wilhelmy balance and a surface potential meter (Kelvin probe). Gramicidin has been shown to shift pressure-area isotherms of phospholipids and to reduce their monolayer surface potentials. Both effects increase with the increase in gramicidin concentration and depend on the kind of phosphatidylcholine used. Application of the dual-wavelength ratiometric fluorescence method using the potential-sensitive dye RH421 has revealed that the addition of gramicidin A to dipalmitoylphosphatidylcholine liposomes leads to a decrease in the fluorescence ratio of RH421. This is similar to the effect of phloretin, which is known to decrease the dipole potential. The comparison of the concentration dependences of the fluorescence ratio for gramicidin and phloretin shows that gramicidin is as potent as phloretin in modifying the membrane dipole potential. PMID:10388758

  6. Molecular dynamics simulations of calcium binding in gramicidin A

    NASA Astrophysics Data System (ADS)

    Baştuğ, Turgut; Kuyucak, Serdar

    2006-06-01

    An important issue in molecular dynamics (MD) simulations of biomolecules is whether membrane proteins can be described using nonpolarizable force fields. To shed further light into this question, we study calcium ion binding and blocking of the gramicidin A channel which has not been investigated in MD simulations before. Potential of mean force calculations for calcium and potassium ions using a nonpolarizable force field reveal that calcium binding to the channel is much weaker compared to potassium, and hence calcium block of potassium current cannot be described. Inclusion of polarization interaction in force fields may help to rectify this problem.

  7. TI-205 nuclear magnetic resonance determination of the thermodynamic parameters for the binding of monovalent cations to gramicidins A and C.

    PubMed Central

    Hinton, J F; Fernandez, J Q; Shungu, D C; Whaley, W L; Koeppe, R E; Millett, F S

    1988-01-01

    Thermodynamic parameters for the binding of the monovalent cations, Li+, Na+, K+, Rb+, Cs+, NH4+, TI+, and Ag+, to gramicidin A and for the binding of TI+ to gramicidin C, incorporated into lysophosphatidylcholine, have been determined using a combination of TI-205 nuclear magnetic resonance spectroscopy and competition binding. The thermodynamic parameters, enthalpy and entropy, are discussed in terms of a process involving the transfer of cations from an aqueous to amide environment. PMID:2462930

  8. Solid-state /sup 15/N NMR of oriented lipid bilayer bound gramicidin A'

    SciTech Connect

    Nicholson, L.K.; Moll, F.; Mixon, T.E.; LoGrasso, P.V.; Lay, J.C.; Cross, T.A.

    1987-10-20

    Highly oriented samples of lipid and gramicidin A' (8:1 molar ratio) have been prepared with the samples extensively hydrated (approximately 70% water v/w). These preparations have been shown to be completely in a bilayer phase with a transition temperature of 28/sup 0/C, and evidence is presented indicating that the gramicidin is in the channel conformation. An estimate of the disorder in the alignment of the bilayers parallel with the glass plates used to align the bilayers can be made from the asymmetry of the nuclear magnetic resonances (NMR). Such an analysis indicates a maximal range of disorder of +-3/sup 0/. Uniformly /sup 15/N-labeled gramicidin has been biosynthesized by Bacillus brevis grown in a media containing /sup 15/N-labeled Escherichia coli cells as the only nitrogen source. When prepared with labeled gramicidin, the oriented samples result in high-resolution /sup 15/N NMR spectra showing 12 resonances for the 20 nitrogen sites of the polypeptide. The frequency of the three major multiple resonance peaks has been interpreted to yield the approximate orientation of the N-H bonds in the peptide linkages with respect to the magnetic field. The bond orientations are only partially consistent with the extant structural models of gramicidin.

  9. Planar lipid bilayers containing gramicidin A as a molecular sensing system based on an integrated current.

    PubMed

    Nishio, Masato; Shoji, Atsushi; Sugawara, Masao

    2012-01-01

    The channel activity of gramicidin A in free-standing planar lipid bilayers with different charges of polar head groups and various lengths of hydrocarbon tails were analyzed in terms of the channel conductance, the lifetime of channel events and the magnitude of integrated currents. The channel activity of gramicidin A in lipid bilayers is tunable by adjusting the membrane composition. The in situ coupling of the anti-BSA antibody as a model protein to the amine moiety of phosphatidylethanolamine (PE) in a lipid bilayer by the amine coupling method allowed us to design an antigen (BSA)-sensitive interface, in which the integrated current, rather than the frequency of channel event, can be used as an analytical signal. The potential of the present system for highly sensitive and selective detection of BSA at 10(-9) g/mL level is demonstrated.

  10. Nonstoichiometric complex of gramicidin D with KI at 0.80 [angstrom] resolution

    SciTech Connect

    Olczak, A.; Glowka, M.L.; Szczesio, M.; Bojarsk, J.; Duax, W.L.; Burkhart, B.M.; Wawrzak, Z.

    2010-03-08

    The crystal structure of a nonstoichiometric complex of gramicidin D (gD) with KI has been determined at 100 K using synchrotron radiation. The final R factor was 0.106 for 83 988 observed reflections (Friedel pairs were not merged) collected to 0.80 {angstrom}. The structure consists of four independent pentadecapeptides and numerous solvent molecules and salt ions. The general architecture of the antiparallel double-stranded gramicidin dimers in the crystal (a right-handed antiparallel DS{beta}H{sub R} form) closely resembles that of previously published cation complexes of gD. However, a significantly different mixture of gramicidin isomers is found in the crystal of the KI complex, including partial occupancy of phenylalanine at position 11. Only three sites in each of the two crystallographically independent channels are partially occupied by potassium cations instead of the commonly observed seven sites. The sum of the partial occupancies of K{sup +} (1.10 per two dimers) is consistent with the sum of the iodide occupancies (1.095 over eight sites), which is also confirmed by the anomalous signal of the iodide. There was a significant asymmetry of the distribution and occupancies of cations in the crystallographically independent gramicidin channels, in contrast to the distribution found in the rubidium chloride complex with gD.

  11. Proton conduction in gramicidin A and in its dioxolane-linked dimer in different lipid bilayers.

    PubMed Central

    Cukierman, S; Quigley, E P; Crumrine, D S

    1997-01-01

    Gramicidin A (gA) molecules were covalently linked with a dioxolane ring. Dioxolane-linked gA dimers formed ion channels, selective for monovalent cations, in planar lipid bilayers. The main goal of this study was to compare the functional single ion channel properties of natural gA and its covalently linked dimer in two different lipid bilayers and HCl concentrations (10-8000 mM). Two ion channels with different gating and conductance properties were identified in bilayers from the product of dimerization reaction. The most commonly observed and most stable gramicidin A dimer is the main object of this study. This gramicidin dimer remained in the open state most of the time, with brief closing flickers (tau(closed) approximately 30 micros). The frequency of closing flickers increased with transmembrane potential, making the mean open time moderately voltage dependent (tau(open) changed approximately 1.43-fold/100 mV). Such gating behavior is markedly different from what is seen in natural gA channels. In PEPC (phosphatidylethanolamine-phosphatidylcholine) bilayers, single-channel current-voltage relationships had an ohmic behavior at low voltages, and a marked sublinearity at relatively higher voltages. This behavior contrasts with what was previously described in GMO (glycerylmonooleate) bilayers. In PEPC bilayers, the linear conductance of single-channel proton currents at different proton concentrations was essentially the same for both natural and gA dimers. g(max) and K(D), obtained from fitting experimental points to a Langmuir adsorption isotherm, were approximately 1500 pS and 300 mM, respectively, for both the natural gA and its dimer. In GMO bilayers, however, proton affinities of gA and the dioxolane-dimer were significantly lower (K(D) of approximately 1 and 1.5 M, respectively), and the g(max) higher (approximately 1750 and 2150 pS, respectively) than in PEPC bilayers. Furthermore, the relationship between single-channel conductance and proton

  12. Proton conduction in gramicidin A and in its dioxolane-linked dimer in different lipid bilayers.

    PubMed

    Cukierman, S; Quigley, E P; Crumrine, D S

    1997-11-01

    Gramicidin A (gA) molecules were covalently linked with a dioxolane ring. Dioxolane-linked gA dimers formed ion channels, selective for monovalent cations, in planar lipid bilayers. The main goal of this study was to compare the functional single ion channel properties of natural gA and its covalently linked dimer in two different lipid bilayers and HCl concentrations (10-8000 mM). Two ion channels with different gating and conductance properties were identified in bilayers from the product of dimerization reaction. The most commonly observed and most stable gramicidin A dimer is the main object of this study. This gramicidin dimer remained in the open state most of the time, with brief closing flickers (tau(closed) approximately 30 micros). The frequency of closing flickers increased with transmembrane potential, making the mean open time moderately voltage dependent (tau(open) changed approximately 1.43-fold/100 mV). Such gating behavior is markedly different from what is seen in natural gA channels. In PEPC (phosphatidylethanolamine-phosphatidylcholine) bilayers, single-channel current-voltage relationships had an ohmic behavior at low voltages, and a marked sublinearity at relatively higher voltages. This behavior contrasts with what was previously described in GMO (glycerylmonooleate) bilayers. In PEPC bilayers, the linear conductance of single-channel proton currents at different proton concentrations was essentially the same for both natural and gA dimers. g(max) and K(D), obtained from fitting experimental points to a Langmuir adsorption isotherm, were approximately 1500 pS and 300 mM, respectively, for both the natural gA and its dimer. In GMO bilayers, however, proton affinities of gA and the dioxolane-dimer were significantly lower (K(D) of approximately 1 and 1.5 M, respectively), and the g(max) higher (approximately 1750 and 2150 pS, respectively) than in PEPC bilayers. Furthermore, the relationship between single-channel conductance and proton

  13. Gramicidin-based fluorescence assay; for determining small molecules potential for modifying lipid bilayer properties.

    PubMed

    Ingólfsson, Helgi I; Sanford, R Lea; Kapoor, Ruchi; Andersen, Olaf S

    2010-10-13

    Many drugs and other small molecules used to modulate biological function are amphiphiles that adsorb at the bilayer/solution interface and thereby alter lipid bilayer properties. This is important because membrane proteins are energetically coupled to their host bilayer by hydrophobic interactions. Changes in bilayer properties thus alter membrane protein function, which provides an indirect way for amphiphiles to modulate protein function and a possible mechanism for "off-target" drug effects. We have previously developed an electrophysiological assay for detecting changes in lipid bilayer properties using linear gramicidin channels as probes. Gramicidin channels are mini-proteins formed by the transbilayer dimerization of two non-conducting subunits. They are sensitive to changes in their membrane environment, which makes them powerful probes for monitoring changes in lipid bilayer properties as sensed by bilayer spanning proteins. We now demonstrate a fluorescence assay for detecting changes in bilayer properties using the same channels as probes. The assay is based on measuring the time-course of fluorescence quenching from fluorophore-loaded large unilamellar vesicles due to the entry of a quencher through the gramicidin channels. We use the fluorescence indicator/quencher pair 8-aminonaphthalene-1,3,6-trisulfonate (ANTS)/Tl(+) that has been successfully used in other fluorescence quenching assays. Tl(+) permeates the lipid bilayer slowly but passes readily through conducting gramicidin channels. The method is scalable and suitable for both mechanistic studies and high-throughput screening of small molecules for bilayer-perturbing, and potential "off-target", effects. We find that results using this method are in good agreement with previous electrophysiological results.

  14. Mechanistic insight into gramicidin-based detection of protein-ligand interactions via sensitized photoinactivation.

    PubMed

    Rokitskaya, Tatyana I; Macrae, Michael X; Blake, Steven; Egorova, Natalya S; Kotova, Elena A; Yang, Jerry; Antonenko, Yuri N

    2010-11-17

    Among the many challenges for the development of ion channel-based sensors is the poor understanding of how to engineer modified transmembrane pores with tailored functionality that can respond to external stimuli. Here, we use the method of sensitized photoinactivation of gramicidin A (gA) channels in planar bilayer lipid membranes to help elucidate the underlying mechanistic details for changes in macroscopic transmembrane ionic current observed upon interaction of C-terminally attached gA ligands with specific proteins in solution. Three different systems were studied: (i) carbonic anhydrase (CA) and gA-sulfonamide, (ii) PSD-95 protein (belonging to the 'PDZ domain-containing protein') and a gA analog carrying the KGGHRRSARYLESSV peptide sequence at the C-terminus, and (iii) an anti-biotin antibody and gA-biotin. The results challenge a previously proposed mechanistic hypothesis suggesting that protein-induced current suppression is due to steric blockage of the ion passage through gA channels, while they reveal new insight for consideration in alternative mechanistic models. Additionally, we demonstrate that the length of a linker between the ligand and the gA channel may be less important for gramicidin-based detection of monovalent compared to multivalent protein-ligand interactions. These studies collectively shed new light on the mechanism of protein-induced current alterations in bilayer recordings of gA derivatives, which may be important in the design of new gramicidin-based sensors.

  15. Mechanistic insight into gramicidin-based detection of protein-ligand interactions via sensitized photoinactivation

    NASA Astrophysics Data System (ADS)

    Rokitskaya, Tatyana I.; Macrae, Michael X.; Blake, Steven; Egorova, Natalya S.; Kotova, Elena A.; Yang, Jerry; Antonenko, Yuri N.

    2010-11-01

    Among the many challenges for the development of ion channel-based sensors is the poor understanding of how to engineer modified transmembrane pores with tailored functionality that can respond to external stimuli. Here, we use the method of sensitized photoinactivation of gramicidin A (gA) channels in planar bilayer lipid membranes to help elucidate the underlying mechanistic details for changes in macroscopic transmembrane ionic current observed upon interaction of C-terminally attached gA ligands with specific proteins in solution. Three different systems were studied: (i) carbonic anhydrase (CA) and gA-sulfonamide, (ii) PSD-95 protein (belonging to the 'PDZ domain-containing protein') and a gA analog carrying the KGGHRRSARYLESSV peptide sequence at the C-terminus, and (iii) an anti-biotin antibody and gA-biotin. The results challenge a previously proposed mechanistic hypothesis suggesting that protein-induced current suppression is due to steric blockage of the ion passage through gA channels, while they reveal new insight for consideration in alternative mechanistic models. Additionally, we demonstrate that the length of a linker between the ligand and the gA channel may be less important for gramicidin-based detection of monovalent compared to multivalent protein-ligand interactions. These studies collectively shed new light on the mechanism of protein-induced current alterations in bilayer recordings of gA derivatives, which may be important in the design of new gramicidin-based sensors.

  16. Residues beyond the selectivity filter of the K+ channel kir2.1 regulate permeation and block by external Rb+ and Cs+.

    PubMed

    Thompson, G A; Leyland, M L; Ashmole, I; Sutcliffe, M J; Stanfield, P R

    2000-07-15

    1. Kir2.1 channels are blocked by Rb+ and Cs+ in a voltage-dependent manner, characteristic of many inward rectifier K+ channels. Mutation of Ser165 in the transmembrane domain M2 to Leu (S165L) abolished Rb+ blockage and lowered Cs+ blocking affinity. At negative voltages Rb+ carried large inward currents. 2. A model of the Kir2.1 channel, built by homology with the structure of the Streptomyces lividans K+ channel KcsA, suggested the existence of an intersubunit hydrogen bond between Ser165 and Thr141 in the channel pore-forming P-region that helps stabilise the structure of this region. However, mutations of Thr141 and Ser165 did not produce effects consistent with a hydrogen bond between these residues being essential for blockage. 3. An alternative alignment between the M2 regions of Kir2.1 and KcsA suggested that Ser165 is itself a pore-lining residue, more directly affecting blockage. We were able to replace Ser165 with a variety of polar and non-polar residues, consistent with this residue being pore lining. Some of these changes affected channel blockage. 4. We tested the hypothesis that Asp172 - a residue implicated in channel gating by polyamines - formed an additional selectivity filter by using the triple mutant T141A/S165L/D172N. Large Rb+ and Cs+ currents were measured in this mutant. 5. We propose that both Thr141 and Ser165 are likely to provide binding sites for monovalent blocking cations in wild-type channels. These residues lie beyond the carbonyl oxygen tunnel thought to form the channel selectivity filter, which the blocking cations must therefore traverse.

  17. Thallous ion interaction with gramicidin incorporated in micelles studies by thallium-205 nuclear magnetic resonance

    SciTech Connect

    Hinton, J.F.; Young, G.; Millett, F.S.

    1982-02-16

    A study has been made of the interaction between the thallous and gramicidin dimers incorporated into micelles by using /sup 205/Tl NMR spectroscopy. The chemical shift data obtained are interpreted in terms of a model in which the dimer has only one tight binding site. The binding constant for this site was determined to be 900 M/sup -1/ at 30omicronC. The thallous ion also appears to be partially solvated in the channel.

  18. Effect of gramicidin on methanogenesis by various methanogenic bacteria.

    PubMed

    Jarrell, K F; Hamilton, E A

    1985-07-01

    Methanogenesis by Methanobacterium thermoautotrophicum strains was extremely sensitive to gramicidin, total inhibition being observed at 0.2 mug/ml. In contrast, methane synthesis by Methanococcus voltae, Methanogenium marisnigri, Methanosarcina mazei, and Methanospirillum hungatei were resistant to the highest concentrations of gramicidin tested (40 mug/ml), although spheroplasts of Methanospirillum hungatei were extremely sensitive. Other species tested showed intermediate sensitivity to gramicidin, methanogenesis inhibition occurring at 4 to 20 mug/ml.

  19. Effect of Gramicidin on Methanogenesis by Various Methanogenic Bacteria

    PubMed Central

    Jarrell, Ken F.; Hamilton, Elizabeth A.

    1985-01-01

    Methanogenesis by Methanobacterium thermoautotrophicum strains was extremely sensitive to gramicidin, total inhibition being observed at 0.2 μg/ml. In contrast, methane synthesis by Methanococcus voltae, Methanogenium marisnigri, Methanosarcina mazei, and Methanospirillum hungatei were resistant to the highest concentrations of gramicidin tested (40 μg/ml), although spheroplasts of Methanospirillum hungatei were extremely sensitive. Other species tested showed intermediate sensitivity to gramicidin, methanogenesis inhibition occurring at 4 to 20 μg/ml. PMID:16346835

  20. Proton conductance by the gramicidin water wire. Model for proton conductance in the F1F0 ATPases?

    PubMed Central

    Akeson, M; Deamer, D W

    1991-01-01

    The gramicidin channel contains a single strand of water molecules associated through hydrogen bonds. Previous work has shown that channels of similar size are formed by association of transmembrane alpha helices of synthetic leucine-serine peptides. Both types of channels translocate protons with considerable selectivity relative to other cations, and it has been proposed that the selectivity arises by proton "hopping" along hydrogen-bonded chains of water, whereas other cations must cross by ordinary diffusion processes. It is possible that a similar mechanism underlies proton transport in the Fo subunit of the F1F0 ATP synthase. Using the gramicidin channel as a model, we have tested whether a single strand of water is kinetically competent to translocate protons at a rate sufficient to support known rates of ATP synthesis. We found that the gramicidin channel saturates at approximately 530 pS of protonic current in 4 M HCl, more than sufficient for typical ATP synthesis rates. It follows that proton diffusion to a putative channel in Fo, rather than the channel itself, may limit ATP synthesis rates. PMID:1715764

  1. Triple-barrel structure of inwardly rectifying K+ channels revealed by Cs+ and Rb+ block in guinea-pig heart cells.

    PubMed

    Matsuda, H; Matsuura, H; Noma, A

    1989-06-01

    1. The hypothesis that the inwardly rectifying K+ channel consists of a triple-barrel structure was investigated. Inward currents were recorded under the blocking effects of external Cs+ or Rb+ in the cell-attached configuration of the patch-clamp technique using single ventricular cells enzymatically isolated from guinea-pig hearts. 2. Cs+ (10-100 microM) or Rb+ (20-100 microM) added to the 150 mM-K+ pipette solution induced rapid open-blocked transitions in the inward open-channel currents. In about 20% of experiments the inward current showed two intermediate current levels equally spaced between the unit amplitude and the zero-conductance level. The current fluctuated between these four levels. In the remaining experiments no obvious sublevels were observed except spontaneous ones, whose amplitudes were not always equal to one-third or two-thirds of the unit amplitude. 3. In experiments showing sublevels, the probability that the open-channel current stayed at each level was measured at various concentrations of blockers and membrane potentials. In both Cs+ and Rb+ block, the distribution of the current levels showed reasonable agreement with the binomial theorem. This finding suggests that the inwardly rectifying K+ channel is composed of three equally conductive subunits and each subunit is independently blocked by Cs+ or Rb+. 4. The dwell-time histogram in each substate was well fitted with a single-exponential function. On the assumption of the binomial model, the blocking (mu) and unblocking (lambda) rate for Cs+ and Rb+ were calculated. The value of mu was linearly proportional to the concentration of the blocking ion at a given membrane potential and increased with hyperpolarization (e-fold increase with a change of -43.5 mV in the Cs+ block). lambda was almost independent of the concentration of the blocking ion and less dependent on the membrane potential than mu. 5. The open and blocked times were calculated in experiments showing no clear sublevels

  2. Screening for small molecules' bilayer-modifying potential using a gramicidin-based fluorescence assay.

    PubMed

    Ingólfsson, Helgi I; Andersen, Olaf S

    2010-08-01

    Many drugs and other small molecules used to modulate biological function are amphiphiles that adsorb at the bilayer/solution interface and thereby alter lipid bilayer properties. This is important because membrane proteins are energetically coupled to their host bilayer by hydrophobic interactions. Changes in bilayer properties thus alter membrane protein function, which provides a possible mechanism for "off-target" drug effects. We have previously shown that channels formed by the linear gramicidins are suitable probes for changes in lipid bilayer properties, as experienced by bilayer-spanning proteins. We now report a gramicidin-based fluorescence assay for changes in bilayer properties. The assay is based on measuring the time course of fluorescence quenching in fluorophore-loaded large unilamellar vesicles, due to entry of a gramicidin channel-permeable quencher. The method is scalable and suitable for both mechanistic studies and high-throughput screening for bilayer-perturbing, potential off-target effects, which we illustrate using capsaicin (Cap) and other compounds.

  3. sup 2 H Nuclear magnetic resonance of the gramicidin A backbone in a phospholipid bilayer

    SciTech Connect

    Prosser, R.S.; Davis, J.H. ); Dahlquist, F.W.; Lindorfer, M.A. )

    1991-05-14

    Solid-state {sup 2}H NMR spectroscopy has been employed to study the channel conformation of gramicidin A (GA) in unoriented 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC) multilayers. Quadrupolar echo spectra were obtained at 44{degree}C and 53{degree}C, from gramicidin A labels in which the proton attached to the {alpha} carbon of residue 3,4,5,10,12, or 14 was replaced with deuterium. Because of the nearly axially symmetric electric field gradient tensor, the quadrupolar splittings obtained from an unoriented multilamellar dispersion of DMPC and singly labeled GA directly yield unambiguous orientational constraints on the C-{sup 2}H bonds. The authors conclude that the helix sense of the channel conformation of GA is right-handed. Assuming that the dominant motions are fast axial diffusion of the gramicidin molecule and reorientation of the diffusion axis with respect to the local bilayer normal, then the theoretical splittings may all be scaled down by a constant motional narrowing factor.

  4. A one-dimensional continuum elastic model for membrane-embedded gramicidin dimer dissociation.

    PubMed

    Stember, Joseph N; Andersen, Olaf

    2011-02-04

    Membrane elastic properties, which are subject to alteration by compounds such as cholesterol, lipid metabolites and other amphiphiles, as well as pharmaceuticals, can have important effects on membrane proteins. A useful tool for measuring some of these effects is the gramicidin A channels, which are formed by transmembrane dimerization of non-conducting subunits that reside in each bilayer leaflet. The length of the conducting channels is less than the bilayer thickness, meaning that channel formation is associated with a local bilayer deformation. Electrophysiological studies have shown that the dimer becomes increasingly destabilized as the hydrophobic mismatch between the channel and the host bilayer increases. That is, the bilayer imposes a disjoining force on the channel, which grows larger with increasing hydrophobic mismatch. The energetic analysis of the channel-bilayer coupling is usually pursued assuming that each subunit, as well as the subunit-subunit interface, is rigid. Here we relax the latter assumption and explore how the bilayer junction responds to changes in this disjoining force using a simple one-dimensional energetic model, which reproduces key features of the bilayer regulation of gramicidin channel lifetimes.

  5. Isolated gramicidin peptides probed by IR spectroscopy.

    PubMed

    Rijs, Anouk M; Kabeláč, Martin; Abo-Riziq, Ali; Hobza, Pavel; de Vries, Mattanjah S

    2011-07-11

    We report double-resonant IR/UV ion-dip spectroscopy of neutral gramicidin peptides in the gas phase. The IR spectra of gramicidin A and C, recorded in both the 1000 cm(-1) to 1800 cm(-1) and the 2700 to 3750 cm(-1) region, allow structural analysis. By studying this broad IR range, various local intramolecular interactions are probed, and complementary IR modes can be accessed. Ab initio quantum chemical calculations are used to support the interpretation of the experimental IR spectra. The comparison of the calculated frequencies with the experimental IR spectrum probed via the strong infrared absorptions of all the amide groups (NH stretch, C=O stretch and NH bend), shows evidence for a helical structure in the gas phase, which is similar to that in the condensed phase. Additionally, we show that to improve the spectral resolution when studying large neutral molecular structures of the size of gramicidin, the use of heavier carrier gas could be advantageous.

  6. Molecular Dynamics Study of Gramicidin A in Lipid Bilayer: Electrostatic Map and Ion Conduction

    NASA Astrophysics Data System (ADS)

    Saito, Hiroaki; Iwayama, Masashi; Kawaguchi, Kazutomo; Mizukami, Taku; Miyakawa, Takeshi; Takasu, Masako; Nagao, Hidemi

    The electrostatic potential (ESP) of gramicidin A (GA) in the DMPC lipid bilayers with/without an external uniform electrostatic field was investigated by molecular dynamics (MD) simulation. We found that the ESP profile with an external electrostatic field became step shape. The water and polar groups of the lipid and GA are rearranged in order to restore a flat ESP in the water bulk and GA channel interior. The reorientation of the polar head group enhances the ESP difference between each hydration regions of the membrane, and this should yield an increase of ion conductance through the GA channel.

  7. Folding simulations of gramicidin A into the beta-helix conformations: Simulated annealing molecular dynamics study.

    PubMed

    Mori, Takaharu; Okamoto, Yuko

    2009-10-28

    Gramicidin A is a linear hydrophobic 15-residue peptide which consists of alternating D- and L-amino acids and forms a unique tertiary structure, called the beta(6.3)-helix, to act as a cation-selective ion channel in the natural conditions. In order to investigate the intrinsic ability of the gramicidin A monomer to form secondary structures, we performed the folding simulation of gramicidin A using a simulated annealing molecular dynamics (MD) method in vacuum mimicking the low-dielectric, homogeneous membrane environment. The initial conformation was a fully extended one. From the 200 different MD runs, we obtained a right-handed beta(4.4)-helix as the lowest-potential-energy structure, and left-handed beta(4.4)-helix, right-handed and left-handed beta(6.3)-helix as local-minimum energy states. These results are in accord with those of the experiments of gramicidin A in homogeneous organic solvent. Our simulations showed a slight right-hand sense in the lower-energy conformations and a quite beta-sheet-forming tendency throughout almost the entire sequence. In order to examine the stability of the obtained right-handed beta(6.3)-helix and beta(4.4)-helix structures in more realistic membrane environment, we have also performed all-atom MD simulations in explicit water, ion, and lipid molecules, starting from these beta-helix structures. The results suggested that beta(6.3)-helix is more stable than beta(4.4)-helix in the inhomogeneous, explicit membrane environment, where the pore water and the hydrogen bonds between Trp side-chains and lipid-head groups have a role to further stabilize the beta(6.3)-helix conformation.

  8. Folding simulations of gramicidin A into the β-helix conformations: Simulated annealing molecular dynamics study

    NASA Astrophysics Data System (ADS)

    Mori, Takaharu; Okamoto, Yuko

    2009-10-01

    Gramicidin A is a linear hydrophobic 15-residue peptide which consists of alternating D- and L-amino acids and forms a unique tertiary structure, called the β6.3-helix, to act as a cation-selective ion channel in the natural conditions. In order to investigate the intrinsic ability of the gramicidin A monomer to form secondary structures, we performed the folding simulation of gramicidin A using a simulated annealing molecular dynamics (MD) method in vacuum mimicking the low-dielectric, homogeneous membrane environment. The initial conformation was a fully extended one. From the 200 different MD runs, we obtained a right-handed β4.4-helix as the lowest-potential-energy structure, and left-handed β4.4-helix, right-handed and left-handed β6.3-helix as local-minimum energy states. These results are in accord with those of the experiments of gramicidin A in homogeneous organic solvent. Our simulations showed a slight right-hand sense in the lower-energy conformations and a quite β-sheet-forming tendency throughout almost the entire sequence. In order to examine the stability of the obtained right-handed β6.3-helix and β4.4-helix structures in more realistic membrane environment, we have also performed all-atom MD simulations in explicit water, ion, and lipid molecules, starting from these β-helix structures. The results suggested that β6.3-helix is more stable than β4.4-helix in the inhomogeneous, explicit membrane environment, where the pore water and the hydrogen bonds between Trp side-chains and lipid-head groups have a role to further stabilize the β6.3-helix conformation.

  9. Comment on ``Free energy simulations of single and double ion occupancy in gramicidin A'' [J. Chem. Phys. 126, 105103 (2007)

    NASA Astrophysics Data System (ADS)

    Roux, Benoît; Andersen, Olaf S.; Allen, Toby W.

    2008-06-01

    In a recent article published by Bastug and Kuyucak [J. Chem. Phys.126, 105103 (2007)] investigated the microscopic factors affecting double ion occupancy in the gramicidin channel. The analysis relied largely on the one-dimensional potential of mean force of ions along the axis of the channel (the so-called free energy profile of the ion along the channel axis), as well as on the calculation of the equilibrium association constant of the ions in the channel binding sites. It is the purpose of this communication to clarify this issue.

  10. Interaction of gramicidin with DPPC/DODAB bilayer fragments.

    PubMed

    Carvalho, Camilla A; Olivares-Ortega, Constanza; Soto-Arriaza, Marco A; Carmona-Ribeiro, Ana M

    2012-12-01

    The interaction between the antimicrobial peptide gramicidin (Gr) and dipalmitoylphosphatidylcholine (DPPC)/dioctadecyldimethylammonium bromide (DODAB) 1:1 large unilamellar vesicles (LVs) or bilayer fragments (BFs) was evaluated by means of several techniques. The major methods were: 1) Gr intrinsic fluorescence and circular dichroism (CD) spectroscopy; 2) dynamic light scattering for sizing and zeta-potential analysis; 3) determination of the bilayer phase transition from extrinsic fluorescence of bilayer probes; 4) pictures of the dispersions for evaluation of coloidal stability over a range of time and NaCl concentration. For Gr in LVs, the Gr dimeric channel conformation is suggested from: 1) CD and intrinsic fluorescence spectra similar to those in trifluoroethanol (TFE); 2) KCl or glucose permeation through the LVs/Gr bilayer. For Gr in BFs, the intertwined dimeric, non-channel Gr conformation is evidenced by CD and intrinsic fluorescence spectra similar to those in ethanol. Both LVs and BFs shield Gr tryptophans against quenching by acrylamide but the Stern-Volmer quenching constant was slightly higher for Gr in BFs confirming that the peptide is more exposed to the water phase in BFs than in LVs. The DPPC/DODAB/Gr supramolecular assemblies may predict the behavior of other antimicrobial peptides in assemblies with lipids.

  11. Large scale conformational transitions in β-structural motif of gramicidin A: kinetic analysis based on CD and FT-IR data.

    PubMed

    Sychev, Sergei V; Ivanov, Vadim T

    2014-08-01

    Gramicidin A (gA) is a polypeptide antibiotic, which forms dimeric channels specific for monovalent cations in artificial and biological membranes. It is a polymorphic molecule that adopts a unique variety of helical conformations, including antiparallel double-stranded ↑↓β5.6 or ↑↓β7.2 helices (number of residues per turn) and a single-stranded β6.3 helix (the 'channel form'). The behavior of gA-Cs(+) complex in the micelles of TX-100 was studied in this work. Transfer of the complex into the micelles activates a cascade of sequential conformational transitions monitored by CD and FT-IR spectroscopy: [Formula: see text] At the first step after Cs(+) removal, the RH ↑↓β5.6 helix is formed, which has been discussed so far only hypothetically. Kinetics of the transitions was measured, and the activation parameters were determined. The activation energies of the ↑↓β5.6 → β-helical monomer transition in dioxane and dioxane/water solutions were also measured for comparison. The presence of water raises the transition rate constant ~10(3) times but does not lead to crucial fall of the activation energy. All activation energies were found in the 20-25 kcal/mol range, i.e. much lower than would be expected for unwinding of the double helix (when 28 H-bonds are broken simultaneously). These results can be accounted for in the light of local unfolding (or 'cracking') model for large scale conformational transitions developed by the P. G.Wolynes team [Miyashita O, Onuchic JN, Wolynes PG. Proc. Natl. Acad. Sci. USA 2003; 100: 12570-12575.].

  12. Electronic control of H+ current in a bioprotonic device with Gramicidin A and Alamethicin

    PubMed Central

    Hemmatian, Zahra; Keene, Scott; Josberger, Erik; Miyake, Takeo; Arboleda, Carina; Soto-Rodríguez, Jessica; Baneyx, François; Rolandi, Marco

    2016-01-01

    In biological systems, intercellular communication is mediated by membrane proteins and ion channels that regulate traffic of ions and small molecules across cell membranes. A bioelectronic device with ion channels that control ionic flow across a supported lipid bilayer (SLB) should therefore be ideal for interfacing with biological systems. Here, we demonstrate a biotic–abiotic bioprotonic device with Pd contacts that regulates proton (H+) flow across an SLB incorporating the ion channels Gramicidin A (gA) and Alamethicin (ALM). We model the device characteristics using the Goldman–Hodgkin–Katz (GHK) solution to the Nernst–Planck equation for transport across the membrane. We derive the permeability for an SLB integrating gA and ALM and demonstrate pH control as a function of applied voltage and membrane permeability. This work opens the door to integrating more complex H+ channels at the Pd contact interface to produce responsive biotic–abiotic devices with increased functionality. PMID:27713411

  13. Response to ``Comment on `Free energy simulations of single and double ion occupancy in gramicidin A' '' [J. Chem. Phys. 128, 227101 (2008)

    NASA Astrophysics Data System (ADS)

    Baştuğ, Turgut; Kuyucak, Serdar

    2008-06-01

    We respond to the criticism that one-dimensional (1D) construction of the potential of mean force (PMF) of ions in channels is flowed. Comparison of the 1D PMF results in the gramicidin A channel with independent free energy difference calculations obtained by using the free energy perturbation and thermodynamic integration methods shows complete agreement, thus providing a justifications for the 1D PMF approximation.

  14. Electric field driven changes of a gramicidin containing lipid bilayer supported on a Au(111) surface.

    PubMed

    Laredo, Thamara; Dutcher, John R; Lipkowski, Jacek

    2011-08-16

    Langmuir-Blodgett and Langmuir-Schaeffer methods were employed to deposit a mixed bilayer consisting of 90% of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 10% of gramicidin (GD), a short 15 residue ion channel forming peptide, onto a Au(111) electrode surface. This architecture allowed us to investigate the effect of the electrostatic potential applied to the electrode on the orientation and conformation of DMPC molecules in the bilayer containing the ion channel. The charge density data were determined from chronocoulometry experiments. The electric field and the potential across the membrane were determined through the use of charge density curves. The magnitudes of potentials across the gold-supported biomimetic membrane were comparable to the transmembrane potential acting on a natural membrane. The information regarding the orientation and conformation of DMPC and GD molecules in the bilayer was obtained from photon polarization modulation infrared reflection absorption spectroscopy (PMIRRAS) measurements. The results show that the bilayer is adsorbed, in direct contact with the metal surface, when the potential across the interface is more positive than -0.4 V and is lifted from the gold surface when the potential across the interface is more negative than -0.4 V. This change in the state of the bilayer has a significant impact on the orientation and conformation of the phospholipid and gramicidin molecules. The potential induced changes in the membrane containing peptide were compared to the changes in the structure of the pure DMPC bilayer determined in earlier studies.

  15. Gramicidin-perforated patch revealed depolarizing effect of GABA in cultured frog melanotrophs

    PubMed Central

    Le Foll, Frank; Castel, Hélène; Soriani, Olivier; Vaudry, Hubert; Cazin, Lionel

    1998-01-01

    In frog pituitary melanotrophs, GABA induces a transient stimulation followed by prolonged inhibition of hormone secretion. This biphasic effect is inconsistent with the elevation of cytosolic calcium and the inhibition of electrical activity also provoked by GABA in single melanotrophs. In the present study, standard patch-clamp configurations and gramicidin-perforated patches were used to investigate the physiological GABAA receptor-mediated response and intracellular chloride concentration ([Cl−]i) in cultured frog melanotrophs. In the gramicidin-perforated patch configuration, 1 μM GABA caused a depolarization associated with an action potential discharge and a slight fall of membrane resistance. In contrast, at a higher concentration (10 μm) GABA elicited a depolarization accompanied by a transient volley of action potentials, followed by a sustained inhibitory plateau and a marked fall of membrane resistance. Isoguvacine mimicked the GABA-evoked responses, indicating a mediation by GABAA receptors. In gramicidin-perforated cells, the depolarizing excitatory effect of 1 μm GABA was converted into a depolarizing inhibitory action when 0.4 μm allopregnanolone was added to the bath solution. After gaining the whole-cell configuration, the amplitude and/or direction of the GABA-evoked current (IGABA) rapidly changed before stabilizing. After stabilization, the reversal potential of IGABA followed the values predicted by the Nernst equation for chloride ions when [Cl−]i was varied. In gramicidin-perforated cells, the steady-state I–V relationships of 10 μm GABA- or isoguvacine-evoked currents yielded reversal potentials of −37.5 ± 1.6 (n= 17) and −38.6 ± 2.0 mV (n= 8), respectively. These values were close to those obtained by using a voltage-ramp protocol in the presence of Na+, K+ and Ca2+ channel blockers. The current evoked by 1 μm GABA also reversed at these potentials. We conclude that, in frog pituitary melanotrophs, chloride is the

  16. Mixed Langmuir monolayers of gramicidin A and fluorinated alcohols.

    PubMed

    Broniatowski, Marcin; Obidowicz, Katarzyna; Vila Romeu, Nuria; Broniatowska, Elzbieta; Dynarowicz-Łatka, Patrycja

    2007-09-15

    Mixed monolayers of gramicidin A (GA) and three alcohols, differing in the degree of fluorination, namely C18OH, F18OH, and F8H10OH have been investigated by means of: surface manometry (pi-A isotherms) and Brewster angle microscopy (BAM) aiming at finding appropriate molecules for incorporating gramicidin A for a biosensor design. Our results proved that only the semifluorinated alcohol is appropriate material for this purpose since it forms miscible and homogeneous monolayers with GA within the whole concentration range. The experimental results have been supported by the calculations of van der Waals energy profiles using the Insight II program. Both the hydrogenated and perfluorinated alcohols were found to aggregate at higher surface pressures, which exclude their application for gramicidin-based biosensor construction.

  17. Gramicidin A-based peptide vector for intracellular protein delivery.

    PubMed

    Stoilova, Tatiana B; Kovalchuk, Sergey I; Egorova, Natalya S; Surovoy, Andrey Y; Ivanov, Vadim T

    2008-10-01

    The development of the peptide-based vectors for the intracellular delivery of biologically active macromolecules has opened new prospects of their application in research and therapy. Earlier the amphipathic cell-penetrating peptide (CPP) Pep-1 was reported to mediate cellular uptake of proteins without covalent binding to them. In this work we studied the ability of a series of membrane-active amphipathic peptides, based on the gramicidin A sequence, to transport a model protein across the eukaryotic cell membrane. Among them the positively charged Cys-containing peptide P10C demonstrated the most effective beta-galactosidase intracellular delivery. Besides, this peptide was shown to form noncovalent associates with beta-galactosidase as judged from electrophoresis and enzymatic activity assays. In addition, a series of new gramicidin analogues were prepared and the effect of N-terminus modification of gramicidin on the protein transduction efficiency was studied.

  18. Optimization of antimicrobial drug gramicidin S dosing regime using biosimulations.

    PubMed

    Smirnov, S; Belashov, A; Demin, O

    2009-01-31

    In this paper we have developed a model of antimicrobial effect of gramicidin S. This model has allowed us to predict the dependence of antimicrobial effect of the drug applied as oral melting tablets on dosage, time of resorption and minimal inhibitory concentration (MIC) of the drug characterizing its ability to kill different bacteria. The model has been employed to optimize dosing regime of gramicidin S containing drug Grammidin. Efficacy of the drug has been studied for the diverse gram-positive and gram-negative bacteria with different MIC. The number of bacteria located in the oral cavity and killed by one-pass administration of the drug (resolution of one tablet) has been calculated under condition of various dosing regimes. Based on the simulation results it has been found that (1) twofold prolongation of prescribed resorption time (from 30 to 60min) of the tablet comprising standard dosage of 3mg of gramicidin S results in 1.5-fold increase in efficacy, (2) 1.5-fold decrease in gramicidin S dosage (from 3 to 2mg per administration) under condition of holding prescribed resorption time (30min) does not lead to any considerable decrease in the efficacy of the drug.

  19. NQRS Data for Gramicidin A (Subst. No. 2507)

    NASA Astrophysics Data System (ADS)

    Chihara, H.; Nakamura, N.

    This document is part of Subvolume B 'Substances Containing C10H16 … Zn' of Volume 48 'Nuclear Quadrupole Resonance Spectroscopy Data' of Landolt-Börnstein - Group III 'Condensed Matter'. It contains an extract of Section '3.2 Data tables' of the Chapter '3 Nuclear quadrupole resonance data' providing the NQRS data for Gramicidin A (Subst. No. 2507)

  20. Molecular dynamics study of electrostatic potential along lipid bilayer with gramicidin A

    NASA Astrophysics Data System (ADS)

    Saito, Hiroaki; Nishimura, Megumi; Takagi, Hiroyuki; Miyakawa, Takeshi; Kawaguchi, Kazutomo; Nagao, Hidemi

    2013-02-01

    The structure and electrostatic potential profile of the DMPC lipid bilayers with a gramicidin A (GA) were studied by molecular dynamics (MD) simulation. The MD simulation reproduced the effect of GA on the membrane structure; the area per lipid decreases and membrane thickness increases, and the observed membrane structures correspond to the experimental data. The polar headgroup of lipid was found to orient toward the membrane normal as the lipid approaches the GA. The observed electrostatic potential map showed that the electrostatic potential around the region of GA gate was lower than the others at the same level of the membrane normal and the values of electrostatic potential in the pore region of GA were negative. These results indicate that a cation in the aqueous region of membrane can be electrostatically led to the GA entrance and penetrate the GA channel following the gradient of ion concentration.

  1. Determining floodplain sedimentation rates using 137Cs in a low fallout environment dominated by channel- and cultivation-derived sediment inputs, central Queensland, Australia.

    PubMed

    Hughes, Andrew O; Olley, Jon M; Croke, Jacky C; Webster, Ian T

    2009-10-01

    Fallout (137)Cs has been widely used to determine floodplain sedimentation rates in temperate environments, particularly in the northern hemisphere. Its application in low fallout, tropical environments in the southern hemisphere has been limited. In this study we assess the utility of (137)Cs for determining rates of floodplain sedimentation in a dry-tropical catchment in central Queensland, Australia. Floodplain and reference site cores were analysed in two centimetre increments, depth profiles were produced and total (137)Cs inventories calculated from the detailed profile data. Information on the rates of (137)Cs migration through local soils was obtained from the reference site soil cores. This data was used in an advection-diffusion model to account of (137)Cs mobility in floodplain sediment cores. This allowed sedimentation rates to be determined without the first year of detection for (137)Cs being known and without having to assume that (137)Cs remains immobile following deposition. Caesium-137 depth profiles in this environment are demonstrated to be an effective way of determining floodplain sedimentation rates. The total (137)Cs inventory approach was found to be less successful, with only one of the three sites analysed being in unequivocal agreement with the depth profile results. The input of sediment from catchment sources that have little, or no, (137)Cs attached results in true depositional sites having total inventories that are not significantly different from those of undisturbed reference sites.

  2. Tryptophan orientations in membrane-bound gramicidin and melittin-a comparative linear dichroism study on transmembrane and surface-bound peptides.

    PubMed

    Svensson, Frida R; Lincoln, Per; Nordén, Bengt; Esbjörner, Elin K

    2011-01-01

    In the search for methods to study structure and function of membrane-associated proteins and peptides flow linear dichroism, LD, spectroscopy has emerged as a promising technique. Using shear-aligned lipid vesicles, conformations and binding geometries of membrane-bound bio-macromolecules can be assessed. Here we investigate anchoring properties and specific orientations of tryptophan relative to the peptide backbone and to the membrane normal for the model peptides gramicidin and melittin. We have monitored the conformational change associated with the refolding of non-channel gramicidin into its channel form, and quantitatively determined the average orientations of its tryptophan transition moments, suggesting that these residues adopt a well-defined orientation at the membrane interface. An important conclusion regards the structural variation of gramicidin between these two distinct transmembrane forms. Whilst circular dichroism (CD) spectra, as has been reported before, vary strongly between the two forms suggesting their structures might be quite different, the LD results clearly evidence both the peptide backbone orientation and tryptophan side-chain positioning to be very similar. The latter are oriented in accord with what is expected from their role to anchor peptide termini to the membrane surface. The variations in CD could be due to, the in LD observed, minor shifts in mutual orientation and distance between neighbouring tryptophans sensitively determining their exciton interactions. Our data dispute that the non-channel form of membrane-bound gramicidin would be any of the intertwined forms often observed in crystal as the positioning of tryptophans along the peptide axis would not be compatible with the strong interfacial positioning observed here. The general role of tryptophans as interfacial anchors is further assessed for melittin whose conformation shows considerable angular spread, consistent with a carpet model of its mechanism for induced

  3. Stabilization of solid-supported phospholipid multilayer against water by gramicidin addition.

    PubMed

    Han, Won Bae; Kim, Yongdeok; An, Hyeun Hwan; Kim, Hee-Soo; Yoon, Chong Seung

    2014-03-20

    It was demonstrated that hydrophobicity of solid supported planar dipalmitoyl phosphatidylcholine (DPPC) phospholipid multilayer can be greatly increased by incorporating a transmembrane protein, gramicidin, into the DPPC membrane. The contact angle of deionized water droplet on the gramicidin-modified DPPC membrane increased from 0° (complete wetting) without gramicidin to 55° after adding 15 mol % gramicidin. The increased hydrophobicity of the gramicidin-modified DPPC membrane allowed the membrane to remain stable at the air/water interface as well as underwater. The Au nanoparticles deposited on the gramicidin-modified DPPC membrane reproduced the characteristic surface plasmon resonance peak after being kept underwater or in phosphate-buffered saline solution for 5 days, attesting to the membrane stability in an aqueous environment. The enhanced underwater stability of the lipid multilayer substantially broadens the potential application of the lipid multilayer which includes biosensing, enzymatic fuel cell, surface enhanced Raman spectroscopy substrate.

  4. Gramicidin conformational changes during riboflavin photosensitized oxidation in solution and the effect of N-methylation of tryptophan residues.

    PubMed

    Fuentealba, Denis; López, Jhon J; Palominos, Marco; Salas, Cristian O; Soto-Arriaza, Marco A

    2015-04-01

    In the present work, we evaluated the role of gramicidin conformation in its photosensitized oxidation in organic solvents when irradiated in the presence of riboflavin. Gramicidin conformation has been described as monomeric in trifluoroethanol and as an intertwined dimer in methanol. Gramicidin showed extensive photo-oxidation upon irradiation in the presence of riboflavin in both solvents, and tryptophan residues were identified to be involved. We synthesized a gramicidin derivative methylated at position 1 of the indole ring of tryptophan to assess its effect on gramicidin conformation and photo-oxidation. Methylated gramicidin showed very similar absorption and emission spectra to gramicidin, but different conformations were identified by circular dichroism spectra. Upon irradiation, N-methylated tryptophan residues in the gramicidin derivative were not easily photo-oxidized by riboflavin compared to gramicidin. Circular dichroism spectra for gramicidin in methanol changed significantly upon irradiation in the presence of riboflavin indicating a change in conformation, while in trifluoroethanol no such changes were observed. Time-resolved fluorescence and anisotropy studies showed that oxidized gramicidin in methanol had shorter fluorescence lifetimes and a shorter rotational correlation time compared to non-irradiated gramicidin. Additionally, SDS-PAGE analysis showed a marked change in the electrophoretic pattern, whereas the high-molecular-weight bands disappeared upon irradiation. We interpret all these results in terms of a riboflavin photosensitized shift in gramicidin conformation from intertwined to monomeric.

  5. Thermodynamic parameters for the binding of divalent cations to gramicidin A incorporated into a lipid environment by Tl-205 nuclear magnetic resonance.

    PubMed Central

    Hinton, J F; Fernandez, J Q; Shungu, D C; Millett, F S

    1989-01-01

    Thermodynamic parameters, enthalpy and entropy, for the binding of the divalent cations, Mg+2, Ca+2, Sr+2, Ba+2, and Cd+2, to gramicidin A, incorporated into lysophosphatidylcholine, have been determined using a combination of Tl-205 nuclear magnetic resonance spectroscopy and competition binding. The binding process is thermodynamically driven by the enthalpy and not the entropy. The enthalpy values are related to the process involving the transfer of cations from an aqueous environment to an amide environment. A comparison is made between the thermodynamic parameters for the binding of monovalent and divalent cations to gramicidin A to illustrate the channel blocking ability of the divalent cations with respect to monovalent cation transport. PMID:2469486

  6. Effect of the addition of alkanols of different topology to dipalmitoyl-phosphatidylcholine vesicles in the presence of gramicidin.

    PubMed

    Soto-Arriaza, M A; Olivares-Ortega, C; Lissi, E A

    2012-11-01

    In the present work, we analyze the effect of incorporation of the nonanol family (e.g., 1-Nonanol (1-N), 5-Nonanol (5-N), and 2,6-Dimethyl-4-Heptanol (2,6-DH)) into DPPC LUVs in the presence of different gramicidin concentrations. The principal aim of this work is to study the effect of alkanols solubilization on the physicochemical properties of lipid bilayers in the presence of peptide trans-membrane channels, that is, the effects of nonanol family in the interface of lipid-peptide region, considering that the study provides the analysis of a ternary system by direct excitation as well as by Fluorescence Resonance Energy Transfer. Fluorescence measurements were carried out at 20°C after direct excitation of the extrinsic probe or by Fluorescence Resonance Energy Transfer (FRET) from the tryptophan group of gramicidin. Alkanol incorporation decreases with increasing gramicidin content and branching of the additives. 1-N generates most important changes in the inner part of the bilayer, where it produces an increase in bulk acyl chain mobility. Similarly, 1-N significantly modifies the properties of the hydrophilic-hydrophobic interface region sensed by Laurdan, increasing the polarity of the probe microenvironment and/or increasing the relaxation time of interfacial water molecules. On the other hand, 1-N produces a decrease in PDA fluorescence lifetime, a result that can be explained by a significant amount of water entrance to the inner part of the bilayer. The same behavior was observed when pseudo-first-order quenching rate constants by oxygen were measured. 1-N produces an increase in mobility/solubility of the oxygen in the lipid membrane, an effect that is more noticeable in the deep region of the bilayer sensed by PDA, in the absence and in the presence of 2 mol% of Gr. 1-N incorporation produces a greater reduction in GP value than 5-N and 2,6-DH when Laurdan was excited by FRET. These results show that 1-N has the greatest effect in the lipidic domains

  7. Anomalous volume change of gramicidin A in ethanol solutions

    NASA Technical Reports Server (NTRS)

    Derechin, M.; Hayashi, D. M.; Jordan, B. E.

    1975-01-01

    Results of studies aimed at clarifying the failure of gramicidin A (GA) to sediment in early experiments are analyzed. In the present work, no sedimentation was observed in pure pentanol or ethanol, while normal sedimentation was observed in ethanol-water mixtures. It is concluded that GA exists in two conformations that differ in volume. Since the apparent specific volume in absolute ethanol sinks to its lowest values on increasing concentration, the GA molecule probably unfolds completely in conditions favorable for dimerization.

  8. Protein stability and conformational rearrangements in lipid bilayers: linear gramicidin, a model system.

    PubMed Central

    Cotten, M; Xu, F; Cross, T A

    1997-01-01

    The replacement of four tryptophans in gramicidin A by four phenylalanines (gramicidin M) causes no change in the molecular fold of this dimeric peptide in a low dielectric isotropic organic solvent, but the molecular folds are dramatically different in a lipid bilayer environment. The indoles of gramicidin A interact with the anisotropic bilayer environment to induce a change in the molecular fold. The double-helical fold of gramicidin M, as opposed to the single-stranded structure of gramicidin A, is not compatible with ion conductance. Gramicidin A/gramicidin M hybrid structures have also been prepared, and like gramicidin M homodimers, these dimeric hybrids appear to have a double-helical fold, suggesting that a couple of indoles are being buried in the bilayer interstices. To achieve this equilibrium structure (i.e., minimum energy conformation), incubation at 68 degrees C for 2 days is required. Kinetically trapped metastable structures may be more common in lipid bilayers than in an aqueous isotropic environment. Structural characterizations in the bilayers were achieved with solid-state NMR-derived orientational constraints from uniformly aligned lipid bilayer samples, and characterizations in organic solvents were accomplished by solution NMR. Images FIGURE 2 FIGURE 3 FIGURE 5 PMID:9251781

  9. Molecular Dynamics Simulated Annealing Study of Gramicidin A in Water and the Hydrophobic Environment

    NASA Astrophysics Data System (ADS)

    Mori, Takaharu; Okamoto, Yuko

    2008-03-01

    Gramicidin A is a hydrophobic 15-residue peptide with alternating D- and L-amino acids, and it forms various conformations depending on its environment. For example, gramicidin A adopts a random coil or helical conformations, such as &4.4circ;-helix, &6.3circ;-helix, and double-stranded helix in organic solvents. To investigate the structural and dynamical properties of gramicidin A in water and the hydrophobic environment, we performed molecular dynamics simulated annealing simulations with implicit solvent based on a generalized Born model. From the simulations, it was found that gramicidin A has a strong tendency to form a random-coil structure in water, while in the hydrophobic environment it becomes compact and can fold into right- and left-handed conformations of β-helix structures. We discuss the folding mechanism of the β-helix conformation of gramicidin A.

  10. Deposition of metal nanoparticles on phospholipid multilayer membranes modified by gramicidin.

    PubMed

    Han, Won Bae; Kim, Yongdeok; An, Hyeun Hwan; Kim, Hee-Soo; Yoon, Chong Seung

    2013-10-29

    A planar dipalmitoyl phosphatidylcholine (DPPC) multilayer phospholipid membrane was structurally modified by introducing a transmembrane protein, gramicidin (up to 25 mol %), to study its effect on the metal nanoparticles deposited on the membrane. Without gramicidin, when 3-nm-thick Ag, Sn, Al, and Au were deposited, the nanoparticles hardly nucleated on the DPPC membrane in rigid gel state (except for Au); however, the gramicidin addition dramatically enhanced the DPPC membrane surface's affinity for metal atoms so that a dense array of metal (Ag, Sn, and Au) or metal-oxide (Al-oxide) nanoparticles was produced on the membrane surface. The particle sizes ranged from 3 to 15 nm depending on the metal and gramicidin concentration, whereas the particle density was strongly dictated by the gramicidin concentration. The proposed method provides a convenient, generally applicable synthesis route for preparing different metal or metal-oxide nanoparticles on a relatively robust biocompatible membrane.

  11. Probing ion channel conformational dynamics using simultaneous single-molecule ultrafast spectroscopy and patch-clamp electric recording

    NASA Astrophysics Data System (ADS)

    Harms, Greg; Orr, Galya; Lu, H. Peter

    2004-03-01

    An approach to probing single-molecule ion channel kinetics and conformational dynamics, patch-clamp confocal fluorescence microscopy (PCCFM), uses simultaneous ultrafast fluorescence spectroscopy and single-channel electric current recording. PCCFM is applied to determine single-channel conformational dynamics by probing single-pair fluorescence resonant energy transfer, fluorescence self-quenching, and anisotropy of the dye-labeled gramicidin ion channel incorporated in an artificial lipid bilayer. Hidden conformational changes were observed, which strongly suggests that multiple intermediate conformation states are involved in gramicidin ion channel dynamics.

  12. Novel gramicidin formulations in cationic lipid as broad-spectrum microbicidal agents.

    PubMed

    Ragioto, Danielle A M T; Carrasco, Letícia D M; Carmona-Ribeiro, Ana M

    2014-01-01

    Dioctadecyldimethylammonium bromide (DODAB) is an antimicrobial lipid that can be dispersed as large closed bilayers (LV) or bilayer disks (BF). Gramicidin (Gr) is an antimicrobial peptide assembling as channels in membranes and increasing their permeability towards cations. In mammalian cells, DODAB and Gr have the drawbacks of Gram-positive resistance and high toxicity, respectively. In this study, DODAB bilayers incorporating Gr showed good antimicrobial activity and low toxicity. Techniques employed were spectroscopy, photon correlation spectroscopy for sizing and evaluation of the surface potential at the shear plane, turbidimetric detection of dissipation of osmotic gradients in LV/Gr, determination of bacterial cell lysis, and counting of colony-forming units. There was quantitative incorporation of Gr and development of functional channels in LV. Gr increased the bilayer charge density in LV but did not affect the BF charge density, with localization of Gr at the BF borders. DODAB/Gr formulations substantially reduce Gr toxicity against eukaryotic cells and advantageously broaden the antimicrobial activity spectrum, effectively killing Escherichia coli and Staphylococcus aureus bacteria with occurrence of cell lysis.

  13. Resonances in the rotational constants of 0_{g}^{-}(6s_{1/2}+6p_{1/2}) Cs2 levels analysed by an improved-Bv-formula associated to a 2-channel model

    NASA Astrophysics Data System (ADS)

    Jelassi, Haikel; Pruvost, Laurence

    2016-08-01

    Spin-orbit coupling in 0g-(6s1/2+6p1/2) long-range molecular levels of Cs2 assigned to resonances in the rotational constant (Bv) spectrum (Lignier L. et al., Phys. Chem. Chem. Phys., 13 (2011) 18910; Pichler M. et al., J. Chem. Phys., 121 (2004) 1796) has been analyzed with a model which associates an improved-B v -formula (established here) and a 2-channel model. The approach explains the Lorentzian shape of the observed resonances superimposing a non-horizontal background and the fit of the B v -spectrum allows us to deduce the spin-orbit coupling parameters. We compare this approach with an analysis of the binding energies using the vibrational quantum defect (VQD). The good agreement between the methods lets one conclude that the improved-B v -formula associated with a 2-channel model provides a pertinent approach for analyzing coupled molecular series.

  14. Conformation of gramicidin in relation to its ability to form bilayers with lysophosphatidylcholine

    SciTech Connect

    Killian, J.A.; Urry, D.W. )

    1988-09-20

    The ability of gramicidin to induce bilayer formation in lysophosphatidylcholine (LPC) systems was investigated as a function of the conformation of the peptide. The conformation was varied by using different solvents to cosolubilize gramicidin and lipid. Using circular dichroism (CD), it was found that when codissolved in trifluoroethanol (TFE), after drying and subsequent hydration, gramicidin is mainly present in the single-stranded {beta}{sup 6.3}-helical configuration, whereas when using chloroform/methanol or ethanol as the solvent, it is proposed that the dominant conformation of gramicidin in the membrane is that of the double-stranded antiparallel dimer. Employing {sup 31}P NMR, the stoichiometry for bilayer formation was found to be 6 to 7 lipid molecules per gramicidin monomer, when samples were prepared from TFE, whereas a stoichiometry of 4 was found when chloroform/methanol or ethanol as the solvent. Upon heating the latter samples, a conversion was observed in the CD pattern toward that indicative of the {beta}{sup 6.3}-helical configuration. This change was accompanied by an increase in the extent of bilayer formation. Next, it was investigated whether the conformation of gramicidin and its ability to induce bilayer formation were dependent on the lipid acyl chain length. The results are discussed in terms of the possible mechanism of bilayer formation in dispersions of gramicidin and LPC in relation to the conformational behavior of the peptide.

  15. Gramicidin S production by Bacillus brevis in simulated microgravity

    NASA Technical Reports Server (NTRS)

    Fang, A.; Pierson, D. L.; Mishra, S. K.; Koenig, D. W.; Demain, A. L.

    1997-01-01

    In a continuing study of microbial secondary metabolism in simulated microgravity, we have examined gramicidin S (GS) production by Bacillus brevis strain Nagano in NASA High Aspect Rotating Vessels (HARVs), which are designed to simulate some aspects of microgravity. Growth and GS production were found to occur under simulated microgravity. When performance under simulated microgravity was compared with that under normal gravity conditions in the bioreactors, GS production was found to be unaffected by simulated microgravity. The repressive effect of glycerol in flask fermentations was not observed in the HARV. Thus the negative effect of glycerol on specific GS formation is dependent on shear and/or vessel geometry, not gravity.

  16. Photosensitizer binding to lipid bilayers as a precondition for the photoinactivation of membrane channels.

    PubMed Central

    Rokitskaya, T I; Block, M; Antonenko, Y N; Kotova, E A; Pohl, P

    2000-01-01

    The photodynamic activity of sulfonated aluminum phthalocyanines (AlPcS(n), 1 gramicidin channels, as revealed by measurements of the electric current across planar lipid bilayers. The increase in the degree of sulfonation of phthalocyanine progressively reduced its affinity for the lipid bilayer as well as its potency of sensitizing gramicidin channel photoinactivation. The portion of photoinactivated gramicidin channels, alpha, increased with rising photosensitizer concentration up to some optimum. The concentration at which alpha was at half-maximum amounted to 80 nM, 30 nM, 200 nM, and 2 microM for AlPcS(1), AlPcS(2), AlPcS(3), and AlPcS(4), respectively. At high concentrations alpha was found to decrease, which was attributed to quenching of reactive oxygen species and self-quenching of the photosensitizer triplet state by its ground state. Fluoride anions were observed to inhibit both AlPcS(n) (2 gramicidin channels. It is concluded that photosensitizer binding to membrane lipids is a prerequisite for the photodynamic inactivation of gramicidin channels. PMID:10777753

  17. How alkali metal ion binding alters the conformation preferences of gramicidin A: a molecular dynamics and ion mobility study.

    PubMed

    Chen, Liuxi; Gao, Yi Qin; Russell, David H

    2012-01-12

    Here, we present a systematic study combing electrospray ionization-ion mobility experiments and an enhanced sampling molecular dynamics, specifically integrated tempering sampling molecular dynamics simulations (ITS-MDS), to explore the conformations of alkali metal ion (Na, K, and Cs) adducts of gramicidin A (GA) in vacuo. Folding simulation is performed to obtain inherent conformational preferences of neutral GA to provide insights about how the binding of metal ions influences the intrinsic conformations of GA. The comparison between conformations of neutral GA and alkali metal ion adducts reveals a high degree of structural similarity, especially between neutral GA and [GA + Na](+); however, the structural similarities decrease as ionic radius of the metal increases. Collision cross section (CCS) profiles for [GA + Na](+) and [GA + Cs](+) ions obtained from by ITS-MDS compare favorably with the experimental CCS, but there are significant differences from CCS profiles for [GA + K](+) ions. Such discrepancies between the calculated and measured CCS profiles for [GA + K](+) are discussed in terms of limitations in the simulation force field as well as possible size-dependent coordination of the [GA + K](+) ion complex.

  18. Synthesis and biological evaluation of asymmetric gramicidin S analogues containing modified D-phenylalanine residues.

    PubMed

    van der Knaap, Matthijs; Engels, Eefje; Busscher, Henk J; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; Mars-Groenendijk, Roos H; Noort, Daan; van der Marel, Gijsbert A; Overkleeft, Herman S; Overhand, Mark

    2009-09-01

    The synthesis of new analogues of the cationic antimicrobial peptide gramicidin S, having a modified D-phenylalanine residue, their antibacterial properties against several gram positive and negative strains, as well as their hemolytic activity is reported.

  19. Ion fluxes through nanopores and transmembrane channels

    NASA Astrophysics Data System (ADS)

    Bordin, J. R.; Diehl, A.; Barbosa, M. C.; Levin, Y.

    2012-03-01

    We introduce an implicit solvent Molecular Dynamics approach for calculating ionic fluxes through narrow nanopores and transmembrane channels. The method relies on a dual-control-volume grand-canonical molecular dynamics (DCV-GCMD) simulation and the analytical solution for the electrostatic potential inside a cylindrical nanopore recently obtained by Levin [Europhys. Lett.EULEEJ0295-507510.1209/epl/i2006-10240-4 76, 163 (2006)]. The theory is used to calculate the ionic fluxes through an artificial transmembrane channel which mimics the antibacterial gramicidin A channel. Both current-voltage and current-concentration relations are calculated under various experimental conditions. We show that our results are comparable to the characteristics associated to the gramicidin A pore, especially the existence of two binding sites inside the pore and the observed saturation in the current-concentration profiles.

  20. Kinetics profiling of gramicidin S synthetase A, a member of nonribosomal peptide synthetases.

    PubMed

    Sun, Xun; Li, Hao; Alfermann, Jonas; Mootz, Henning D; Yang, Haw

    2014-12-23

    Nonribosomal peptide synthetases (NRPS) incorporate assorted amino acid substrates into complex natural products. The substrate is activated via the formation of a reactive aminoacyl adenylate and is subsequently attached to the protein template via a thioester bond. The reactive nature of such intermediates, however, leads to side reactions that also break down the high-energy anhydride bond. The off-pathway kinetics or their relative weights compared to that of the on-pathway counterpart remains generally elusive. Here, we introduce multiplatform kinetics profiling to quantify the relative weights of on- and off-pathway reactions. Using the well-defined stoichiometry of thioester formation, we integrate a mass spectrometry (MS) kinetics assay, a high-performance liquid chromatography (HPLC) assay, and an ATP-pyrophosphate (PPi) exchange assay to map out a highly efficient on-pathway kinetics profile of the substrate activation and intermediate uploading (>98% relative weight) for wide-type gramicidin S synthetase A (GrsA) and a 87% rate profile for a cysteine-free GrsA mutant. Our kinetics profiling approach complements the existing enzyme-coupled byproduct-release assays, unraveling new mechanistic insights of substrate activation/channeling in NRPS enzymes.

  1. Computational Investigation of the Effect of Lipid Membranes on Ion Permeation in Gramicidin A

    PubMed Central

    Setiadi, Jeffry; Kuyucak, Serdar

    2016-01-01

    Membrane proteins are embedded in a lipid bilayer and interact with the lipid molecules in subtle ways. This can be studied experimentally by examining the effect of different lipid bilayers on the function of membrane proteins. Understanding the causes of the functional effects of lipids is difficult to dissect experimentally but more amenable to a computational approach. Here we perform molecular dynamics simulations and free energy calculations to study the effect of two lipid types (POPC and NODS) on the conductance of the gramicidin A (gA) channel. A larger energy barrier is found for the K+ potential of mean force in gA embedded in POPC compared to that in NODS, which is consistent with the enhanced experimental conductance of cations in gA embedded in NODS. Further analysis of the contributions to the potential energy of K+ reveals that gA and water molecules in gA make similar contributions in both bilayers but there are significant differences between the two bilayers when the lipid molecules and interfacial waters are considered. It is shown that the stronger dipole moments of the POPC head groups create a thicker layer of interfacial waters with better orientation, which ultimately is responsible for the larger energy barrier in the K+ PMF in POPC. PMID:26999229

  2. 'Inverted' analogs of the antibiotic gramicidin S with an improved biological profile.

    PubMed

    Kapoerchan, Varsha V; Knijnenburg, Annemiek D; Keizer, Peter; Spalburg, Emile; de Neeling, Albert J; Mars-Groenendijk, Roos H; Noort, Daan; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; van der Marel, Gijs A; Overkleeft, Herman S; Overhand, Mark

    2012-10-15

    A series of gramicidin S derivatives 4-15 are presented that have four ornithine residues as polar protonated side chains and two central hydrophobic amino acids with unaltered turn regions. These peptides were screened against human erthrocytes and our standard panel of Gram negative- and Gram positive bacteria, including four MRSA strains. Based on the antibacterial- and hemolytic data, peptides 13 and 14 have an improved biological profile compared to the clinically applied topical antibiotic gramicidin S.

  3. The Antimicrobial Activity of Gramicidin A Is Associated with Hydroxyl Radical Formation

    PubMed Central

    Liou, Je-Wen; Hung, Yu-Jiun; Yang, Chin-Hao; Chen, Yi-Cheng

    2015-01-01

    Gramicidin A is an antimicrobial peptide that destroys gram-positive bacteria. The bactericidal mechanism of antimicrobial peptides has been linked to membrane permeation and metabolism disruption as well as interruption of DNA and protein functions. However, the exact bacterial killing mechanism of gramicidin A is not clearly understood. In the present study, we examined the antimicrobial activity of gramicidin A on Staphylococcus aureus using biochemical and biophysical methods, including hydroxyl radical and NAD+/NADH cycling assays, atomic force microscopy, and Fourier transform infrared spectroscopy. Gramicidin A induced membrane permeabilization and changed the composition of the membrane. The morphology of Staphylococcus aureus during gramicidin A destruction was divided into four stages: pore formation, water permeability, bacterial flattening, and lysis. Changes in membrane composition included the destruction of membrane lipids, proteins, and carbohydrates. Most interestingly, we demonstrated that gramicidin A not only caused membrane permeabilization but also induced the formation of hydroxyl radicals, which are a possible end product of the transient depletion of NADH from the tricarboxylic acid cycle. The latter may be the main cause of complete Staphylococcus aureus killing. This new finding may provide insight into the underlying bactericidal mechanism of gA. PMID:25622083

  4. The antimicrobial activity of gramicidin A is associated with hydroxyl radical formation.

    PubMed

    Liou, Je-Wen; Hung, Yu-Jiun; Yang, Chin-Hao; Chen, Yi-Cheng

    2015-01-01

    Gramicidin A is an antimicrobial peptide that destroys gram-positive bacteria. The bactericidal mechanism of antimicrobial peptides has been linked to membrane permeation and metabolism disruption as well as interruption of DNA and protein functions. However, the exact bacterial killing mechanism of gramicidin A is not clearly understood. In the present study, we examined the antimicrobial activity of gramicidin A on Staphylococcus aureus using biochemical and biophysical methods, including hydroxyl radical and NAD+/NADH cycling assays, atomic force microscopy, and Fourier transform infrared spectroscopy. Gramicidin A induced membrane permeabilization and changed the composition of the membrane. The morphology of Staphylococcus aureus during gramicidin A destruction was divided into four stages: pore formation, water permeability, bacterial flattening, and lysis. Changes in membrane composition included the destruction of membrane lipids, proteins, and carbohydrates. Most interestingly, we demonstrated that gramicidin A not only caused membrane permeabilization but also induced the formation of hydroxyl radicals, which are a possible end product of the transient depletion of NADH from the tricarboxylic acid cycle. The latter may be the main cause of complete Staphylococcus aureus killing. This new finding may provide insight into the underlying bactericidal mechanism of gA.

  5. Channel

    NASA Technical Reports Server (NTRS)

    2006-01-01

    [figure removed for brevity, see original site] Context image for PIA03693 Channel

    This channel is located south of Iani Chaos.

    Image information: VIS instrument. Latitude -10.9N, Longitude 345.5E. 17 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  6. Deuterium NMR of 2HCO-Val1. gramicidin A and 2HCO-Val1-D-Leu2. gramicidin A in oriented DMPC bilayers

    SciTech Connect

    Hing, A.W.; Adams, S.P.; Silbert, D.F.; Norberg, R.E. )

    1990-05-01

    Deuterium NMR is used to study the structure and dynamics of the formyl C-2H bond in selectively deuterated gramicidin molecules. Specifically, the functionally different analogues 2HCO-Val1...gramicidin A and 2HCO-Val1-D-Leu2...gramicidin A are studied by 2H NMR so that any conformational or dynamical differences between the two analogues can be correlated with their difference in lifetime. These analogues are first synthesized, purified, and characterized and then incorporated into oriented bilayers of dimyristoylphosphatidylcholine sandwiched between glass coverslips. Phosphorous NMR line shapes obtained from these samples are consistent with the presence of the bilayer phase and indicate that the disorder exhibited by the lipid matrix is approximately of the same type and degree for both analogues. Deuterium NMR line shapes obtained from these samples indicate that the motional axis of the formyl group of gramicidin is parallel to the coverslip normal, that the distribution of motional axis orientations has a width of 7-9 degrees, and that a similar, major conformational and dynamical state exists for the formyl C-2H bond of both analogues. In this state, if the only motion present is fast axial rotation, then the experimentally derived angle between the formyl C-2H bond and the motional axis is consistent with the presence of a right-handed, single-stranded, beta 6.3 helical dimer but is not consistent with the presence of a left-handed, single-stranded, beta 6.3 helical dimer. However, if fast axial rotation is not the only motion present, then the left-handed, single-stranded, beta 6.3 helical dimer cannot be absolutely excluded as a possibility. Also, a second, minor conformational and dynamical state appears to be present in the spectrum of 2HCO-Val1-D-Leu2...gramicidin A but is not observed in the spectrum of 2HCO-Val1...gramicidin A.

  7. A combination of curcumin with either gramicidin or ouabain selectively kills cells that express the multidrug resistance-linked ABCG2 transporter.

    PubMed

    Rao, Divya K; Liu, Haiyan; Ambudkar, Suresh V; Mayer, Michael

    2014-11-01

    This paper introduces a strategy to kill selectively multidrug-resistant cells that express the ABCG2 transporter (also called breast cancer resistance protein, or BCRP). The approach is based on specific stimulation of ATP hydrolysis by ABCG2 transporters with subtoxic doses of curcumin combined with stimulation of ATP hydrolysis by Na(+),K(+)-ATPase with subtoxic doses of gramicidin A or ouabain. After 72 h of incubation with the drug combinations, the resulting overconsumption of ATP by both pathways inhibits the efflux activity of ABCG2 transporters, leads to depletion of intracellular ATP levels below the viability threshold, and kills resistant cells selectively over cells that lack ABCG2 transporters. This strategy, which was also tested on a clinically relevant human breast adenocarcinoma cell line (MCF-7/FLV1), exploits the overexpression of ABCG2 transporters and induces caspase-dependent apoptotic cell death selectively in resistant cells. This work thus introduces a novel strategy to exploit collateral sensitivity (CS) with a combination of two clinically used compounds that individually do not exert CS. Collectively, this work expands the current knowledge on ABCG2-mediated CS and provides a potential strategy for discovery of CS drugs against drug-resistant cancer cells.

  8. Gramicidin D enhances the antibacterial activity of fluoride.

    PubMed

    Nelson, James W; Zhou, Zhiyuan; Breaker, Ronald R

    2014-07-01

    Fluoride is a toxic anion found in many natural environments. One of the major bacterial defenses against fluoride is the cell envelope, which limits passage of the membrane-impermeant fluoride anion. Accordingly, compounds that enhance the permeability of bacterial membranes to fluoride should also enhance fluoride toxicity. In this study, we demonstrate that the pore-forming antibiotic gramicidin D increases fluoride uptake in Bacillus subtilis and that the antibacterial activity of this compound is potentiated by fluoride. Polymyxin B, another membrane-targeting antibiotic with a different mechanism of action, shows no such improvement. These results, along with previous findings, indicate that certain compounds that destabilize bacterial cell envelopes can enhance the toxicity of fluoride.

  9. Simultaneous Patch-Clamp Recording and Single-Molecule Imaging Study of Single-Molecule Ion Channel Dynamics

    NASA Astrophysics Data System (ADS)

    Harms, Greg; Orr, Galya; Thrall, Brian; Montal, Mauricio; Colson, Steve; Lu, H. Peter

    2002-03-01

    By combining real-time single-molecule fluorescence imaging measurements with real-time single-channel current measurements in membranes of lipid bilayers or in living cells, we are now able to probe single ion-channel-protein conformational changes simultaneously correlated with single ion-channel current trajectories, providing an understanding at the molecular-level of the dynamics and mechanisms of ion-channel proteins. This technical innovation has been used to gain an understanding of how ion-channel activities are regulated by conformational change dynamics and assembly mechanisms of the dye-labeled gramicidin channels, which has revealed that the gramicidin channel activity is regulated by complex gramicidin dimer conformational changes. A new multiple-state model for gramicidin ion-channel dynamics, more complex than the classic two-state diffusion model, has been postulated based on our experimental results. The single-channel activity of recombinant NMDA receptors transiently expressed in a mammalian cell-line has been recorded when tetramethylrhodamine-labeled cysteine (TMR-cysteine) and glycine were introduced into the patch-pipette in cell-attached or inside-out patches. Correlated images of a single TMR-cysteine and single NMDA channel recordings have been investigated in real-time, which begins to shed light on the molecular-level understanding of the ligand-receptor interaction dynamics in NMDA receptors in living cells.

  10. Conformation of gramicidin A in Triton X-100 micelles from CD and FTIR data: a clean example of antiparallel double β5.6 helix formation.

    PubMed

    Sychev, Sergei V; Barsukov, Leonid I; Ivanov, Vadim T

    2013-07-01

    The linear peptide gramicidin A (gA) forms prototypical ion channels specific for monovalent cations and has been extensively used to study the organization and dynamics of membrane channels. This polymorphic peptide can adopt two different types of structures, the helical dimer β6.3 ('channel state') and the double helical structure with two intertwined monomers. The structure of gA in micelles of detergent Triton X-100 has been studied using CD, Fourier transform infrared, and fluorescence spectroscopy. The results obtained demonstrate that only one thermodynamically stable gA structure, the antiparallel left-handed double helix β5.6, is formed in this membrane-mimetic environment. The position of the tryptophan fluorescence maximum at 332 nm is the same as that in phospholipid membranes. The causative factors governing the double helix formation in the micellar medium are discussed on the basis of known physicochemical properties of Triton X-100.

  11. Influence of Hydrophobic Mismatch on Structures and Dynamics of Gramicidin A and Lipid Bilayers

    PubMed Central

    Kim, Taehoon; Lee, Kyu Il; Morris, Phillip; Pastor, Richard W.; Andersen, Olaf S.; Im, Wonpil

    2012-01-01

    Gramicidin A (gA) is a 15-amino-acid antibiotic peptide with an alternating L-D sequence, which forms (dimeric) bilayer-spanning, monovalent cation channels in biological membranes and synthetic bilayers. We performed molecular dynamics simulations of gA dimers and monomers in all-atom, explicit dilauroylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), and 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) bilayers. The variation in acyl chain length among these different phospholipids provides a way to alter gA-bilayer interactions by varying the bilayer hydrophobic thickness, and to determine the influence of hydrophobic mismatch on the structure and dynamics of both gA channels (and monomeric subunits) and the host bilayers. The simulations show that the channel structure varied little with changes in hydrophobic mismatch, and that the lipid bilayer adapts to the bilayer-spanning channel to minimize the exposure of hydrophobic residues. The bilayer thickness, however, did not vary monotonically as a function of radial distance from the channel. In all simulations, there was an initial decrease in thickness within 4–5 Å from the channel, which was followed by an increase in DOPC and POPC or a further decrease in DLPC and DMPC bilayers. The bilayer thickness varied little in the monomer simulations—except one of three independent simulations for DMPC and all three DLPC simulations, where the bilayer thinned to allow a single subunit to form a bilayer-spanning water-permeable pore. The radial dependence of local lipid area and bilayer compressibility is also nonmonotonic in the first shell around gA dimers due to gA-phospholipid interactions and the hydrophobic mismatch. Order parameters, acyl chain dynamics, and diffusion constants also differ between the lipids in the first shell and the bulk. The lipid behaviors in the first shell around gA dimers are more complex than predicted from a simple mismatch

  12. Influence of hydrophobic mismatch on structures and dynamics of gramicidin a and lipid bilayers.

    PubMed

    Kim, Taehoon; Lee, Kyu Il; Morris, Phillip; Pastor, Richard W; Andersen, Olaf S; Im, Wonpil

    2012-04-01

    Gramicidin A (gA) is a 15-amino-acid antibiotic peptide with an alternating L-D sequence, which forms (dimeric) bilayer-spanning, monovalent cation channels in biological membranes and synthetic bilayers. We performed molecular dynamics simulations of gA dimers and monomers in all-atom, explicit dilauroylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), and 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) bilayers. The variation in acyl chain length among these different phospholipids provides a way to alter gA-bilayer interactions by varying the bilayer hydrophobic thickness, and to determine the influence of hydrophobic mismatch on the structure and dynamics of both gA channels (and monomeric subunits) and the host bilayers. The simulations show that the channel structure varied little with changes in hydrophobic mismatch, and that the lipid bilayer adapts to the bilayer-spanning channel to minimize the exposure of hydrophobic residues. The bilayer thickness, however, did not vary monotonically as a function of radial distance from the channel. In all simulations, there was an initial decrease in thickness within 4-5 Å from the channel, which was followed by an increase in DOPC and POPC or a further decrease in DLPC and DMPC bilayers. The bilayer thickness varied little in the monomer simulations-except one of three independent simulations for DMPC and all three DLPC simulations, where the bilayer thinned to allow a single subunit to form a bilayer-spanning water-permeable pore. The radial dependence of local lipid area and bilayer compressibility is also nonmonotonic in the first shell around gA dimers due to gA-phospholipid interactions and the hydrophobic mismatch. Order parameters, acyl chain dynamics, and diffusion constants also differ between the lipids in the first shell and the bulk. The lipid behaviors in the first shell around gA dimers are more complex than predicted from a simple mismatch model

  13. Gramicidin-induced hexagonal H/sub II/ phase formation in erythrocyte membranes

    SciTech Connect

    Tournois, H.; Leunissen-Bijvelt, J.; Haest, C.W.M.; de Gier, J.; Kruij, B.

    1987-10-20

    Using /sup 31/P nuclear magnetic resonance (NMR), small-angle X-ray scattering (SAXS), and freeze-fracture electron microscopic (FFEM) techniques, it is shown that gramicidin induces a hexagonal H/sub II/ phase not only in liposomes prepared from total lipids extracted from human erythrocytes but also in isolated human erythrocyte membranes (white ghosts). A 37/sup 0/C, H/sub II/ phase formation is detected at a gramicidin to phospholipid molar ratio exceeding 1:80. The gramicidin-induced H/sub II/ phase exhibits a very small /sup 31/P chemical shift anisotropy indicating decreased head-group order, and it displays a temperature-dependent increase in tube diameter from 60.2 A at 4/sup 0/C to 64.2 A at 37/sup 0/C in ghosts and from 62.8 to 69.4 A at 37/sup 0/C in total lipid extracts, both in the presence of 1 mol of gramicidin/10 mol of phospholipid. /sup 31/P NMR data indicate that the H/sub II/ phase formation by gramicidin is temperature dependent and show the gradual disappearance of the H/sub II/ phase at low temperatures resulting in a bilayer type of /sup 31/P NMR line shape at 4/sup 0/C, whereas SAXS and FFEM data suggest equal amounts of H/sub II/ phases at all temperatures. The induction of the H/sub II/ phase by gramicidin is specific in that N-formylation of the four tryptophan residues of gramicidin completely blocks the hexagonal (H/sub II/) phase inducing ability of the peptide. The striking parallel between hexagonal H/sub II/ phase induction and the enhancement of lysophosphatidylcholine and palmitoylcarnitine transbilayer reorientation by gramicidin as well the lack of effect of the formylated gramicidin strongly suggests the H/sub II/ phase formation and flip enhancement are mechanistically related phenomena. It is suggested that the formation of gramicidin aggregates of specific structure which are intermediates in H/sub II/ phase formation leads to enhancement of transbilayer reorientation of phospholipids.

  14. Gated Ion Channel-Based Biosensor Device

    NASA Astrophysics Data System (ADS)

    Separovic, Frances; Cornell, Bruce A.

    A biosensor device based on the ion channel gramicidin A (gA) incorporated into a bilayer membrane is described. This generic immunosensing device utilizes gA coupled to an antibody and assembled in a lipid membrane. The membrane is chemically tethered to a gold electrode, which reports on changes in the ionic conduction of the lipid bilayer. Binding of a target molecule in the bathing solution to the antibody causes the gramicidin channels to switch from predominantly conducting dimers to predominantly nonconducting monomers. Conventional a.c. impedance spectroscopy between the gold and a counter electrode in the bathing solution is used to measure changes in the ionic conductivity of the membrane. This approach permits the quantitative detection of a range of target species, including bacteria, proteins, toxins, DNA sequences, and drug molecules.

  15. CS Informativeness Governs CS-US Associability

    PubMed Central

    Ward, Ryan D.; Gallistel, C. R.; Jensen, Greg; Richards, Vanessa L.; Fairhurst, Stephen; Balsam, Peter D

    2012-01-01

    In a conditioning protocol, the onset of the conditioned stimulus (CS) provides information about when to expect reinforcement (the US). There are two sources of information from the CS in a delay conditioning paradigm in which the CS-US interval is fixed. The first depends on the informativeness, the degree to which CS onset reduces the average expected time to onset of the next US. The second depends only on how precisely a subject can represent a fixed-duration interval (the temporal Weber fraction). In three experiments with mice, we tested the differential impact of these two sources of information on rate of acquisition of conditioned responding (CS-US associability). In Experiment 1, we show that associability (the inverse of trials to acquisition) increases in proportion to informativeness. In Experiment 2, we show that fixing the duration of the US-US interval or the CS-US interval or both has no effect on associability. In Experiment 3, we equated the increase in information produced by varying the C̅/T̅ ratio with the increase produced by fixing the duration of the CS-US interval. Associability increased with increased informativeness, but, as in Experiment 2, fixing the CS-US duration had no effect on associability. These results are consistent with the view that CS-US associability depends on the increased rate of reward signaled by CS onset. The results also provide further evidence that conditioned responding is temporally controlled when it emerges. PMID:22468633

  16. Invariance of single-file water mobility in gramicidin-like peptidic pores as function of pore length.

    PubMed

    Portella, Guillem; Pohl, Peter; de Groot, Bert L

    2007-06-01

    We investigated the structural and energetic determinants underlying water permeation through peptidic nanopores, motivated by recent experimental findings that indicate that water mobility in single-file water channels displays nonlinear length dependence. To address the molecular mechanism determining the observed length dependence, we studied water permeability in a series of designed gramicidin-like channels of different length using atomistic molecular dynamics simulations. We found that within the studied range of length the osmotic water permeability is independent of pore length. This result is at variance with textbook models, where the relationship is assumed to be linear. Energetic analysis shows that loss of solvation rather than specific water binding sites in the pore form the main energetic barrier for water permeation, consistent with our dynamics results. For this situation, we propose a modified expression for osmotic permeability that fully takes into account water motion collectivity and does not depend on the pore length. Different schematic barrier profiles are discussed that explain both experimental and computational interpretations, and we propose a set of experiments aimed at validation of the presented results. Implications of the results for the design of peptidic channels with desired permeation characteristics are discussed.

  17. Effects of cholesterol or gramicidin on slow and fast motions of phospholipids in oriented bilayers

    SciTech Connect

    Peng, Z.Y.; Simplaceanu, V.; Dowd, S.R.; Ho, C. )

    1989-11-01

    Nuclear spin-lattice relaxation both in the rotating frame and in the laboratory frame is used to investigate the slow and fast molecular motions of phospholipids in oriented bilayers in the liquid crystalline phase. The bilayers are prepared from a perdeuterated phospholipid labeled with a pair of {sup 19}F atoms at the 7 position of the 2-sn acyl chain. Phospholipid-cholesterol or phospholipid-gramicidin interactions are characterized by measuring the relaxation rates as a function of the bilayer orientation, the locking field, and the temperature. These studies show that cholesterol or gramicidin can specifically enhance the relaxation due to slow motions in phospholipid bilayers with correlation times {tau}{sub s} longer than 10{sup {minus}8} sec. The perturbations of the geometry of the slow motions induced by cholesterol are qualitatively different from those induced by gramicidin. In contrast, the presence of cholesterol or gramicidin slightly suppresses the fast motions with correlation times {tau}{sub f} = 10{sup {minus}9} to 10{sup {minus}10} sec without significantly affecting their geometry. Weak locking-field and temperature dependences are observed for both pure lipid bilayers and bilayers containing either cholesterol or gramicidin, suggesting that the motions of phospholipid acyl chains may have dispersed correlation times.

  18. Kinetics for development of gramicidin-induced ion permeabiliity in unilamellar phospholipid vesicles

    SciTech Connect

    Clement, N.R.; Gould, J.M.

    1981-01-01

    The kinetics for the development of gramicidin-dependent cation permeability in small unilamellar vesicles have been studied by using a vesicle-entrapped, pH-sensitive fluorescence probe to continuously report changes in intravesicular pH. The incorporation of 4 to 5 gramicidin dimers/vesicle was sufficient to increase the proton and counterion permeabilty of that vesicle by several orders of magnitude, so that ionic equilibration following a perturbation of the external medium pH occurred in <1 s. Once a functional gramicidin dimer has become incorporated into one vesicle, it does not readily exchange into another, so that the effects of gramicidin with regard to an individual vesicle can be considered to be essentially all or none. The rate at which transmembrane ion permeability develops in a vesicle suspension was found to depend upon the degree of fluidity of the membrane hydrocarbon interior, being much lower at low temperatures or when cholesterol was present in the bilayer. Low temperatures and increasing bilayer cholesterol content also decreased the number of vesicles affected by a given gramicidin concentration, indicating a decreased membrane solubility for the ionophore at low bilayer fluidities.

  19. Computer Simulation Studies of Ion Channels at High Temperatures

    NASA Astrophysics Data System (ADS)

    Song, Hyun Deok

    The gramicidin channel is the smallest known biological ion channel, and it exhibits cation selectivity. Recently, Dr. John Cuppoletti's group at the University of Cincinnati showed that the gramicidin channel can function at high temperatures (360 ˜ 380K) with significant currents. This finding may have significant implications for fuel cell technology. In this thesis, we have examined the gramicidin channel at 300K, 330K, and 360K by computer simulation. We have investigated how the temperature affects the current and differences in magnitude of free energy between the two gramicidin forms, the helical dimer (HD) and the double helix (DH). A slight decrease of the free energy barrier inside the gramicidin channel and increased diffusion at high temperatures result in an increase of current. An applied external field of 0.2V/nm along the membrane normal results in directly observable ion transport across the channels at high temperatures for both HD and DH forms. We found that higher temperatures also affect the probability distribution of hydrogen bonds, the bending angle, the distance between dimers, and the size of the pore radius for the helical dimer structure. These findings may be related to the gating of the gramicidin channel. Methanococcus jannaschii (MJ) is a methane-producing thermophile, which was discovered at a depth of 2600m in a Pacific Ocean vent in 1983. It has the ability to thrive at high temperatures and high pressures, which are unfavorable for most life forms. There have been some experiments to study its stability under extreme conditions, but still the origin of the stability of MJ is not exactly known. MJ0305 is the chloride channel protein from the thermophile MJ. After generating a structure of MJ0305 by homology modeling based on the Ecoli ClC templates, we examined the thermal stability, and the network stability from the change of network entropy calculated from the adjacency matrices of the protein. High temperatures increase the

  20. N-Terminally Glutamate-Substituted Analogue of Gramicidin A as Protonophore and Selective Mitochondrial Uncoupler

    PubMed Central

    Sorochkina, Alexandra I.; Plotnikov, Egor Y.; Rokitskaya, Tatyana I.; Kovalchuk, Sergei I.; Kotova, Elena A.; Sychev, Sergei V.; Zorov, Dmitry B.; Antonenko, Yuri N.

    2012-01-01

    Limited uncoupling of oxidative phosphorylation could be beneficial for cells by preventing excessive generation of reactive oxygen species. Typical uncouplers are weak organic acids capable of permeating across membranes with a narrow gap between efficacy and toxicity. Aimed at designing a nontoxic uncoupler, the protonatable amino acid residue Glu was substituted for Val at the N-terminus of the pentadecapeptide gramicidin A (gA). The modified peptide [Glu1]gA exhibited high uncoupling activity in isolated mitochondria, in particular, abolishing membrane potential at the inner mitochondrial membrane with the same or even larger efficacy as gA. With mitochondria in cell culture, the depolarizing activity of [Glu1]gA was observed at concentrations by an order of magnitude lower than those of gA. On the contrary, [Glu1]gA was much less potent in forming proton channels in planar lipid bilayers than gA. Remarkably, at uncoupling concentrations, [Glu1]gA did not alter cell morphology and was nontoxic in MTT test, in contrast to gA showing high toxicity. The difference in the behavior of [Glu1]gA and gA in natural and artificial membranes could be ascribed to increased capability of [Glu1]gA to permeate through membranes and/or redistribute between different membranes. Based on the protective role of mild uncoupling, [Glu1]gA and some other proton-conducting gA analogues may be considered as prototypes of prospective therapeutic agents. PMID:22911866

  1. N-terminally glutamate-substituted analogue of gramicidin A as protonophore and selective mitochondrial uncoupler.

    PubMed

    Sorochkina, Alexandra I; Plotnikov, Egor Y; Rokitskaya, Tatyana I; Kovalchuk, Sergei I; Kotova, Elena A; Sychev, Sergei V; Zorov, Dmitry B; Antonenko, Yuri N

    2012-01-01

    Limited uncoupling of oxidative phosphorylation could be beneficial for cells by preventing excessive generation of reactive oxygen species. Typical uncouplers are weak organic acids capable of permeating across membranes with a narrow gap between efficacy and toxicity. Aimed at designing a nontoxic uncoupler, the protonatable amino acid residue Glu was substituted for Val at the N-terminus of the pentadecapeptide gramicidin A (gA). The modified peptide [Glu1]gA exhibited high uncoupling activity in isolated mitochondria, in particular, abolishing membrane potential at the inner mitochondrial membrane with the same or even larger efficacy as gA. With mitochondria in cell culture, the depolarizing activity of [Glu1]gA was observed at concentrations by an order of magnitude lower than those of gA. On the contrary, [Glu1]gA was much less potent in forming proton channels in planar lipid bilayers than gA. Remarkably, at uncoupling concentrations, [Glu1]gA did not alter cell morphology and was nontoxic in MTT test, in contrast to gA showing high toxicity. The difference in the behavior of [Glu1]gA and gA in natural and artificial membranes could be ascribed to increased capability of [Glu1]gA to permeate through membranes and/or redistribute between different membranes. Based on the protective role of mild uncoupling, [Glu1]gA and some other proton-conducting gA analogues may be considered as prototypes of prospective therapeutic agents.

  2. Hydrophobic mismatch in gramicidin A prime /lecithin systems

    SciTech Connect

    Watnick, P.I.; Chan, S.I. ); Dea, P. )

    1990-07-03

    Gramicidin A{prime} (GA{prime}) has been added to three lipid systems of varying hydrophobic thickness: dimyristoyllecithin (DML), dipalmitoyllecithin (DPL), and distearoyllecithin (DSL). The similarity in length between the hydrophobic portion of GA{prime} and the hydrocarbon chains of the lipid bilayers has been studied by using {sup 31}P and {sup 2}H NMR. Hydrophobic mismatch has been found to be most severe in the DML bilayer system and minimal in the case of DSL. In addition, the effects of hydrophobic mismatch on the cooperative properties of the bilayer have been obtained from {sup 2}H NMR relaxation measurements. The results indicate that incorporation of the peptide into the bilayer disrupts the cooperative director fluctuations characteristic of pure multilamellar lipid dispersions. Finally, the GA{prime}/lecithin ratio at which the well-known transformation from bilayer to reverse hexagonal (H{sub II}) phase occurs is shown to depend on the acyl chain length of the phospholipid. A rationale is proposed for this chain length dependence.

  3. Interaction of polypeptide antibiotic gramicidin S with platelets.

    PubMed

    Hackl, Ellen V; Berest, Vladimir P; Gatash, Sergey V

    2012-12-01

    Gramicidin S (GS) is a cyclic decapeptide antibiotic active against both Gram-positive and Gram-negative bacteria as well as against several pathogenic fungi. However, clinical application of GS is limited because of GS hemolytic activity. The large number of GS analogues with potentially attenuated hemolytic activity has been developed over the last two decades. For all new GS derivatives, the antimicrobial test is accompanied with the hemolytic activity assay. At the same time, neither GS nor its analogues were tested against other blood cells. In the present work, the effects of GS on platelets and platelet aggregates have been studied. GS interaction with platelets is concentration dependent and leads either to platelet swelling or platelet shape change. Effect of GS on platelets is independent of platelet aggregation mechanism. GS induces disaggregation of platelet aggregates formed in the presence of aggregation agonists. The rate of the GS interaction with platelet membranes depends on membrane lipid mobility and significantly increases with temperature. The interaction of GS with the platelet membranes depends strongly on the state of the membrane lipids. Factors affecting the membrane lipids (temperature, lipid peroxidation and ionising irradiation) modify GS interaction with platelets. Our results show that GS is active not only against erythrocytes but also against other blood cells (platelets). The estimated numbers of GS molecules per 1 µm2 of a blood cell required to induce erythrocyte hemolysis and disaggregation of platelet aggregates are comparable. This must be considered when developing new antimicrobial GS analogues with improved hemolytic properties.

  4. 5-A Fourier map of gramicidin A phased by deuterium-hydrogen solvent difference neutron diffraction

    SciTech Connect

    Koeppe, R.E. II; Schoenborn, B.P.

    1984-03-01

    Crystals of ion-free gramicidin A (P2/sub 1/2/sub 1/2/sub 1/: a = 24.61, b = 32.28, c = 32.52) have been investigated using neutron diffraction. A difference analysis of crystals soaked in ethanol/H/sub 2/O as opposed to ethanol-d/sub 6//D/sub 2/O has lead to single isomorphous replacement Fourier projections of the structure at 5-A resolution. The gramicidin dimer appears to be a 32-A-long cylinder oriented parallel to the c-axis in these crystals.

  5. Thallium-205 nuclear magnetic resonance study of the thallium(I)-gramicidin A association in trifluoroethanol

    SciTech Connect

    Turner, G.L.; Hinton, J.F.; Millett, F.S.

    1982-02-16

    Association of the thallous ion with gramicidin in 2,2,2-trifluoroethanol has been investigated by thallium-205 NMR spectroscopy. The data obtained suggest that the gramicidin dimer has two strong binding sites and one or more weak binding sites. Association constants for the strong binding sites were found to have the same value. From the temperature dependence of the strong binding site association constants, values for the association enthalpy and entropy of -2.13 +/- 0.12 kcal/mol and +5.45 +/- 0.04 eu, respectively, were obtained.

  6. Changes in liposomes permeability induced by gramicidin D after microwave irradiation

    SciTech Connect

    Yova, D.; Boudouris, G.; Loukas, S.

    1984-08-01

    The response of model membranes (liposomes) to microwave irradiation (9.2 GHz, energy absorption 20 mW/g, continuous 1.5-h exposure in an orthogonal waveguide) was determined spectrophotometrically by recording after irradiation the gramicidin D-induced cation permeability. The irradiation modified the gramicidin D-induced permeability to the cations K/sup +/, Na/sup +/, and Rb/sup +/ through the liposomes and seemed to facilitate the movement of Na/sup +/ and Rb/sup +/. These results are discussed in relation with the hypothesis that microwave radiation may induce changes to the structure of liposomes.

  7. Vertical profile of 137Cs in soil.

    PubMed

    Krstić, D; Nikezić, D; Stevanović, N; Jelić, M

    2004-12-01

    In this paper, a vertical distribution of 137Cs in undisturbed soil was investigated experimentally and theoretically. Soil samples were taken from the surroundings of the city of Kragujevac in central Serbia during spring-summer of 2001. The sampling locations were chosen in such a way that the influence of soil characteristics on depth distribution of 137Cs in soil could be investigated. Activity of 137Cs in soil samples was measured using a HpGe detector and multi-channel analyzer. Based on vertical distribution of 137Cs in soil which was measured for each of 10 locations, the diffusion coefficient of 137Cs in soil was determined. In the next half-century, 137Cs will remain as the source of the exposure. Fifteen years after the Chernobyl accident, and more than 30 years after nuclear probes, the largest activity of 137Cs is still within 10 cm of the upper layer of the soil. This result confirms that the penetration of 137Cs in soil is a very slow process. Experimental results were compared with two different Green functions and no major differences were found between them. While both functions fit experimental data well in the upper layer of soil, the fitting is not so good in deeper layers. Although the curves obtained by these two functions are very close to each other, there are some differences in the values of parameters acquired by them. PMID:15388151

  8. Salivary levels of gramicidin after use of a tyrothricin lozenge and a tyrothricin gargle/mouth-wash.

    PubMed

    Kreuzig, F; Nahler, G

    1983-01-01

    Salivary levels and total salivary recovery of gramicidin were determined by high performance liquid chromatography in healthy volunteers after administration of a tyrothricin lozenge and a tyrothricin gargle/mouth-wash. Average peak values of gramicidin after sucking the lozenge were 37.5 mg/l and were usually reached within the five minutes. After use of the lozenge mean total recovery of gramicidin in saliva was 44% within the first 30 minutes. After application of the gargle/mouth-wash about 11% was retained in the oral cavity and excreted with saliva within the following 30 minutes. The implications of these findings are discussed. PMID:6207119

  9. Cyclic decapeptide gramicidin S derivatives containing phosphines: novel ligands for asymmetric catalysis.

    PubMed

    Guisado-Barrios, Gregorio; Muñoz, Bianca K; Kamer, Paul C J; Lastdrager, Bas; van der Marel, Gijs; Overhand, Mark; Vega-Vázquez, Marino; Martin-Pastor, Manuel

    2013-02-14

    The cyclic peptide gramicidin S was used as a rigid template to provide novel peptide-based bisphosphine ligands for transition metal catalysis. Two bisphosphine-coordinated Rh(I) complexes allowed asymmetric hydrogenation with 10-52% ee and the corresponding Pd(II) analogues catalysed asymmetric allylic alkylation with 13-15% ee.

  10. Solvent-free thermocyclization of the unactivated linear gramicidin S precursor and analogues.

    PubMed

    An, Lin-Kun; Li, Run-Lin; Zuo, Ying-Lin; Gu, Lian-Quan

    2011-01-01

    A convenient thermocyclization of the linear gramicidin S precursor and its analogues is demonstrated. With the preorganized β-sheet conformation, the unactivated linear precursors can cyclize into the corresponding head-to-tail cyclic products in high yield after being heated under solvent-free conditions.

  11. NMR study of the interactions of polymyxin B, gramicidin S, and valinomycin with dimyristoyllecithin bilayers

    SciTech Connect

    Zidovetzki, R.; Banerjee, U.; Harrington, D.W.; Chan, S.I.

    1988-07-26

    The interactions of three polypeptide antibiotics (polymyxin B, gramicidin S, and valinomycin) with artificial lecithin membranes were studied by nuclear magnetic resonance (NMR). Combination of /sup 31/P and /sup 2/H NMR allowed observation of perturbations of the bilayer membrane structure induced by each of the antibiotics in the regions of the polar headgroups and acyl side chains of the phospholipids. The comparative study of the effects of these membrane-active antibiotics and the lipid bilayer structure demonstrated distinct types of antibiotic-membrane interactions in each case. Thus, the results showed the absence of interaction of polymyxin B with the dimyristoyllecithin membranes. In contrast, gramicidin S exhibited strong interaction with the lipid above the gel to liquid-crystalline phase transition temperature: disordering of the acyl side chains was evident. Increasing the concentration of gramicidin S led to disintegration of the bilayer membrane structure. At a molar ratio of 1:16 of gramicidin S to lecithin, the results are consistent with coexistence of gel and liquid-crystalline phases of the phospholipids near the phase transition temperature. Valinomycin decreased the phase transition temperature of the lipids and increased the order parameters of the lipid side chains. Such behavior is consistent with penetration of the valinomycin molecule into the interior of the lipid bilayers.

  12. Collaborative study for the establishment of the second international standard for gramicidin.

    PubMed

    Rautmann, G; Daas, A; Buchheit, K-H

    2010-10-01

    An international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) to establish the WHO second International Standard (IS) for gramicidin as the stocks of the 1st IS, established in 1964, were close to depletion. The candidate material did not show any sign of potency loss when kept at elevated temperatures of + 4 °C, + 20 °C, + 37 °C and + 45 °C for 3 months. Six laboratories from 5 countries as well as the EDQM laboratory participated in the collaborative study. Potencies of the candidate material were estimated by microbiological assays with sensitive micro-organisms. To ensure continuity between consecutive batches, the 1st IS for gramicidin was used as standard. Based on the results of the study, the 2nd IS for gramicidin was adopted at the meeting of the WHO Expert Committee on Biological Standardization (ECBS) in 2008 with an assigned potency of 1070 International Units per mg (IU/mg). The 2nd IS for gramicidin is available from the EDQM.

  13. Folding and membrane insertion of the pore-forming peptide gramicidin occur as a concerted process.

    PubMed

    Hicks, Matthew R; Damianoglou, Angeliki; Rodger, Alison; Dafforn, Timothy R

    2008-11-01

    Many antibiotic peptides function by binding and inserting into membranes. Understanding this process provides an insight into the fundamentals of both membrane protein folding and antibiotic peptide function. For the first time, in this work, flow-aligned linear dichroism (LD) is used to study the folding of the antibiotic peptide gramicidin. LD provides insight into the combined processes of peptide folding and insertion and has the advantage over other similar techniques of being insensitive to off-membrane aggregation events. By combining LD data with conventional measurements of protein fluorescence and circular dichroism, the mechanism of gramicidin insertion is elucidated. The mechanism consists of five separately assignable steps that include formation of a water-insoluble gramicidin aggregate, dissociation from the aggregate, partitioning of peptide to the membrane surface, oligomerisation on the surface and concerted insertion and folding of the peptide to the double-helical form of gramicidin. Measurement of the rates of each step shows that although changes in the fluorescence signal cease 10 s after the initiation of the process, the insertion of the peptide into the membrane is actually not complete for a further 60 min. This last membrane insertion phase is only apparent by measurement of LD and circular dichroism signal changes. In summary, this study demonstrates the importance of multi-technique approaches, including LD, in studies of membrane protein folding.

  14. Synchrotron radiation linear dichroism spectroscopy of the antibiotic peptide gramicidin in lipid membranes.

    PubMed

    Hicks, Matthew R; Dafforn, Timothy R; Damianoglou, Angeliki; Wormell, Paul; Rodger, Alison; Hoffmann, Søren V

    2009-08-01

    We have developed synchrotron radiation linear dichroism (SRLD) to measure the insertion of peptides into lipid bilayers, significantly improving both signal-to-noise and wavelength range over existing methods. Our wavelength cut-off is currently determined by the quality of quartz in the cell, rather than the light source, with signal quality still high at the cut-off. We demonstrate the use of a lipid probe to measure the orientation of the lipid bilayers under flow and describe the way in which this can be used to further interpret SRLD data. The antibiotic peptide gramicidin is shown to exhibit drastically different kinetic and equilibrium behaviour when interacting with lipid membranes with different properties. The charge on the membrane is of interest because of differences in charge between human and bacterial membranes. For this reason we increased the negative charge on the membrane by changing the lipid composition. Increasing negative charge in the gel phase stabilises the liposomes but changes the kinetics of peptide folding. In a gel phase with no negatively charged lipids, gramicidin does not fold well and gives a small signal that indicates a change in orientation of the tryptophan side chains over time. In the fluid phase with no negatively charged lipids, there is initially >10-fold greater peptide signal relative to the gel phase indicating a highly folded and ordered gramicidin backbone. This is followed by liposome disruption. In the gel phase with negatively charged lipids the liposomes are resistant to disruption by gramicidin and exhibit different folding kinetics depending on membrane composition. In the fluid phase with negatively charged lipids there is little signal from either the peptide or the lipid probe indicating that the liposomes have been disrupted by the gramicidin in the time it takes to make the first measurement.

  15. Observation of interspecies 6Li-133Cs Feshbach resonances

    NASA Astrophysics Data System (ADS)

    Repp, M.; Pires, R.; Ulmanis, J.; Heck, R.; Kuhnle, E. D.; Weidemüller, M.; Tiemann, E.

    2013-01-01

    We report on the observation of 19 interspecies Feshbach resonances in an optically trapped ultracold Bose-Fermi mixture of 133Cs and 6Li in the two energetically lowest spin states. We assign the resonances to s- and p-wave molecular channels by a coupled-channels calculation, resulting in an accurate determination of LiCs ground-state potentials. Fits of the resonance position based on the undressed asymptotic bound state model do not provide the same level of accuracy as the coupled-channels calculation. Several broad s-wave resonances provide prospects to create fermionic LiCs molecules with a large dipole moment via Feshbach association followed by stimulated Raman passage. Two of the s-wave resonances overlap with a zero crossing of the Cs scattering length, which offers prospects for the investigation of polarons in an ultracold Li-Cs mixture.

  16. Permeation in ion channels: the interplay of structure and theory.

    PubMed

    Miloshevsky, Gennady V; Jordan, Peter C

    2004-06-01

    Combined with high-resolution atomic-level crystal structures of channel forming peptides, theory has become a powerful tool for illuminating factors influencing permeation. Here, advantages and limitations of the more familiar continuum and molecular modeling techniques are briefly outlined. These methods are applied to issues of permeation in two different channel families: gramicidin and K(+) channels. Using structural data, theory provides verifiable atomic-level insights into permeation dynamics, channel conductance and molecular selectivity mechanisms. Not only can theory confirm experimental inference, it can also sometimes provide structural perspectives in advance of experiment.

  17. Preparation of the multienzyme system gramicidin S-synthetase 2 with an aqueous three-phase system.

    PubMed

    Kirchner, A; Simonis, M; von Döhren, H

    1987-06-19

    The distribution of gramicidin S-synthetase activity from disrupted cells suspended in aqueous two- and three-phase systems was investigated. An optimized three-phase system containing 5% dextran, 8% Ficoll, 11% PEG and 6.7% disrupted cells was found to be effective in extracting gramicidin S-synthetase activity. The activity yield achieved was higher in comparison to other preparation methods, and the subsequent purification steps were greatly facilitated. The time needed for the preparation of the labile gramicidin S-synthetase was considerably reduced. The combination of the aqueous phase extraction with chromatographic methods yielded 19 mg gramicidin S-synthetase 2 in essentially pure form from 30 g (wet weight) of cells.

  18. Inhibiting and reversing amyloid-β peptide (1-40) fibril formation with gramicidin S and engineered analogues.

    PubMed

    Luo, Jinghui; Otero, José M; Yu, Chien-Hung; Wärmländer, Sebastian K T S; Gräslund, Astrid; Overhand, Mark; Abrahams, Jan Pieter

    2013-12-16

    In Alzheimer's disease, amyloid-β (Aβ) peptides aggregate into extracellular fibrillar deposits. Although these deposits may not be the prime cause of the neurodegeneration that characterizes this disease, inhibition or dissolution of amyloid fibril formation by Aβ peptides is likely to affect its development. ThT fluorescence measurements and AFM images showed that the natural antibiotic gramicidin S significantly inhibited Aβ amyloid formation in vitro and could dissolve amyloids that had formed in the absence of the antibiotic. In silico docking suggested that gramicidin S, a cyclic decapeptide that adopts a β-sheet conformation, binds to the Aβ peptide hairpin-stacked fibril through β-sheet interactions. This may explain why gramicidin S reduces fibril formation. Analogues of gramicidin S were also tested. An analogue with a potency that was four-times higher than that of the natural product was identified.

  19. Selective and ATP-driven transport of ions across supported membranes into nanoporous carriers using gramicidin A and ATP synthase.

    PubMed

    Oliynyk, Vitaliy; Mille, Christian; Ng, Jovice B S; von Ballmoos, Christoph; Corkery, Robert W; Bergström, Lennart

    2013-02-28

    We report a robust and versatile membrane protein based system for selective uptake and release of ions from nanoporous particles sealed with ion-tight lipid bilayers of various compositions that is driven by the addition of ATP or a chemical potential gradient. We have successfully incorporated both a passive ion channel-type peptide (gramicidin A) and a more complex primary sodium ion transporter (ATP synthase) into the supported lipid bilayers on solid nanoporous silica particles. Protein-mediated controlled release/uptake of sodium ions across the ion-tight lipid bilayer seal from or into the nanoporous silica carrier was imaged in real time using a confocal laser scanning microscope and the intensity changes were quantified. ATP-driven transport of sodium ions across the supported lipid bilayer against a chemical gradient was demonstrated. The possibility of designing durable carriers with tight lipid membranes, containing membrane proteins for selective ion uptake and release, offers new possibilities for functional studies of single or cascading membrane protein systems and could also be used as biomimetic microreactors for controlled synthesis of inorganic multicomponent materials.

  20. Peptide-induced membrane leakage by lysine derivatives of gramicidin A in liposomes, planar bilayers, and erythrocytes.

    PubMed

    Sorochkina, Alexandra I; Kovalchuk, Sergei I; Omarova, Elena O; Sobko, Alexander A; Kotova, Elena A; Antonenko, Yuri N

    2013-11-01

    Introducing a charged group near the N-terminus of gramicidin A (gA) is supposed to suppress its ability to form ion channels by restricting its head-to-head dimerization. The present study dealt with the activity of [Lys1]gA, [Lys3]gA, [Glu1]gA, [Glu3]gA, [Lys2]gA, and [Lys5]gA in model membrane systems (planar lipid bilayers and liposomes) and erythrocytes. In contrast to the Glu-substituted peptides, the lysine derivatives of gA caused non-specific liposomal leakage monitored by fluorescence dequenching of lipid vesicles loaded with carboxyfluorescein or other fluorescent dyes. Measurements of electrical current through a planar lipid membrane revealed formation of giant pores by Lys-substituted analogs, which depended on the presence of solvent in the bilayer lipid membrane. The efficacy of unselective pore formation in liposomes depended on the position of the lysine residue in the amino acid sequence, increasing in the row: [Lys2]gA<[Lys5]gA<[Lys1]gA<[Lys3]gA. The similar series of potency was exhibited by the Lys-substituted gA analogs in facilitating erythrocyte hemolysis, whereas the Glu-substituted analogs showed negligible hemolytic activity. Oligomerization of the Lys-substituted peptides is suggested to be involved in the process of nonselective pore formation.

  1. Multiphoton dissociative ionization of CS+

    NASA Astrophysics Data System (ADS)

    Rajput, Jyoti; Jochim, Bethany; Zohrabi, M.; Betsch, K. J.; Ablikim, U.; Berry, Ben; Severt, T.; Summers, A. M.; Armstrong, G. S. J.; Esry, B. D.; Carnes, K. D.; Ben-Itzhak, I.

    2015-05-01

    We have studied the dissociative photoionization of a CS+ molecular ion beam in the strong-field regime using <50 fs IR laser pulses (λ ~ 790 nm) from a 10 kHz, ~2 mJ (per pulse) Ti:Sapphire laser system. A coincidence three-dimensional momentum imaging method was used to measure all ions and neutrals formed during this multiphoton process. Two prominent channels were observed: charge-symmetric dissociation, yielding C+ + S+, and charge-asymmetric dissociation, yielding C + S2+. The differences between these two channels with reference to their relative production probability, energetics, and angular distributions is the focus of this work. This work was supported by the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy. BJ is also supported by DOE-SCGF (DE-AC05-06OR23100).

  2. Dynamics of an integral membrane peptide: a deuterium NMR relaxation study of gramicidin.

    PubMed Central

    Prosser, R S; Davis, J H

    1994-01-01

    Solid state deuterium (2H) NMR inversion-recovery and Jeener-Broekaert relaxation experiments were performed on oriented multilamellar dispersions consisting of 1,2-dilauroyl-sn-glycero-3-phosphatidylcholine and 2H exchange-labeled gramicidin D, at a lipid to protein molar ratio (L/P) of 15:1, in order to study the dynamics of the channel conformation of the peptide in a liquid crystalline phase. Our dynamic model for the whole body motions of the peptide includes diffusion of the peptide around its helix axis and a wobbling diffusion around a second axis perpendicular to the local bilayer normal in a simple Maier-Saupe mean field potential. This anisotropic diffusion is characterized by the correlation times, tau R parallel and tau R perpendicular. Aligning the bilayer normal perpendicular to the magnetic field and graphing the relaxation rate, 1/T1Z, as a function of (1-S2N-2H), where S2N-2H represents the orientational order parameter, wer were able to estimate the correlation time, tau R parallel, for rotational diffusion. Although in the quadrupolar splitting, which varies as (3 cos2 theta D-1), has in general two possible solutions to theta D in the range 0 < or = theta D < or = 90 degrees, the 1/T1Z vs. (1-S2N-2H) curve can be used to determine a single value of theta D in this range. Thus, the 1/T1Z vs. (1-S2N-2H) profile can be used both to define the axial diffusion rate and to remove potential structural ambiguities in the splittings. The T1Z anisotropy permits us to solve for the two correlation times (tau R parallel = 6.8 x 10(-9) s and tau R perpendicular = 6 x 10(-6) s). The simulated parameters were corroborated by a Jeener-Broekaert experiment where the bilayer normal was parallel to the principal magnetic field. At this orientation the ratio, J2(2 omega 0)/J1(omega 0) was obtained in order to estimate the strength of the restoring potential in a model-independent fashion. This measurement yields the rms angle, 1/2 (= 16 +/- 2 degrees at

  3. Equilibrium binding constants for Tl+ with gramicidins A, B and C in a lysophosphatidylcholine environment determined by 205Tl nuclear magnetic resonance spectroscopy.

    PubMed Central

    Hinton, J F; Koeppe, R E; Shungu, D; Whaley, W L; Paczkowski, J A; Millett, F S

    1986-01-01

    Nuclear Magnetic Resonance (NMR) 205Tl spectroscopy has been used to monitor the binding of Tl+ to gramicidins A, B, and C packaged in aqueous dispersions of lysophosphatidylcholine. For 5 mM gramicidin dimer in the presence of 100 mM lysophosphatidylcholine, only approximately 50% or less of the gramicidin appears to be accessible to Tl+. Analysis of the 205Tl chemical shift as a function of Tl+ concentration over the 0.65-50 mM range indicates that only one Tl+ ion can be bound by gramicidin A, B, or C under these experimental conditions. In this system, the Tl+ equilibrium binding constant is 582 +/- 20 M-1 for gramicidin 1949 +/- 100 M-1 for gramicidin B, and 390 +/- 20 M-1 for gramicidin C. Gramicidin B not only binds Tl+ more strongly but it is also in a different conformational state than that of A and C, as shown by Circular Dichroism spectroscopy. The 205Tl NMR technique can now be extended to determinations of binding constants of other cations to gramicidin by competition studies using a 205Tl probe. PMID:2420383

  4. The pH-dependent induction of lipid membrane ionic permeability by N-terminally lysine-substituted analogs of gramicidin A.

    PubMed

    Rokitskaya, Tatyana I; Sorochkina, Alexandra I; Kovalchuk, Sergey I; Egorova, Natalya S; Kotova, Elena A; Sychev, Sergey V; Antonenko, Yuri N

    2012-02-01

    Insertion of charged groups at the N-terminus of the gramicidin A (gA) amino acid sequence is considered to be fatal for peptide channel-forming activity because of hindrance to the head-to-head dimer formation. Here the induction of ionic conductivity in planar bilayer lipid membranes (BLM) was studied with gA analogs having lysine either in the first ([Lys1]gA) or the third ([Lys3]gA) position. If added to the bathing solution at neutral or acidic pH, these analogs, being protonated and thus positively charged, were unable to induce ionic current across BLM. By contrast, at pH 11 the induction of BLM conductivity was observed with both lysine-substituted analogs. Based on the dependence of the macroscopic current on the side of the peptide addition, sensitivity to calcium ions and susceptibility to sensitized photoinactivation, as well as on the single-channel properties of the analogs, we surmise that at alkaline pH [Lys1]gA formed channels with predominantly single-stranded structure of head-to-head helical dimers, whereas [Lys3]gA open channels had the double-stranded helical structure. CD spectra of the lysine-substituted analogs in liposomes were shown to be pH-dependent.

  5. Gramicidin S and polymyxins: the revival of cationic cyclic peptide antibiotics.

    PubMed

    Mogi, Tatsushi; Kita, Kiyoshi

    2009-12-01

    Gramicidin S and polymyxins are small cationic cyclic peptides and act as potent antibiotics against Gram-negative and Gram-positive bacteria by perturbing integrity of the bacterial membranes. Screening of a natural antibiotics library with bacterial membrane vesicles identified gramicidin S as an inhibitor of cytochrome bd quinol oxidase and an alternative NADH dehydrogenase (NDH-2) and polymyxin B as an inhibitor of NDH-2 and malate: quinone oxidoreductase. Our studies showed that cationic cyclic peptide antibiotics have novel molecular targets in the membrane and interfere ligand binding on the hydrophobic surface of enzymes. Improvement of the toxicity and optimization of the structures and clinical uses are urgently needed for their effective application in combating drug-resistant bacteria.

  6. The Gramicidin Dimer Shows Both EX1 and EX2 Mechanisms of H/D Exchange

    PubMed Central

    Chitta, Raghu K.; Rempel, Don L.; Gross, Michael L.

    2009-01-01

    We describe the use of H/D amide exchange and electrospray ionization mass spectrometry to study, in organic solvents, the pentadecapeptide gramicidin as a model for protein self association. In methanol-OD, all active H’s in the peptide exchange for D within 5 min, indicating a monomer/dimer equilibrium that is shifted towards the fast-exchanging monomer. H/D exchange in n-propanol-OD, however, showed a partially protected gramicidin that slowly converts to a second species that exchanges nearly all the active hydrogens, indicating EX1 kinetics for the H/D exchange. We propose that this behavior is the result of slower rate of unfolding in n-propanol compared to that in methanol. The rate constant for the unfolding of the dimer is the rate of disappearance of the partially protected species, and it agrees within a factor of two with a value reported in literature. The rate constant of dimer refolding can be determined from the ratio of the rate constant for unfolding and the affinity constant for the dimer, which we determined in an earlier study. The unfolding activation energy is 20 kcal mol-1, determined by performing the exchange experiments as a function of temperature. To study gramicidin in an even more hydrophobic medium than n-propanol, we measured its H/D exchange kinetics in a phospholipids vesicle and found a different H/D amide exchange behavior. Gramicidin is an unusual peptide dimer that can exhibit both EX1 and EX2 mechanisms for its H/D exchange, depending on the solvent. PMID:19631556

  7. The gramicidin dimer shows both EX1 and EX2 mechanisms of H/D exchange.

    PubMed

    Chitta, Raghu K; Rempel, Don L; Gross, Michael L

    2009-10-01

    We describe the use of H/D amide exchange and electrospray ionization mass spectrometry to study, in organic solvents, the pentadecapeptide gramicidin as a model for protein self association. In methanol-OD, all active H's in the peptide exchange for D within 5 min, indicating a monomer/dimer equilibrium that is shifted towards the fast-exchanging monomer. H/D exchange in n-propanol-OD, however, showed a partially protected gramicidin that slowly converts to a second species that exchanges nearly all the active hydrogens, indicating EX1 kinetics for the H/D exchange. We propose that this behavior is the result of the slower rate of unfolding in n-propanol compared with that in methanol. The rate constant for the unfolding of the dimer is the rate of disappearance of the partially protected species, and it agrees within a factor of two with a value reported in literature. The rate constant of dimer refolding can be determined from the ratio of the rate constant for unfolding and the affinity constant for the dimer, which we determined in an earlier study. The unfolding activation energy is 20 kcal mol(-1), determined by performing the exchange experiments as a function of temperature. To study gramicidin in an even more hydrophobic medium than n-propanol, we measured its H/D exchange kinetics in a phospholipids vesicle and found a different H/D amide exchange behavior. Gramicidin is an unusual peptide dimer that can exhibit both EX1 and EX2 mechanisms for its H/D exchange, depending on the solvent.

  8. Distinguishing gramicidin D conformers through two-dimensional infrared spectroscopy of vibrational excitons

    PubMed Central

    Tokmakoff, Andrei

    2015-01-01

    Gramicidin D is a short peptide which dimerizes to form helical pores, adopting one of two conformations in the process. These conformations differ primarily in number of residues per turn and the hydrogen-bond registry between rungs of the helix. Using amide I 2D infrared (IR) and FTIR, we have demonstrated that it is possible to distinguish between the different conformers of gramicidin D in solution. We show that the spectra observed for this helical peptide bear no resemblance to the spectra of α- or 310-helices and that while the FTIR spectra appear similar to spectra of β-sheets, 2D IR reveals that the observed resonances arise from vibrational modes unlike those observed in β-sheets. We also present an idealized model which reproduces the experimental data with high fidelity. This model is able to explain the polarization-dependence of the experimental 2D IR data. Using this model, we show the coupling between the rungs of the helix dominates the spectra, and as a consequence of this, the number of residues per turn can greatly influence the amide I spectra of gramicidin D. PMID:26049444

  9. Oxygen-dependent inactivation of gramicidin S synthetase in Bacillus brevis.

    PubMed Central

    Friebel, T E; Demain, A L

    1977-01-01

    Incorporation of L-[14C]ornithine into gramicidin S by crude, unfractionated lysozyme extracts of Bacillus brevis ATCC 9999 was shown to represent the activity of the gramicidin synthetase complex. Frozen-thawed cells were the source of active extracts, but when cells were shaken in air at 37 degrees C, they rapidly lost activity in a first-order reaction with a half-life of 13 min. Protease inhibitors and inhibitors of energy metabolism had no effect on the inactivation process in frozen-thawed cells. Stabilization was achieved when the cells were shaken in nitrogen or helium instead of air. The addition of dithiothreitol produced a moderate degree of stabilization. The L-ornithine- and D-phenylalanine-activating activities of the gramicidin S synthetase complex were also lost during aeration of the cells. Crude cell-free extracts also lost activity when they were shaken in oxygen, but, in this case, inactivation was slower (half-life of 80 min). Nitrogen also stabilized these cell-free extracts. PMID:68033

  10. Gramicidin Alters the Lipid Compositions of Liquid-Ordered and Liquid-Disordered Membrane Domains

    NASA Astrophysics Data System (ADS)

    Hassan-Zadeh, Ebrahim; Huang, Juyang

    2012-10-01

    The effects of adding 1 mol % of gramicidin A to the well-known DOPC/DSPC/cholesterol lipid mixtures were investigated. 4-component giant unilamellar vesicles (GUV) were prepared using our recently developed Wet-Film method. The phase boundary of liquid-ordered and liquid-disordered (Lo-Ld) coexisting region was determined using video fluorescence microscopy. We found that if cares were not taken, light-induced domain artifacts could significantly distort the measured phase boundary. After testing several fluorescence dyes, we found that the emission spectrum of Nile Red is quite sensitive to membrane composition. By fitting the Nile Red emission spectra at the phase boundary to the spectra in the Lo-Ld coexisting region, the thermodynamic tie-lines were determined. As an active component of lipid membranes, gramicidin not only partitions favorably into the liquid-disordered (Ld) phase, it also alters the phase boundary and thermodynamic tie-lines. Even at as low as 1 mol %, gramicidin decreases the cholesterol mole fraction of Ld phase and increases the area of Lo phase.

  11. Distinguishing gramicidin D conformers through two-dimensional infrared spectroscopy of vibrational excitons.

    PubMed

    Stevenson, Paul; Tokmakoff, Andrei

    2015-06-01

    Gramicidin D is a short peptide which dimerizes to form helical pores, adopting one of two conformations in the process. These conformations differ primarily in number of residues per turn and the hydrogen-bond registry between rungs of the helix. Using amide I 2D infrared (IR) and FTIR, we have demonstrated that it is possible to distinguish between the different conformers of gramicidin D in solution. We show that the spectra observed for this helical peptide bear no resemblance to the spectra of α- or 310-helices and that while the FTIR spectra appear similar to spectra of β-sheets, 2D IR reveals that the observed resonances arise from vibrational modes unlike those observed in β-sheets. We also present an idealized model which reproduces the experimental data with high fidelity. This model is able to explain the polarization-dependence of the experimental 2D IR data. Using this model, we show the coupling between the rungs of the helix dominates the spectra, and as a consequence of this, the number of residues per turn can greatly influence the amide I spectra of gramicidin D.

  12. Distinguishing gramicidin D conformers through two-dimensional infrared spectroscopy of vibrational excitons

    NASA Astrophysics Data System (ADS)

    Stevenson, Paul; Tokmakoff, Andrei

    2015-06-01

    Gramicidin D is a short peptide which dimerizes to form helical pores, adopting one of two conformations in the process. These conformations differ primarily in number of residues per turn and the hydrogen-bond registry between rungs of the helix. Using amide I 2D infrared (IR) and FTIR, we have demonstrated that it is possible to distinguish between the different conformers of gramicidin D in solution. We show that the spectra observed for this helical peptide bear no resemblance to the spectra of α- or 310-helices and that while the FTIR spectra appear similar to spectra of β-sheets, 2D IR reveals that the observed resonances arise from vibrational modes unlike those observed in β-sheets. We also present an idealized model which reproduces the experimental data with high fidelity. This model is able to explain the polarization-dependence of the experimental 2D IR data. Using this model, we show the coupling between the rungs of the helix dominates the spectra, and as a consequence of this, the number of residues per turn can greatly influence the amide I spectra of gramicidin D.

  13. EXTRAGALACTIC CS SURVEY

    SciTech Connect

    Bayet, E.; Viti, S.; Aladro, R.; MartIn, S.; MartIn-Pintado, J.

    2009-12-10

    We present a coherent and homogeneous multi-line study of the CS molecule in nearby (D < 10 Mpc) galaxies. We include, from the literature, all the available observations from the J = 1-0 to the J = 7-6 transitions toward NGC 253, NGC 1068, IC 342, Henize 2-10, M 82, the Antennae Galaxies, and M 83. We have, for the first time, detected the CS(7-6) line in NGC 253, M 82 (both in the northeast and southwest molecular lobes), NGC 4038, M 83 and tentatively in NGC 1068, IC 342, and Henize 2-10. We use the CS molecule as a tracer of the densest gas component of the interstellar medium in extragalactic star-forming regions, following previous theoretical and observational studies by Bayet et al. In this first paper out of a series, we analyze the CS data sample under both local thermodynamical equilibrium (LTE) and non-LTE (large velocity gradient) approximations. We show that except for M 83 and Overlap (a shifted gas-rich position from the nucleus NGC 4039 in the Antennae Galaxies), the observations in NGC 253, IC 342, M 82-NE, M 82-SW, and NGC 4038 are not well reproduced by a single set of gas component properties and that, at least, two gas components are required. For each gas component, we provide estimates of the corresponding kinetic temperature, total CS column density, and gas density.

  14. On resin amino acid side chain attachment strategy for the head to tail synthesis of new glutamine containing gramicidin-S analogs and their antimicrobial activity.

    PubMed

    Derbal, Safa; Hensler, Mary; Fang, Weiqin; Nizet, Victor; Ghedira, Kamel; Nefzi, Adel

    2010-10-01

    The alarming increase in infections caused by multiple drug resistant bacteria including methicillin-resistant Staphylococcus aureus has prompted a desperate search for new antimicrobials. Augmenting the discoveries of completely new scaffolds with antimicrobial activity are efforts aimed at modifying existing molecules to optimize activity or reduce toxicity. We report herein the parallel solid-phase synthesis of analogues of the cationic antimicrobial peptide gramicidin S (GS) using amino acid side chain attachment strategy. The ornithine (Orn) residues were replaced by glutamine (Gln) and the aromatic D-phenylalanine (Phe) were replaced by different aromatic D-amino acids. Additional Gln containing GS analogues with all the possible combinations of the hydrophobic amino acids valine and leucine were also synthesized. In this work we also report the antibacterial activity of these analogs against several clinically-important drug-resistant Gram-positive and Gram-negative pathogens.

  15. Structure and dynamics of one-dimensional ionic solutions in biological transmembrane channels.

    PubMed Central

    Skerra, A; Brickmann, J

    1987-01-01

    The structure and dynamics of solvated alkali metal cations in transmembrane channels are treated using the molecular dynamics simulation technique. The simulations are based on a modified Fischer-Brickmann model (Fischer, W., and J. Brickmann, 1983, Biophys. Chem., 18:323-337) for gramicidin A-type channels. The trajectories of all particles in the channel as well as two-dimensional pair correlation functions are analyzed. It is found from the analysis of the stationary simulation state that one-dimensional solvation complexes are formed and that the number of water molecules in the channel varies for different alkali metal cations. PMID:2440485

  16. Lipid bilayer array for simultaneous recording of ion channel activities

    NASA Astrophysics Data System (ADS)

    Hirano-Iwata, Ayumi; Nasu, Tomohiro; Oshima, Azusa; Kimura, Yasuo; Niwano, Michio

    2012-07-01

    This paper describes an array of stable and reduced-solvent bilayer lipid membranes (BLMs) formed in microfabricated silicon chips. BLMs were first vertically formed simultaneously and then turned 90° in order to realize a horizontal BLM array. Since the present BLMs are mechanically stable and robust, the BLMs survive this relatively tough process. Typically, a ˜60% yield in simultaneous BLM formation over 9 sites was obtained. Parallel recordings of gramicidin channel activities from different BLMs were demonstrated. The present system has great potential as a platform of BLM-based high throughput drug screening for ion channel proteins.

  17. Tuning hydrophobicity of highly cationic tetradecameric Gramicidin S analogues using adamantane amino acids.

    PubMed

    Knijnenburg, Annemiek D; Kapoerchan, Varsha V; Spalburg, Emile; de Neeling, Albert J; Mars-Groenendijk, Roos H; Noort, Daan; van der Marel, Gijs A; Overkleeft, Herman S; Overhand, Mark

    2010-12-01

    Ring extended Gramicidin S analogues containing adamantane amino acids and six cationic residues were designed and evaluated. Systematic replacement of the hydrophobic residues with adamantane amino acids resulted in a small set of compounds with varying amphipathic character. It was found that the amphipathicity of these compounds is correlated to their biological activity. Several bacterial strains including MRSA strains were shown to be killed by the novel peptides. The most potent antibacterial peptides are tetradecameric GS analogues containing six positives charges and two adamantane moieties.

  18. Tetrahydrofuran amino acid-containing gramicidin S analogues with improved biological profiles.

    PubMed

    Pal, Sudip; Singh, Gajendra; Singh, Shyam; Tripathi, Jitendra Kumar; Ghosh, Jimut Kanti; Sinha, Sudhir; Ampapathi, Ravi Sankar; Chakraborty, Tushar Kanti

    2015-06-28

    Gramicidin S (GS) is a cyclic cationic antimicrobial peptide (CAP) with a wide spectrum of antibiotic activities whose usage has been limited to topical applications owing to its cytotoxic side effects. We have synthesized tetrahydrofuran amino acid (Taa)-containing GS analogues, and we have carried out conformational analysis and explored their structure activity relationships by evaluating their antitubercular, antibacterial and cytotoxic properties. Two of these analogues showed impressive as well as selective activity against Mycobacterium tuberculosis (MTB) without toxicity towards mammalian Vero cells or human RBCs, and are promising as potential leads.

  19. Inhibiting mild steel corrosion from sulfate-reducing and iron-oxidizing bacteria using gramicidin-S-producing biofilms.

    PubMed

    Zuo, Rongjun; Wood, Thomas K

    2004-11-01

    A gramicidin-S-producing Bacillus brevis 18-3 biofilm was shown to reduce corrosion rates of mild steel by inhibiting both the sulfate-reducing bacterium Desulfosporosinus orientis and the iron-oxidizing bacterium Leptothrix discophora SP-6. When L. discophora SP-6 was introduced along with D. orientis to a non-antimicrobial-producing biofilm control, Paenibacillus polymyxa ATCC 10401, a corrosive synergy was created and mild steel coupons underwent more severe corrosion than when only D. orientis was present, showing a 2.3-fold increase via electrochemical impedance spectroscopy (EIS) and a 1.8-fold difference via mass-loss measurements. However, when a gramicidin-S-producing, protective B. brevis 18-3 biofilm was established on mild steel, the metal coupons were protected against the simultaneous attack of D. orientis and L. discophora SP-6. EIS data showed that the protective B. brevis 18-3 biofilm decreased the corrosion rate about 20-fold compared with the non-gramicidin-producing P. polymyxa ATCC 10401 biofilm control. The mass loss for the protected mild steel coupons was also significantly lower than that for the unprotected ones (4-fold decrease). Scanning electron microscope images corroborated the corrosion inhibition by the gramicidin-S-producing B. brevis biofilm on mild steel by showing that the metal surface remained untarnished, i.e., the polishing grooves were still visible after exposure to the simultaneous attack of the sulfate-reducing bacterium and the iron-oxidizing bacterium. PMID:15278311

  20. Inhibiting mild steel corrosion from sulfate-reducing and iron-oxidizing bacteria using gramicidin-S-producing biofilms.

    PubMed

    Zuo, Rongjun; Wood, Thomas K

    2004-11-01

    A gramicidin-S-producing Bacillus brevis 18-3 biofilm was shown to reduce corrosion rates of mild steel by inhibiting both the sulfate-reducing bacterium Desulfosporosinus orientis and the iron-oxidizing bacterium Leptothrix discophora SP-6. When L. discophora SP-6 was introduced along with D. orientis to a non-antimicrobial-producing biofilm control, Paenibacillus polymyxa ATCC 10401, a corrosive synergy was created and mild steel coupons underwent more severe corrosion than when only D. orientis was present, showing a 2.3-fold increase via electrochemical impedance spectroscopy (EIS) and a 1.8-fold difference via mass-loss measurements. However, when a gramicidin-S-producing, protective B. brevis 18-3 biofilm was established on mild steel, the metal coupons were protected against the simultaneous attack of D. orientis and L. discophora SP-6. EIS data showed that the protective B. brevis 18-3 biofilm decreased the corrosion rate about 20-fold compared with the non-gramicidin-producing P. polymyxa ATCC 10401 biofilm control. The mass loss for the protected mild steel coupons was also significantly lower than that for the unprotected ones (4-fold decrease). Scanning electron microscope images corroborated the corrosion inhibition by the gramicidin-S-producing B. brevis biofilm on mild steel by showing that the metal surface remained untarnished, i.e., the polishing grooves were still visible after exposure to the simultaneous attack of the sulfate-reducing bacterium and the iron-oxidizing bacterium.

  1. Influence of an electric field on oriented films of DMPC/gramicidin bilayers: a circular dichroism study.

    PubMed

    Fiche, J B; Laredo, T; Tanchak, O; Lipkowski, J; Dutcher, J R; Yada, R Y

    2010-01-19

    A film of oriented bilayers containing a mixture of gramicidin and dimyristoylphosphatidylcholine (DMPC) has been deposited on a fused-silica window coated with a 10 nm thick gold layer. The thin layer of gold allows the application of an electric potential across the film and the study of its influence on the structure and integrity of the bilayers. Electrochemical measurements, ellipsometry, and far-UV circular dichroism (CD) were employed to characterize the properties of the film of bilayers as a function of the potential applied to the gold electrode. For potentials across the film that are within the range approximately +300 to -150 mV the oriented film of bilayers is stable, and no change in the CD spectra of gramicidin molecule is observed. At more negative potentials, an increase in the film thickness and water content measured by ellipsometry indicated that the film swells and incorporates water, which causes a change in the circular dichroism spectrum of gramicidin molecules in the film. This transformation was interpreted as a change in the average orientation of gramicidin molecules within the film due to a decrease in the ordering of the molecules upon swelling.

  2. Pantetheine-Linked Peptide Intermediates in Gramicidin S and Tyrocidine Biosynthesis

    PubMed Central

    Kleinkauf, Horst; Roskoski, Robert; Lipmann, Fritz

    1971-01-01

    To study the function of pantetheine in gramicidin S and tyrocidine biosynthesis, pepsin digests of the polymerizing enzymes, of which only the heavy ones contain pantetheine, were analyzed. The digests of gramicidin S enzymes charged with either [14C]proline or with D-phenylalanyl-[14C]proline, were analyzed by thin-layer chromatography; only the dipeptide showed a derivative associated with pantetheine. Similar results were obtained from the heavy tyrocidine enzyme charged with either [14C]asparagine alone or with the pentapeptide D-Phe-Pro-Phe-D-Phe-[14C]Asn. Several radioactive products appeared on the thin-layer chromatograms of both these digests; association with pantetheine was found only in the case of the pentapeptide. Exposure of the chromatogram from the pentapeptide-labeled digest to performic acid and development in a second direction separated the peptide from pantetheine, indicating that a nascent peptide was originally linked to the cofactor by a thioester bond. The connection of pantetheine only with peptide residues appears to confirm its role in transpeptidation during peptide chain growth. Images PMID:4943784

  3. Influence of gramicidin on the dynamics of DMPC studied by incoherent elastic neutron scattering

    NASA Astrophysics Data System (ADS)

    Wanderlingh, U.; D'Angelo, G.; Conti Nibali, V.; Gonzalez, M.; Crupi, C.; Mondelli, C.

    2008-03-01

    By using the fixed energy window method in incoherent elastic neutron scattering, molecular motions in the 150 ps timescale in highly oriented multilayers of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) membranes in excess of water (D2O) have been studied as a function of temperature, in the range from 27 to 325 K. The same system in partially deuterated form and with the addition of a pore-forming peptide (gramicidin) has also been investigated. By proper orientation of the membrane plane with respect to the scattering wavevector Q, information on in plane and out of plane motions of lipid membranes have been derived. Two relevant dynamical transitions were observed at T = 297 K and at T = 270 K. The former is related to the structural main transition from gel to liquid phase of the phospholipid bilayer, while the latter is related to a transition of the aqueous solvent. The inclusion of gramicidin shifts the main transition down to 294 K and the second transition up to 276 K. In both cases the observed dynamical transitions show an enhanced mobility in the direction normal to the membrane plane.

  4. Near-yrast structure of Cs142 and Cs144

    NASA Astrophysics Data System (ADS)

    Rząca-Urban, T.; Genevey, J.; Materna, T.; Urban, W.; Smith, A. G.; Pinston, J. A.; Simpson, G. S.; Sadowski, M. P.; Köster, U.; Faust, H.; Bail, A.; Mathieu, L.; Serot, O.; Michel-Sendis, F.; Ahmad, I.

    2009-12-01

    Excited states in Cs142 and Cs144, populated in the spontaneous fission of Cm248 and Cf252 and in thermal neutron-induced fission of U235 and Am242 were studied by means of γ spectroscopy using the EUROGAM2 and Gammasphere multidetector Ge arrays and the LOHENGRIN fission-fragment separator, respectively. In Cs142, a band and an isomer with a half-life of T1/2=11(3) ns have been identified. Spins and parities have been proposed for excited levels in this nucleus. In Cs144 excited levels have been observed. A T1/2=1.1(1) μs isomer was found with a γ cascade, which probably feeds this isomer. There is also an indication of a nanosecond isomer in Cs144. Quasiparticle-rotor model calculations done in this work allowed proton-neutron configurations to be proposed for levels in Cs142 and Cs144.

  5. Deuterium NMR of Val1. (2-2H)Ala3. gramicidin A in oriented DMPC bilayers

    SciTech Connect

    Hing, A.W.; Adams, S.P.; Silbert, D.F.; Norberg, R.E. )

    1990-05-01

    Deuterium NMR is used to study the selectively labeled Val1...(2-2H)Ala3...gramicidin A molecule to investigate the structure and dynamics of the C alpha-2H bond in the Ala3 residue of gramicidin. Val1...(2-2H)Ala3...gramicidin A is synthesized, purified, and characterized and then incorporated into oriented bilayers of dimyristoylphosphatidylcholine sandwiched between glass coverslips. Phosphorus NMR line shapes obtained from this sample are consistent with the presence of the bilayer phase and indicate that no nonbilayer phases are present in significant amounts. Deuterium NMR line shapes obtained from this sample indicate that the motional axis of the gramicidin Ala3 residue is parallel to the coverslip normal, that the distribution of motional axis orientations has a width of 2 degrees, and that only one major conformational and dynamical state of the Ala3 C alpha-2H bond is observed on the NMR time scale. Furthermore, the Ala3 C alpha-2H bond angle relative to the motional axis is 19-20 degrees if fast axial rotation is assumed to be the only motion present but is less than or equal to 19-20 degrees in the absence of such an assumption. This result indicates that various double-stranded, helical dimer models are very unlikely to represent the structure of gramicidin in the sample studied but that the single-stranded, beta 6.3 helical dimer models are consistent with the experimental data. However, a definitive distinction between the left-handed, single-stranded, beta 6.3 helical dimer model and the right-handed, single-stranded, beta 6.3 helical dimer model cannot be made on the basis of the experimental data obtained in this study.

  6. Differential scanning calorimetry and /sup 2/H NMR studies of the phase behavior of gramicidin-phosphatidylcholine mixtures

    SciTech Connect

    Morrow, M.R.; Davis, J.H.

    1988-03-22

    The extents of two-phase coexistence in the phase diagrams of mixtures of gramicidin with 1,2-bis(perdeuteriopalmitoyl)-sn-glycero-3-phosphocholine (DPPC-d62) and with 1,2-bis(perdeuteriomyristoyl)-sn-glycero-3-phosphocholine (DMPC-d54) mixtures have been explored with differential scanning calorimetry (DSC) and deuterium nuclear magnetic resonance (/sup 2/H NMR). For both systems, increased gramicidin content causes a decrease in transition enthalpy and a broadening of the peak in excess heat capacity at the transition. In DMPC-d54-based mixtures, the broadening is roughly symmetric about the pure lipid transition temperature. Addition of gramicidin to DPPC-d62 extends the excess heat capacity peak on the low-temperature side, resulting in a slightly asymmetric scan. Deuterium NMR spectra showing a superposition of gel and liquid-crystalline components, observed for both mixtures, indicate the presence of two-phase coexistence. For the DPPC-d62-based mixtures, two-phase coexistence is restricted to an approximately 2 degrees C temperature range below the pure transition temperature. For DMPC-d54-based mixtures, the region of two-phase coexistence is even narrower. For both mixtures, beyond a gramicidin mole fraction of 2%, distinct gel and liquid-crystal contributions to the spectra cannot be distinguished. Along with the broad featureless nature of the DSC scan in this region, this is taken to indicate that the transition has been replaced by a continuous phase change. These results are consistent with the existence of a closed two-phase region having a critical concentration of gramicidin below 2 mol%.

  7. Generalized Langevin models of molecular dynamics simulations with applications to ion channels

    NASA Astrophysics Data System (ADS)

    Gordon, Dan; Krishnamurthy, Vikram; Chung, Shin-Ho

    2009-10-01

    We present a new methodology, which combines molecular dynamics and stochastic dynamics, for modeling the permeation of ions across biological ion channels. Using molecular dynamics, a free energy profile is determined for the ion(s) in the channel, and the distribution of random and frictional forces is measured over discrete segments of the ion channel. The parameters thus determined are used in stochastic dynamics simulations based on the nonlinear generalized Langevin equation. We first provide the theoretical basis of this procedure, which we refer to as "distributional molecular dynamics," and detail the methods for estimating the parameters from molecular dynamics to be used in stochastic dynamics. We test the technique by applying it to study the dynamics of ion permeation across the gramicidin pore. Given the known difficulty in modeling the conduction of ions in gramicidin using classical molecular dynamics, there is a degree of uncertainty regarding the validity of the MD-derived potential of mean force (PMF) for gramicidin. Using our techniques and systematically changing the PMF, we are able to reverse engineer a modified PMF which gives a current-voltage curve closely matching experimental results.

  8. Complete Genome Sequence of Aneurinibacillus migulanus E1, a Gramicidin S- and d-Phenylalanyl-l-Propyl Diketopiperazine-Deficient Mutant

    PubMed Central

    Alenezi, Faizah N.; Luptakova, Lenka; Rateb, Mostafa E.; Woodward, Steve

    2015-01-01

    We report here the complete genome sequence of the Aneurinibacillus migulanus E1 mutant deficient in gramicidin S (GS) and d-phenylalanyl-l-propyl diketopiperazine (DKP) formation. The genome consists of a circular chromosome (6,301,904 bp, 43.20% G+C content) without any plasmid. The complete genome sequence enables further investigation of the biosynthetic mechanism and the biological function of gramicidin S. PMID:26679577

  9. Charge Fluctuations and Boundary Conditions of Biological Ion Channels: Effect on the Ionic Transition Rate

    NASA Astrophysics Data System (ADS)

    Tindjong, R.; Luchinsky, D. G.; McClintock, P. V. E.; Kaufman, I.; Eisenberg, R. S.

    2009-04-01

    A self-consistent solution is derived for the Poisson-Nernst-Planck (PNP) equation, valid both inside a biological ion channel and in the adjacent bulk fluid. An iterative procedure is used to match the two solutions together at the channel mouth. Charge fluctuations at the mouth are modeled as shot noise flipping the height of the potential barrier at the selectivity site. The resultant estimates of the conductivity of the ion channel are in good agreement with Gramicidin experimental measurements and they reproduce the observed current saturation with increasing concentration.

  10. Charge Fluctuations and Boundary Conditions of Biological Ion Channels: Effect on the Ionic Transition Rate

    SciTech Connect

    Tindjong, R.; McClintock, P. V. E.; Luchinsky, D. G.; Kaufman, I.; Eisenberg, R. S.

    2009-04-23

    A self-consistent solution is derived for the Poisson-Nernst-Planck (PNP) equation, valid both inside a biological ion channel and in the adjacent bulk fluid. An iterative procedure is used to match the two solutions together at the channel mouth. Charge fluctuations at the mouth are modeled as shot noise flipping the height of the potential barrier at the selectivity site. The resultant estimates of the conductivity of the ion channel are in good agreement with Gramicidin experimental measurements and they reproduce the observed current saturation with increasing concentration.

  11. High temperature synthesis of two open-framework uranyl silicates with ten-ring channels: Cs{sub 2}(UO{sub 2}){sub 2}Si{sub 8}O{sub 19} and Rb{sub 2}(UO{sub 2}){sub 2}Si{sub 5}O{sub 13}

    SciTech Connect

    Babo, Jean-Marie; Albrecht-Schmitt, Thomas E.

    2013-01-15

    The uranyl silicates Cs{sub 2}(UO{sub 2}){sub 2}Si{sub 8}O{sub 19} and Rb{sub 2}(UO{sub 2}){sub 2}Si{sub 5}O{sub 13} were obtained by mixing stoichiometric amounts of uranium metal, tellurium dioxide, silicon dioxide, and an excess of correspondent alkali metal halide flux. These compounds crystallize in the orthorhombic space groups Pnma and C222 with eight and two units per unit cell, respectively. Their crystal structures are dominated by zippered pentagonal bipyramidal chains of UO{sub 7} and silicates layer that are further connected into 3D frameworks. The cesium compound has silicate double layers while rubidium has a single layer. Six-ring voids and ten-ring channels are found in both compounds. - Graphical abstract: A view of the three-dimensional network structure of Cs{sub 2}(UO{sub 2}){sub 2}Si{sub 8}O{sub 19}. Highlights: Black-Right-Pointing-Pointer Three-dimensional uranium silicates. Black-Right-Pointing-Pointer Analogs of natural uranyl silicate minerals. Black-Right-Pointing-Pointer Complexity and symmetry ambiguity of uranyl silicates.

  12. Solution conformation of gramicidin S: An intramolecular nuclear Overhauser effect study.

    PubMed

    Huang, D H; Walter, R; Glickson, J D; Krishna, N R

    1981-02-01

    The solution conformation of gramicidin S in deuterated dimethyl sulfoxide was investigated by using the intramolecular nuclear Overhauser effect experiment. Experimental Overhauser enhancements were compared with predicted values for each of the nine most-stable conformations (M1-M9) calculated by Dygert et al. on the basis of energy-minimization procedures [Dygert, M., Gō, N. & Scheraga, H. A. (1975) Macromolecules 8, 750-761]. By using statistical hypothesis testing, the three lowest-energy conformations (M1, M2, and M3) were shown to give the best fit with the experimental data. All other conformations (M4-M9) were found to be inconsistent with the experimental data. PMID:16592969

  13. Synthesis, biological activity and solution structure of new analogues of the antimicrobial Gramicidin S.

    PubMed

    Kamysz, Elżbieta; Mickiewicz, Beata; Kamysz, Wojciech; Bielińska, Sylwia; Rodziewicz-Motowidło, Sylwia; Ciarkowski, Jerzy

    2011-03-01

    Gramicidin S (GS) is a cyclo-decapeptide antibiotic isolated from Bacillus brevis. The structural studies have shown that GS forms a two-stranded antiparallel β-sheet imposed by two II' β-turns. Despite its wide Gram+ and Gram- antimicrobial spectrum, GS is useless in therapy because of its high hemotoxicity in humans. It was found, however, that the analogues of GS-14 (GS with 14 amino acid residues) attained a better antimicrobial selectivity when their amphipatic moments were perturbed. In this study, we report effects of similar perturbations imposed on GS cyclo-decapeptide analogues. Having solved their structures by NMR/molecular dynamics and having tested their activities/selectivities, we have concluded that the idea of perturbation of the amphipatic moment does not work for GS-10_0 analogues. An innovative approach to the synthesis of head-to-tail cyclopeptides was used.

  14. Polycationic gramicidin S analogues with both high antibiotic activity and very low hemolytic activity.

    PubMed

    Tamaki, Makoto; Harada, Takuji; Fujinuma, Kenta; Takanashi, Kazumasa; Shindo, Mitsuno; Kimura, Masahiro; Uchida, Yoshiki

    2012-01-01

    The substitution of each constituent amino acid residue of gramicidin S (GS), cyclo(-Val(1,1')-Orn(2,2')-Leu(3,3')-D-Phe(4,4')-Pro(5,5')-)(2) with Lys residue indicated that each side chain structure of the constituent amino acid residues affect largely the antibiotic activity and hemolytic activity of GS. Further, the substitution of D-Phe(4,4') and Pro(5,5') residues with basic amino acid residues as a Lys residue results the high antibiotic activity and the very low hemolytic activity. Thus, we have found novel positions on the scaffold of GS at D-Phe(4,4') and Pro(5,5') residues whose modification will significantly increase the therapeutic index.

  15. Electron transport, uptake of protons, and photophosphorylation in chloroplasts: effects of gramicidin d at various ph values

    SciTech Connect

    Gil'miyarova, S.G.; Blymentel'd, L.A.; Puuge, E.K.; Tikhonov, A.N.

    1986-02-01

    A comparative investigation is presented of the photoinduced uptake of protons by bean chloroplasts, electron transport, and photophosphorylation, with the results of previous work on the subject. The study suggests that the dual nature of the action of gramicidin D--effective uncoupling action at low concentrations and it ''inability'' to suppress proton uptake and the formation of ..delta..ph--may be a consequence of the morphological and functional heterogeneity of the proton acceptor groups localized in the thyalkoid membrane.

  16. /sup 2/H nuclear magnetic resonance of exchange-labeled gramicidin in an oriented lyotropic nematic phase

    SciTech Connect

    Davis, J.H.

    1988-01-12

    Lyotropic nematic liquid-crystalline phases, such as that formed by potassium laurate/decanol/KCl/water, are found to accept readily large amphiphilic solute molecules. Since these phases spontaneously orient in high magnetic fields, it becomes possible to obtain NMR spectra of biologically interesting solutes in an oriented axially symmetric environment. The amide hydrogens of the peptide backbone of gramicidin D (Dubos) were exchanged for deuterium, and the gramicidin was incorporated into a lyotropic nematic phase made with deuteriated buffer in place of water. /sup 2/H NMR spectra of oriented, exchange-labeled gramicidin were then obtained. The strong water signal from the deuteriated buffer was eliminated by using selective excitation and a polynomial subtraction procedure. The /sup 2/H NMR spectra at high temperature consist of twelve major quadrupolar doublets. The splittings observed are largely independent of temperature, suggesting a highly rigid backbone structure. Two of the doublets, which are chemically shifted relative to the others, show stronger temperature dependence. These two probably arise from the exchangeable amino hydrogens on the tryptophan indole moieties of the peptide. While we cannot yet assign all of the doublets, the spectra and nuclear magnetic relaxation data are consistent with a rigid slightly distorted ..beta../sub LD//sup 6.3/ helix undergoing axially symmetric reorientation about the director of the liquid-crystalline phase. The correlation time for the axially symmetric reorientation is determined by relaxation measurements to be about 10/sup -7/s.

  17. Single-Molecule Ion Channel Conformational Dynamics in Living Cells

    NASA Astrophysics Data System (ADS)

    Lu, H. Peter

    2014-03-01

    Stochastic and inhomogeneous conformational changes regulate the function and dynamics of ion channels that are crucial for cell functions, neuronal signaling, and brain functions. Such complexity makes it difficult, if not impossible, to characterize ion channel dynamics using conventional electrical recording alone since that the measurement does not specifically interrogate the associated conformational changes but rather the consequences of the conformational changes. Recently, new technology developments on single-molecule spectroscopy, and especially, the combined approaches of using single ion channel patch-clamp electrical recording and single-molecule fluorescence imaging have provided us the capability of probing ion channel conformational changes simultaneously with the electrical single channel recording. By combining real-time single-molecule fluorescence imaging measurements with real-time single-channel electric current measurements in artificial lipid bilayers and in living cell membranes, we were able to probe single ion-channel-protein conformational changes simultaneously, and thus providing an understanding the dynamics and mechanism of ion-channel proteins at the molecular level. The function-regulating and site-specific conformational changes of ion channels are now measurable under physiological conditions in real-time, one molecule at a time. We will focus our discussion on the new development and results of real-time imaging of the dynamics of gramicidin, colicin, and NMDA receptor ion channels in lipid bilayers and living cells. Our results shed light on new perspectives of the intrinsic interplay of lipid membrane dynamics, solvation dynamics, and the ion channel functions.

  18. Bandhead Energies in 125Cs

    NASA Astrophysics Data System (ADS)

    Sun, Ji; Hu, Xue-Yuan; Ma, Ying-Jun; Liu, Yun-Zuo; Tetsuro, Komatsubara; Kohei, Furuno; Zhang, Yu-Hu; Zhou, Wen-Ping; Wang, Shou-Yu

    Excited states in 125Cs have been studied with the fusion-evaporation-reaction 116Cd(14N,5n)125Cs at 65 MeV beam energy, using the Nordball-multidetector-system at the Niels-Bohr-Institute in Denmark. The level scheme of 125Cs was extended with the addition of more than 40 new γ-transitions. Moreover, the bandhead excitation energies of the previously known g9/2 and h11/2 bands were unambiguously corrected with plenty of hard evidence.

  19. Evaluation of readily accessible azoles as mimics of the aromatic ring of D-phenylalanine in the turn region of gramicidin S.

    PubMed

    van der Knaap, Matthijs; Lageveen, Lianne T; Busscher, Henk J; Mars-Groenendijk, Roos; Noort, Daan; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; van der Marel, Gijsbert A; Overkleeft, Herman S; Overhand, Mark

    2011-05-01

    The influence of replacing the d-phenylalanine residue with substituted and unsubstituted azoles on the structure and biological activity of the antibiotic gramicidin S was investigated against a representative panel of Gram-positive and Gram-negative bacteria strains. Substituted triazole derivatives, obtained using a convergent synthetic strategy, are as active as gramicidin S, provided that any substituent on the triazole moiety is not too large. The unsubstituted triazole derivative was biologically less active than the parent natural product, gramicidin S. In general for the triazole series, the hemolytic activity could be correlated with the antibacterial activity, that is, the higher the antibacterial activity, the higher the toxicity towards blood cells. Interestingly, its imidazole counterpart showed high antibacterial activity, combined with significantly diminished hemolytic activity.

  20. Gramicidin A Mutants with Antibiotic Activity against Both Gram-Positive and Gram-Negative Bacteria.

    PubMed

    Zerfas, Breanna L; Joo, Yechaan; Gao, Jianmin

    2016-03-17

    Antimicrobial peptides (AMPs) have shown potential as alternatives to traditional antibiotics for fighting infections caused by antibiotic-resistant bacteria. One promising example of this is gramicidin A (gA). In its wild-type sequence, gA is active by permeating the plasma membrane of Gram-positive bacteria. However, gA is toxic to human red blood cells at similar concentrations to those required for it to exert its antimicrobial effects. Installing cationic side chains into gA has been shown to lower its hemolytic activity while maintaining the antimicrobial potency. In this study, we present the synthesis and the antibiotic activity of a new series of gA mutants that display cationic side chains. Specifically, by synthesizing alkylated lysine derivatives through reductive amination, we were able to create a broad selection of structures with varied activities towards Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). Importantly, some of the new mutants were observed to have an unprecedented activity towards important Gram-negative pathogens, including Escherichia coli, Klebsiella pneumoniae and Psuedomonas aeruginosa. PMID:26918268

  1. Interaction of Gramicidin S and its Aromatic Amino-Acid Analog with Phospholipid Membranes

    PubMed Central

    Jelokhani-Niaraki, Masoud; Hodges, Robert S.; Meissner, Joseph E.; Hassenstein, Una E.; Wheaton, Laura

    2008-01-01

    To investigate the mechanism of interaction of gramicidin S-like antimicrobial peptides with biological membranes, a series of five decameric cyclic cationic β-sheet-β-turn peptides with all possible combinations of aromatic D-amino acids, Cyclo(Val-Lys-Leu-D-Ar1-Pro-Val-Lys-Leu-D-Ar2-Pro) (Ar ≡ Phe, Tyr, Trp), were synthesized. Conformations of these cyclic peptides were comparable in aqueous solutions and lipid vesicles. Isothermal titration calorimetry measurements revealed entropy-driven binding of cyclic peptides to POPC and POPE/POPG lipid vesicles. Binding of peptides to both vesicle systems was endothermic—exceptions were peptides containing the Trp-Trp and Tyr-Trp pairs with exothermic binding to POPC vesicles. Application of one- and two-site binding (partitioning) models to binding isotherms of exothermic and endothermic binding processes, respectively, resulted in determination of peptide-lipid membrane binding constants (Kb). The Kb1 and Kb2 values for endothermic two-step binding processes corresponded to high and low binding affinities (Kb1 ≥ 100 Kb2). Conformational change of cyclic peptides in transferring from buffer to lipid bilayer surfaces was estimated using fluorescence resonance energy transfer between the Tyr-Trp pair in one of the peptide constructs. The cyclic peptide conformation expands upon adsorption on lipid bilayer surface and interacts more deeply with the outer monolayer causing bilayer deformation, which may lead to formation of nonspecific transient peptide-lipid porelike zones causing membrane lysis. PMID:18621820

  2. Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S.

    PubMed Central

    Conti, E; Stachelhaus, T; Marahiel, M A; Brick, P

    1997-01-01

    The non-ribosomal synthesis of the cyclic peptide antibiotic gramicidin S is accomplished by two large multifunctional enzymes, the peptide synthetases 1 and 2. The enzyme complex contains five conserved subunits of approximately 60 kDa which carry out ATP-dependent activation of specific amino acids and share extensive regions of sequence similarity with adenylating enzymes such as firefly luciferases and acyl-CoA ligases. We have determined the crystal structure of the N-terminal adenylation subunit in a complex with AMP and L-phenylalanine to 1.9 A resolution. The 556 amino acid residue fragment is folded into two domains with the active site situated at their interface. Each domain of the enzyme has a similar topology to the corresponding domain of unliganded firefly luciferase, but a remarkable relative domain rotation of 94 degrees occurs. This conformation places the absolutely conserved Lys517 in a position to form electrostatic interactions with both ligands. The AMP is bound with the phosphate moiety interacting with Lys517 and the hydroxyl groups of the ribose forming hydrogen bonds with Asp413. The phenylalanine substrate binds in a hydrophobic pocket with the carboxylate group interacting with Lys517 and the alpha-amino group with Asp235. The structure reveals the role of the invariant residues within the superfamily of adenylate-forming enzymes and indicates a conserved mechanism of nucleotide binding and substrate activation. PMID:9250661

  3. Therapeutic Potential of Gramicidin S in the Treatment of Root Canal Infections

    PubMed Central

    Berditsch, Marina; Lux, Hannah; Babii, Oleg; Afonin, Sergii; Ulrich, Anne S.

    2016-01-01

    An intrinsic clindamycin-resistant Enterococcus faecalis, the most common single species present in teeth after failed root canal therapy, often possesses acquired tetracycline resistance. In these cases, root canal infections are commonly treated with Ledermix® paste, which contains demeclocycline, or the new alternative endodontic paste Odontopaste, which contains clindamycin; however, these treatments are often ineffective. We studied the killing activity of the cyclic antimicrobial peptide gramicidin S (GS) against planktonic and biofilm cells of tetracycline-resistant clinical isolates of E. faecalis. The high therapeutic potential of GS for the topical treatment of problematic teeth is based on the rapid bactericidal effect toward the biofilm-forming, tetracycline-resistant E. faecalis. GS reduces the cell number of planktonic cells within 20–40 min at a concentration of 40–80 μg/mL. It kills the cells of pre-grown biofilms at concentrations of 100–200 μg/mL, such that no re-growth is possible. The translocation of the peptide into the cell interior and its complexation with intracellular nucleotides, including the alarmon ppGpp, can explain its anti-biofilm effect. The successful treatment of persistently infected root canals of two volunteers confirms the high effectiveness of GS. The broad GS activity towards resistant, biofilm-forming E. faecalis suggests its applications for approval in root canal medication. PMID:27618065

  4. Therapeutic Potential of Gramicidin S in the Treatment of Root Canal Infections.

    PubMed

    Berditsch, Marina; Lux, Hannah; Babii, Oleg; Afonin, Sergii; Ulrich, Anne S

    2016-01-01

    An intrinsic clindamycin-resistant Enterococcus faecalis, the most common single species present in teeth after failed root canal therapy, often possesses acquired tetracycline resistance. In these cases, root canal infections are commonly treated with Ledermix(®) paste, which contains demeclocycline, or the new alternative endodontic paste Odontopaste, which contains clindamycin; however, these treatments are often ineffective. We studied the killing activity of the cyclic antimicrobial peptide gramicidin S (GS) against planktonic and biofilm cells of tetracycline-resistant clinical isolates of E. faecalis. The high therapeutic potential of GS for the topical treatment of problematic teeth is based on the rapid bactericidal effect toward the biofilm-forming, tetracycline-resistant E. faecalis. GS reduces the cell number of planktonic cells within 20-40 min at a concentration of 40-80 μg/mL. It kills the cells of pre-grown biofilms at concentrations of 100-200 μg/mL, such that no re-growth is possible. The translocation of the peptide into the cell interior and its complexation with intracellular nucleotides, including the alarmon ppGpp, can explain its anti-biofilm effect. The successful treatment of persistently infected root canals of two volunteers confirms the high effectiveness of GS. The broad GS activity towards resistant, biofilm-forming E. faecalis suggests its applications for approval in root canal medication. PMID:27618065

  5. Gramicidin A Mutants with Antibiotic Activity against Both Gram-Positive and Gram-Negative Bacteria.

    PubMed

    Zerfas, Breanna L; Joo, Yechaan; Gao, Jianmin

    2016-03-17

    Antimicrobial peptides (AMPs) have shown potential as alternatives to traditional antibiotics for fighting infections caused by antibiotic-resistant bacteria. One promising example of this is gramicidin A (gA). In its wild-type sequence, gA is active by permeating the plasma membrane of Gram-positive bacteria. However, gA is toxic to human red blood cells at similar concentrations to those required for it to exert its antimicrobial effects. Installing cationic side chains into gA has been shown to lower its hemolytic activity while maintaining the antimicrobial potency. In this study, we present the synthesis and the antibiotic activity of a new series of gA mutants that display cationic side chains. Specifically, by synthesizing alkylated lysine derivatives through reductive amination, we were able to create a broad selection of structures with varied activities towards Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). Importantly, some of the new mutants were observed to have an unprecedented activity towards important Gram-negative pathogens, including Escherichia coli, Klebsiella pneumoniae and Psuedomonas aeruginosa.

  6. Synthesis and evaluation of strand and turn modified ring-extended gramicidin S derivatives.

    PubMed

    Knijnenburg, Annemiek D; Kapoerchan, Varsha V; Grotenbreg, Gijsbert M; Spalburg, Emile; de Neeling, Albert J; Mars-Groenendijk, Roos H; Noort, Daan; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; Ravensbergen, Bep; Nibbering, Peter H; van der Marel, Gijs A; Overkleeft, Herman S; Overhand, Mark

    2011-06-01

    In this paper, we describe the crystal structure of previously reported ring-extended gramicidin S (GS) derivative 2 (GS14K4), containing a d-amino acid residue in one of the β-strand regions. This structure is in agreement with a previously reported modeling study of the same molecule. The polar side chain of the additional d-amino acid residue is positioned at the same face of the molecule as the hydrophobic side chains, and we believe that because of this compound 2 is considerably less hydrophobic than extended GS derivatives in which the strand regions are exclusively composed of l-amino acids. Using this backbone structure as our benchmark we prepared a small series of ring-extended GS analogues featuring sugar amino acid dipeptide isosteres of varied hydrophobicity at the turn region. We show that via this approach hydrophobicity of extended GS analogues can be tuned without affecting the secondary structure (as observed from NMR and CD spectra). Biological evaluation reveals that hydrophobicity correlates to cell toxicity, but still bacteriolysis is induced with GS analogues that are too hydrophilic to efficiently lyse human red blood cells.

  7. Optical Waveguide Lightmode Spectroscopic Techniques for Investigating Membrane-Bound Ion Channel Activities

    PubMed Central

    Székács, Inna; Kaszás, Nóra; Gróf, Pál; Erdélyi, Katalin; Szendrő, István; Mihalik, Balázs; Pataki, Ágnes; Antoni, Ferenc A.; Madarász, Emilia

    2013-01-01

    Optical waveguide lightmode spectroscopic (OWLS) techniques were probed for monitoring ion permeation through channels incorporated into artificial lipid environment. A novel sensor set-up was developed by depositing liposomes or cell-derived membrane fragments onto hydrophilic polytetrafluoroethylene (PTFE) membrane. The fibrous material of PTFE membrane could entrap lipoid vesicles and the water-filled pores provided environment for the hydrophilic domains of lipid-embedded proteins. The sensor surface was kept clean from the lipid holder PTFE membrane by a water- and ion-permeable polyethylene terephthalate (PET) mesh. The sensor set-up was tested with egg yolk lecithin liposomes containing gramicidin ion channels and with cell-derived membrane fragments enriched in GABA-gated anion channels. The method allowed monitoring the move of Na+ and organic cations through gramicidin channels and detecting the Cl–-channel functions of the (α5β2γ2) GABAA receptor in the presence or absence of GABA and the competitive GABA-blocker bicuculline. PMID:24339925

  8. Functional and structural insights on self-assembled nanofiber-based novel antibacterial ointment from antimicrobial peptides, bacitracin and gramicidin S.

    PubMed

    Mandal, Santi M; Roy, Anupam; Mahata, Denial; Migliolo, Ludovico; Nolasco, Diego O; Franco, Octavio L

    2014-11-01

    A novel antibacterial ointment using bacitracin, specific for Gram-positive bacteria, and gramicidin S, a highly toxic antibacterial peptide, was here developed showing broad-spectrum antibacterial activities against pathogenic strains with less toxicity after self-assembly into nanofiber structures. Such structures were confirmed with scanning electron microscopy and CD analyses. In addition, in silico studies using docking associated with molecular dynamics were carried out to obtain information about fiber structural oligomerization. Thus, the bacitracin and gramicidin S-based self-assembled nanopeptide ribbon may be a successful ointment formulation for bacterial infection control.

  9. Channel estimation in DCT-based OFDM.

    PubMed

    Wang, Yulin; Zhang, Gengxin; Xie, Zhidong; Hu, Jing

    2014-01-01

    This paper derives the channel estimation of a discrete cosine transform-(DCT-) based orthogonal frequency-division multiplexing (OFDM) system over a frequency-selective multipath fading channel. Channel estimation has been proved to improve system throughput and performance by allowing for coherent demodulation. Pilot-aided methods are traditionally used to learn the channel response. Least square (LS) and mean square error estimators (MMSE) are investigated. We also study a compressed sensing (CS) based channel estimation, which takes the sparse property of wireless channel into account. Simulation results have shown that the CS based channel estimation is expected to have better performance than LS. However MMSE can achieve optimal performance because of prior knowledge of the channel statistic.

  10. Amide-I and -II vibrations of the cyclic beta-sheet model peptide gramicidin S in the gas phase.

    PubMed

    Kupser, Peter; Pagel, Kevin; Oomens, Jos; Polfer, Nick; Koksch, Beate; Meijer, Gerard; von Helden, Gert

    2010-02-17

    In the condensed phase, the peptide gramicidin S is often considered as a model system for a beta-sheet structure. Here, we investigate gramicidin S free of any influences of the environment by measuring the mid-IR spectra of doubly protonated (deuterated) gramicidin S in the gas phase. In the amide I (i.e., C=O stretch) region, the spectra show a broad split peak between 1580 and 1720 cm(-1). To deduce structural information, the conformational space has been searched using molecular dynamics methods and several structural candidates have been further investigated at the density functional level. The calculations show the importance of the interactions of the charged side-chains with the backbone, which is responsible for the lower frequency part of the amide I peak. When this interaction is inhibited via complexation with two 18-crown-6 molecules, the amide I peak narrows and shows two maxima at 1653 and 1680 cm(-1). A comparison to calculations shows that for this complexed ion, four C=O groups are in an antiparallel beta-sheet arrangement. Surprisingly, an analysis of the calculated spectra shows that these beta-sheet C=O groups give rise to the vibrations near 1680 cm(-1). This is in sharp contrast to expectations based on values for the condensed phase, where resonances of beta-sheet sections are thought to occur near 1630 cm(-1). The difference between those values might be caused by interactions with the environment, as the condensed phase value is mostly deduced for beta-sheet sections that are embedded in larger proteins, that interact strongly with solvent or that are part of partially aggregated species.

  11. Salivary levels of gramicidin after use of a tyrothricin-containing gargle/mouth-wash and tyrothricin lozenges.

    PubMed

    Matula, C; Nahler, G; Kreuzig, F

    1988-01-01

    Three pharmaceutical preparations for the disinfection of the oropharynx were tested with regard to the gramicidin concentration obtained in the saliva after their appropriate application. Peak values followed a dose-concentration relationship and were highest after sucking a lozenge containing 10 mg tyrothricin (mean 109.3 mg/L) followed by that of a gargle/mouth-wash containing 667 mg tyrothricin/L (mean 21.1 mg/L) and of a lozenge containing 4 mg tyrothricin (mean 14.4 mg/L). PMID:2460410

  12. An adamantyl amino acid containing gramicidin S analogue with broad spectrum antibacterial activity and reduced hemolytic activity.

    PubMed

    Kapoerchan, Varsha V; Knijnenburg, Annemiek D; Niamat, Miquel; Spalburg, Emile; de Neeling, Albert J; Nibbering, Peter H; Mars-Groenendijk, Roos H; Noort, Daan; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; van der Marel, Gijs A; Overkleeft, Herman S; Overhand, Mark

    2010-10-25

    The cyclic cationic antimicrobial peptide gramicidin S (GS) is an effective topical antibacterial agent that is toxic for human red blood cells (hemolysis). Herein, we present a series of amphiphilic derivatives of GS with either two or four positive charges and characteristics ranging between very polar and very hydrophobic. Screening of this series of peptide derivatives identified a compound that combines effective antibacterial activity with virtually no toxicity within the same concentration range. This peptide acts against both Gram-negative and Gram-positive bacteria, including several MRSA strains, and represents an interesting lead for the development of a broadly applicable antibiotic.

  13. Interaction of gramicidin S and its aromatic amino-acid analog with phospholipid membranes.

    PubMed

    Jelokhani-Niaraki, Masoud; Hodges, Robert S; Meissner, Joseph E; Hassenstein, Una E; Wheaton, Laura

    2008-10-01

    To investigate the mechanism of interaction of gramicidin S-like antimicrobial peptides with biological membranes, a series of five decameric cyclic cationic beta-sheet-beta-turn peptides with all possible combinations of aromatic D-amino acids, Cyclo(Val-Lys-Leu-D-Ar1-Pro-Val-Lys-Leu-D-Ar2-Pro) (Ar identical with Phe, Tyr, Trp), were synthesized. Conformations of these cyclic peptides were comparable in aqueous solutions and lipid vesicles. Isothermal titration calorimetry measurements revealed entropy-driven binding of cyclic peptides to POPC and POPE/POPG lipid vesicles. Binding of peptides to both vesicle systems was endothermic-exceptions were peptides containing the Trp-Trp and Tyr-Trp pairs with exothermic binding to POPC vesicles. Application of one- and two-site binding (partitioning) models to binding isotherms of exothermic and endothermic binding processes, respectively, resulted in determination of peptide-lipid membrane binding constants (K(b)). The K(b1) and K(b2) values for endothermic two-step binding processes corresponded to high and low binding affinities (K(b1) >or= 100 K(b2)). Conformational change of cyclic peptides in transferring from buffer to lipid bilayer surfaces was estimated using fluorescence resonance energy transfer between the Tyr-Trp pair in one of the peptide constructs. The cyclic peptide conformation expands upon adsorption on lipid bilayer surface and interacts more deeply with the outer monolayer causing bilayer deformation, which may lead to formation of nonspecific transient peptide-lipid porelike zones causing membrane lysis.

  14. Novel cycloundecapeptides related to gramicidin S with both high antibiotic activity and low hemolytic activity.

    PubMed

    Tamaki, Makoto; Takanashi, Kazumasa; Harada, Takuji; Fujinuma, Kenta; Shindo, Mitsuno; Kimura, Masahiro; Uchida, Yoshiki

    2011-01-01

    To find candidates with high antimicrobial and low hemolytic activities, many gramicidin S (GS) analogs of various ring sizes have been designed and synthesized. However, syntheses of antimicrobially active analogues of GS having a disordered symmetry structure from C(2) have almost never been reported, because the stable, amphiphilic β-sheet structure of GS with C(2) symmetry is considered essential for its strong antibacterial activity. In the present studies, novel thirteen cycloundecapeptides 1-13 related to GS were synthesized and examined. Among them, cyclo(-Va1(1)-Orn(2)-Leu(3)-D-Phe(4)-X(5)-Pro(6)-Val(7)-Orn(8)-Leu(9)-D-Phe(10)-Pro(11)-) (X=Lys (10), Orn (11), Arg (12) and Lys(Lys) (13)) resulted in high antibiotic activity against both Gram-positive and Gram-negative microorganisms tested. In addition, 11 showed low toxicity against sheep blood cells compared with that of GS. Further, circular dichroism (CD) spectra of 10-13 had a curve similar to each other, suggesting that the conformations of these analogues in methanol are similar to each other. However, CD spectra of 10-13 were different from that of GS in the 190-210 nm region. These results suggest that the presences of one added amino acid residue at position 5 of 10-13 might be partially effective through a structural change in the biological activity of 10-13. In addition, the structural modifications at position 5 lower the undesirable hemolytic activity and enhance the desirable antibiotic activity.

  15. Reactive barriers for 137Cs retention.

    PubMed

    Krumhansl, J L; Brady, P V; Anderson, H L

    2001-02-01

    137Cs was dispersed globally by cold war activities and, more recently, by the Chernobyl accident. Engineered extraction of 137Cs from soils and groundwaters is exceedingly difficult. Because the half-life of 137Cs is only 30.2 years, remediation might be more effective (and less costly) if 137Cs bioavailability could be demonstrably limited for even a few decades by use of a reactive barrier. Essentially permanent isolation must be demonstrated in those few settings where high nuclear level wastes contaminated the environment with 135Cs (half-life 2.3 x 10(6) years) in addition to 137Cs. Clays are potentially a low-cost barrier to Cs movement, though their long-term effectiveness remains untested. To identify optimal clays for Cs retention, Cs desorption was measured for five common clays: Wyoming Montmorillonite (SWy-1), Georgia Kaolinites (KGa-1 and KGa-2), Fithian Illite (F-Ill), and K-Metabentonite (K-Mbt). Exchange sites were pre-saturated with 0.16 M CsCl for 14 days and readily exchangeable Cs was removed by a series of LiNO3 and LiCl washes. Washed clays were then placed into dialysis bags and the Cs release to the deionized water outside the bags measured. Release rates from 75 to 139 days for SWy-1, K-Mbt and F-Ill were similar; 0.017% to 0.021% sorbed Cs released per day. Both kaolinites released Cs more rapidly (0.12% to 0.05% of the sorbed Cs per day). In a second set of experiments, clays were Cs-doped for 110 days and subjected to an extreme and prolonged rinsing process. All the clays exhibited some capacity for irreversible Cs uptake. However, the residual loading was greatest on K-Mbt (approximately 0.33 wt.% Cs). Thus, this clay would be the optimal material for constructing artifical reactive barriers. PMID:11288579

  16. Reactive barriers for 137Cs retention.

    PubMed

    Krumhansl, J L; Brady, P V; Anderson, H L

    2001-02-01

    137Cs was dispersed globally by cold war activities and, more recently, by the Chernobyl accident. Engineered extraction of 137Cs from soils and groundwaters is exceedingly difficult. Because the half-life of 137Cs is only 30.2 years, remediation might be more effective (and less costly) if 137Cs bioavailability could be demonstrably limited for even a few decades by use of a reactive barrier. Essentially permanent isolation must be demonstrated in those few settings where high nuclear level wastes contaminated the environment with 135Cs (half-life 2.3 x 10(6) years) in addition to 137Cs. Clays are potentially a low-cost barrier to Cs movement, though their long-term effectiveness remains untested. To identify optimal clays for Cs retention, Cs desorption was measured for five common clays: Wyoming Montmorillonite (SWy-1), Georgia Kaolinites (KGa-1 and KGa-2), Fithian Illite (F-Ill), and K-Metabentonite (K-Mbt). Exchange sites were pre-saturated with 0.16 M CsCl for 14 days and readily exchangeable Cs was removed by a series of LiNO3 and LiCl washes. Washed clays were then placed into dialysis bags and the Cs release to the deionized water outside the bags measured. Release rates from 75 to 139 days for SWy-1, K-Mbt and F-Ill were similar; 0.017% to 0.021% sorbed Cs released per day. Both kaolinites released Cs more rapidly (0.12% to 0.05% of the sorbed Cs per day). In a second set of experiments, clays were Cs-doped for 110 days and subjected to an extreme and prolonged rinsing process. All the clays exhibited some capacity for irreversible Cs uptake. However, the residual loading was greatest on K-Mbt (approximately 0.33 wt.% Cs). Thus, this clay would be the optimal material for constructing artifical reactive barriers.

  17. Automatable lipid bilayer formation for ion channel studies

    NASA Astrophysics Data System (ADS)

    Poulos, Jason L.; Bang, Hyunwoo; Jeon, Tae-Joon; Schmidt, Jacob J.

    2008-08-01

    Transmembrane proteins and ion channels are important drug targets and have been explored as single molecule sensors. For these proteins to function normally they must be integrated within lipid bilayers; however, the labor and skill required to create artificial lipid bilayers have the limited the possible applications utilizing these proteins. In order to reduce the complexity and cost of lipid bilayer formation and measurement, we have modified a previously published lipid bilayer formation technique using mechanically contacted monolayers so that the process is automatable, requiring minimal operator input. Measurement electronics are integrated with the fluid handling system, greatly reducing the time and operator feedback characteristically required of traditional bilayer experiments. To demonstrate the biological functionality of the resultant bilayers and the system's capabilities as a membrane platform, the ion channel gramicidin A was incorporated and measured with this system.

  18. Lysophospholipids modulate channel function by altering the mechanical properties of lipid bilayers

    PubMed Central

    1994-01-01

    Lipid metabolites, free fatty acids and lysophospholipids, modify the function of membrane proteins including ion channels. Such alterations can occur through signal transduction pathways, but may also result from "direct" effects of the metabolite on the protein. To investigate possible mechanisms for such direct effects, we examined the alterations of gramicidin channel function by lysophospholipids (LPLs): lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), lysophosphatidylserine (LPS), and lysophosphatidylinositol (LPI). The experiments were done on planar bilayers formed by diphytanoylphosphatidylcholine in n-decane a system where receptor- mediated effects can be excluded. At aqueous concentrations below the critical micelle concentration (CMC), LPLs can increase the dimerization constant for membrane-bound gramicidin up to 500-fold (at 2 microM). The relative potency increases as a function of the size of the polar head group, but does not seem to vary as a function of head group charge. The increased dimerization constant results primarily from an increase in the rate constant for channel formation, which can increase more than 100-fold (in the presence of LPC and LPI), whereas the channel dissociation rate constant decreases only about fivefold. The LPL effect cannot be ascribed to an increased membrane fluidity, which would give rise to an increased channel dissociation rate constant. The ability of LPC to decrease the channel dissociation rate constant varies as a function of channel length (which is always less than the membrane's equilibrium thickness): as the channel length is decreased, the potency of LPC is increased. LPC has no effect on membrane thickness or the surface tension of monolayers at the air/electrolyte interface. The bilayer-forming glycerolmonooleate does not decrease the channel dissociation rate constant. These results show that LPLs alter gramicidin channel function by altering the membrane deformation energy, and

  19. Theoretical and computational models of biological ion channels

    NASA Astrophysics Data System (ADS)

    Roux, Benoit

    2004-03-01

    A theoretical framework for describing ion conduction through biological molecular pores is established and explored. The framework is based on a statistical mechanical formulation of the transmembrane potential (1) and of the equilibrium multi-ion potential of mean forces through selective ion channels (2). On the basis of these developments, it is possible to define computational schemes to address questions about the non-equilibrium flow of ions through ion channels. In the case of narrow channels (gramicidin or KcsA), it is possible to characterize the ion conduction in terms of the potential of mean force of the ions along the channel axis (i.e., integrating out the off-axis motions). This has been used for gramicidin (3) and for KcsA (4,5). In the case of wide pores (i.e., OmpF porin), this is no longer a good idea, but it is possible to use a continuum solvent approximations. In this case, a grand canonical monte carlo brownian dynamics algorithm was constructed for simulating the non-equilibrium flow of ions through wide pores. The results were compared with those from the Poisson-Nernst-Planck mean-field electrodiffusion theory (6-8). References; 1. B. Roux, Biophys. J. 73:2980-2989 (1997); 2. B. Roux, Biophys. J. 77, 139-153 (1999); 3. Allen, Andersen and Roux, PNAS (2004, in press); 4. Berneche and Roux. Nature, 414:73-77 (2001); 5. Berneche and Roux. PNAS, 100:8644-8648 (2003); 6. W. Im and S. Seefeld and B. Roux, Biophys. J. 79:788-801 (2000); 7. W. Im and B. Roux, J. Chem. Phys. 115:4850-4861 (2001); 8. W. Im and B. Roux, J. Mol. Biol. 322:851-869 (2002).

  20. Solid-phase peptide synthesis and solid-state NMR spectroscopy of (Ala/sup 3/-/sup 15/N)(Val/sup 1/)gramicidin A

    SciTech Connect

    Fields, G.B.; Fields, C.G.; Petefish, J.; Van Wart, H.E.; Cross, T.A.

    1988-03-01

    (Ala/sup 3-15/N)(Val/sup 1/)Gramicidin A has been prepared by solid-phase peptide synthesis and studied by solid-state /sup 15/N nuclear magnetic resonance spectroscopy. The synthesis of desformyl(Ala/sup 3-15/N)(Val/sup 1/)gramicidin A employed N-hydroxysuccinimide esters of 9-fluorenylmethoxycarbonyl-N/sup ..cap alpha../-amino acids and completely avoided the use of acid. Since deblocking was done with piperidine and the peptide was removed from the resin by treatment with ethanolamine, this synthetic protocol prevented oxidation of the indole rings of this tryptophan-rich peptide and reduced truncations produced by acid hydrolysis. After formylation and purification by anion-exchange and high-pressure liquid chromatography, the peptide was obtained in an overall yield of 30%. Solid-state /sup 15/N nuclear magnetic resonance spectra of this peptide and uniformly labeled (/sup 15/N)gramicidin A' oriented in hydrated lipid bilayers have been obtained, allowing unambiguous assignment of the (/sup 15/N)Ala/sup 3/ resonance in the latter. The solid-state /sup 15/N nuclear magnetic resonance experiments provide evidence that (Val/sup 1/)gramicidin A is rotating about an axis that is perpendicular to the plane of the lipid bilayer and that the N-H axis is nearly parallel with the rotational axis. This study demonstrates that site-specifically labeled (/sup 15/N)gramicidin A analogs prepared by solid-phase peptide synthesis are valuable tools in the study of the solid-state nuclear magnetic resonance spectra of samples in oriented lipid bilayers.

  1. CS2SAT Desktop Tool

    2006-03-15

    The Idaho National Laboratory (INL) has developed a Control System Cyber Security Self-Assessment Tool (CS2SAT) desktop tool that provides a repeatable and systematic approach for control system users to assess the cyber security posture of their control system networks. The tool assists users in identifying the cyber security parameters of their systems and then offers security objectives, in the form of requirements, for improving the security of their specific network. Each requirement is linked tomore » a series of associated recommendations for compliance dependent upon the desired level of security protection. Each requirement is supported by links to the original standards document and recommendations are supported by links to whitepapers and other help documents. Package also includes two back-end supporting codes: CS2SAT Requirements Matrix and Control System Security Information System.« less

  2. Cs diffusion in cubic silicon carbide

    NASA Astrophysics Data System (ADS)

    Shrader, David; Szlufarska, Izabela; Morgan, Dane

    2012-02-01

    Undesired release of Cs through a silicon carbide coating of nuclear fuel is a significant concern for the design of the Very High Temperature Reactor (VHTR). However, mechanisms of Cs transport are currently unclear. To better understand the possible mechanisms of Cs release here we use density functional theory to study diffusion of Cs in crystalline bulk SiC. Cs point defects and Cs - vacancy clusters have been investigated for stability and structure. The most stable state for the Cs impurity in SiC, under n-type doping conditions, is found to be a negatively charged Cs atom substituting for a C atom and bound to two Si vacancies ( Cs-2VSi3-). Bulk diffusion coefficients are estimated for several Cs impurity states. The Cs-2VSi3- defect structure is found to have the lowest overall activation energy for diffusion with a value of approximately 5.14 eV. This activation energy agrees well with diffusion activation energies estimated for Cs in SiC based on high temperature integral release experiments.

  3. Gramicidin A disassembles large conductive clusters of its lysine-substituted derivatives in lipid membranes.

    PubMed

    Antonenko, Yuri N; Gluhov, Grigory S; Firsov, Alexander M; Pogozheva, Irina D; Kovalchuk, Sergey I; Pechnikova, Evgeniya V; Kotova, Elena A; Sokolova, Olga S

    2015-07-14

    N-terminally substituted lysine derivatives of gramicidin A (gA), [Lys1]gA and [Lys3]gA, but not glutamate- or aspartate-substituted peptides have been previously shown to cause the leakage of carboxyfluorescein from liposomes. Here, the leakage induction was also observed for [Arg1]gA and [Arg3]gA, while [His1]gA and [His3]gA were inactive at neutral pH. The Lys3-containing analogue with all tryptophans replaced by isoleucines did not induce liposome leakage, similar to gA. This suggests that the presence of both tryptophans and N-terminal cationic residues is critical for pore formation. Remarkably, the addition of gA blocked the leakage induced by [Lys3]gA. By examining with fluorescence correlation spectroscopy the peptide-induced leakage of fluorescent markers from liposomes, we estimated the diameter of pores responsible for the leakage to be about 1.6 nm. Transmission electron cryo-microscopy imaging of liposomes with [Lys3]gA showed that the liposomal membranes contained high electron density particles with a size of about 40 Å, suggesting the formation of peptide clusters. No such clusterization was observed in liposomes incorporating gA or a mixture of gA with [Lys3]gA. Three-dimensional reconstruction of the clusters was compatible with their pentameric arrangement. Based on experimental data and computational modeling, we suggest that the large pore formed by [Lys3]gA represents a barrel-stave oligomeric cluster formed by antiparallel double-stranded helical dimers (DH). In a tentative model, the pentamer of dimers may be stabilized by aromatic Trp-Trp and cation-π Trp-Lys interactions between the neighboring DHs. The inhibiting effect of gA on the [Lys3]gA-induced leakage can be attributed to breaking of cation-π interactions, which prevents peptide clusterization and pore formation.

  4. Cs based photocathodes for gaseous detectors

    SciTech Connect

    Borovick-Romanov, A.; Peskov, V.

    1993-08-01

    We demonstrated that some standard photocathodes SbCs, GaAs(Cs), Au(Cs) can easily be manufactured for use inside gaseous detectors. When filed with clean quenched gases such detectors have a quantum efficiency of a few percent in the visible region of the spectra and can operate at a gain >10{sup 3}. We tried to make these photocathodes more air stable by protecting their surfaces with a thin layer of CsI or liquid TMAE. The most air stable were photocathodes with a CsI protective layer. A wavelengths {le}185 nm such photocathodes have the highest quantum efficiency among all known air stable photocathodes, including CsI. Gaseous detectors with such photocathodes can operate at a gain of 10{sup 5}. Results of first tests of doped CsI photocathode are also presented. Possible fields of application of new photocathodes are discussed.

  5. Therapeutic index of gramicidin S is strongly modulated by d-phenylalanine analogues at the β-turn

    PubMed Central

    Solanas, Concepción; de la Torre, Beatriz G.; Fernández-Reyes, María; Santiveri, Clara M.; Jiménez, M. Ángeles; Rivas, Luis; Jiménez, Ana I.; Andreu, David; Cativiela, Carlos

    2009-01-01

    Analogues of the cationic antimicrobial peptide gramicidin S (GS), cyclo(Val-Orn-Leu-d-Phe-Pro)2, with d-Phe residues replaced by different (restricted mobility, mostly) surrogates have been synthesized and used in SAR studies against several pathogenic bacteria. While all d-Phe substitutions are shown by NMR to preserve the overall β-sheet conformation, they entail subtle structural alterations that lead to significant modifications in biological activity. In particular, the analogue incorporating d-Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) shows a modest but significant increase in therapeutic index, mostly due to a sharp decrease in hemolytic effect. The fact that NMR data show a shortened distance between the d-Tic aromatic ring and the Orn δ-amino group may help explain the improved antibiotic profile of this analogue. PMID:19132829

  6. Sequence inversion and phenylalanine surrogates at the β-turn enhance the antibiotic activity of gramicidin S

    PubMed Central

    Solanas, Concepción; de la Torre, Beatriz G.; Fernández-Reyes, María; Santiveri, Clara M.; Jiménez, M. Ángeles; Rivas, Luis; Jiménez, Ana I.; Andreu, David; Cativiela, Carlos

    2010-01-01

    A series of gramicidin S (GS) analogs have been synthesized where the Phe (i+1) and Pro (i+2) residues of the β-turn have been swapped while the respective chiralities (D-, L-) at each position are preserved, and Phe is replaced by surrogates with aromatic side chains of diverse size, orientation and flexibility. Although most analogs preserve the β-sheet structure, as assessed by NMR, their antibiotic activities turn out to be highly dependent on the bulkiness and spatial arrangement of the aromatic side chain. Significant increases in microbicidal potency against both Gram-positive and Gram-negative pathogens are observed for several analogs, resulting in improved therapeutic profiles. Data indicate that seemingly minor replacements at the GS β-turn can have significant impact on antibiotic activity, highlighting this region as a hot spot for modulating GS plasticity and activity. PMID:20411945

  7. Sequence inversion and phenylalanine surrogates at the beta-turn enhance the antibiotic activity of gramicidin S.

    PubMed

    Solanas, Concepción; de la Torre, Beatriz G; Fernández-Reyes, María; Santiveri, Clara M; Jiménez, M Angeles; Rivas, Luis; Jiménez, Ana I; Andreu, David; Cativiela, Carlos

    2010-05-27

    A series of gramicidin S (GS) analogues have been synthesized where the Phe (i + 1) and Pro (i + 2) residues of the beta-turn have been swapped while the respective chiralities (D-, L-) at each position are preserved, and Phe is replaced by surrogates with aromatic side chains of diverse size, orientation, and flexibility. Although most analogues preserve the beta-sheet structure, as assessed by NMR, their antibiotic activities turn out to be highly dependent on the bulkiness and spatial arrangement of the aromatic side chain. Significant increases in microbicidal potency against both Gram-positive and Gram-negative pathogens are observed for several analogues, resulting in improved therapeutic profiles. Data indicate that seemingly minor replacements at the GS beta-turn can have significant impact on antibiotic activity, highlighting this region as a hot spot for modulating GS plasticity and activity.

  8. Detection of single ion channel activity with carbon nanotubes

    PubMed Central

    Zhou, Weiwei; Wang, Yung Yu; Lim, Tae-Sun; Pham, Ted; Jain, Dheeraj; Burke, Peter J.

    2015-01-01

    Many processes in life are based on ion currents and membrane voltages controlled by a sophisticated and diverse family of membrane proteins (ion channels), which are comparable in size to the most advanced nanoelectronic components currently under development. Here we demonstrate an electrical assay of individual ion channel activity by measuring the dynamic opening and closing of the ion channel nanopores using single-walled carbon nanotubes (SWNTs). Two canonical dynamic ion channels (gramicidin A (gA) and alamethicin) and one static biological nanopore (α-hemolysin (α-HL)) were successfully incorporated into supported lipid bilayers (SLBs, an artificial cell membrane), which in turn were interfaced to the carbon nanotubes through a variety of polymer-cushion surface functionalization schemes. The ion channel current directly charges the quantum capacitance of a single nanotube in a network of purified semiconducting nanotubes. This work forms the foundation for a scalable, massively parallel architecture of 1d nanoelectronic devices interrogating electrophysiology at the single ion channel level. PMID:25778101

  9. Detection of single ion channel activity with carbon nanotubes.

    PubMed

    Zhou, Weiwei; Wang, Yung Yu; Lim, Tae-Sun; Pham, Ted; Jain, Dheeraj; Burke, Peter J

    2015-01-01

    Many processes in life are based on ion currents and membrane voltages controlled by a sophisticated and diverse family of membrane proteins (ion channels), which are comparable in size to the most advanced nanoelectronic components currently under development. Here we demonstrate an electrical assay of individual ion channel activity by measuring the dynamic opening and closing of the ion channel nanopores using single-walled carbon nanotubes (SWNTs). Two canonical dynamic ion channels (gramicidin A (gA) and alamethicin) and one static biological nanopore (α-hemolysin (α-HL)) were successfully incorporated into supported lipid bilayers (SLBs, an artificial cell membrane), which in turn were interfaced to the carbon nanotubes through a variety of polymer-cushion surface functionalization schemes. The ion channel current directly charges the quantum capacitance of a single nanotube in a network of purified semiconducting nanotubes. This work forms the foundation for a scalable, massively parallel architecture of 1d nanoelectronic devices interrogating electrophysiology at the single ion channel level.

  10. Detection of single ion channel activity with carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Zhou, Weiwei; Wang, Yung Yu; Lim, Tae-Sun; Pham, Ted; Jain, Dheeraj; Burke, Peter J.

    2015-03-01

    Many processes in life are based on ion currents and membrane voltages controlled by a sophisticated and diverse family of membrane proteins (ion channels), which are comparable in size to the most advanced nanoelectronic components currently under development. Here we demonstrate an electrical assay of individual ion channel activity by measuring the dynamic opening and closing of the ion channel nanopores using single-walled carbon nanotubes (SWNTs). Two canonical dynamic ion channels (gramicidin A (gA) and alamethicin) and one static biological nanopore (α-hemolysin (α-HL)) were successfully incorporated into supported lipid bilayers (SLBs, an artificial cell membrane), which in turn were interfaced to the carbon nanotubes through a variety of polymer-cushion surface functionalization schemes. The ion channel current directly charges the quantum capacitance of a single nanotube in a network of purified semiconducting nanotubes. This work forms the foundation for a scalable, massively parallel architecture of 1d nanoelectronic devices interrogating electrophysiology at the single ion channel level.

  11. Role of Epithelium Sodium Channel in Bone Formation

    PubMed Central

    Wang, Ruo-Yu; Yang, Shu-Hua; Xu, Wei-Hua

    2016-01-01

    Objective: To review the recent developments in the mechanisms of epithelium sodium channels (ENaCs) induced bone formation and regulation. Data Sources: Studies written in English or Chinese were searched using Medline, PubMed and the index of Chinese-language literature with time restriction from 2005 to 2014. Keywords included ENaC, bone, bone formation, osteonecrosis, estrogen, and osteoporosis. Data from published articles about the structure of ENaC, mechanism of ENaC in bone formation in recent domestic and foreign literature were selected. Study Selection: Abstract and full text of all studies were required to obtain. Studies those were not accessible and those did not focus on the keywords were excluded. Results: ENaCs are tripolymer ion channels which are assembled from homologous α, β, and γ subunits. Crystal structure of ENaCs suggests that ENaC has a central ion-channel located in the central symmetry axis of the three subunits. ENaCs are protease sensitive channels whose iron-channel activity is regulated by the proteolytic reaction. Channel opening probability of ENaCs is regulated by proteinases, mechanical force, and shear stress. Several molecules are involved in regulation of ENaCs in bone formation, including nitride oxide synthases, voltage-sensitive calcium channels, and cyclooxygenase-2. Conclusion: The pathway of ENaC involved in shear stress has an effect on stimulating osteoblasts even bone formation by estrogen interference. PMID:26904995

  12. Reactive barriers for {sup 137}Cs retention

    SciTech Connect

    KRUMHANSL,JAMES L.; BRADY,PATRICK V.; ANDERSON,HOWARD L.

    2000-05-19

    {sup 137}Cs was dispersed globally by cold war activities and, more recently, by the Chernobyl accident. Engineered extraction of {sup 137}Cs from soils and groundwaters is exceedingly difficult. Because the half life of {sup 137}Cs is only 30.2 years, remediation might be more effective (and less costly) if {sup 137}Cs bioavailability could be demonstrably limited for even a few decades by use of a reactive barrier. Essentially permanent isolation must be demonstrated in those few settings where high nuclear level wastes contaminated the environment with {sup 135}Cs (half life 2.3x10{sup 6} years) in addition to {sup 137}Cs. Clays are potentially a low-cost barrier to Cs movement, though their long-term effectiveness remains untested. To identify optimal clays for Cs retention Cs resorption was measured for five common clays: Wyoming Montmorillonite (SWy-1), Georgia Kaolinites (KGa-1 and KGa-2), Fithian Illite (F-Ill), and K-Metabentonite (K-Mbt). Exchange sites were pre-saturated with 0.16 M CsCl for 14 days and readily exchangeable Cs was removed by a series of LiNO{sub 3} and LiCl washes. Washed clay were then placed into dialysis bags and the Cs release to the deionized water outside the bags measured. Release rates from 75 to 139 days for SWy-1, K-Mbt and F- 111 were similar; 0.017 to 0.021% sorbed Cs released per day. Both kaolinites released Cs more rapidly (0.12 to 0.05% of the sorbed Cs per day). In a second set of experiments, clays were doped for 110 days and subjected to an extreme and prolonged rinsing process. All the clays exhibited some capacity for irreversible Cs uptake so most soils have some limited ability to act as a natural barrier to Cs migration. However, the residual loading was greatest on K-Mbt ({approximately} 0.33 wt% Cs). Thus, this clay would be the optimal material for constructing artificial reactive barriers.

  13. Comparison of the effects of internal TEA+ and Cs+ on potassium current in squid giant axons.

    PubMed Central

    Clay, J R

    1985-01-01

    Internal tetraethylammonium (TEA) and cesium ions block outward potassium current in nerve membrane in a voltage-dependent manner. Blockade with Cs+ occurs virtually instantaneously after membrane depolarization, whereas blockade with TEA+ occurs after a delay. The latter result suggested to Armstrong (1966, J. Gen. Physiol., 50:279-293; 1969, J. Gen. Physiol., 54:553-575) that potassium channels must open before TEA+ blockade can occur, which is in contrast to Cs+ blockade, which appears to be independent of channel gating. The results in this study concerning the effect of TEA+ on inward (tail) current argue against the Armstrong model. Specifically, TEA+ (partially) blocks inward current without altering the tail current time constant. This result indicates that TEA+ can occupy its binding site within the channel whether or not the channel gates are open. This alternative hypothesis can describe both the steady-state and time-dependent components of TEA+ blockade. PMID:2418889

  14. Formation of Stoichiometric CsFn Compounds

    NASA Astrophysics Data System (ADS)

    Zhu, Qiang; Oganov, Artem R.; Zeng, Qingfeng

    2015-01-01

    Alkali halides MX, have been viewed as typical ionic compounds, characterized by 1:1 ratio necessary for charge balance between M+ and X-. It was proposed that group I elements like Cs can be oxidized further under high pressure. Here we perform a comprehensive study for the CsF-F system at pressures up to 100 GPa, and find extremely versatile chemistry. A series of CsFn (n >= 1) compounds are predicted to be stable already at ambient pressure. Under pressure, 5p electrons of Cs atoms become active, with growing tendency to form Cs (III) and (V) valence states at fluorine-rich conditions. Although Cs (II) and (IV) are not energetically favoured, the interplay between two mechanisms (polyfluoride anions and polyvalent Cs cations) allows CsF2 and CsF4 compounds to be stable under pressure. The estimated defluorination temperatures of CsFn (n = 2,3,5) compounds at atmospheric pressure (218°C, 150°C, -15°C, respectively), are attractive for fluorine storage applications.

  15. P-wave [cs][cs] tetraquark state: Y(4260) or Y(4660)?

    SciTech Connect

    Zhang Jianrong; Huang Mingqiu

    2011-02-01

    The mass of a P-wave cs-scalar-diquark cs-scalar-antidiquark state is computed in the framework of QCD sum rules. The result 4.69{+-}0.36 GeV is in good agreement with the experimental value of Y(4660) but higher than Y(4260)'s, which supports the P-wave [cs][cs] configuration for Y(4660) while disfavors the interpretation of Y(4260) as the P-wave [cs][cs] state. In the same picture, the mass of P-wave [bs][bs] is predicted to be 11.19{+-}0.49 GeV.

  16. Gramicidin S biosynthesis operon containing the structural genes grsA and grsB has an open reading frame encoding a protein homologous to fatty acid thioesterases.

    PubMed Central

    Krätzschmar, J; Krause, M; Marahiel, M A

    1989-01-01

    The DNA sequence of about 5.9 kilobase pairs (kbp) of the gramicidin S biosynthesis operon (grs) was determined. Three open reading frames were identified; the corresponding genes, called grsT, grsA, and grsB, were found to be organized in one transcriptional unit, not two as previously reported (M. Krause and M. A. Marahiel, J. Bacteriol. 170:4669-4674, 1988). The entire nucleotide sequence of grsA, coding for the 126.663-kilodalton gramicidin S synthetase 1, grsT, encoding a 29.191-kilodalton protein of unknown function, and 732 bp of the 5' end of grsB, encoding the gramicidin S synthetase 2, were determined. A single initiation site of transcription 81 bp upstream of the grsT initiation condon GTG was identified by high-resolution S1 mapping studies. The sequence of the grsA gene product showed a high degree of homology to the tyrocidine synthetase 1 (TycA protein), and that of grsT exhibited a significant degree of homology to vertebrate fatty acid thioesterases. Images PMID:2477357

  17. Theoretical study of the CS2+ dication

    NASA Astrophysics Data System (ADS)

    Šedivcová, T.; Špirko, V.; Fišer, J.

    2006-10-01

    The potential energy and spin-orbit coupling functions of 11 lowest electronic states of CS2+ dication have been calculated using internally contracted multireference configuration method. Using these functions, the positions and widths of the corresponding vibronic levels have been evaluated by means of the stabilization and log-phase-amplitude methods. The states governing the second step in the sequential pathway CS23+→ S++CS2+→S++C++S+ of the overall three-body Coulomb explosion of CS23+ have been determined.

  18. Microstructure of Cs-implanted zirconia: Role of temperature

    SciTech Connect

    Vincent, L.; Thome, L.; Garrido, F.; Kaitasov, O.; Houdelier, F.

    2008-12-01

    The aim of this study was to identify experimentally the phase which includes cesium in yttria stabilized zirconia (YSZ). The solubility and retention of cesium in YSZ were studied at high temperature (HT). Cesium was ion implanted (at 300 keV) into YSZ at room temperature (RT), 750 deg. C, or 900 deg. C at fluences up to 5x10{sup 16} cm{sup -2}. The temperature dependence of the radiation-induced damage and of the cesium distribution in YSZ single crystals was investigated by Rutherford backscattering spectrometry and ion channeling. Transmission electron microscopy (TEM) studies were performed in order to determine the damage nature and search for a predicted ternary phase of cesium zirconate. Whatever the implantation temperature, the thickness of the damaged layer increases inwards with ion fluence. At RT, amorphization occurs, caused by the high Cs concentration (7 at. %). In situ TEM during postannealing shows recrystallization of cubic zirconia after release of cesium. A high implantation temperature has a significant influence on the nature of radiation defects and on the retained Cs concentration. At HT, dislocation loops and voids are formed but no amorphization is observed whereas polygonization occurs at high fluence. The implanted cesium concentration reaches a saturation value of 1.5 at. % above which Cs can no longer be retained in the matrix and is then released at the surface. At that concentration, cesium forms a solid solution in YSZ; no other phase is formed, neither during irradiation nor after thermal annealing.

  19. 134Cs: 137Cs and 106Ru: 137Cs ratios in intertidal sediments from the Cumbria and Lancashire coasts England

    NASA Astrophysics Data System (ADS)

    Stanners, D. A.; Aston, S. R.

    1981-10-01

    The distributions of 134Cs, 137Cs and 106Ru in intertidal surface sediments from the coasts of Cumbria and Lancashire, north-west England, are reported. The ratios of 134Cs: 137Cs and 106Ru: 137Cs activities have been used together with the isotopic composition of the Windscale radioactive effluents to examine the contamination history of sediments. Distinct differences between the activities and time of contamination of muds, silts and sands are found, and the apparent lag times of transport of radioactive wastes to different sediment localities are estimated. The relatively high activities in fine sediments reflect recent discharges indicating a rapid response to discharge, while the sands contain low levels of older contamination. Apparent lag times of up to 6 years are estimated for the study area; the transport to the south is generally more rapid than to the north. These results have consequences for the operation and interpretation of radiological monitoring in coastal areas.

  20. Cs(2)UPd(3)Se(6).

    PubMed

    Oh, George N; Ibers, James A

    2011-01-12

    Dicaesium uranium(IV) tripalladium(II) hexa-selenide, Cs(2)UPd(3)Se(6), crystallizes in the space group Fmmm in the Ba(2)NaCu(3)O(6) structure type. The asymmetric unit comprises the following atoms with site symmetries as shown: U1 (mm2), Cs1 (222), Cs2 (m2m), Pd1 (.m.), Pd2 (2mm), Se1 (m..), and Se2 (1). This layered structure contains six edge-sharing square-planar [PdSe(4)] units that form a hexa-gon. These, in turn, edge-share with [USe(6)] trigonal-prismatic units, forming an extended layer parallel to (010). The layers are stacked along [010]. They are staggered, and are separated by the Cs atoms. The Cs atoms are either coordinated in a square anti-prism of Se atoms or are ten-coordinate, with one square face and the opposite face hexa-gonal.

  1. Numerical methods for a Poisson-Nernst-Planck-Fermi model of biological ion channels.

    PubMed

    Liu, Jinn-Liang; Eisenberg, Bob

    2015-07-01

    Numerical methods are proposed for an advanced Poisson-Nernst-Planck-Fermi (PNPF) model for studying ion transport through biological ion channels. PNPF contains many more correlations than most models and simulations of channels, because it includes water and calculates dielectric properties consistently as outputs. This model accounts for the steric effect of ions and water molecules with different sizes and interstitial voids, the correlation effect of crowded ions with different valences, and the screening effect of polarized water molecules in an inhomogeneous aqueous electrolyte. The steric energy is shown to be comparable to the electrical energy under physiological conditions, demonstrating the crucial role of the excluded volume of particles and the voids in the natural function of channel proteins. Water is shown to play a critical role in both correlation and steric effects in the model. We extend the classical Scharfetter-Gummel (SG) method for semiconductor devices to include the steric potential for ion channels, which is a fundamental physical property not present in semiconductors. Together with a simplified matched interface and boundary (SMIB) method for treating molecular surfaces and singular charges of channel proteins, the extended SG method is shown to exhibit important features in flow simulations such as optimal convergence, efficient nonlinear iterations, and physical conservation. The generalized SG stability condition shows why the standard discretization (without SG exponential fitting) of NP equations may fail and that divalent Ca(2+) may cause more unstable discrete Ca(2+) fluxes than that of monovalent Na(+). Two different methods-called the SMIB and multiscale methods-are proposed for two different types of channels, namely, the gramicidin A channel and an L-type calcium channel, depending on whether water is allowed to pass through the channel. Numerical methods are first validated with constructed models whose exact solutions are

  2. Numerical methods for a Poisson-Nernst-Planck-Fermi model of biological ion channels

    NASA Astrophysics Data System (ADS)

    Liu, Jinn-Liang; Eisenberg, Bob

    2015-07-01

    Numerical methods are proposed for an advanced Poisson-Nernst-Planck-Fermi (PNPF) model for studying ion transport through biological ion channels. PNPF contains many more correlations than most models and simulations of channels, because it includes water and calculates dielectric properties consistently as outputs. This model accounts for the steric effect of ions and water molecules with different sizes and interstitial voids, the correlation effect of crowded ions with different valences, and the screening effect of polarized water molecules in an inhomogeneous aqueous electrolyte. The steric energy is shown to be comparable to the electrical energy under physiological conditions, demonstrating the crucial role of the excluded volume of particles and the voids in the natural function of channel proteins. Water is shown to play a critical role in both correlation and steric effects in the model. We extend the classical Scharfetter-Gummel (SG) method for semiconductor devices to include the steric potential for ion channels, which is a fundamental physical property not present in semiconductors. Together with a simplified matched interface and boundary (SMIB) method for treating molecular surfaces and singular charges of channel proteins, the extended SG method is shown to exhibit important features in flow simulations such as optimal convergence, efficient nonlinear iterations, and physical conservation. The generalized SG stability condition shows why the standard discretization (without SG exponential fitting) of NP equations may fail and that divalent Ca2 + may cause more unstable discrete Ca2 + fluxes than that of monovalent Na+. Two different methods—called the SMIB and multiscale methods—are proposed for two different types of channels, namely, the gramicidin A channel and an L-type calcium channel, depending on whether water is allowed to pass through the channel. Numerical methods are first validated with constructed models whose exact solutions are

  3. Memory Is Not Extinguished along with CS Presentation but within a Few Seconds after CS-Offset

    ERIC Educational Resources Information Center

    Perez-Cuesta, Luis Maria; Hepp, Yanil; Pedreira, Maria Eugenia; Maldonado, Hector

    2007-01-01

    Prior work with the crab's contextual memory model showed that CS-US conditioned animals undergoing an unreinforced CS presentation would either reconsolidate or extinguish the CS-US memory, depending on the length of the reexposure to the CS. Either memory process is only triggered once the CS is terminated. Based on these results, the following…

  4. Potential value of Cs-137 capsules

    SciTech Connect

    Bloomster, C.H.; Brown, D.R.; Bruno, G.A.; Hazelton, R.F.; Hendrickson, P.L.; Lezberg, A.J.; Tingey, G.L.; Wilfert, G.L.

    1985-04-01

    We determined the value of Cs-137 compared to Co-60 as a source for the irradiation of fruit (apples and cherries), pork and medical supplies. Cs-137, in the WESF capsule form, had a value of approximately $0.40/Ci as a substitute for Co-60 priced at approximately $1.00/Ci. The comparison was based on the available curies emitted from the surface of each capsule. We developed preliminary designs for fourteen irradiation facilities; seven were based on Co-60 and seven were based on Cs-137. These designs provided the basis for estimating capital and operating costs which, in turn, provided the basis for determining the value of Cs-137 relative to Co-60 in these applications. We evaluated the effect of the size of the irradiation facility on the value of Cs-137. The cost of irradiation is low compared to the value of the product. Irradiation of apples for disinfestation costs $.01 to .02 per pound. Irradiation for trichina-safe pork costs $.02 per pound. Irradiation of medical supplies for sterilization costs $.07 to .12 per pound. The cost of the irradiation source, either Co-60 or Cs-137, contributed only a minor amount to the total cost of irradiation, about 5% for the fruit and hog cases and about 20% for the medical supply cases. We analyzed the sensitivity of the irradiation costs and Cs-137 value to several key assumptions.

  5. Second harmonic generation from tryptophan-rich short peptides: W(n)K(m) and gramicidin A.

    PubMed

    Duboisset, J; Matar, G; Besson, F; Ficheux, D; Benichou, E; Russier-Antoine, I; Jonin, Ch; Brevet, P F

    2014-09-01

    We report the first hyperpolarizability of a series of tryptophan-rich short peptides with the respective sequence KWK, KWWK, KWWWK, KWWKWWK, where W and K stand for tryptophan and lysine. The measurements were performed with the technique of hyper-Rayleigh scattering in the bulk of an aqueous Tris buffer solution at a pH of 8.5 and a salt concentration of 150 mM at the non-resonant fundamental wavelength of 784 nm. The first hyperpolarizability of the different peptides follows a simple additive model scaling with the number of tryptophan residues contained in the peptide. However, it appears that the first hyperpolarizability response of a single tryptophan residue in the peptide strongly differs from that of an isolated tryptophan. Hence, it is therefore demonstrated that the local environment of the tryptophan residues within the peptide strongly influences its nonlinear optical response. A comparison with the first hyperpolarizability of the natural peptide gramicidin A measured in trifluoroethanol (TFE) further confirms the key role of the local environment on the first hyperpolarizability of tryptophan residues in peptides.

  6. An experimental study of the solvent-dependent self-assembly/disassembly and conformer preferences of gramicidin A.

    PubMed

    Chen, Liuxi; Chen, Shu-Hua; Russell, David H

    2013-08-20

    The solvent dependence of self-assembly/disassembly kinetics and conformer preferences of the gramicidin A (GA) dimer is investigated using a combination of techniques, viz., electrospray ionization-ion mobility-mass spectrometry (IM-MS), collision-induced dissociation (CID), and hydrogen/deuterium exchange (HDX)-MS. IM-MS measurements reveal that there are possibly three distinct GA dimeric species, detected as sodium ion adduct ions [2GA + 2Na](2+), and these are assigned as the parallel β-helix, antiparallel β-helix, and head-to-head dimer. The monomerization kinetics and equilibrium abundances of the dimer ions depend upon solvent polarity. The antiparallel β-helix was the thermodynamically preferred species in less polar solvents. HDX measurements and collision-induced dissociation (CID) of the intermediate complex confirm the well-protected dimer geometry with strong intermolecular hydrogen bonds. This combined IM-HDX-CID methodology provides a comprehensive view of GA self-assembly/disassembly in low dielectric solutions, showing its potential utility in solving solution-phase protein self-assembly/disassembly kinetics and providing structural information of the multimers at the same time.

  7. Direct surfactin-gramicidin S antagonism supports detoxification in mixed producer cultures of Bacillus subtilis and Aneurinibacillus migulanus.

    PubMed

    Rautenbach, Marina; Eyéghé-Bickong, Hans André; Vlok, Nicolas Maré; Stander, Marietjie; de Beer, Abré

    2012-12-01

    Antibiotic production as a defence mechanism is a characteristic of a wide variety of organisms. In natural evolutionary adaptation, cellular events such as sporulation, biofilm formation and resistance to antibiotics enable some micro-organisms to survive environmental and antibiotic stress conditions. The two antimicrobial cyclic peptides in this study, gramicidin S (GS) from Aneurinibacillus migulanus and the lipopeptide surfactin (Srf) from Bacillus subtilis, have been shown to affect both membrane and intercellular components of target organisms. Many functions, other than that of antimicrobial activity, have been assigned to Srf. We present evidence that an additional function may exist for Srf, namely that of a detoxifying agent that protects its producer from the lytic activity of GS. We observed that Srf producers were more resistant to GS and could be co-cultured with the GS producer. Furthermore, exogenous Srf antagonized the activity of GS against both Srf-producing and non-producing bacterial strains. A molecular interaction between the anionic Srf and the cationic GS was observed with circular dichroism and electrospray MS. Our results indicate that the formation of an inactive complex between GS and Srf supports resistance towards GS, with the anionic Srf forming a chemical barrier to protect its producer. This direct detoxification combined with the induction of protective stress responses in B. subtilis by Srf confers resistance toward GS from A. migulanus and allows survival in mixed cultures.

  8. A venom-derived neurotoxin, CsTx-1, from the spider Cupiennius salei exhibits cytolytic activities.

    PubMed

    Kuhn-Nentwig, Lucia; Fedorova, Irina M; Lüscher, Benjamin P; Kopp, Lukas S; Trachsel, Christian; Schaller, Johann; Vu, Xuan Lan; Seebeck, Thomas; Streitberger, Kathrin; Nentwig, Wolfgang; Sigel, Erwin; Magazanik, Lev G

    2012-07-20

    CsTx-1, the main neurotoxic acting peptide in the venom of the spider Cupiennius salei, is composed of 74 amino acid residues, exhibits an inhibitory cysteine knot motif, and is further characterized by its highly cationic charged C terminus. Venom gland cDNA library analysis predicted a prepropeptide structure for CsTx-1 precursor. In the presence of trifluoroethanol, CsTx-1 and the long C-terminal part alone (CT1-long; Gly-45-Lys-74) exhibit an α-helical structure, as determined by CD measurements. CsTx-1 and CT1-long are insecticidal toward Drosophila flies and destroys Escherichia coli SBS 363 cells. CsTx-1 causes a stable and irreversible depolarization of insect larvae muscle cells and frog neuromuscular preparations, which seem to be receptor-independent. Furthermore, this membranolytic activity could be measured for Xenopus oocytes, in which CsTx-1 and CT1-long increase ion permeability non-specifically. These results support our assumption that the membranolytic activities of CsTx-1 are caused by its C-terminal tail, CT1-long. Together, CsTx-1 exhibits two different functions; as a neurotoxin it inhibits L-type Ca(2+) channels, and as a membranolytic peptide it destroys a variety of prokaryotic and eukaryotic cell membranes. Such a dualism is discussed as an important new mechanism for the evolution of spider venomous peptides.

  9. Engineered ion channels as emerging tools for chemical biology.

    PubMed

    Mayer, Michael; Yang, Jerry

    2013-12-17

    Over the last 25 years, researchers have developed exogenously expressed, genetically engineered, semi-synthetic, and entirely synthetic ion channels. These structures have sufficient fidelity to serve as unique tools that can reveal information about living organisms. One of the most exciting success stories is optogenetics: the use of light-gated channels to trigger action potentials in specific neurons combined with studies of the response from networks of cells or entire live animals. Despite this breakthrough, the use of molecularly engineered ion channels for studies of biological systems is still in its infancy. Historically, researchers studied ion channels in the context of their own function in single cells or in multicellular signaling and regulation. Only recently have researchers considered ion channels and pore-forming peptides as responsive tools to report on the chemical and physical changes produced by other biochemical processes and reactions. This emerging class of molecular probes has a number of useful characteristics. For instance, these structures can greatly amplify the signal of chemical changes: the binding of one molecule to a ligand-gated ion channel can result in flux of millions of ions across a cell membrane. In addition, gating occurs on sub-microsecond time scales, resulting in fast response times. Moreover, the signal is complementary to existing techniques because the output is ionic current rather than fluorescence or radioactivity. And finally, ion channels are also localized at the membrane of cells where essential processes such as signaling and regulation take place. This Account highlights examples, mostly from our own work, of uses of ion channels and pore-forming peptides such as gramicidin in chemical biology. We discuss various strategies for preparing synthetically tailored ion channels that range from de novo designed synthetic molecules to genetically engineered or simply exogenously expressed or reconstituted wild

  10. The "Seven Cs" for Employee Retention.

    ERIC Educational Resources Information Center

    Taguchi, Sherrie Gong

    2001-01-01

    Defines the "Seven Cs," traditional yet effective business fundamentals used to engage employees. Discusses how many companies are leveraging the basics of good employee relations in order to inspire staff productivity and loyalty. (GCP)

  11. 135Cs activity and 135Cs/137Cs atom ratio in environmental samples before and after the Fukushima Daiichi Nuclear Power Plant accident

    NASA Astrophysics Data System (ADS)

    Yang, Guosheng; Tazoe, Hirofumi; Yamada, Masatoshi

    2016-04-01

    135Cs/137Cs is a potential tracer for radiocesium source identification. However, due to the challenge to measure 135Cs, there were no 135Cs data available for Japanese environmental samples before the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. It was only 3 years after the accident that limited 135Cs values could be measured in heavily contaminated environmental samples. In the present study, activities of 134Cs, 135Cs, and 137Cs, along with their ratios in 67 soil and plant samples heavily and lightly contaminated by the FDNPP accident were measured by combining γ spectrometry with ICP-MS/MS. The arithmetic means of the 134Cs/137Cs activity ratio (1.033 ± 0.006) and 135Cs/137Cs atom ratio (0.334 ± 0.005) (decay corrected to March 11, 2011), from old leaves of plants collected immediately after the FDNPP accident, were confirmed to represent the FDNPP derived radiocesium signature. Subsequently, for the first time, trace 135Cs amounts before the FDNPP accident were deduced according to the contribution of global and FDNPP accident-derived fallout. Apart from two soil samples with a tiny global fallout contribution, contributions of global fallout radiocesium in other soil samples were observed to be 0.338%–52.6%. The obtained 135Cs/137Cs database will be useful for its application as a geochemical tracer in the future.

  12. 135Cs activity and 135Cs/137Cs atom ratio in environmental samples before and after the Fukushima Daiichi Nuclear Power Plant accident.

    PubMed

    Yang, Guosheng; Tazoe, Hirofumi; Yamada, Masatoshi

    2016-04-07

    (135)Cs/(137)Cs is a potential tracer for radiocesium source identification. However, due to the challenge to measure (135)Cs, there were no (135)Cs data available for Japanese environmental samples before the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. It was only 3 years after the accident that limited (135)Cs values could be measured in heavily contaminated environmental samples. In the present study, activities of (134)Cs, (135)Cs, and (137)Cs, along with their ratios in 67 soil and plant samples heavily and lightly contaminated by the FDNPP accident were measured by combining γ spectrometry with ICP-MS/MS. The arithmetic means of the (134)Cs/(137)Cs activity ratio (1.033 ± 0.006) and (135)Cs/(137)Cs atom ratio (0.334 ± 0.005) (decay corrected to March 11, 2011), from old leaves of plants collected immediately after the FDNPP accident, were confirmed to represent the FDNPP derived radiocesium signature. Subsequently, for the first time, trace (135)Cs amounts before the FDNPP accident were deduced according to the contribution of global and FDNPP accident-derived fallout. Apart from two soil samples with a tiny global fallout contribution, contributions of global fallout radiocesium in other soil samples were observed to be 0.338%-52.6%. The obtained (135)Cs/(137)Cs database will be useful for its application as a geochemical tracer in the future.

  13. 135Cs activity and 135Cs/137Cs atom ratio in environmental samples before and after the Fukushima Daiichi Nuclear Power Plant accident

    PubMed Central

    Yang, Guosheng; Tazoe, Hirofumi; Yamada, Masatoshi

    2016-01-01

    135Cs/137Cs is a potential tracer for radiocesium source identification. However, due to the challenge to measure 135Cs, there were no 135Cs data available for Japanese environmental samples before the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. It was only 3 years after the accident that limited 135Cs values could be measured in heavily contaminated environmental samples. In the present study, activities of 134Cs, 135Cs, and 137Cs, along with their ratios in 67 soil and plant samples heavily and lightly contaminated by the FDNPP accident were measured by combining γ spectrometry with ICP-MS/MS. The arithmetic means of the 134Cs/137Cs activity ratio (1.033 ± 0.006) and 135Cs/137Cs atom ratio (0.334 ± 0.005) (decay corrected to March 11, 2011), from old leaves of plants collected immediately after the FDNPP accident, were confirmed to represent the FDNPP derived radiocesium signature. Subsequently, for the first time, trace 135Cs amounts before the FDNPP accident were deduced according to the contribution of global and FDNPP accident-derived fallout. Apart from two soil samples with a tiny global fallout contribution, contributions of global fallout radiocesium in other soil samples were observed to be 0.338%–52.6%. The obtained 135Cs/137Cs database will be useful for its application as a geochemical tracer in the future. PMID:27052481

  14. 135Cs activity and 135Cs/137Cs atom ratio in environmental samples before and after the Fukushima Daiichi Nuclear Power Plant accident.

    PubMed

    Yang, Guosheng; Tazoe, Hirofumi; Yamada, Masatoshi

    2016-01-01

    (135)Cs/(137)Cs is a potential tracer for radiocesium source identification. However, due to the challenge to measure (135)Cs, there were no (135)Cs data available for Japanese environmental samples before the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. It was only 3 years after the accident that limited (135)Cs values could be measured in heavily contaminated environmental samples. In the present study, activities of (134)Cs, (135)Cs, and (137)Cs, along with their ratios in 67 soil and plant samples heavily and lightly contaminated by the FDNPP accident were measured by combining γ spectrometry with ICP-MS/MS. The arithmetic means of the (134)Cs/(137)Cs activity ratio (1.033 ± 0.006) and (135)Cs/(137)Cs atom ratio (0.334 ± 0.005) (decay corrected to March 11, 2011), from old leaves of plants collected immediately after the FDNPP accident, were confirmed to represent the FDNPP derived radiocesium signature. Subsequently, for the first time, trace (135)Cs amounts before the FDNPP accident were deduced according to the contribution of global and FDNPP accident-derived fallout. Apart from two soil samples with a tiny global fallout contribution, contributions of global fallout radiocesium in other soil samples were observed to be 0.338%-52.6%. The obtained (135)Cs/(137)Cs database will be useful for its application as a geochemical tracer in the future. PMID:27052481

  15. Fast CsI-phoswich detector

    DOEpatents

    Langenbrunner, James R.

    1996-01-01

    An improved phoswich radiation detector used pure CsI crystal and a fast plastic scintillator and a single photomultiplier tube. The plastic is arranged to receive incident radiation, and that which passed through then strikes the CsI crystal. Scintillation light from both the plastic and CsI crystal are applied to the photomultiplier tube, with the light from the plastic passing through the crystal without absorption therein. Electronics are provided for analyzing the output of the photomultiplier tube to discriminate responses due to the plastic and the CsI crystal, through short gate and long gate integration, to produce results which are indicative of the characteristics of the different types of incident radiation, even in the presence of large amounts of radiation. The phoswich detector has excellent timing resolution. The scintillators of the CsI- phoswich were chosen for their fast risetimes, of about 3 ns for NE102A, and 30 ns for the pure CsI.

  16. Fast CsI-phoswich detector

    DOEpatents

    Langenbrunner, J.R.

    1996-05-07

    An improved phoswich radiation detector used pure CsI crystal and a fast plastic scintillator and a single photomultiplier tube. The plastic is arranged to receive incident radiation, and that which passed through then strikes the CsI crystal. Scintillation light from both the plastic and CsI crystal are applied to the photomultiplier tube, with the light from the plastic passing through the crystal without absorption therein. Electronics are provided for analyzing the output of the photomultiplier tube to discriminate responses due to the plastic and the CsI crystal, through short gate and long gate integration, to produce results which are indicative of the characteristics of the different types of incident radiation, even in the presence of large amounts of radiation. The phoswich detector has excellent timing resolution. The scintillators of the CsI- phoswich were chosen for their fast risetimes, of about 3 ns for NE102A, and 30 ns for the pure CsI. 5 figs.

  17. Storable droplet interface lipid bilayers for cell-free ion channel studies.

    PubMed

    Jung, Sung-Ho; Choi, Sangbaek; Kim, Young-Rok; Jeon, Tae-Joon

    2012-01-01

    An artificially created lipid bilayer is an important platform in studying ion channels and engineered biosensor applications. However, a lipid bilayer created using conventional techniques is fragile and short-lived, and the measurement of ion channels requires expertise and laborious procedures, precluding practical applications. Here, we demonstrate a storable droplet lipid bilayer precursor frozen with ion channels, resulting in a droplet interface bilayer upon thawing. A small vial with an aqueous droplet in organic solution was flash frozen in -80 °C methanol immediately after an aqueous droplet was introduced into the organic solution and gravity draws the droplet down to the interface upon thawing. A lipid bilayer created along the interface using this method had giga-ohm resistance and typical specific capacitance values. The noise level of this system is favorably comparable to the conventional system. The subsequent incorporation of ion channels, alpha-hemolysin and gramicidin A, showed typical conductance values consistent with those in previous literatures. This novel system to create a lipid bilayer as a whole can be automated from its manufacture to use and indefinitely stored when frozen. As a result, ion channel measurements can be carried out in any place, increasing the accessibility of ion channel studies as well as a number of applications, such as biosensors, ion channel drug screening, and biophysical studies. PMID:21909672

  18. Distribution of radioactive cesium ((134)Cs plus (137)Cs) in rice fractions during polishing and cooking.

    PubMed

    Hachinohe, Mayumi; Okunishi, Tomoya; Hagiwara, Shoji; Todoriki, Setsuko; Kawamoto, Shinichi; Hamamatsu, Shioka

    2015-03-01

    We investigated the distribution of cesium-134 ((134)Cs) and cesium-137 ((137)Cs) during polishing and cooking of rice to obtain their processing factors (Pf) and food processing retention factors (Fr) to make the information available for an adequate understanding of radioactive Cs dynamics. Polishing brown rice resulted in a decreased radioactive Cs concentration of the polished rice, but the bran and germ (outer layers) exhibited higher concentrations than brown rice. The Pf values for 100% polished rice and outer layers ranged from 0.47 to 0.48 and 6.5 to 7.8, respectively. The Fr values for 100% polished rice and outer layers were 0.43 and 0.58 to 0.60, respectively. The distribution of radioactive Cs in polished rice and outer layers was estimated at approximately 40 and 60%, respectively. On the other hand, cooked rice showed significantly lower levels of radioactive Cs than polished rice, and transfer of radioactive Cs into wash water was observed. The Pf and Fr values for cooked rice were 0.28 and 0.65 to 0.66, respectively. From these results, we can calculate that if the radioactive Cs concentration in brown rice is 100 Bq/kg, the concentrations of Cs in polished rice and cooked rice will be 47 to 48 Bq/kg and 13 Bq/kg, respectively. PMID:25719881

  19. Distribution of radioactive cesium ((134)Cs plus (137)Cs) in rice fractions during polishing and cooking.

    PubMed

    Hachinohe, Mayumi; Okunishi, Tomoya; Hagiwara, Shoji; Todoriki, Setsuko; Kawamoto, Shinichi; Hamamatsu, Shioka

    2015-03-01

    We investigated the distribution of cesium-134 ((134)Cs) and cesium-137 ((137)Cs) during polishing and cooking of rice to obtain their processing factors (Pf) and food processing retention factors (Fr) to make the information available for an adequate understanding of radioactive Cs dynamics. Polishing brown rice resulted in a decreased radioactive Cs concentration of the polished rice, but the bran and germ (outer layers) exhibited higher concentrations than brown rice. The Pf values for 100% polished rice and outer layers ranged from 0.47 to 0.48 and 6.5 to 7.8, respectively. The Fr values for 100% polished rice and outer layers were 0.43 and 0.58 to 0.60, respectively. The distribution of radioactive Cs in polished rice and outer layers was estimated at approximately 40 and 60%, respectively. On the other hand, cooked rice showed significantly lower levels of radioactive Cs than polished rice, and transfer of radioactive Cs into wash water was observed. The Pf and Fr values for cooked rice were 0.28 and 0.65 to 0.66, respectively. From these results, we can calculate that if the radioactive Cs concentration in brown rice is 100 Bq/kg, the concentrations of Cs in polished rice and cooked rice will be 47 to 48 Bq/kg and 13 Bq/kg, respectively.

  20. The nature of ion and water barrier crossings in a simulated ion channel.

    PubMed Central

    Chiu, S. W.; Novotny, J. A.; Jakobsson, E.

    1993-01-01

    Using a combination of techniques, including molecular dynamics, time-correlation analysis, stochastic dynamics, and fitting of continuum diffusion theory to electrophysiological data, a characterization is made of thermally driven sodium, water, and D2O motion within the gramicidin A channel. Since the channel contents are constrained to move in a single-file fashion, the motion that corresponds to experimentally measurable rates of permeation of the membrane is the motion of the center of mass of the channel contents. We therefore emphasize channel contents center-of-mass motion in our analysis of molecular dynamics computations. The usual free energy calculation techniques would be of questionable validity when applied to such motion. As an alternative to those techniques, we postulate a periodic sinusoidal free energy profile (related to the periodic structure of the helical channel) and deduce the fluid dynamic diffusion coefficient and the height and spacing of the free energy barriers from the form of the mean-square-deviation function, using stochastic computations. The fluid dynamic friction in each case appears similar to that for aqueous solution. However, the diffusive motions are modulated by a spatially periodic free energy profile with a periodicity characteristic of an L-D pair of amino acids in the gramicidin helix, approximately 1.7 A in the model we use. The barrier height depends on which substance is moving in the channel, but in each case is several times thermal energy. For barriers of this width and height, the motion is intermediate between the low-friction (transition-state) and high-friction (Brownian) limits. Thus, neither of these formalisms that have been used commonly to describe membrane permeation gives an accurate picture of the underlying physical process (although the Brownian description seems closer to correct). The non-Markovian Langevin equation must be solved to describe properly the statistics of the process. The "channel

  1. Channel-forming bacterial toxins in biosensing and macromolecule delivery.

    PubMed

    Gurnev, Philip A; Nestorovich, Ekaterina M

    2014-08-21

    To intoxicate cells, pore-forming bacterial toxins are evolved to allow for the transmembrane traffic of different substrates, ranging from small inorganic ions to cell-specific polypeptides. Recent developments in single-channel electrical recordings, X-ray crystallography, protein engineering, and computational methods have generated a large body of knowledge about the basic principles of channel-mediated molecular transport. These discoveries provide a robust framework for expansion of the described principles and methods toward use of biological nanopores in the growing field of nanobiotechnology. This article, written for a special volume on "Intracellular Traffic and Transport of Bacterial Protein Toxins", reviews the current state of applications of pore-forming bacterial toxins in small- and macromolecule-sensing, targeted cancer therapy, and drug delivery. We discuss the electrophysiological studies that explore molecular details of channel-facilitated protein and polymer transport across cellular membranes using both natural and foreign substrates. The review focuses on the structurally and functionally different bacterial toxins: gramicidin A of Bacillus brevis, α-hemolysin of Staphylococcus aureus, and binary toxin of Bacillus anthracis, which have found their "second life" in a variety of developing medical and technological applications.

  2. Channel-Forming Bacterial Toxins in Biosensing and Macromolecule Delivery

    PubMed Central

    Gurnev, Philip A.; Nestorovich, Ekaterina M.

    2014-01-01

    To intoxicate cells, pore-forming bacterial toxins are evolved to allow for the transmembrane traffic of different substrates, ranging from small inorganic ions to cell-specific polypeptides. Recent developments in single-channel electrical recordings, X-ray crystallography, protein engineering, and computational methods have generated a large body of knowledge about the basic principles of channel-mediated molecular transport. These discoveries provide a robust framework for expansion of the described principles and methods toward use of biological nanopores in the growing field of nanobiotechnology. This article, written for a special volume on “Intracellular Traffic and Transport of Bacterial Protein Toxins”, reviews the current state of applications of pore-forming bacterial toxins in small- and macromolecule-sensing, targeted cancer therapy, and drug delivery. We discuss the electrophysiological studies that explore molecular details of channel-facilitated protein and polymer transport across cellular membranes using both natural and foreign substrates. The review focuses on the structurally and functionally different bacterial toxins: gramicidin A of Bacillus brevis, α-hemolysin of Staphylococcus aureus, and binary toxin of Bacillus anthracis, which have found their “second life” in a variety of developing medical and technological applications. PMID:25153255

  3. Three-Dimensional Stress Field around a Membrane Protein: Atomistic and Coarse-Grained Simulation Analysis of Gramicidin A

    PubMed Central

    Yoo, Jejoong; Cui, Qiang

    2013-01-01

    Using both atomistic and coarse-grained (CG) models, we compute the three-dimensional stress field around a gramicidin A (gA) dimer in lipid bilayers that feature different degrees of negative hydrophobic mismatch. The general trends in the computed stress field are similar at the atomistic and CG levels, supporting the use of the CG model for analyzing the mechanical features of protein/lipid/water interfaces. The calculations reveal that the stress field near the protein-lipid interface exhibits a layered structure with both significant repulsive and attractive regions, with the magnitude of the stress reaching 1000 bar in certain regions. Analysis of density profiles and stress field distributions helps highlight the Trp residues at the protein/membrane/water interface as mechanical anchors, suggesting that similar analysis is useful for identifying tension sensors in other membrane proteins, especially membrane proteins involved in mechanosensation. This work fosters a connection between microscopic and continuum mechanics models for proteins in complex environments and makes it possible to test the validity of assumptions commonly made in continuum mechanics models for membrane mediated processes. For example, using the calculated stress field, we estimate the free energy of membrane deformation induced by the hydrophobic mismatch, and the results for regions beyond the annular lipids are in general consistent with relevant experimental data and previous theoretical estimates using elasticity theory. On the other hand, the assumptions of homogeneous material properties for the membrane and a bilayer thickness at the protein/lipid interface being independent of lipid type (e.g., tail length) appear to be oversimplified, highlighting the importance of annular lipids of membrane proteins. Finally, the stress field analysis makes it clear that the effect of even rather severe hydrophobic mismatch propagates to only about two to three lipid layers, thus putting a

  4. Three-dimensional stress field around a membrane protein: atomistic and coarse-grained simulation analysis of gramicidin A.

    PubMed

    Yoo, Jejoong; Cui, Qiang

    2013-01-01

    Using both atomistic and coarse-grained (CG) models, we compute the three-dimensional stress field around a gramicidin A (gA) dimer in lipid bilayers that feature different degrees of negative hydrophobic mismatch. The general trends in the computed stress field are similar at the atomistic and CG levels, supporting the use of the CG model for analyzing the mechanical features of protein/lipid/water interfaces. The calculations reveal that the stress field near the protein-lipid interface exhibits a layered structure with both significant repulsive and attractive regions, with the magnitude of the stress reaching 1000 bar in certain regions. Analysis of density profiles and stress field distributions helps highlight the Trp residues at the protein/membrane/water interface as mechanical anchors, suggesting that similar analysis is useful for identifying tension sensors in other membrane proteins, especially membrane proteins involved in mechanosensation. This work fosters a connection between microscopic and continuum mechanics models for proteins in complex environments and makes it possible to test the validity of assumptions commonly made in continuum mechanics models for membrane mediated processes. For example, using the calculated stress field, we estimate the free energy of membrane deformation induced by the hydrophobic mismatch, and the results for regions beyond the annular lipids are in general consistent with relevant experimental data and previous theoretical estimates using elasticity theory. On the other hand, the assumptions of homogeneous material properties for the membrane and a bilayer thickness at the protein/lipid interface being independent of lipid type (e.g., tail length) appear to be oversimplified, highlighting the importance of annular lipids of membrane proteins. Finally, the stress field analysis makes it clear that the effect of even rather severe hydrophobic mismatch propagates to only about two to three lipid layers, thus putting a

  5. Potential role of CS2 photooxidation in tropospheric sulfur chemistry

    NASA Technical Reports Server (NTRS)

    Wine, P. H.; Chameides, W. L.; Ravishankara, A. R.

    1981-01-01

    Absorption cross section measurements and model calculations indicate that CS2 photooxidation may be an important tropospheric sink for the CS2, giving a lifetime on the order of a week or two. If background CS2 levels are 10-20 pptv, then CS2 photooxidation may be an important global source of OCS as well.

  6. A biokinetic model for {sup 137}Cs

    SciTech Connect

    Melo, D.R.; Lipsztein, J.L.; Oliveira, C.A.N.; Lundgren, D.L.

    1997-08-01

    An improved biokinetic model for {sup 137}Cs in humans was developed based on an analysis of data obtained from individuals internally contaminated during an accident in Goiania, Brazil, and other data. Seventeen children (ten girls and seven boys 1-10 y old), ten adolescents (four females and six males), and thirty adults, (fifteen females and fifteen males) contaminated in the accident in Goiania contributed to this study. {sup 137}Cs retention was determined through periodic measurements in a whole-body counter. In addition to the data on {sup 137}Cs retention from these individuals, data from a study on the metabolism of {sup 137}Cs in immature, adult, and aged Beagle dogs and data from the literature were used in the formulation of the {sup 137}Cs biokinetic model presented. Mathematically, the retention of cesium is described by three exponential terms, and the retention model is based on a step function of body weight. When the ICRP Publication 56 model for cesium was compared to the model suggested in this paper, it was determined that the ICRP model predicts lower effective doses in 5-y-old children and higher effective doses in infants, adolescents, and adults.

  7. CS Unplugged and Middle-School Students' Views, Attitudes, and Intentions regarding CS

    ERIC Educational Resources Information Center

    Taub, Rivka; Armoni, Michal; Ben-Ari, Mordechai

    2012-01-01

    Many students hold incorrect ideas and negative attitudes about computer science (CS). In order to address these difficulties, a series of learning activities called Computer Science Unplugged was developed by Tim Bell and his colleagues. These activities expose young people to central concepts in CS in an entertaining way without requiring a…

  8. TRP Channels

    NASA Astrophysics Data System (ADS)

    Voets, Thomas; Owsianik, Grzegorz; Nilius, Bernd

    The TRP superfamily represents a highly diverse group of cation-permeable ion channels related to the product of the Drosophila trp (transient receptor potential) gene. The cloning and characterization of members of this cation channel family has experienced a remarkable growth during the last decade, uncovering a wealth of information concerning the role of TRP channels in a variety of cell types, tissues, and species. Initially, TRP channels were mainly considered as phospholipase C (PLC)-dependent and/or store-operated Ca2+-permeable cation channels. More recent research has highlighted the sensitivity of TRP channels to a broad array of chemical and physical stimuli, allowing them to function as dedicated biological sensors involved in processes ranging from vision to taste, tactile sensation, and hearing. Moreover, the tailored selectivity of certain TRP channels enables them to play key roles in the cellular uptake and/or transepithelial transport of Ca2+, Mg2+, and trace metal ions. In this chapter we give a brief overview of the TRP channel superfamily followed by a survey of current knowledge concerning their structure and activation mechanisms.

  9. Lactosylated gramicidin-based lipid nanoparticles (Lac-GLN) for targeted delivery of anti-miR-155 to hepatocellular carcinoma.

    PubMed

    Zhang, Mengzi; Zhou, Xiaoju; Wang, Bo; Yung, Bryant C; Lee, Ly J; Ghoshal, Kalpana; Lee, Robert J

    2013-06-28

    Lactosylated gramicidin-containing lipid nanoparticles (Lac-GLN) were developed for delivery of anti-microRNA-155 (anti-miR-155) to hepatocellular carcinoma (HCC) cells. MiR-155 is an oncomiR frequently elevated in HCC. The Lac-GLN formulation contained N-lactobionyl-dioleoyl phosphatidylethanolamine (Lac-DOPE), a ligand for the asialoglycoprotein receptor (ASGR), and an antibiotic peptide gramicidin A. The nanoparticles exhibited a mean particle diameter of 73 nm, zeta potential of +3.5mV, anti-miR encapsulation efficiency of 88%, and excellent colloidal stability at 4°C. Lac-GLN effectively delivered anti-miR-155 to HCC cells with a 16.1- and 4.1-fold up-regulation of miR-155 targets C/EBPβ and FOXP3 genes, respectively, and exhibited significant greater efficiency over Lipofectamine 2000. In mice, intravenous injection of Lac-GLN containing Cy3-anti-miR-155 led to preferential accumulation of the anti-miR-155 in hepatocytes. Intravenous administration of 1.5 mg/kg anti-miR-155 loaded Lac-GLN resulted in up-regulation of C/EBPβ and FOXP3 by 6.9- and 2.2-fold, respectively. These results suggest potential application of Lac-GLN as a liver-specific delivery vehicle for anti-miR therapy.

  10. Tear gasses CN, CR, and CS are potent activators of the human TRPA1 receptor

    SciTech Connect

    Brone, Bert; Peeters, Pieter J.; Marrannes, Roger; Mercken, Marc; Nuydens, Ronny; Meert, Theo; Gijsen, Harrie J.M.

    2008-09-01

    The TRPA1 channel is activated by a number of pungent chemicals, such as allylisothiocyanate, present in mustard oil and thiosulfinates present in garlic. Most of the known activating compounds contain reactive, electrophilic chemical groups, reacting with cysteine residues in the active site of the TRPA1 channel. This covalent modification results in activation of the channel and has been shown to be reversible for several ligands. Commonly used tear gasses CN, CR and CS are also pungent chemicals, and in this study we show that they are extremely potent and selective activators of the human TRPA1 receptor. To our knowledge, these are the most potent TRPA1 agonists known to date. The identification of the molecular target for these tear gasses may open up possibilities to alleviate the effects of tear gasses via treatment with TRPA1 antagonists. In addition these results may contribute to the basic knowledge of the TRPA1 channel that is gaining importance as a pharmacological target.

  11. 135Cs/137Cs isotopic composition of environmental samples across Europe: Environmental transport and source term emission applications

    DOE PAGESBeta

    Snow, Mathew S.; Snyder, Darin C.

    2015-11-02

    135Cs/137Cs isotopic analyses represent an important tool for studying the fate and transport of radiocesium in the environment; in this work the 135Cs/137Cs isotopic composition in environmental samples taken from across Europe is reported. Surface soil and vegetation samples from western Russia, Ukraine, Austria, and Hungary show consistent aged thermal fission product 135Cs/137Cs isotope ratios of 0.58 ± 0.01 (age corrected to 1/1/15), with the exception of one sample of soil-moss from Hungary which shows an elevated 135Cs/137Cs ratio of 1.78 ± 0.12. With the exception of the outlier sample from Hungary, surface soil/vegetation data are in quantitative agreement withmore » values previously reported for soils within the Chernobyl exclusion zone, suggesting that radiocesium at these locations is primarily composed of homogenous airborne deposition from Chernobyl. Seawater samples taken from the Irish Sea show 135Cs/137Cs isotope ratios of 1.22 ± 0.11 (age corrected to 1/1/15), suggesting aged thermal fission product Cs discharged from Sellafield. Furthermore, the differences in 135Cs/137Cs isotope ratios between Sellafield, Chernobyl, and global nuclear weapons testing fallout indicate that 135Cs/137Cs isotope ratios can be utilized to discriminate between and track radiocesium transport from different nuclear production source terms, including major emission sources in Europe.« less

  12. (135)Cs/(137)Cs isotopic composition of environmental samples across Europe: Environmental transport and source term emission applications.

    PubMed

    Snow, Mathew S; Snyder, Darin C

    2016-01-01

    (135)Cs/(137)Cs isotopic analyses represent an important tool for studying the fate and transport of radiocesium in the environment; in this work the (135)Cs/(137)Cs isotopic composition in environmental samples taken from across Europe is reported. Surface soil and vegetation samples from western Russia, Ukraine, Austria, and Hungary show consistent aged thermal fission product (135)Cs/(137)Cs isotope ratios of 0.58 ± 0.01 (age corrected to 1/1/15), with the exception of one sample of soil-moss from Hungary which shows an elevated (135)Cs/(137)Cs ratio of 1.78 ± 0.12. With the exception of the outlier sample from Hungary, surface soil/vegetation data are in quantitative agreement with values previously reported for soils within the Chernobyl exclusion zone, suggesting that radiocesium at these locations is primarily composed of homogenous airborne deposition from Chernobyl. Seawater samples taken from the Irish Sea show (135)Cs/(137)Cs isotope ratios of 1.22 ± 0.11 (age corrected to 1/1/15), suggesting aged thermal fission product Cs discharged from Sellafield. The differences in (135)Cs/(137)Cs isotope ratios between Sellafield, Chernobyl, and global nuclear weapons testing fallout indicate that (135)Cs/(137)Cs isotope ratios can be utilized to discriminate between and track radiocesium transport from different nuclear production source terms, including major emission sources in Europe.

  13. (135)Cs/(137)Cs isotopic composition of environmental samples across Europe: Environmental transport and source term emission applications.

    PubMed

    Snow, Mathew S; Snyder, Darin C

    2016-01-01

    (135)Cs/(137)Cs isotopic analyses represent an important tool for studying the fate and transport of radiocesium in the environment; in this work the (135)Cs/(137)Cs isotopic composition in environmental samples taken from across Europe is reported. Surface soil and vegetation samples from western Russia, Ukraine, Austria, and Hungary show consistent aged thermal fission product (135)Cs/(137)Cs isotope ratios of 0.58 ± 0.01 (age corrected to 1/1/15), with the exception of one sample of soil-moss from Hungary which shows an elevated (135)Cs/(137)Cs ratio of 1.78 ± 0.12. With the exception of the outlier sample from Hungary, surface soil/vegetation data are in quantitative agreement with values previously reported for soils within the Chernobyl exclusion zone, suggesting that radiocesium at these locations is primarily composed of homogenous airborne deposition from Chernobyl. Seawater samples taken from the Irish Sea show (135)Cs/(137)Cs isotope ratios of 1.22 ± 0.11 (age corrected to 1/1/15), suggesting aged thermal fission product Cs discharged from Sellafield. The differences in (135)Cs/(137)Cs isotope ratios between Sellafield, Chernobyl, and global nuclear weapons testing fallout indicate that (135)Cs/(137)Cs isotope ratios can be utilized to discriminate between and track radiocesium transport from different nuclear production source terms, including major emission sources in Europe. PMID:26540258

  14. Properties of CsI and CsI-TMAE photocathodes

    SciTech Connect

    Anderson, D.F.; Kwan, S.; Peskov, V.; Hoeneisen, B.

    1992-06-01

    The importance of heating the CsI or CsI-TMAE photocathodes during preparation, as well as the importance of the gas environment on the quantum efficiency is presented. The dependence of the aging characteristics of these photocathodes on the operating temperature, on the presence of gas, and on the charge amplification of the chamber is also discussed. For CsI photocathodes charges in excess of 2{times}10{sup 14} e{sup {minus}}/mm{sup 2} can be collected with little degradation of performance. A timing resolution of 0.55 ns is also achieved for single photoelectrons suggesting a possible time-of-flight detector.

  15. Isolated-patch recording from liposomes containing functionally reconstituted chloride channels from Torpedo electroplax.

    PubMed Central

    Tank, D W; Miller, C; Webb, W W

    1982-01-01

    Small unilamellar vesicles formed from purified phospholids by detergent/dialysis methods may be enlarged to 30-microns diameter by freezing and thawing. Very-high-resistance seals were formed by applying a glass micropipette to the surface of these large liposomes, and single bilayer "patches" of membrane were isolated from the liposome surface while remaining sealed to the micropipette. The exogenous channel-forming peptides gramicidin and alamethicin induced characteristic single-channel fluctuation behavior in these excised patches held under voltage-clamp conditions. Large liposomes were formed from the small unilamellar vesicles made from cholate extracts of Torpedo electroplax plasma membrane vesicles. Isolated patches formed from these reconstituted membranes displayed current fluctuations due to single voltage-gated Cl- channels from non-innervated-face membranes; the properties of these Cl- channels are identical to those observed in planar bilayer membranes after direct insertion from native membranes. This liposome-patch method combines the advantages of membrane protein incorporation into liposomes with high-resolution electrical recording methods and may provide a generally applicable approach to the study of integral membrane channel proteins after solubilization and reconstitution. Images PMID:6296849

  16. A Low-Noise Transimpedance Amplifier for BLM-Based Ion Channel Recording.

    PubMed

    Crescentini, Marco; Bennati, Marco; Saha, Shimul Chandra; Ivica, Josip; de Planque, Maurits; Morgan, Hywel; Tartagni, Marco

    2016-01-01

    High-throughput screening (HTS) using ion channel recording is a powerful drug discovery technique in pharmacology. Ion channel recording with planar bilayer lipid membranes (BLM) is scalable and has very high sensitivity. A HTS system based on BLM ion channel recording faces three main challenges: (i) design of scalable microfluidic devices; (ii) design of compact ultra-low-noise transimpedance amplifiers able to detect currents in the pA range with bandwidth >10 kHz; (iii) design of compact, robust and scalable systems that integrate these two elements. This paper presents a low-noise transimpedance amplifier with integrated A/D conversion realized in CMOS 0.35 μm technology. The CMOS amplifier acquires currents in the range ±200 pA and ±20 nA, with 100 kHz bandwidth while dissipating 41 mW. An integrated digital offset compensation loop balances any voltage offsets from Ag/AgCl electrodes. The measured open-input input-referred noise current is as low as 4 fA/√Hz at ±200 pA range. The current amplifier is embedded in an integrated platform, together with a microfluidic device, for current recording from ion channels. Gramicidin-A, α-haemolysin and KcsA potassium channels have been used to prove both the platform and the current-to-digital converter. PMID:27213382

  17. A Low-Noise Transimpedance Amplifier for BLM-Based Ion Channel Recording

    PubMed Central

    Crescentini, Marco; Bennati, Marco; Saha, Shimul Chandra; Ivica, Josip; de Planque, Maurits; Morgan, Hywel; Tartagni, Marco

    2016-01-01

    High-throughput screening (HTS) using ion channel recording is a powerful drug discovery technique in pharmacology. Ion channel recording with planar bilayer lipid membranes (BLM) is scalable and has very high sensitivity. A HTS system based on BLM ion channel recording faces three main challenges: (i) design of scalable microfluidic devices; (ii) design of compact ultra-low-noise transimpedance amplifiers able to detect currents in the pA range with bandwidth >10 kHz; (iii) design of compact, robust and scalable systems that integrate these two elements. This paper presents a low-noise transimpedance amplifier with integrated A/D conversion realized in CMOS 0.35 μm technology. The CMOS amplifier acquires currents in the range ±200 pA and ±20 nA, with 100 kHz bandwidth while dissipating 41 mW. An integrated digital offset compensation loop balances any voltage offsets from Ag/AgCl electrodes. The measured open-input input-referred noise current is as low as 4 fA/√Hz at ±200 pA range. The current amplifier is embedded in an integrated platform, together with a microfluidic device, for current recording from ion channels. Gramicidin-A, α-haemolysin and KcsA potassium channels have been used to prove both the platform and the current-to-digital converter. PMID:27213382

  18. Low level detection of Cs-135 and Cs-137 in environmental samples by ICP-MS

    SciTech Connect

    Liezers, Martin; Farmer, Orville T.; Thomas, Linda MP

    2009-10-01

    The measurement of the fission product cesium isotopes 135Cs and 137Cs at low femtogram (fg) 10-15 levels in ground water by Inductively Coupled Plasma-Mass Spectrometry ICP-MS is reported. To eliminate the potential natural barium isobaric interference on the cesium isotopes, in-line chromatographic separation of the cesium from barium was performed followed by high sensitivity ICP-MS analysis. A high efficiency desolvating nebulizer system was employed to maximize ICP-MS sensitivity ~10cps/femtogram. The three sigma detection limit measured for 135Cs was 2fg/ml (0.1uBq/ml) and for 137Cs 0.9fg/ml (0.0027Bq/ml) with analysis time of less than 30 minutes/sample. Cesium detection and 135/137 isotope ratio measurement at very low femtogram levels using this method in a ground water matrix is also demonstrated.

  19. Determination of ¹³⁵Cs and ¹³⁷Cs in environmental samples: A review.

    PubMed

    Russell, B C; Croudace, Ian W; Warwick, Phil E

    2015-08-26

    Radionuclides of caesium are environmentally important since they are formed as significant high yield fission products ((135)Cs and (137)Cs) and activation products ((134)Cs and (136)Cs) during nuclear fission. They originate from a range of nuclear activities such as weapons testing, nuclear reprocessing and nuclear fuel cycle discharges and nuclear accidents. Whilst (137)Cs, (134)Cs and (136)Cs are routinely measurable at high sensitivity by gamma spectrometry, routine detection of long-lived (135)Cs by radiometric methods is challenging. This measurement is, however, important given its significance in long-term nuclear waste storage and disposal. Furthermore, the (135)Cs/(137)Cs ratio varies with reactor, weapon and fuel type, and accurate measurement of this ratio can therefore be used as a forensic tool in identifying the source(s) of nuclear contamination. The shorter-lived activation products (134)Cs and (136)Cs have a limited application but provide useful early information on fuel irradiation history and have importance in health physics. Detection of (135)Cs (and (137)Cs) is achievable by mass spectrometric techniques; most commonly inductively coupled plasma mass spectrometry (ICP-MS), as well as thermal ionisation (TIMS), accelerator (AMS) and resonance ionisation (RIMS) techniques. The critical issues affecting the accuracy and detection limits achievable by this technique are effective removal of barium to eliminate isobaric interferences arising from (135)Ba and (137)Ba, and elimination of peak tailing of stable (133)Cs on (135)Cs. Isobaric interferences can be removed by chemical separation, most commonly ion exchange chromatography, and/or instrumental separation using an ICP-MS equipped with a reaction cell. The removal of the peak tailing interference is dependent on the instrument used for final measurement. This review summarizes and compares the analytical procedures developed for determination of (135)Cs/(137)Cs, with particular focus on

  20. Determination of ¹³⁵Cs and ¹³⁷Cs in environmental samples: A review.

    PubMed

    Russell, B C; Croudace, Ian W; Warwick, Phil E

    2015-08-26

    Radionuclides of caesium are environmentally important since they are formed as significant high yield fission products ((135)Cs and (137)Cs) and activation products ((134)Cs and (136)Cs) during nuclear fission. They originate from a range of nuclear activities such as weapons testing, nuclear reprocessing and nuclear fuel cycle discharges and nuclear accidents. Whilst (137)Cs, (134)Cs and (136)Cs are routinely measurable at high sensitivity by gamma spectrometry, routine detection of long-lived (135)Cs by radiometric methods is challenging. This measurement is, however, important given its significance in long-term nuclear waste storage and disposal. Furthermore, the (135)Cs/(137)Cs ratio varies with reactor, weapon and fuel type, and accurate measurement of this ratio can therefore be used as a forensic tool in identifying the source(s) of nuclear contamination. The shorter-lived activation products (134)Cs and (136)Cs have a limited application but provide useful early information on fuel irradiation history and have importance in health physics. Detection of (135)Cs (and (137)Cs) is achievable by mass spectrometric techniques; most commonly inductively coupled plasma mass spectrometry (ICP-MS), as well as thermal ionisation (TIMS), accelerator (AMS) and resonance ionisation (RIMS) techniques. The critical issues affecting the accuracy and detection limits achievable by this technique are effective removal of barium to eliminate isobaric interferences arising from (135)Ba and (137)Ba, and elimination of peak tailing of stable (133)Cs on (135)Cs. Isobaric interferences can be removed by chemical separation, most commonly ion exchange chromatography, and/or instrumental separation using an ICP-MS equipped with a reaction cell. The removal of the peak tailing interference is dependent on the instrument used for final measurement. This review summarizes and compares the analytical procedures developed for determination of (135)Cs/(137)Cs, with particular focus on

  1. Monitoring 137Cs and 134Cs at marine coasts in Indonesia between 2011 and 2013.

    PubMed

    Suseno, Heny; Prihatiningsih, Wahyu Retno

    2014-11-15

    Environmental samples (seawater, sediments and biota) were collected along the eastern and western Indonesian coasts between 2011 and 2013 to anticipate the possible impacts of the Fukushima radioactive releases in Indonesia. On the eastern coasts (south and north Sulawesi), the (137)Cs concentrations in the seawater and sediments were 0.12-0.32 Bq m(-3) and 0.10-1.03 Bq kg(-1), respectively. On the western coasts (West Sumatra, Bangka Island, North Java, South Java and Madura island), the (137)Cs concentrations in the seawater and sediments were 0.12-0.66 Bq m(-3) and 0.19-1.64 Bq kg(-1), respectively. In general, the (137)Cs concentrations in the fish from several Indonesian coasts were Cs concentrations in mollusk, crab and prawn were 10.65-38.78, 4.02 and 6.16 mBq kg(-1), respectively. (134)Cs was not detected in the seawater, sediments or biota. Thus, it was concluded that (137)Cs on the eastern and western Indonesian coasts originated from global fallout.

  2. In-situ Dehydration Studies of Fully K- Rb- and Cs-exchanged Natrolites

    SciTech Connect

    Y Lee; D Seoung; D Liu; M Park; S Hong; H Chen; J Bai; C Kao; T Vogt; Y Lee

    2011-12-31

    In-situ synchrotron X-ray powder diffraction studies of K-, Rb-, and Cs-exchanged natrolites between room temperature and 425 C revealed that the dehydrated phases with collapsed frameworks start to form at 175, 150, and 100 C, respectively. The degree of the framework collapse indicated by the unit-cell volume contraction depends on the size of the non-framework cation: K-exchanged natrolite undergoes an 18.8% unit-cell volume contraction when dehydrated at 175 C, whereas Rb- and Cs-exchanged natrolites show unit-cell volume contractions of 18.5 and 15.2% at 150 and 100 C, respectively. In the hydrated phases, the dehydration-induced unit-cell volume reduction diminishes as the cation size increases and reveals increasingly a negative slope as smaller cations are substituted into the pores of the natrolite structure. The thermal expansion of the unit-cell volumes of the dehydrated K-, Rb-, and Cs-phases have positive thermal expansion coefficients of 8.80 x 10{sup -5} K{sup -1}, 1.03 x 10{sup -4} K{sup 01}, and 5.06 x 10{sup -5} K{sup -1}, respectively. Rietveld structure refinements of the dehydrated phases at 400 C reveal that the framework collapses are due to an increase of the chain rotation angles, {Psi}, which narrow the channels to a more elliptical shape. Compared to their respective hydrated structures at ambient conditions, the dehydrated K-exchanged natrolite at 400 C shows a 2.2-fold increase in {Psi}, whereas the dehydrated Rb- and Cs-natrolites at 400 C reveal increases of {Psi} by ca. 3.7 and 7.3 times, respectively. The elliptical channel openings of the dehydrated K-, Rb-, to Cs-phases become larger as the cation size increases. The disordered non-framework cations in the hydrated K-, Rb-, and Cs-natrolite order during dehydration and the subsequent framework collapse. The dehydrated phases of Rb- and Cs-natrolite can be stabilized at ambient conditions.

  3. English Channel

    NASA Technical Reports Server (NTRS)

    1984-01-01

    The cloud covered earthscape of Northern Europe demonstrates the difficulty of photographing this elusive subject from space. The English Channel (51.0N, 1.5E) separating the British Islands from Europe is in the center of the scene. The white cliffs of Dover on the SE coast of the UK, the Thames River estuary and a partial view of the city of London can be seen on the north side of the Channel while the Normandy coast of France is to the south.

  4. IR-Improved DGLAP-CS Theory

    DOE PAGESBeta

    Ward, B. F. L.

    2008-01-01

    We show that it is possible to improve the infrared aspects of the standard treatment of the DGLAP-CS evolution theory to take into account a large class of higher-order corrections that significantly improve the precision of the theory for any given level of fixed-order calculation of its respective kernels. We illustrate the size of the effects we resum using the moments of the parton distributions.

  5. Communications satellite no. 2 (CS-2)

    NASA Technical Reports Server (NTRS)

    1982-01-01

    The purpose of the Japanese CS-2 satellite is to provide national communications and industrial communications, such as special emergency and remote communications, and to contribute to the development of technology pertaining to communications satellites. Description and operating parameters of the following satellite components are presented: structure, communications system, telemetry/command system, electric power system, attitude and antenna control system, secondary propulsion system, apogee motor, framework, and heat control system.

  6. Teaching Algorithm Efficiency at CS1 Level: A Different Approach

    ERIC Educational Resources Information Center

    Gal-Ezer, Judith; Vilner, Tamar; Zur, Ela

    2004-01-01

    Realizing the importance of teaching efficiency at early stages of the program of study in computer science (CS) on one hand, and the difficulties encountered when introducing this concept on the other, we advocate a different didactic approach in the introductory CS course (CS1). This paper describes the approach as it is used at the Open…

  7. Developments towards detection of 135Cs at VERA

    NASA Astrophysics Data System (ADS)

    Lachner, Johannes; Kasberger, Magdalena; Martschini, Martin; Priller, Alfred; Steier, Peter; Golser, Robin

    2015-10-01

    Radioisotopes produced in natural or anthropogenic fission are widely used for tracer studies of environmental processes, in nuclear forensics, and are important for nuclear waste disposal. Besides the well-known 137Cs, the longer-lived sister isotope 135Cs (T1/2 = 2.3 Myr) is also produced, and the combined measurement of the two isotopes would allow for assessment of contaminating sources. The insufficient suppression of the stable isobar 135Ba presently prevents AMS measurements down to expected natural levels of 135Cs/133Cs ≈ 10-11. Via the difference in electron affinities between Cs and Ba further isobar suppression should be achievable after the installation of the Ion-Laser-Interaction System (ILIAS) at VERA. We present a preparatory study on the performance of the 3 MV VERA AMS facility for 135Cs concerning ion formation, transmission and detection. Since the usual Cs sputtering would obscure the 135Cs/133Cs ratio of a sample, Rb sputtering was successfully applied and tested also for various other typical AMS elements. Partial suppression of 135Ba is possible with the extraction of Cs- and negative Cs-fluorides. Cs- currents of several 10 nA were extracted over hours from mg amounts of Cs2SO4 material. The transmission to various charge states was tested with gas (Ar, He) and foil stripping. Experiments showed that no suppression in the detection system is possible at high beam energies with the VERA facility. For this reason, gas stripping to low charge states (2+, 3+) with transmissions up to 30% is favorable to guarantee optimal beam transport to the detector. In the present setup, utilizing a simple Bragg-type detector, the blank 135Cs/133Cs ratios from chemically pure samples are determined by the 135Ba background to a value of (4.0 ± 1.3)·10-9.

  8. Pressure-induced metathesis reaction to sequester Cs.

    PubMed

    Im, Junhyuck; Seoung, Donghoon; Lee, Seung Yeop; Blom, Douglas A; Vogt, Thomas; Kao, Chi-Chang; Lee, Yongjae

    2015-01-01

    We report here a pressure-driven metathesis reaction where Ag-exchanged natrolite (Ag16Al16Si24O80·16H2O, Ag-NAT) is pressurized in an aqueous CsI solution, resulting in the exchange of Ag(+) by Cs(+) in the natrolite framework forming Cs16Al16Si24O80·16H2O (Cs-NAT-I) and, above 0.5 GPa, its high-pressure polymorph (Cs-NAT-II). During the initial cation exchange, the precipitation of AgI occurs. Additional pressure and heat at 2 GPa and 160 °C transforms Cs-NAT-II to a pollucite-related, highly dense, and water-free triclinic phase with nominal composition CsAlSi2O6. At ambient temperature after pressure release, the Cs remains sequestered in a now monoclinic pollucite phase at close to 40 wt % and a favorably low Cs leaching rate under back-exchange conditions. This process thus efficiently combines the pressure-driven separation of Cs and I at ambient temperature with the subsequent sequestration of Cs under moderate pressures and temperatures in its preferred waste form suitable for long-term storage at ambient conditions. The zeolite pollucite CsAlSi2O6·H2O has been identified as a potential host material for nuclear waste remediation of anthropogenic (137)Cs due to its chemical and thermal stability, low leaching rate, and the large amount of Cs it can contain. The new water-free pollucite phase we characterize during our process will not display radiolysis of water during longterm storage while maintaining the Cs content and low leaching rate. PMID:25515673

  9. Free energy of formation of Cs 3PuCl 6 and CsPu 2Cl 7

    NASA Astrophysics Data System (ADS)

    Williamson, M. A.; Kleinschmidt, P. D.

    The free energy, enthalpy and entropy of formation of the compounds Cs 3PuCl 6 and CsPu 2Cl 7 have been determined by measuring the sublimation pressures for the reactions CsCl( s) / aiCsCl( g), {2}/{5}Cs 3PuCl 6(s) /ai {1}/{5}CsPu 2Cl 7(s) + CsCl(g) , and CsPu2Cl7( s) / ai 2 PuCl3( s) + CsCl( g). The pressures are measured using Knudsen effusion mass spectrometry over the temperature range 600 to 850 K. For the formation of Cs 3PuCl 6 from CsCl and PuCl 3, ΔG0298 = -77.3 ± 8.5 kJ/ mol, ΔH0298 = -82.1 ± 7.8 kJ/ mol, and ΔS0298 = -16.2 ± 10.9 J/ Kmol. For CsPu 2Cl 7, ΔG0298 = -39.4 ± 3.5 kJ/ mol, ΔH0298 = -40.8 ± 3.2 kJ/ mol, and ΔS0298 = -4.6 ± 4.2 J/ Kmol.

  10. Charging the Quantum Capacitance of Graphene with a Single Biological Ion Channel

    PubMed Central

    2015-01-01

    The interaction of cell and organelle membranes (lipid bilayers) with nanoelectronics can enable new technologies to sense and measure electrophysiology in qualitatively new ways. To date, a variety of sensing devices have been demonstrated to measure membrane currents through macroscopic numbers of ion channels. However, nanoelectronic based sensing of single ion channel currents has been a challenge. Here, we report graphene-based field-effect transistors combined with supported lipid bilayers as a platform for measuring, for the first time, individual ion channel activity. We show that the supported lipid bilayers uniformly coat the single layer graphene surface, acting as a biomimetic barrier that insulates (both electrically and chemically) the graphene from the electrolyte environment. Upon introduction of pore-forming membrane proteins such as alamethicin and gramicidin A, current pulses are observed through the lipid bilayers from the graphene to the electrolyte, which charge the quantum capacitance of the graphene. This approach combines nanotechnology with electrophysiology to demonstrate qualitatively new ways of measuring ion channel currents. PMID:24754625

  11. Electrochemical immobilization of Cs in single-crystalline SYNROC

    NASA Astrophysics Data System (ADS)

    Abe, Hideki; Satoh, Akira; Nishida, Kenji; Abe, Eiji; Naka, Takashi; Imai, Motoharu; Kitazawa, Hideaki

    2006-05-01

    The development of a disposal technique for the radiotoxic 137Cs in nuclear wastes is one of the most urgent issues in nuclear fuel technology. An effective disposal method of 137Cs is to immobilize it in a synthetic rock (SYNROC) material: cesium titanate hollandite, 137Cs xTi 8O 16 ( I4/ m, a=10.2866(3) Å, c=2.9669(1) Å). Practical applications of 137Cs xTi 8O 16 have been restricted so far because the conventional synthetic method requires strong chemical reducers and reaction temperatures higher than 1250 °C. In this report, we present a milder preparation method of Cs xTi 8O 16 by electrolysis of a mixture of Cs 2MoO 4 and TiO 2 in ambient atmosphere at 900 °C. The Cs content in the resultant single-crystalline Cs 1.35Ti 8O 16 is competitive with the highest value in polycrystalline Cs 1.36±0.03Ti 8O 16 prepared by the conventional synthetic method. The electrochemical preparation of Cs 1.35Ti 8O 16 is a promising way to immobilize a high quantity of 137Cs ions in a stable form of single-crystalline SYNROC.

  12. Bacterial CS2 hydrolases from Acidithiobacillus thiooxidans strains are homologous to the archaeal catenane CS2 hydrolase.

    PubMed

    Smeulders, Marjan J; Pol, Arjan; Venselaar, Hanka; Barends, Thomas R M; Hermans, John; Jetten, Mike S M; Op den Camp, Huub J M

    2013-09-01

    Carbon disulfide (CS(2)) and carbonyl sulfide (COS) are important in the global sulfur cycle, and CS(2) is used as a solvent in the viscose industry. These compounds can be converted by sulfur-oxidizing bacteria, such as Acidithiobacillus thiooxidans species, to carbon dioxide (CO(2)) and hydrogen sulfide (H2S), a property used in industrial biofiltration of CS(2)-polluted airstreams. We report on the mechanism of bacterial CS(2) conversion in the extremely acidophilic A. thiooxidans strains S1p and G8. The bacterial CS(2) hydrolases were highly abundant. They were purified and found to be homologous to the only other described (archaeal) CS(2) hydrolase from Acidianus strain A1-3, which forms a catenane of two interlocked rings. The enzymes cluster in a group of β-carbonic anhydrase (β-CA) homologues that may comprise a subclass of CS(2) hydrolases within the β-CA family. Unlike CAs, the CS(2) hydrolases did not hydrate CO(2) but converted CS(2) and COS with H(2)O to H(2)S and CO(2). The CS(2) hydrolases of A. thiooxidans strains G8, 2Bp, Sts 4-3, and BBW1, like the CS(2) hydrolase of Acidianus strain A1-3, exist as both octamers and hexadecamers in solution. The CS(2) hydrolase of A. thiooxidans strain S1p forms only octamers. Structure models of the A. thiooxidans CS(2) hydrolases based on the structure of Acidianus strain A1-3 CS(2) hydrolase suggest that the A. thiooxidans strain G8 CS(2) hydrolase may also form a catenane. In the A. thiooxidans strain S1p enzyme, two insertions (positions 26 and 27 [PD] and positions 56 to 61 [TPAGGG]) and a nine-amino-acid-longer C-terminal tail may prevent catenane formation.

  13. The channels of Mars

    NASA Technical Reports Server (NTRS)

    Baker, Victor R.

    1988-01-01

    The geomorphology of Mars is discussed, focusing on the Martian channels. The great flood channels of Mars, the processes of channel erosion, and dendritic channel networks, are examined. The topography of the Channeled Scabland region of the northwestern U.S. is described and compared to the Martian channels. The importance of water in the evolution of the channel systems is considered.

  14. Crystal structure of CsCrAs2O7, a new member of the diarsenate family

    PubMed Central

    Bouhassine, Mohamad Alem; Boughzala, Habib

    2015-01-01

    Caesium chromium(III) diarsenate(V), CsCrAs2O7, was prepared by solid-state reactions. The title structure consists of isolated CrO6 octa­hedra and As2O7 diarsenate groups, sharing corners to build up a three-dimensional [CrAs2O7]− anionic framework. In this framework, channels extending parallel to [001] are present in which the ten-coordinate Cs+ ions reside. CsCrAs2O7 is isotypic with the monoclinic A I M III X 2O7 (A I = alkali metal; M III = Al, Cr, Fe; X = As, P) type I family of compounds crystallizing in the space group P21/c. PMID:26090138

  15. Plasma Formation During Operation of a Diode Pumped Alkali Laser (DPAL) in Cs

    NASA Astrophysics Data System (ADS)

    Babaeva, Natalia Yu.; Zatsarinny, Oleg; Bartschat, Klaus; Kushner, Mark J.

    2014-10-01

    Diode pumped Alkali Lasers (DPALs) produce laser action on the resonant lines of alkali atoms. Diode lasers resonantly pump the 2P3/2 state of the alkali atom which is collisionally relaxed to the 2P3/2 state which then lases to the ground state 2S1/2. The low optical quality of high power semiconductor diode lasers is converted into high optical quality laser radiation from the alkali vapor. The Cs DPAL system using Ar/Cs/C2H6 mixtures has shown promising results. (C2H6 is the collisional relaxant.) In other studies, resonant excitation of alkali vapor by low power lasers has been used to produce highly ionized channels, initiated through associative ionization and superelastic electron heating. The issue then arises if plasma formation occurs during DPAL by similar mechanisms which would be detrimental to laser performance. In this paper, we report on results from a computational study of a DPAL using Cs vapor. The global model addresses quasi-cw pumping of the Cs(2P3/2) state by laser diodes, and includes a full accounting of the resulting electron kinetics. To enable this study, the B-spline R-matrix (BSR) with pseudostates method was employed to calculate electron impact cross sections for Cs. We found that for pump rates of many to 10 kW/cm2, plasma densities approaching 1013 cm-3 occur during laser oscillation with higher values in the absence of laser oscillation. Supported by DoD High Energy Laser Mult. Res. Initiative and NSF.

  16. Nonlinear channelizer.

    PubMed

    In, Visarath; Longhini, Patrick; Kho, Andy; Neff, Joseph D; Leung, Daniel; Liu, Norman; Meadows, Brian K; Gordon, Frank; Bulsara, Adi R; Palacios, Antonio

    2012-12-01

    The nonlinear channelizer is an integrated circuit made up of large parallel arrays of analog nonlinear oscillators, which, collectively, serve as a broad-spectrum analyzer with the ability to receive complex signals containing multiple frequencies and instantaneously lock-on or respond to a received signal in a few oscillation cycles. The concept is based on the generation of internal oscillations in coupled nonlinear systems that do not normally oscillate in the absence of coupling. In particular, the system consists of unidirectionally coupled bistable nonlinear elements, where the frequency and other dynamical characteristics of the emergent oscillations depend on the system's internal parameters and the received signal. These properties and characteristics are being employed to develop a system capable of locking onto any arbitrary input radio frequency signal. The system is efficient by eliminating the need for high-speed, high-accuracy analog-to-digital converters, and compact by making use of nonlinear coupled systems to act as a channelizer (frequency binning and channeling), a low noise amplifier, and a frequency down-converter in a single step which, in turn, will reduce the size, weight, power, and cost of the entire communication system. This paper covers the theory, numerical simulations, and some engineering details that validate the concept at the frequency band of 1-4 GHz.

  17. Size-induced effect on cathode luminescence spectra of CsI(Na) and CsI(Tl) crystals

    NASA Astrophysics Data System (ADS)

    Liu, Fang; Ouyang, XiaoPing; Liu, Bin; Liu, JinLiang; Chen, Liang; Zhang, ZhongBing; Zhang, XueBin; Feng, Yi

    2014-09-01

    We investigated the cathode luminescence characteristics of CsI(Na) and CsI(Tl) crystals by the spectrum and structure properties at room temperature. We fabricated three different sizes of CsI(Na) and CsI(Tl) crystals and measured their luminescence spectra under cathode rays. We found that CsI(Na) cathode luminescence peaks appear at 420 and 305 nm, and CsI(Tl) cathode luminescence peaks are 540 and 410 nm, the grain size affects CsI(Na) luminescence significantly, and the Na-related 420 nm luminescence intensified relatively when the average grain size reaches ˜20 μm, which becomes weak when the grain size is down to nano-scale. But the cathode luminescence spectra of CsI(Tl) crystals with different size have no obvious changes. Our explanations for these phenomena are that the different impurities in the same host material CsI lead to different luminescence mechanisms. These cathode luminescence characteristics indicate the suitability of CsI(Na) and CsI(Tl) crystals to match photomultiplier tube for large area crystal detector development.

  18. (135)Cs/(137)Cs isotopic ratio as a new tracer of radiocesium released from the Fukushima nuclear accident.

    PubMed

    Zheng, Jian; Tagami, Keiko; Bu, Wenting; Uchida, Shigeo; Watanabe, Yoshito; Kubota, Yoshihisa; Fuma, Shoichi; Ihara, Sadao

    2014-05-20

    Since the Fukushima Daiichi nuclear power plant (FDNPP) accident in 2011, intensive studies of the distribution of released fission products, in particular (134)Cs and (137)Cs, in the environment have been conducted. However, the release sources, that is, the damaged reactors or the spent fuel pools, have not been identified, which resulted in great variation in the estimated amounts of (137)Cs released. Here, we investigated heavily contaminated environmental samples (litter, lichen, and soil) collected from Fukushima forests for the long-lived (135)Cs (half-life of 2 × 10(6) years), which is usually difficult to measure using decay-counting techniques. Using a newly developed triple-quadrupole inductively coupled plasma tandem mass spectrometry method, we analyzed the (135)Cs/(137)Cs isotopic ratio of the FDNPP-released radiocesium in environmental samples. We demonstrated that radiocesium was mainly released from the Unit 2 reactor. Considering the fact that the widely used tracer for the released Fukushima accident-sourced radiocesium in the environment, the (134)Cs/(137)Cs activity ratio, will become unavailable in the near future because of the short half-life of (134)Cs (2.06 years), the (135)Cs/(137)Cs isotopic ratio can be considered as a new tracer for source identification and long-term estimation of the mobility of released radiocesium in the environment.

  19. Antibodies derived from an enterotoxigenic Escherichia coli (ETEC) adhesin tip MEFA (multiepitope fusion antigen) against adherence of nine ETEC adhesins: CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA.

    PubMed

    Nandre, Rahul M; Ruan, Xiaosai; Duan, Qiangde; Sack, David A; Zhang, Weiping

    2016-06-30

    Diarrhea continues to be a leading cause of death in children younger than 5 years in developing countries. Enterotoxigenic Escherichia coli (ETEC) is a leading bacterial cause of children's diarrhea and travelers' diarrhea. ETEC bacteria initiate diarrheal disease by attaching to host receptors at epithelial cells and colonizing in small intestine. Therefore, preventing ETEC attachment has been considered the first line of defense against ETEC diarrhea. However, developing vaccines effectively against ETEC bacterial attachment encounters challenge because ETEC strains produce over 23 immunologically heterogeneous adhesins. In this study, we applied MEFA (multiepitope fusion antigen) approach to integrate epitopes from adhesin tips or adhesive subunits of CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA adhesins and to construct an adhesin tip MEFA peptide. We then examined immunogenicity of this tip MEFA in mouse immunization, and assessed potential application of this tip MEFA for ETEC vaccine development. Data showed that mice intraperitoneally immunized with this adhesin tip MEFA developed IgG antibody responses to all nine ETEC adhesins. Moreover, ETEC and E. coli bacteria expressing these nine adhesins, after incubation with serum of the immunized mice, exhibited significant reduction in attachment to Caco-2 cells. These results indicated that anti-adhesin antibodies induced by this adhesin tip MEFA blocked adherence of the most important ETEC adhesins, suggesting this multivalent tip MEFA may be useful for developing a broadly protective anti-adhesin vaccine against ETEC diarrhea. PMID:27228947

  20. The pore properties of human nociceptor channel TRPA1 evaluated in single channel recordings

    PubMed Central

    Bobkov, Y.V.; Corey, E.A.; Ache, B.W.

    2011-01-01

    TRPA channels detect stimuli of different sensory modalities, including a broad spectrum of chemosensory stimuli, noxious stimuli associated with tissue damage and inflammation, mechanical stimuli, and thermal stimuli. Despite a growing understanding of potential modulators, agonists, and antagonists for these channels, the exact mechanisms of channel regulation and activation remain mostly unknown or controversial and widely debated. Relatively little is also known about the basic biophysical parameters of both native and heterologously expressed TRPA channels. Here we use conventional single channel inside-out and outside-out patch recording from the human TRPA1 channel transiently expressed in human embryonic kidney 293T cells to characterize the selectivity of the channel for inorganic mono-/divalent and organic monovalent cations in the presence of Allylisothiocyanate (AITC). We show the relative permeability of the hTRPA1 channel to inorganic cations to be: Ca2+(5.1)>Ba2+(3.5)>Mg2+(2.8)>NH4+(1.5)>Li+(1.2)>Na+(1.0)≥K+(0.98)≥Rb+(0.98)>Cs+(0.95); and to organic cations: Na+(1.0)≥Dimethylamine(0.99)>Trimethylamine(0.7)>Tetramethylammonium(0.4)>N-methyl-d-glucamine(0.1). Activation of the hTRPA1 channels by AITC appears to recruit the channels to a conformational state with an increased permeability to large organic cations. The pore of the channels in this state can be characterized as dilated by approximately 1–2.5A. These findings provide important insight into the basic fundamental properties and function of TRPA1 channels in general and human TRPA1 channel in particular. PMID:21195050

  1. Results of computer network experiment via the Japanese communication satellite CS - Performance evaluation of communication protocols

    NASA Astrophysics Data System (ADS)

    Ito, A.; Kakinuma, Y.; Uchida, K.; Matsumoto, K.; Takahashi, H.

    1984-03-01

    Computer network experiments have been performed by using the Japanese communication satellite CS. The network is of a centralized (star) type, consisting of one center station and many user stations. The protocols are determined taking into consideration the long round trip delay of a satellite channel. This paper treats the communication protocol aspects of the experiments. Performances of the burst level and the link protocols (which correspond nearly to data link layer of OSI 7 layer model) are evaluated. System performances of throughput, delay, link level overhead are measured by using the statistically generated traffic.

  2. Epithelial Sodium and Chloride Channels and Asthma

    PubMed Central

    Wang, Wen; Ji, Hong-Long

    2015-01-01

    Objective: To focus on the asthmatic pathogenesis and clinical manifestations related to epithelial sodium channel (ENaC)/chlorine ion channel. Data Sources: The data analyzed in this review were the English articles from 1980 to 2015 from journal databases, primarily PubMed and Google Scholar. The terms used in the literature search were: (1) ENaCs; cystic fibrosis (CF) transmembrane conductance regulator (CFTR); asthma/asthmatic, (2) ENaC/sodium salt; CF; asthma/asthmatic, (3) CFTR/chlorine ion channels; asthma/asthmatic, (4) ENaC/sodium channel/scnn1a/scnn1b/scnn1g/scnn1d/amiloride-sensitive/amiloride-inhibtable sodium channels/sodium salt; asthma/asthmatic, lung/pulmonary/respiratory/tracheal/alveolar, and (5) CFTR; CF; asthma/asthmatic (ti). Study Selection: These studies included randomized controlled trials or studies covering asthma pathogenesis and clinical manifestations related to ENaC/chlorine ion channels within the last 25 years (from 1990 to 2015). The data involving chronic obstructive pulmonary disease and CF obtained from individual studies were also reviewed by the authors. Results: Airway surface liquid dehydration can cause airway inflammation and obstruction. ENaC and CFTR are closely related to the airway mucociliary clearance. Ion transporters may play a critical role in pathogenesis of asthmatic exacerbations. Conclusions: Ion channels have been the center of many studies aiming to understand asthmatic pathophysiological mechanisms or to identify therapeutic targets for better control of the disease. PMID:26265620

  3. Quantum efficiencies of imaging detectors with alkali halide photocathodes. I - Microchannel plates with separate and integral CsI photocathodes

    NASA Technical Reports Server (NTRS)

    Carruthers, George R.

    1987-01-01

    Measurements and comparisons have been made of the quantum efficiencies of microchannel plate (MCP) detectors in the far-UV (below 2000-A) wavelength range using CsI photocathodes (a) deposited on the front surfaces of microchannel plates and (b) deposited on solid substrates as opaque photocathodes with the resulting photoelectrons input to microchannel plates. The efficiences were measured in both pulse-counting and photodiode modes of operation. Typical efficiencies are about 15 percent at 1216 A for a CsI-coated MCP compared with 65 percent for an opaque CsI photocathode MCP detector. Special processing has yielded an efficiency as high as 20 percent for a CsI-coated MCP. This may possibly be further improved by optimization of the tilt angle of the MCP channels relative to the front face of the MCP and incident radiation. However, at present there still remains a factor of at least 3 quantum efficiency advantage in the separate opaque CsI photocathode configuration.

  4. A pore segment in DEG/ENaC Na(+) channels.

    PubMed

    Snyder, P M; Olson, D R; Bucher, D B

    1999-10-01

    DEG/ENaC Na(+) channels have diverse functions, including Na(+) absorption, neurotransmission, and sensory transduction. The ability of these channels to discriminate between different ions is critical for their normal function. Several findings suggest that DEG/ENaC channels have a pore structure similar to K(+) channels. To test this hypothesis, we examined the accessibility of native and introduced cysteines in the putative P loop of ENaC. We identified residues that span a barrier that excludes amiloride as well as anionic and large methanethiosulfonate reagents from the pore. This segment contains a structural element ((S/G)CS) involved in selectivity of ENaC. The results are not consistent with predictions from the K(+) channel pore, suggesting that DEG/ENaC Na(+) channels have a novel pore structure. PMID:10497211

  5. Thermionic work function of /Cs/ZnO

    NASA Technical Reports Server (NTRS)

    Sommer, A. H.; Briere, T. R.

    1976-01-01

    The collector electrode of a thermionic converter requires a material having a low thermionic work function and chemical stability in a Cs atmosphere in the 800-K range. This letter reports that ZnO with an adsorbed Cs film meets these requirements. The work function is approximately 1.3 eV. Various methods of preparing the ZnO film are described as well as an experiment in which Cs was replaced by K.

  6. Electrical conductivity of Cs2CuCl4 crystals

    NASA Astrophysics Data System (ADS)

    Sorokin, N. I.

    2016-05-01

    The electrical conductivity of Cs2CuCl4 single crystals, synthesized by crystallization from aqueous solutions in the CsCl-CuCl2-H2O system, has been investigated. The temperature dependence of the electrical conductivity of crystals in a temperature range of 338-584 K exhibits no anomalies. The electrical transfer activation enthalpy is Δ H σ = 0.72 ± 0.05 eV and the conductivity is σ = 3 × 10-4 S/cm at 584 K. The most likely carriers in Cs2CuCl4 are Cs+ cations, which transfer electric charge according to the vacancy mechanism.

  7. Improved TV-CS Approaches for Inverse Scattering Problem

    PubMed Central

    Bevacqua, M. T.; Di Donato, L.

    2015-01-01

    Total Variation and Compressive Sensing (TV-CS) techniques represent a very attractive approach to inverse scattering problems. In fact, if the unknown is piecewise constant and so has a sparse gradient, TV-CS approaches allow us to achieve optimal reconstructions, reducing considerably the number of measurements and enforcing the sparsity on the gradient of the sought unknowns. In this paper, we introduce two different techniques based on TV-CS that exploit in a different manner the concept of gradient in order to improve the solution of the inverse scattering problems obtained by TV-CS approach. Numerical examples are addressed to show the effectiveness of the method. PMID:26495420

  8. Microhydration of caesium compounds: Cs, CsOH, CsI and Cs₂I₂ complexes with one to three H₂O molecules of nuclear safety interest.

    PubMed

    Sudolská, Mária; Cantrel, Laurent; Cernušák, Ivan

    2014-04-01

    Structure and thermodynamic properties (standard enthalpies of formation and Gibbs free energies) of hydrated caesium species of nuclear safety interest, Cs, CsOH, CsI and its dimer Cs₂I₂, with one up to three water molecules, are calculated to assess their possible existence in severe accident occurring to a pressurized water reactor. The calculations were performed using the coupled cluster theory including single, double and non-iterative triple substitutions (CCSD(T)) in conjunction with the basis sets (ANO-RCC) developed for scalar relativistic calculations. The second-order spin-free Douglas-Kroll-Hess Hamiltonian was used to account for the scalar relativistic effects. Thermodynamic properties obtained by these correlated ab initio calculations (entropies and thermal capacities at constant pressure as a function of temperature) are used in nuclear accident simulations using ASTEC/SOPHAEROS software. Interaction energies, standard enthalpies and Gibbs free energies of successive water molecules addition determine the ordering of the complexes. CsOH forms the most hydrated stable complexes followed by CsI, Cs₂I₂, and Cs. CsOH still exists in steam atmosphere even at quite high temperature, up to around 1100 K.

  9. Standardization of 134Cs by three methods.

    PubMed

    García-Toraño, E; Rodríguez, Barquero L; Roteta, M

    2002-01-01

    The nuclide 34Cs decays by beta-emission followed by gamma-deexcitation to 134Ba with a half-life T(1/2) = 2.065 a. It has been standardized by three methods: liquid scintillation counting (LSC), 4pi beta-gamma coincidence counting and 4pi gamma counting. In the LSC measurements, the CIEMAT/NIST method was used to calculate the efficiency. For the coincidence measurements, a conventional 4pi beta (proportional counter)-gamma(NaI) system was used. For the 4pi gamma standardization, a well-type Nal(Tl) detector was modeled with the Monte Carlo package PENELOPE, and the counting efficiency obtained by calculation. Results of the three methods agree within 0.65%.

  10. Thermal ionization of Cs Rydberg states

    NASA Astrophysics Data System (ADS)

    Glukhov, I. L.; Ovsiannikov, V. D.

    2009-01-01

    Rates Pnl of photoionization from Rydberg ns-, np-, nd-states of a valence electron in Cs, induced by black-body radiation, were calculated on the basis of the modified Fues model potential method. The numerical data were approximated with a three-term expression which reproduces in a simple analytical form the dependence of Pnl on the ambient temperature T and on the principal quantum number n. The comparison between approximate and exactly calculated values of the thermal ionization rate demonstrates the applicability of the proposed approximation for highly excited states with n from 20 to 100 in a wide temperature range of T from 100 to 10,000 K. We present coefficients of this approximation for the s-, p- and d-series of Rydberg states.

  11. CS-Studio Scan System Parallelization

    SciTech Connect

    Kasemir, Kay; Pearson, Matthew R

    2015-01-01

    For several years, the Control System Studio (CS-Studio) Scan System has successfully automated the operation of beam lines at the Oak Ridge National Laboratory (ORNL) High Flux Isotope Reactor (HFIR) and Spallation Neutron Source (SNS). As it is applied to additional beam lines, we need to support simultaneous adjustments of temperatures or motor positions. While this can be implemented via virtual motors or similar logic inside the Experimental Physics and Industrial Control System (EPICS) Input/Output Controllers (IOCs), doing so requires a priori knowledge of experimenters requirements. By adding support for the parallel control of multiple process variables (PVs) to the Scan System, we can better support ad hoc automation of experiments that benefit from such simultaneous PV adjustments.

  12. The Mechanism Responsible for Extraordinary Cs-ion Selectivity in Crystalline Silicotitanate

    SciTech Connect

    Celestian,A.; Kubicki, J.; Hanson, J.; Clearfield, A.; Parise, J.

    2008-01-01

    Combining information from time-resolved X-ray and neutron scattering with theoretical calculations has revealed the elegant mechanism whereby hydrogen crystalline silicotitanate (H-CST; H2Ti2SiO7{center_dot}1.5H2O) achieves its remarkable ion-exchange selectivity for cesium. Rather than a simple ion-for-ion displacement reaction into favorable sites, which has been suggested by static structural studies of ion-exchanged variants of CST, Cs+ exchange proceeds via a two-step process mediated by conformational changes in the framework. Similar to the case of ion channels in proteins, occupancy of the most favorable site does not occur until the first lever, cooperative repulsive interactions between water and the initial Cs-exchange site, repels a hydrogen lever on the silicotitanate framework. Here we show that these interactions induce a subtle conformational rearrangement in CST that unlocks the preferred Cs site and increases the overall capacity and selectivity for ion exchange.

  13. Coulomb explosion of CS2 molecule under an intense femtosecond laser field

    NASA Astrophysics Data System (ADS)

    Xiao, Wang; Jian, Zhang; Shi-An, Zhang; Zhen-Rong, Sun

    2016-05-01

    We experimentally demonstrate the Coulomb explosion process of CS2 molecule under a near-infrared (800 nm) intense femtosecond laser field by a DC-sliced ion imaging technique. We obtain the DC-sliced images of these fragment ions S+, S2+, CS+, and CS2+ by breaking one C–S bond, and assign their Coulomb explosion channels by considering their kinetic energy release and angular distribution. We also numerically simulate the dissociation dynamics of parent ions by a Coulomb potential approximation, and obtain the time evolution of Coulomb energy and kinetic energy release, which indicates that the dissociation time of parent ions decreases with the increase of the charge number k. These experimental and theoretical results can serve as a useful benchmark for those researchers who work in the related area. Project supported by the National Natural Science Foundation of China (Grant Nos. 51132004 and 11474096), and the Science and Technology Commission of Shanghai Municipality, China (Grant No. 14JC1401500). We acknowledge the support of the NYU-ECNU Institute of Physics at NYU Shanghai, China.

  14. Multiple band structures of {sup 131}Cs

    SciTech Connect

    Sihotra, S.; Palit, R.; Naik, Z.; Joshi, P. K.; Deo, A. Y.; Jain, H. C.; Singh, K.; Goswamy, J.; Mehta, D.; Singh, N.; Malik, S. S.; Praharaj, C. R.

    2008-09-15

    Excited states in {sup 131}Cs were investigated through in-beam {gamma}-ray spectroscopic techniques following its population in the {sup 124}Sn({sup 11}B, 4n) fusion-evaporation reaction at a beam energy of 46 MeV. The previously known level scheme has been substantially extended up to {approx}9 MeV excitation energy and 49/2({Dirac_h}/2{pi}) spin with the addition of seven new band structures. The present level scheme consisting of 15 bands exhibits a variety of collective features in this nucleus at intermediate spin. The excitation energies of the observed levels in different bands and the corresponding ratios of transition strengths, i.e., B(M1)/B(E2), have been compared with the results of projected deformed Hartree-Fock calculations based on various quasiparticle configurations. A strongly coupled band has been reassigned a high-K three-quasiparticle {pi}h{sub 11/2} x {nu}(h{sub 11/2}d{sub 3/2}) configuration based on the properties of this band and that of its new coupled side band. The configurations of these bands are also discussed in the framework of tilted-axis cranking model calculations and the systematics of the odd-A Cs isotopes. Additional three energetically closely placed coupled bands have been assigned different unpaired three-quasiparticle configurations. {gamma}-vibrational bands coupled to the {pi}h{sub 11/2} and {pi}g{sub 7/2} single-particle configurations have been reported in this nucleus. Observation of new E1 transitions linking the opposite-parity {pi}h{sub 11/2} and {pi}d{sub 5/2} bands provides fingerprints of possible octupole correlations.

  15. Further studies on the distribution of 137Cs in British coastal waters—I. Irish Sea

    NASA Astrophysics Data System (ADS)

    Jefferies, D. F.; Steele, A. K.; Preston, A.

    1982-06-01

    The concentrations of 137Cs, discharged to the northeast Irish Sea from the British Nuclear Fuels Limited (BNFL) Windscale Works, Cumbria, have been measured in the Irish Sea and its approaches during 1970 to 1978. Seawater labelled by 137Cs has been used to study the variability of local and distant dispersion and to estimate residence times and volume transports. In the immediate vicinity of the outfall a release rate of 1 Ci d -1 produces a mean annual concentration of 4.2 pCi l -1 (2.6 to 6.3 pCi l -1) with an approximate 30-fold reduction to 0.15 pCi l -1 in the northern exit to the Irish Sea and at least a 300-fold reduction at the southern entrance. The release rate ranged from 60 to 400 Ci d -1 (annual average) during the study period. The main flow patterns are deduced and show a substantial change before and after 1976. Using estimated inventories of the radionuclide in various areas, the rate of input into the system, and a simple exponential model, the residence 'half time' of water east of a line south from Scotland through the Isle of Man to Anglesey is estimated to be 200 days. The estimate for the whole northern Irish Sea was 1 year between 1970 and 1976, with a sharp reduction thereafter. The change is confirmed by estimating the volume transport through the North Channel from data from the Stranraer-Larne, Fishguard-Rosslare, and Swansea-Cork ferries. The mean flow through the Irish Sea for the period October 1971 to May 1978 was 5 km 3 d -1 with mean flows of 3.4 km 3 d -1 prior to January 1976 and 7.9 km 3 d -1 thereafter to May 1978. Transit times from the discharge point to the North Channel have also been estimated from the change in concentration of the radionuclides 137Cs( t1/2 = 30.1 y) and 134Cs( t1/2 = 2.1 y) and from serial correlation of rates of discharge and concentrations of 137Cs at distance.

  16. Dual channel self-oscillating optical magnetometer

    SciTech Connect

    Belfi, J.; Bevilacqua, G.; Biancalana, V.; Dancheva, Y.; Khanbekyan, K.; Moi, L.; Cartaleva, S.

    2009-05-15

    We report on a two-channel magnetometer based on nonlinear magneto-optical rotation in a Cs glass cell with buffer gas. The Cs atoms are optically pumped and probed by free running diode lasers tuned to the D{sub 2} line. A wide frequency modulation of the pump laser is used to produce both synchronous Zeeman optical pumping and hyperfine repumping. The magnetometer works in an unshielded environment, and a spurious signal from distant magnetic sources is rejected by means of differential measurement. In this regime the magnetometer simultaneously gives the magnetic field modulus and the field difference. Rejection of the common-mode noise allows for high-resolution magnetometry with a sensitivity of 2 pT/{radical}(Hz). This sensitivity, in conjunction with long-term stability and a large bandwidth, makes it possible to detect water proton magnetization and its free induction decay in a measurement volume of 5 cm{sup 3}.

  17. Memory is not extinguished along with CS presentation but within a few seconds after CS-offset.

    PubMed

    Pérez-Cuesta, Luis María; Hepp, Yanil; Pedreira, María Eugenia; Maldonado, Héctor

    2007-01-01

    Prior work with the crab's contextual memory model showed that CS-US conditioned animals undergoing an unreinforced CS presentation would either reconsolidate or extinguish the CS-US memory, depending on the length of the reexposure to the CS. Either memory process is only triggered once the CS is terminated. Based on these results, the following questions are raised. First, when is extinction memory acquired, if not along extinction training, and how long does it take? Second, can acquisition and consolidation of extinction memory be pharmacologically dissected? Here we address these questions performing three series of experiments: a first one aimed to study systematically the relationship between extinction and increasing periods of unreinforced CS presentations, a second one to determine the time boundaries of the extinction memory acquisition, and the third one to assay the requirement for protein synthesis and NMDA-like receptors of acquisition and consolidation of extinction memory. Our results confirm that it is CS-offset and not the mere retrieval (CS-onset) that triggers acquisition of extinction memory and that it is completed in less than 45 sec after CS-offset. In addition, protein synthesis is required for consolidation but not for acquisition of this memory and, conversely, NMDA-like receptor activity is required for its acquisition but not for its consolidation. Finally, we offer an interpretative scheme of our results and we discuss to what extent it could apply to multitrial extinction.

  18. Transfer of Chernobyl 134Cs and 137Cs in cows from silage to milk.

    PubMed

    Voors, P I; Van Weers, A W

    1989-09-01

    The transfer of Chernobyl Cs-134 and Cs-137 to milk in dairy cows from a diet, predominated by silage, was studied under normal farming conditions. The study was carried out in the period December 1986 till August 1987 and comprised three periods with different levels of radioactive contamination of the silage. Transfer coefficients for both radionuclides were derived from measured radiocaesium concentrations in the diet components and in mixed milk samples. Average silage intake was calculated on basis of total need which depends on milk production. The transfer coefficients calculated for Cs-134 and Cs-137 after a 6 weeks feeding period of high-level post-Chernobyl silage were 0.26 and 0.27% d/l, respectively. These results indicate that a transfer coefficient of 1.2% d/l, from tracer and fallout studies and recommended for predictive purposes, is a conservative estimate in view of the post-Chernobyl experience. The transfer of caesium isotopes from silage to milk in dairy cows is dynamically described with a simple two-compartment model of Pelletier and Voilleque. The most important choices of parameter values to be made in matching calculated and measured concentrations in milk, are those for F and u/m. F is the fraction of ingested activity that is excreted in milk plus urine, and u/m is the ratio between the transfer rates to urine and milk, respectively. The study was carried out with financial support from the Dutch Ministry of Agriculture and Fishery.

  19. Small Column Ion Exchange Testing of Superlig 644 for Removal of 137Cs from Hanford Tank Waste Envelope C (Tank 241-AN-107)

    SciTech Connect

    DE Kurath; DL Blanchard; JR Bontha

    2000-06-28

    The current BNFL Inc. flowsheet for the pretreatment of the Hanford high-level tank wastes includes the use of Superlig{reg_sign} materials for removing {sup 137}Cs from the aqueous fraction of the waste. The Superlig materials applicable to cesium removal include the cesium-selective Superlig 632and Superlig 644. These materials have been developed and supplied by IBC Advanced Technologies, Inc., American Fork, Utah. This report describes the testing of the Superlig 644 ion exchange material in a small dual-column system. The bed volume of the lead column was 18.6 mL (L/D = 7), and the bed volume of the lag column was 15.9 mL (L/D = 6) during the loading phase. The sample processed was approximately 1.6 L of diluted waste ([Na{sup +}] = 4.84 M) from Tank 241-AN-107 (Envelope C). This sample had been previously treated for removal of Sr/transuranic (TRU) values and clarified in a single tube cross-flow filtration unit. All ion exchange process steps were tested, including resin-bed preparation, loading, feed displacement, water rinse, elution, eluant rinse, and resin regeneration. A summary of performance measures for both columns is shown in Table S1. The Cs {lambda} values represent a measure of the effective capacity of the SL-644 resin. The Cs {lambda} of 20 for the lead column is much lower than the estimated 150 obtained by the Savannah River Technology Center during Phase 1A testing. Equilibrium data obtained with batch contacts using the AN-107 Cs IX feed predicts a Cs {lambda} of 183. A Cs {lambda} for the lag column could not be determined due to insufficient breakthrough, but it appeared to work well and removed nearly all of the cesium not removed by the lead column. The low value for the lead column indicates that it did not perform as expected. This may have been due to air or gas in the bed that caused fluid channeling or blinding of the resin. The maximum decontamination factor (DF) for {sup 137}Cs listed in Table S1 is based on {sup 137}Cs

  20. Spectroscopic [correction of eSpectroscopic] and structural properties of valine gramicidin A in monolayers at the air-water interface.

    PubMed Central

    Lavoie, Hugo; Blaudez, Daniel; Vaknin, David; Desbat, Bernard; Ocko, Benjamin M; Salesse, Christian

    2002-01-01

    Monomolecular films of valine gramicidin A (VGA) were investigated in situ at the air-water interface by x-ray reflectivity and x-ray grazing incidence diffraction as well as polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS). These techniques were combined to obtain information on the secondary structure and the orientation of VGA and to characterize the shoulder observed in its pi-A isotherm. The thickness of the film was obtained by x-ray reflectivity, and the secondary structure of VGA was monitored using the frequency position of the amide I band. The PM-IRRAS spectra were compared with the simulated ones to identify the conformation adopted by VGA in monolayer. At large molecular area, VGA shows a disordered secondary structure, whereas at smaller molecular areas, VGA adopts an anti-parallel double-strand intertwined beta(5.6) helical conformation with 30 degrees orientation with respect to the normal with a thickness of 25 A. The interface between bulk water and the VGA monolayer was investigated by x-ray reflectivity as well as by comparing the experimental and the simulated PM-IRRAS spectra on D(2)O and H(2)O, which suggested the presence of oriented water molecules between the bulk and the monolayer. PMID:12496123

  1. Narrating Data Structures: The Role of Context in CS2

    ERIC Educational Resources Information Center

    Yarosh, Svetlana; Guzdial, Mark

    2008-01-01

    Learning computing with respect to the context of its use has been linked in previous reports to student motivation in introductory Computer Science (CS) courses. In this report, we consider the role of context in a second course. We present a case study of a CS2 data structures class that uses a media computation context. In this course, students…

  2. Occasion Setting Is Specific to the CS-US Association

    ERIC Educational Resources Information Center

    Bonardi, Charlotte

    2007-01-01

    In Experiment 1, rats were trained on a discrimination in which one occasion setter, A, signaled that one cue (conditioned stimulus, CS), x, would be followed by one outcome, p (unconditioned stimulus, US), and a second CS, y, by a different outcome, q (x [right arrow] p and y [right arrow] q); a second occasion setter, B signalled the reverse…

  3. Methacrylate Polymer Scaffolding Enhances the Stability of Suspended Lipid Bilayers for Ion Channel Recordings and Biosensor Development

    PubMed Central

    Bright, Leonard K.; Baker, Christopher A.; Bränström, Robert; Saavedra, S. Scott; Aspinwall, Craig A.

    2016-01-01

    Black lipid membranes (BLMs) provide a synthetic environment that facilitates measurement of ion channel activity in diverse analytical platforms. The limited electrical, mechanical and temporal stabilities of BLMs pose a significant challenge to development of highly stable measurement platforms. Here, ethylene glycol dimethacrylate (EGDMA) and butyl methacrylate (BMA) were partitioned into BLMs and photopolymerized to create a cross-linked polymer scaffold in the bilayer lamella that dramatically improved BLM stability. The commercially available methacrylate monomers provide a simple, low cost, and broadly accessible approach for preparing highly stabilized BLMs useful for ion channel analytical platforms. When prepared on silane-modified glass microapertures, the resulting polymer scaffold-stabilized (PSS)-BLMs exhibited significantly improved lifetimes of 23 ± 9 to 40 ± 14 h and > 10-fold increase in mechanical stability, with breakdown potentials > 2000 mV attainable, depending on surface modification and polymer cross-link density. Additionally, the polymer scaffold exerted minimal perturbations to membrane electrical integrity as indicated by mean conductance measurements. When gramicidin A and α-hemolysin were reconstituted into PSS-BLMs, the ion channels retained function comparable to conventional BLMs. This approach is a key advance in the formation of stabilized BLMs and should be amenable to a wide range of receptor and ion channel functionalized platforms. PMID:26925461

  4. Determination of 135Cs by accelerator mass spectrometry

    NASA Astrophysics Data System (ADS)

    MacDonald, C. M.; Charles, C. R. J.; Zhao, X.-L.; Kieser, W. E.; Cornett, R. J.; Litherland, A. E.

    2015-10-01

    The ratio of anthropogenic 135Cs and 137Cs isotopes is characteristic of a uranium fission source. This research evaluates the technique of isotope dilution (yield tracing) for the purpose of quantifying 135Cs by accelerator mass spectrometry with on-line isobar separation. Interferences from Ba, Zn2, and isotopes of equal mass to charge ratios were successfully suppressed. However, some sample crosstalk from source contamination remains. The transmission and di-fluoride ionization efficiencies of Cs isotopes were found to be 8 × 10-3 and 1.7 × 10-7 respectively. This quantification of 135Cs using yield tracing by accelerator mass spectrometry shows promise for future environmental sample analysis once the issues of sample crosstalk and low efficiency can be resolved.

  5. CS (5-4) survey towards nearby infrared bright galaxies

    NASA Astrophysics Data System (ADS)

    Wang, Junzhi; Zhang, Zhiyu; Shi, Yong

    2011-09-01

    With the observations of the CS (5-4) line towards a sample of 24 infrared bright galaxies using Heinrich Hertz Submillimeter Telescope (HHSMT), we detected CS (5-4) emission in 14 galaxies, including 12 ultraluminous infrared galaxies (ULIRGs)/luminous infrared galaxies (LIRGs) and two nearby normal galaxies. As a good dense gas tracer, which has been well used for studying star formation in the Milky Way, CS (5-4) can trace the active star-forming gas in galaxies. The correlation between CS (5-4) luminosity, which is estimated with detected CS (5-4) line emission, and the infrared luminosity in these 14 galaxies, is fitted with a correlation coefficient of 0.939 and a slope close to unity. This correlation confirms that dense gas, which is closely linked to star formation, is very important for understanding star formation in galaxies.

  6. Transformation of Sintered CsPbBr3 Nanocrystals to Cubic CsPbI3 and Gradient CsPbBrxI3-x through Halide Exchange.

    PubMed

    Hoffman, Jacob B; Schleper, A Lennart; Kamat, Prashant V

    2016-07-13

    All-inorganic cesium lead halide (CsPbX3, X = Br(-), I(-)) perovskites could potentially provide comparable photovoltaic performance with enhanced stability compared to organic-inorganic lead halide species. However, small-bandgap cubic CsPbI3 has been difficult to study due to challenges forming CsPbI3 in the cubic phase. Here, a low-temperature procedure to form cubic CsPbI3 has been developed through a halide exchange reaction using films of sintered CsPbBr3 nanocrystals. The reaction was found to be strongly dependent upon temperature, featuring an Arrhenius relationship. Additionally, film thickness played a significant role in determining internal film structure at intermediate reaction times. Thin films (50 nm) showed only a small distribution of CsPbBrxI3-x species, while thicker films (350 nm) exhibited much broader distributions. Furthermore, internal film structure was ordered, featuring a compositional gradient within film. Transient absorption spectroscopy showed the influence of halide exchange on the excited state of the material. In thicker films, charge carriers were rapidly transferred to iodide-rich regions near the film surface within the first several picoseconds after excitation. This ultrafast vectorial charge-transfer process illustrates the potential of utilizing compositional gradients to direct charge flow in perovskite-based photovoltaics. PMID:27322132

  7. 137Cs Results and Interpretation of Cesium Soil Data on the Upper Fortymile Wash Alluvial Fan, Amargosa Valley, Nevada.

    NASA Astrophysics Data System (ADS)

    Harrington, C.

    2004-12-01

    Studies using 137Cs were used to produce soil Cs profiles and to use them to determine erosion rates on interchannel divides of the Fortymile Wash alluvial fan over the last 50 years. Sample locations whose 137Cs profiles most resemble the reference-sample (stable surface) profiles are located on interchannel divide areas between distributary channels. These profiles are similar to the reference profiles that have low 137Cs values (in the range of 0.02 to 0.08 pCi/g) in the 3 to 6 cm layers. However, the surface layers (1-3 cm depth) typically have values much less than the reference samples from equivalent depths (range from 0.251 to 0.421 pCi/g). The data indicate that many of these interchannel divide areas have had part of the upper layer removed. Interchannel divide areas have the least likelihood of having been submerged during floods over the last fifty years. Thus, the loss of material from these otherwise stable surfaces appears to be due to eolian processes. Erosion of an interchannel divide area with little evidence of recent water movement is most easily explained by eolian removal. Evidence for wind erosion as the predominant process on the interchannel divide areas includes the lack of new or developing stream channels and the presence of modern coppice dunes near channels on interchannel divides. The presence of nearby Big Dune and other eolian deposits provides strong support for eolian erosion and transport. The amount of material removed from the interchannel divide areas was estimated by comparing the 137Cs value of the upper 3 cm layer to that of the reference value and calculating the thickness of the layer that would have to be removed to obtain the lower value. Applying this method across the interchannel divide sample locations indicates 1 to 2 cm of material has been removed from the interchannel divide surfaces in the last 50 years. This results in erosion rates that range from 0.02 to 0.04 cm/yr. These rates are similar to erosion rates

  8. Epiphytic fruticose lichens as biomonitors for retrospective evaluation of the (134)Cs/(137)Cs ratio in Fukushima fallout.

    PubMed

    Ramzaev, V; Barkovsky, A; Gromov, A; Ivanov, S; Kaduka, M

    2014-12-01

    In 2011-2013, sampling of epiphytic fruticose lichens of the genera Usnea, Bryoria and Alectoria was carried out on Sakhalin and Kuril Islands (the Sakhalin region, Russia) to investigate contamination of these organisms with the Fukushima-derived (134)Cs and (137)Cs. Activities of the radionuclides were determined in all 56 samples of lichens taken for the analysis. After correction for radioactive decay (on 15 March 2011), the activity concentrations ranged from 2.1 Bq kg(-1) (d.w.) to 52 Bq kg(-1) for (134)Cs and from 2.3 Bq kg(-1) to 52 Bq kg(-1) for (137)Cs. Cesium-134 and (137)Cs activities for the whole set of lichens (n = 56) were strongly positively correlated; Spearman's rank correlation coefficient was calculated as 0.991 (P < 0.01). The activity concentrations of (134)Cs and (137)Cs in Usnea lichens from the Sakhalin and Kunashir islands declined with a factor of three in the period from 2011 to 2013. The average biological half-time for both cesium radionuclides in lichens of the genus Usnea is estimated as 1.3 y. The mean of 0.99 ± 0.10 and median of 0.99 were calculated for the decay corrected (134)Cs/(137)Cs activities ratios in the lichens (n = 56). The radionuclides ratio in the lichens did not depend on location of sampling site, species and the time that had passed after the Fukushima accident. The regression analysis has shown the background pre-Fukushima level of (137)Cs of 0.4 ± 0.3 Bq kg(-1), whereas the ratio between the Fukushima-borne (134)Cs and (137)Cs in the lichens was estimated as 1.04. The (134)Cs/(137)Cs activities ratio in lichens from the Sakhalin region is consistent with the ratios reported by others for the heavy contaminated areas on Honshu Island in Japan following the Fukushima accident. The activity concentrations of natural (7)Be in lichens from the Sakhalin region varied between 100 Bq kg(-1) and 600 Bq kg(-1); the activity concentrations did not exhibit temporal variations during a 2y

  9. Volunteer challenge with enterotoxigenic Escherichia coli that express intestinal colonization factor fimbriae CS17 and CS19.

    PubMed

    McKenzie, Robin; Porter, Chad K; Cantrell, Joyce A; Denearing, Barbara; O'Dowd, Aisling; Grahek, Shannon L; Sincock, Stephanie A; Woods, Colleen; Sebeny, Peter; Sack, David A; Tribble, David R; Bourgeois, A Louis; Savarino, Stephen J

    2011-07-01

    Human challenges with enterotoxigenic Escherichia coli (ETEC) have broadened our understanding of this important enteropathogen. We report findings from the first challenge studies using ETEC-expressing colonization factor fimbria CS17 and CS19. LSN03-016011/A (LT, CS17) elicited a dose-dependent effect, with the upper dose (6 × 10(9) organisms) causing diarrhea in 88% of recipients. WS0115A (LTSTp, CS19) also showed a dose response, with a 44% diarrhea rate at 9 × 10(9) organisms. Both strains elicited homologous antifimbrial and anti-LT antibody seroconversion. These studies establish the relative pathogenicity of ETEC expressing newer class 5 fimbriae and suggest suitability of the LT|CS17-ETEC challenge model for interventional trials. PMID:21628659

  10. 137Cs in the western South Pacific Ocean.

    PubMed

    Yamada, Masatoshi; Wang, Zhong-Liang

    2007-09-01

    The 137Cs activities were determined for seawater samples from the East Caroline, Coral Sea, New Hebrides, South Fiji and Tasman Sea (two stations) Basins of the western South Pacific Ocean by gamma spectrometry using a low background Ge detector. The 137Cs activities ranged from 1.4 to 2.3 Bq m(-3) over the depth interval 0-250 m and decreased exponentially from the subsurface to 1000 m depth. The distribution profiles of 137Cs activity at these six western South Pacific Ocean stations did not differ from each other significantly. There was a remarkable difference for the vertical profiles of 137Cs activity between the East Caroline Basin station in this study and the GEOSECS (Geochemical Ocean Sections Study) station at the same latitude in the Equatorial Pacific Ocean; the 137Cs inventory over the depth interval 100-1000 m increased from 400+/-30 Bq m(-2) to 560+/-30 Bq m(-2) during the period from 1973 to 1992. The total 137Cs inventories in the western South Pacific Ocean ranged from 850+/-70 Bq m(-2) in the Coral Sea Basin to 1270+/-90 Bq m(-2) in the South Fiji Basin. Higher 137Cs inventories were observed at middle latitude stations in the subtropical gyre than at low latitude stations. The 137Cs inventories were 1.9-4.5 times (2.9+/-0.7 on average) and 1.7-4.3 times (3.1+/-0.7 on average) higher than that of the expected deposition density of atmospheric global fallout at the same latitude and that of the estimated 137Cs deposition density in 10 degrees latitude by 10 degrees longitude grid data obtained by Aoyama et al. [Aoyama M, Hirose K, Igarashi Y. Re-construction and updating our understanding on the global weapons tests 137Cs fallout. J Environ Monit 2006;8:431-438], respectively. The possible processes for higher 137Cs inventories in the western South Pacific Ocean than that of the expected deposition density of atmospheric global fallout may be attributable to the inter-hemisphere dispersion of the atmospheric nuclear weapons testing 137Cs from

  11. 137Cs in the western South Pacific Ocean.

    PubMed

    Yamada, Masatoshi; Wang, Zhong-Liang

    2007-09-01

    The 137Cs activities were determined for seawater samples from the East Caroline, Coral Sea, New Hebrides, South Fiji and Tasman Sea (two stations) Basins of the western South Pacific Ocean by gamma spectrometry using a low background Ge detector. The 137Cs activities ranged from 1.4 to 2.3 Bq m(-3) over the depth interval 0-250 m and decreased exponentially from the subsurface to 1000 m depth. The distribution profiles of 137Cs activity at these six western South Pacific Ocean stations did not differ from each other significantly. There was a remarkable difference for the vertical profiles of 137Cs activity between the East Caroline Basin station in this study and the GEOSECS (Geochemical Ocean Sections Study) station at the same latitude in the Equatorial Pacific Ocean; the 137Cs inventory over the depth interval 100-1000 m increased from 400+/-30 Bq m(-2) to 560+/-30 Bq m(-2) during the period from 1973 to 1992. The total 137Cs inventories in the western South Pacific Ocean ranged from 850+/-70 Bq m(-2) in the Coral Sea Basin to 1270+/-90 Bq m(-2) in the South Fiji Basin. Higher 137Cs inventories were observed at middle latitude stations in the subtropical gyre than at low latitude stations. The 137Cs inventories were 1.9-4.5 times (2.9+/-0.7 on average) and 1.7-4.3 times (3.1+/-0.7 on average) higher than that of the expected deposition density of atmospheric global fallout at the same latitude and that of the estimated 137Cs deposition density in 10 degrees latitude by 10 degrees longitude grid data obtained by Aoyama et al. [Aoyama M, Hirose K, Igarashi Y. Re-construction and updating our understanding on the global weapons tests 137Cs fallout. J Environ Monit 2006;8:431-438], respectively. The possible processes for higher 137Cs inventories in the western South Pacific Ocean than that of the expected deposition density of atmospheric global fallout may be attributable to the inter-hemisphere dispersion of the atmospheric nuclear weapons testing 137Cs from

  12. Accumulation and transfer of 137Cs and 90Sr in the plants of the forest ecosystem near the Ignalina Nuclear Power Plant.

    PubMed

    Lukšienė, B; Marčiulionienė, D; Gudelienė, I; Schönhofer, F

    2013-02-01

    The radioecological state of the forest ecosystem in the vicinity of the Ignalina Power Plant prior to decommissioning was analysed with specific emphasis on (137)Cs and (90)Sr activity concentrations in plant species growing in two reference sampling sites (Tilze and Grikiniskes). In the period of 1996-2008 the mean contamination of plants with (137)Cs was from 45 to 119 Bq/kg and with (90)Sr - from 3 to 42 Bq/kg. Measured (137)Cs TF values for soil-root transfer mainly ranged between 1.0-1.4, except for Calamagrostis arundinacea which had a TF value of 0.1. On average, the (137)Cs TF value from root to shoot was 1.7 fold higher than for soil to root transfer. (90)Sr TF values (soil-root) were in the range of 1.2-1.8 but for Calluna vulgaris it was 0.2. The mean root to shoot TF value for (90)Sr was 7.7 fold higher. These results indicate the higher (90)Sr bioavailability than that of (137)Cs in the forested area. The Grikiniskes reference site is located nearby the Ignalina NPP, specifically the heated water outlet channel, which results in altered microclimatic conditions. These specific microclimatic conditions result in relationships between (137)Cs TF (soil-root) values and pH, moisture and organic matter content in the soil at Grikiniskes which appear to be different to those at the Tilze reference sampling site. PMID:23085187

  13. Accumulation and transfer of 137Cs and 90Sr in the plants of the forest ecosystem near the Ignalina Nuclear Power Plant.

    PubMed

    Lukšienė, B; Marčiulionienė, D; Gudelienė, I; Schönhofer, F

    2013-02-01

    The radioecological state of the forest ecosystem in the vicinity of the Ignalina Power Plant prior to decommissioning was analysed with specific emphasis on (137)Cs and (90)Sr activity concentrations in plant species growing in two reference sampling sites (Tilze and Grikiniskes). In the period of 1996-2008 the mean contamination of plants with (137)Cs was from 45 to 119 Bq/kg and with (90)Sr - from 3 to 42 Bq/kg. Measured (137)Cs TF values for soil-root transfer mainly ranged between 1.0-1.4, except for Calamagrostis arundinacea which had a TF value of 0.1. On average, the (137)Cs TF value from root to shoot was 1.7 fold higher than for soil to root transfer. (90)Sr TF values (soil-root) were in the range of 1.2-1.8 but for Calluna vulgaris it was 0.2. The mean root to shoot TF value for (90)Sr was 7.7 fold higher. These results indicate the higher (90)Sr bioavailability than that of (137)Cs in the forested area. The Grikiniskes reference site is located nearby the Ignalina NPP, specifically the heated water outlet channel, which results in altered microclimatic conditions. These specific microclimatic conditions result in relationships between (137)Cs TF (soil-root) values and pH, moisture and organic matter content in the soil at Grikiniskes which appear to be different to those at the Tilze reference sampling site.

  14. Waste forms based on Cs-loaded silicotitanates

    SciTech Connect

    Balmer, M.L.; Bunker, B.C.

    1995-04-01

    Silicotitanate ion exchange materials are being considered for removal of radioactive Cs and Sr from tank wastes at the Hanford site. The phase evolution as a function of heat treatment temperature for several sol gel derived compositions within the Cs{sub 2}O-SiO{sub 2}-TiO{sub 2} system was investigated, in order to determine if an adequate waste form can be achieved by direct thermal conversion. The Cs leach rates and Cs loss during heat treatment of select materials were measured. Some compositions which contain large amounts of Ti melt to form a glass with reasonably low aqueous leach rates. A new Cs-silicotitanate material with a structure isomorphous to pollucite was discovered. This material forms at low temperatures (700--800 C) where Cs volatility is negligible. The silicotitanate-pollucite exhibits extremely low leach rates (1.42 g/m{sup 2}day ) at 90 C, and has been identified as a promising waste form for Cs containment.

  15. Radioactive Cs capture in the early solar system

    PubMed Central

    Hidaka, Hiroshi; Yoneda, Shigekazu

    2013-01-01

    Barium isotopic compositions of primitive materials in the solar system are generally affected by s- and r-process nucleosynthetic components that hide the contribution of the isotopic excess of 135Ba formed by decay of radioactive 135Cs. However, the Ba isotopic composition of the chemical separates from chondrules in the Sayama CM2 chondrite shows an excess of 135Ba isotopic abundance up to (0.33 ± 0.06)%, which is independent of the isotopic components from s- and r-process nucleosyntheses. The isotopic excesses of 135Ba correlate with the elemental abundance of Ba relative to Cs, providing chemical and isotopic evidence for the existence of the presently extinct radionuclide 135Cs (t1/2 = 2.3 million years) in the early solar system. The estimated abundance of 135Cs/133Cs = (6.8 ± 1.9) × 10−4 is more than double that expected from the uniform production model of the short-lived radioisotopes, suggesting remobilization of Cs including 135Cs in the chondrules of the meteorite parent body. PMID:23435551

  16. Antimicrobial activity of CS-940, a new trifluorinated quinolone.

    PubMed Central

    Biedenbach, D J; Sutton, L D; Jones, R N

    1995-01-01

    The antimicrobial activity of CS-940, a new trifluorinated quinolone drug, was tested against 761 clinical isolates. CS-940 activity against members of the family Enterobacteriaceae was most similar to that of ciprofloxacin and ofloxacin, with a large range of MICs inhibiting 90% of isolates tested (MIC90S) of 0.015 to 16 micrograms/ml (median MIC90, 0.06 micrograms/ml). CS-940 had greater activity than ciprofloxacin or ofloxacin when they were tested against Acinetobacter spp. (MIC90S, 0.03 micrograms/ml) and Stenotrophomonas (Xanthomonas) maltophilia (MIC90S, 2 micrograms/ml). CS-940 demonstrated a high degree of potency against Haemophilus influenzae, Moraxella catarrhalis, and Neisseria spp. (MIC90S, < or = 0.06 micrograms/ml). CS-940 was two- to eightfold more active than ciprofloxacin or ofloxacin against oxacillin-susceptible Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and coagulase-negative Staphylococcus spp. CS-940 was also very active against Streptococcus spp. and enterococci, for which MIC90S were < or = 2 micrograms/ml; for Enterococcus faecium, however, the MIC90 was 4 micrograms/ml. CS-940 was generally less active than a comparison investigational fluoroquinolone, clinafloxacin. This compound appears promising by in vitro test analysis and warrants further in vivo trials. PMID:8619590

  17. The Density Structure of UCHII Regions: CS Observations

    NASA Astrophysics Data System (ADS)

    Butner, H. M.; Lauwers, T. L.

    2000-12-01

    Ultra-compact HII (UCHII) regions are sites of massive star formation. As yet, the physical conditions inside these regions are still poorly known. Submillimeter observations offer one way of probing the gas density and temperature. As part of an extensive study of the chemistry and physical conditions inside UCHII regions, we undertook a project to map several UCHII regions in CS. CS is an abundant molecule, and is an excellent density probe. Using the Submillimeter Telescope Observatory (SMTO), we are mapping ten UCHII regions in detail at a variety of CS transitions, including the CS 5-4, 7-6 and C34S 5-4 and 7-6 lines. The regions chosen also have far-infrared data and submm data available, so we will be able to compare the dust and gas properties. We report the first results of the CS mapping program for CS 5-4 and CS 7-6 lines. We compare our conclusions with other studies of these regions. T. L. Lauwers was supported by the University of Arizona/NASA Space Grant Undergraduate Research Internship Program.

  18. Radioactive Cs capture in the early solar system.

    PubMed

    Hidaka, Hiroshi; Yoneda, Shigekazu

    2013-01-01

    Barium isotopic compositions of primitive materials in the solar system are generally affected by s- and r-process nucleosynthetic components that hide the contribution of the isotopic excess of (135)Ba formed by decay of radioactive (135)Cs. However, the Ba isotopic composition of the chemical separates from chondrules in the Sayama CM2 chondrite shows an excess of (135)Ba isotopic abundance up to (0.33 ± 0.06)%, which is independent of the isotopic components from s- and r-process nucleosyntheses. The isotopic excesses of (135)Ba correlate with the elemental abundance of Ba relative to Cs, providing chemical and isotopic evidence for the existence of the presently extinct radionuclide (135)Cs (t(1/2) = 2.3 million years) in the early solar system. The estimated abundance of (135)Cs/(133)Cs = (6.8 ± 1.9) × 10(-4) is more than double that expected from the uniform production model of the short-lived radioisotopes, suggesting remobilization of Cs including (135)Cs in the chondrules of the meteorite parent body. PMID:23435551

  19. New type of borophosphate anionic radical in the crystal structure of CsAl2BP6O20

    NASA Astrophysics Data System (ADS)

    Shvanskaya, L. V.; Yakubovich, O. V.; Belik, V. I.

    2016-09-01

    The crystal structure of a new borophosphate CsAl2BP6O20 obtained by spontaneous crystallization in a multicomponent Cs-Cu-B-P-O system is determined by X-ray diffraction ( a = 11.815(2), b = 10.042(2), and c = 26.630(4) Å; space group Pbca, Z = 8, V = 3159.5(10) Å3; R 1 = 0.043). A new type of borophosphate anionic 2D radical characterized by the lowest B: P = 1: 6 ratio and containing P3O10 phosphate groups is found in the compound. A mixed-type anionic framework consisting of vertex-sharing BO4 and PO4 tetrahedra and AlO6 octahedra is distinguished in the structure. Large cesium atoms are located in the channels of the framework. Topological relationships are revealed between the structures of the CsAl3(P3O10)2 and CsAl2BP6O20 phases having different cationic compositions. These compounds can be considered quasi-polytypic phases.

  20. A Kv3-like persistent, outwardly rectifying, Cs+-permeable, K+ current in rat subthalamic nucleus neurones

    PubMed Central

    Wigmore, Mark A; Lacey, Michael G

    2000-01-01

    A persistent outward K+ current (IPO), activated by depolarization from resting potential, has been identified and characterized in rat subthalamic nucleus (SThN) neurones using whole-cell voltage-clamp recording in brain slices.IPO both rapidly activated (τ= 8 ms at +5 mV) and deactivated (τ= 2 ms at −68 mV), while showing little inactivation. Tail current reversal potentials varied with extracellular K+ concentration in a Nernstian manner.Intracellular Cs+ did not alter either IPO amplitude or the voltage dependence of activation, but blocked transient (A-like) outward currents activated by depolarization. When extracellular K+ was replaced with Cs+, IPO tail current reversal potentials were dependent upon the extracellular Cs+ concentration, indicating an ability to conduct Cs+, as well as K+.IPO was blocked by Ba2+ (1 mm), 4-aminopyridine (1 mm) and tetraethylammonium (TEA; 20 mm), with an IC50 for TEA of 0.39 mm.The IPO conductance appeared maximal (38 nS) at around +27 mV, half-maximal at −13 mV, with the threshold for activation at around −38 mV.TEA (1 mm) blocked the action potential after-hyperpolarization and permitted accommodation of action potential firing at frequencies greater than around 200 Hz.We conclude that IPO, which shares many characteristics of currents attributable to Kv3.1 K+ channels, enables high-frequency spike trains in SThN neurones. PMID:10990536

  1. VizieR Online Data Catalog: TW Hya CO (2-1), CN (2-1) and CS (5-4) data cubes (Teague+, 2016)

    NASA Astrophysics Data System (ADS)

    Teague, R.; Guilloteau, S.; Semenov, D.; Henning, T.; Dutrey, A.; Pietu, V.; Birnstiel, T.; Chapillon, E.; Hollenbach, D.; Gorti, U.

    2016-07-01

    The observations were performed using ALMA on May 13, 2015 under excellent weather conditions (Cycle 2, 2013.1.00387.S). The receivers were tuned to cover CO J=(2-1), CS J=(5-4) and all strong hyperfine components of CN N=(2-1) simultaneously. The correlator was configured to deliver very high spectral resolution, with a channel spacing of 15kHz (and an effective velocity resolution of 40m/s) for the CO J=(2-1) and CS J=(5-4) lines, and 30kHz (80m/s) for the CN N=(2-1) transition. (2 data files).

  2. First Test Results on ITER CS Model Coil and CS Insert

    SciTech Connect

    Martovetsky, N; Michael, P; Minervini, J; Radovinsky, A; Takayasu, M; Thome, R; Ando, T; Isono, T; Kato, T; Nakajima, H; Nishijima, G; Nunoya, Y; Sugimoto, M; Takahashi, Y; Tsuji, H; Bessette, D; Okuno, K; Ricci, M; Maix, R

    2000-10-12

    The Inner and Outer modules of the Central Solenoid Model Coil (CSMC) were built by US and Japanese home teams in collaboration with European and Russian teams to demonstrate the feasibility of a superconducting Central Solenoid for ITER and other large tokamak reactors. The CSMC mass is about 120 t, OD is about 3.6 m and the stored energy is 640 MJ at 46 kA and peak field of 13 T. Testing of the CSMC and the CS Insert took place at the Japan Atomic Energy Research Institute (JAERI) from mid March until mid August 2000. This paper presents the main results of the tests performed.

  3. Influence of water management and fertilizer application on (137)Cs and (133)Cs uptake in paddy rice fields.

    PubMed

    Wakabayashi, Shokichi; Itoh, Sumio; Kihou, Nobuharu; Matsunami, Hisaya; Hachinohe, Mayumi; Hamamatsu, Shioka; Takahashi, Shigeru

    2016-06-01

    Cesium-137 derived from the Tokyo Electric Power Company's Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident contaminated large areas of agricultural land in Eastern Japan. Previous studies before the accident have indicated that flooding enhances radiocesium uptake in rice fields. We investigated the influence of water management in combination with fertilizers on (137)Cs concentrations in rice plants at two fields in southern Ibaraki Prefecture. Stable Cs ((133)Cs) in the plants was also determined as an analogue for predicting (137)Cs behavior after long-term aging of soil (137)Cs. The experimental periods comprised 3 y starting from 2012 in one field, and 2 y from 2013 in another field. These fields were divided into three water management sections: a long-flooding section without midsummer drainage, and medial-flooding, and short-flooding sections with one- or two-week midsummer drainage and earlier end of flooding than the long-flooding section. Six or four types of fertilizer subsections (most differing only in potassium application) were nested in each water management section. Generally, the long-flooding treatment led to higher (137)Cs and (133)Cs concentrations in both straw and brown rice than medial- and short-flooding treatments, although there were some notable exceptions in the first experimental year at each site. Effects of differing potassium fertilizer treatments were cumulative; the effects on (137)Cs and (133)Cs concentrations in rice plants were not obvious in 2012 and 2013, but in 2014, these concentrations were highest where potassium fertilizer had been absent and lowest where basal dressings of K had been tripled. The relationship between (137)Cs and (133)Cs in rice plants was not correlative in the first experimental year at each site, but correlation became evident in the subsequent year(s). This study demonstrates a novel finding that omitting midsummer drainage and/or delaying drainage during the grain-filling period enhances

  4. Physics Design for ARIES-CS

    SciTech Connect

    L.P. Ku, P.R. Garabedian, J. Lyon, A. Turnbull, A. Grossman, T.K. Mau, M. Zarnstorff, and the ARIES Team

    2007-10-10

    Novel stellarator configurations have been developed for ARIES-CS. These configurations are optimized to provide good plasma confinement and flux surface integrity at high beta. Modular coils have been designed for them in which the space needed for the breeding blanket and radiation shielding was specifically targeted such that reactors generating GW electrical powers would require only moderate major radii (<10 m). These configurations are quasi-axially symmetric in the magnetic field topology and have small number of field periods (≤3) and low aspect ratios (≤6). The baseline design chosen for detailed systems and power plant studies has 3 field periods, aspect ratio 4.5 and major radius 7.5 m operating at β~6.5% to yield 1 GW electric power. The shaping of the plasma accounts for ≥75% of the rotational transform. The effective helical ripples are very small (< 0.6% everywhere) and the energy loss of alpha particles is calculated to be ≤5% when operating in high density regimes. An interesting feature in this configuration is that instead of minimizing all residues in the magnetic spectrum, we preferentially retained a small amount of the non-axisymmetric mirror field. The presence of this mirror and its associated helical field alters the ripple distribution, resulting in the reduced ripple-trapped loss of alpha particles despite the long connection length in a tokamak-like field structure. Additionally, we discuss two other potentially attractive classes of configurations, both quasi-axisymmetric: one with only two field periods, very low aspect ratios (~2.5), and less complex coils, and the other with the plasma shaping designed to produce low shear rotational transform so as to assure the robustness and integrity of flux surfaces when operating at high β.

  5. Production of low radioactivity CsI(Tl) crystals for WIMP searches

    SciTech Connect

    Kim, Y.D.; Kang, W.G.; Lee, H.S.; Lee, J.I.; Lee, J.Y.; Hahn, I.S.; Lee, E.K.; Hwang, M.J.; Kwon, Y.J.; Jin, Li; Kim, H.J.; Kim, S.C.; Kim, S.K.; Kim, S.Y.; Kim, T.Y.; Kwak, J.W.; Lee, M.J.; Myung, S.S.; Yang, H.Y.; Park, H.

    2005-09-08

    We have extensively studied the mechanism of the 137Cs and 134Cs contamination in CsI compounds. The nuclei 137Cs and 134Cs in various samples have been studied using a HPGe detector at deep underground. The main source of the 137Cs contamination in CsI powder was desolved 137Cs in the water used in the chemical process of cesium extraction at company side. We also found that 134Cs was produced mainly from 133Cs by neutron capture. We have successfully produced CsI powder containing no more than 2 mBq/kg of 137Cs by using only ultra-pure water for whole extraction process from raw ore(pollucite)

  6. Altimeter Products for the Sentinel-6/Jason-CS Mission

    NASA Astrophysics Data System (ADS)

    Scharroo, Remko; Bonekamp, Hans; Ponsard, Christelle; Nogueira Loddo, Carolina

    2015-12-01

    The Sentinel-6 mission will be developed and implemented through a partnership between the EU, ESA, EUMETSAT and NOAA . Its aim is to secure the continuity until 2030+ of critical high precision observations of ocean surface topography beyond Jason-3. The European contribution will be implemented through the combination of the ESA Copernicus Space Component, the EUMETSAT Jason-CS optional programme, and the EU Copernicus programme, for the joint benefits of the meteorological and Copernicus user communities in Europe. NASA and CNES will be supporting partners. The mission will start with the launch of Jason-CS A in 2020, followed by Jason-CS B in 2025.

  7. The transfer of {sup 137}Cs from barley to beer

    SciTech Connect

    Proehl, G.; Mueller, H.; Voigt, G.

    1997-01-01

    Beer has been brewed from barley contaminated with {sup 137}Cs as a consequence of the Chernobyl accident. The {sup 137}Cs activity has been measured in all intermediate steps and in the by-products of the production process. About 35 % of the {sup 137}Cs in barley were recovered in beer. Processing factors defined as the concentration ratio of processed and raw products were determined to be 0.61, 3.3, 0.1 and 0.11 for malt, malt germs, spent grains and beer, respectively. 4 refs., 2 tabs.

  8. The KTeV Pure CsI Calorimeter

    SciTech Connect

    Ray, R.E.

    1994-12-01

    KTeV is currently building a state-of-the-art pure CsI electromagnetic calorimeter with a sophisticated digital readout. The CsI array is expected to have better than 1% resolution over a dynamic range of 2--64 GeV. The design of the CsI array is driven by the difficult physics goal of attempting to measure the CP violation parameter Re({epsilon}{prime}/{epsilon}) to 1 part in 10000 in a high-rate neutral beam environment. The physics requirements and their impact on the final design will be discussed.

  9. Low temperature phase transition and crystal structure of CsMgPO{sub 4}

    SciTech Connect

    Orlova, Maria; Khainakov, Sergey; Michailov, Dmitriy; Perfler, Lukas; Langes, Christoph; Kahlenberg, Volker; Orlova, Albina

    2015-01-15

    CsMgPO{sub 4} doped with radioisotopes is a promising compound for usage as a radioactive medical source. However, a low temperature phase transition at temperatures close to ambient conditions (∼−40 °C) was observed. Information about such kind of structural changes is important in order to understand whether it can cause any problem for medical use of this compound. The phase transition has been investigated in detail using synchrotron powder diffraction, Raman spectroscopy and DFT calculations. The structure undergoes a transformation from an orthorhombic modification, space group Pnma (RT phase) to a monoclinic polymorph, space group P2{sub 1}/n (LT phase). New LT modification adopts similar to RT but slightly distorted unit cell: a=9.58199(2) Å, b=8.95501(1) Å, c=5.50344(2) Å, β=90.68583(1)°, V=472.198(3) Å{sup 3}. CsMgPO{sub 4} belongs to the group of framework compounds and is made up of strictly alternating MgO{sub 4}- and PO{sub 4}-tetrahedra sharing vertices. The cesium counter cations are located in the resulting channel-like cavities. Upon the transformation a combined tilting of the tetrahedra is observed. A comparison with other phase transitions in ABW-type framework compounds is given. - Graphical abstract: Structural behavior of β-tridymite-type phosphate CsMgPO{sub 4}, considered as potential chemical form for radioactive Cs-source has been studied at near ambient temperatures. A phase transition at (∼−40 °C) has been found and investigated. It has been established that the known orthorhombic RT modification, space group Pnma, adopts a monoclinic cell with space group P2{sub 1}/n at low temperatures. In this paper, we present results of structural analysis of changes accompanying this phase transition and discuss its possible impact on the application properties. - Highlights: • β-Tridymite type phosphate CsMgPO{sub 4} undergoes so called translationengleiche phase transition of index 2 at −40 °C. • The structure

  10. Status and prospect of the Swiss continuous Cs fountain FoCS-2

    NASA Astrophysics Data System (ADS)

    Jallageas, A.; Devenoges, L.; Petersen, M.; Morel, J.; Bernier, L.-G.; Thomann, P.; Südmeyer, T.

    2016-06-01

    The continuous cesium fountain clock FoCS-2 at METAS presents many unique characteristics and challenges in comparison with standard pulsed fountain clocks. For several years FoCS-2 was limited by an unexplained frequency sensitivity on the velocity of the atoms, in the range of 140 • 10-15. Recent experiments allowed us to identify the origin of this problem as undesirable microwave surface currents circulating on the shield of the coaxial cables that feed the microwave cavity. A strong reduction of this effect was obtained by adding microwave absorbing coatings on the coaxial cables and absorbers inside of the vacuum chamber. This breakthrough opens the door to a true metrological validation of the fountain. A series of simulation tools have already been developed and proved their efficiency in the evaluation of some of the uncertainties of the continuous fountain. With these recent improvements, we are confident in the future demonstration of an uncertainty budget at the 10-15 level and below.

  11. Kinetic Monte Carlo Model of Scintillation Mechanisms in CsI and CsI(Tl)

    SciTech Connect

    Kerisit, Sebastien N.; Rosso, Kevin M.; Cannon, Bret D.

    2008-06-01

    We have developed a computational model of energy transfer processes in scintillators using the kinetic Monte Carlo (KMC) approach. In this publication, we focus on the alkali halide compound CsI both pure and doped with a range of thallium concentrations. The KMC model makes use of an explicit atomistic representation of the crystal lattice, activator sites, defect sites, and individual electron-hole pairs. The probability of individual diffusion, recombination, and scintillation events is calculated from rate equations parameterized with data published in the literature. Scintillation decay curves, relative intensities of emission peaks, and light yields are computed and found to be in good agreement with experimental data for a range of temperatures and thallium concentrations. This demonstrates that the KMC scintillation model is capable of reproducing both the kinetics and the efficiency of the scintillation process in CsI. In addition, novel predictions emerge from our simulations such as the diffusion distance distributions of self-trapped holes and excitons. Finally, the KMC scintillation model provides a framework for probing possible physical processes responsible for the nonlinear relationship between scintillation light yield and incident gamma-ray energy.

  12. Computer Simulation of Electron Thermalization in CsI and CsI(Tl)

    SciTech Connect

    Wang, Zhiguo; Xie, YuLong; Cannon, Bret D.; Campbell, Luke W.; Gao, Fei; Kerisit, Sebastien N.

    2011-09-15

    A Monte Carlo (MC) model was developed and implemented to simulate the thermalization of electrons in inorganic scintillator materials. The model incorporates electron scattering with both longitudinal optical and acoustic phonons. In this paper, the MC model was applied to simulate electron thermalization in CsI, both pure and doped with a range of thallium concentrations. The inclusion of internal electric fields was shown to increase the fraction of recombined electron-hole pairs and to broaden the thermalization distance and thermalization time distributions. The MC simulations indicate that electron thermalization, following {gamma}-ray excitation, takes place within approximately 10 ps in CsI and that electrons can travel distances up to several hundreds of nanometers. Electron thermalization was studied for a range of incident {gamma}-ray energies using electron-hole pair spatial distributions generated by the MC code NWEGRIM (NorthWest Electron and Gamma Ray Interaction in Matter). These simulations revealed that the partition of thermalized electrons between different species (e.g., recombined with self-trapped holes or trapped at thallium sites) vary with the incident energy. Implications for the phenomenon of nonlinearity in scintillator light yield are discussed.

  13. Measurement of dissolved Cs-137 in stream water, soil water and groundwater at Headwater Forested Catchment in Fukushima after Fukushima Dai-ichi Nuclear Power Plant Accident

    NASA Astrophysics Data System (ADS)

    Iwagami, Sho; Tsujimura, Maki; Onda, Yuichi; Sakakibara, Koichi; Konuma, Ryohei; Sato, Yutaro

    2016-04-01

    Radiocesium migration from headwater forested catchment is important perception as output from the forest which is also input to the subsequent various land use and downstream rivers after Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident. In this study, dissolved Cs-137 concentration of stream water, soil water and groundwater were measured. Observations were conducted at headwater catchment in Yamakiya district, located 35 km northwest of FDNPP from April 2014 to November 2015. Stream water discharge was monitored and stream water samples were taken at main channel and sub channel. Stream water discharge was monitored by combination of parshallflume and v-notch weir. Stream water was sampled manually at steady state condition in 3-4 month interval and also intense few hours interval sampling were conducted during rainfall events using automated water sampler. Around the sub channel, it is found that there is a regularly saturated area at the bottom of the slope, temporary saturated area which saturate during the rainy season in summer and regularly dry area. 6 interval cameras were installed to monitor the changing situation of saturated area. Suction lysimeters were installed at three areas (regularly saturated area, temporary saturated area and dry area) for sampling soil water in depth of 0.1 m and 0.3 m. Boreholes were installed at three points along the sub channel. Three boreholes with depth of 3 m, 5 m and 10 m were installed at temporary saturated area, 20 m upstream of sub channel weir. Another three boreholes with depth of 3 m, 5 m and 10 m were installed at dry area, 40 m upstream of sub channel weir. And a borehole with depth of 20 m was installed at ridge of sub catchment, 52 m upstream of sub channel weir. Groundwater was sampled by electrically powered pump and groundwater level was monitored. Also suction-free lysimeter was installed at temporary saturated area for sampling the near surface subsurface water. Soil water samples were collected

  14. Damage of the bacterial cell envelope by antimicrobial peptides gramicidin S and PGLa as revealed by transmission and scanning electron microscopy.

    PubMed

    Hartmann, Mareike; Berditsch, Marina; Hawecker, Jacques; Ardakani, Mohammad Fotouhi; Gerthsen, Dagmar; Ulrich, Anne S

    2010-08-01

    Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to examine the ultrastructural changes in bacteria induced by antimicrobial peptides (AMPs). Both the beta-stranded gramicidin S and the alpha-helical peptidyl-glycylleucine-carboxyamide (PGLa) are cationic amphiphilic AMPs known to interact with bacterial membranes. One representative Gram-negative strain, Escherichia coli ATCC 25922, and one representative Gram-positive strain, Staphylococcus aureus ATCC 25923, were exposed to the AMPs at sub-MICs and supra-MICs in salt-free medium. SEM revealed a shortening and swelling of the E. coli cells, and multiple blisters and bubbles formed on their surface. The S. aureus cells seemed to burst upon AMP exposure, showing open holes and deep craters in their envelope. TEM revealed the formation of intracellular membranous structures in both strains, which is attributed to a lateral expansion of the lipid membrane upon peptide insertion. Also, some morphological alterations in the DNA region were detected for S. aureus. After E. coli was incubated with AMPs in medium with low ionic strength, the cells appeared highly turgid compared to untreated controls. This observation suggests that the AMPs enhance osmosis through the inner membrane, before they eventually cause excessive leakage of the cellular contents. The adverse effect on the osmoregulatory capacity of the bacteria is attributed to the membrane-permeabilizing action of the amphiphilic peptides, even at low (sub-MIC) AMP concentrations. Altogether, the results demonstrate that both TEM and SEM, as well as appropriate sample preparation protocols, are needed to obtain detailed mechanistic insights into peptide function. PMID:20530225

  15. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis

    PubMed Central

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian

    2015-01-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly 13C/15N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive 13C/15N-labeled amino acids. The most cost-effective production of 13C/15N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% 13C-glycerol and 0.5% 15N-ammonium sulfate, supplemented with only 0.025% of 13C/15N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state. PMID:25795666

  16. Synergistic effect of membrane-active peptides polymyxin B and gramicidin S on multidrug-resistant strains and biofilms of Pseudomonas aeruginosa.

    PubMed

    Berditsch, Marina; Jäger, Thomas; Strempel, Nikola; Schwartz, Thomas; Overhage, Jörg; Ulrich, Anne S

    2015-09-01

    Multidrug-resistant Pseudomonas aeruginosa is a major cause of severe hospital-acquired infections. Currently, polymyxin B (PMB) is a last-resort antibiotic for the treatment of infections caused by Gram-negative bacteria, despite its undesirable side effects. The delivery of drug combinations has been shown to reduce the required therapeutic doses of antibacterial agents and thereby their toxicity if a synergistic effect is present. In this study, we investigated the synergy between two cyclic antimicrobial peptides, PMB and gramicidin S (GS), against different P. aeruginosa isolates, using a quantitative checkerboard assay with resazurin as a growth indicator. Among the 28 strains that we studied, 20 strains showed a distinct synergistic effect, represented by a fractional inhibitory concentration index (FICI) of ≤0.5. Remarkably, several clinical P. aeruginosa isolates that grew as small-colony variants revealed a nonsynergistic effect, as indicated by FICIs between >0.5 and ≤0.70. In addition to inhibiting the growth of planktonic bacteria, the peptide combinations significantly decreased static biofilm growth compared with treatment with the individual peptides. There was also a faster and more prolonged effect when the combination of PMB and GS was used compared with single-peptide treatments on the metabolic activity of pregrown biofilms. The results of the present study define a synergistic interaction between two cyclic membrane-active peptides toward 17 multidrug-resistant P. aeruginosa and biofilms of P. aeruginosa strain PAO1. Thus, the application of PMB and GS in combination is a promising option for a topical medication and in the prevention of acute and chronic infections caused by multidrug-resistant or biofilm-forming P. aeruginosa. PMID:26077259

  17. Synergistic Effect of Membrane-Active Peptides Polymyxin B and Gramicidin S on Multidrug-Resistant Strains and Biofilms of Pseudomonas aeruginosa

    PubMed Central

    Berditsch, Marina; Jäger, Thomas; Strempel, Nikola; Schwartz, Thomas; Overhage, Jörg

    2015-01-01

    Multidrug-resistant Pseudomonas aeruginosa is a major cause of severe hospital-acquired infections. Currently, polymyxin B (PMB) is a last-resort antibiotic for the treatment of infections caused by Gram-negative bacteria, despite its undesirable side effects. The delivery of drug combinations has been shown to reduce the required therapeutic doses of antibacterial agents and thereby their toxicity if a synergistic effect is present. In this study, we investigated the synergy between two cyclic antimicrobial peptides, PMB and gramicidin S (GS), against different P. aeruginosa isolates, using a quantitative checkerboard assay with resazurin as a growth indicator. Among the 28 strains that we studied, 20 strains showed a distinct synergistic effect, represented by a fractional inhibitory concentration index (FICI) of ≤0.5. Remarkably, several clinical P. aeruginosa isolates that grew as small-colony variants revealed a nonsynergistic effect, as indicated by FICIs between >0.5 and ≤0.70. In addition to inhibiting the growth of planktonic bacteria, the peptide combinations significantly decreased static biofilm growth compared with treatment with the individual peptides. There was also a faster and more prolonged effect when the combination of PMB and GS was used compared with single-peptide treatments on the metabolic activity of pregrown biofilms. The results of the present study define a synergistic interaction between two cyclic membrane-active peptides toward 17 multidrug-resistant P. aeruginosa and biofilms of P. aeruginosa strain PAO1. Thus, the application of PMB and GS in combination is a promising option for a topical medication and in the prevention of acute and chronic infections caused by multidrug-resistant or biofilm-forming P. aeruginosa. PMID:26077259

  18. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis.

    PubMed

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian; Ulrich, Anne S

    2015-06-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly (13)C/(15)N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive (13)C/(15)N-labeled amino acids. The most cost-effective production of (13)C/(15)N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% (13)C-glycerol and 0.5% (15)N-ammonium sulfate, supplemented with only 0.025% of (13)C/(15)N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state.

  19. Effects of imidazolium-based ionic liquids on the stability and dynamics of gramicidin A and lipid bilayers at different salt concentrations.

    PubMed

    Lee, Hwankyu; Kim, Sun Min; Jeon, Tae-Joon

    2015-09-01

    Gramicidin A (gA) dimers with bilayers, which consist of phospholipids and ionic liquids (ILs) at different molar ratios, were simulated at different salt concentrations of 0.15 and 1M NaCl. Bilayer thickness is larger than the length of a gA dimer, and hence lipids around the gA dimer are significantly disordered to adapt to the gA dimer, yielding membrane curvature. As the IL concentration increases, the bilayer thickness decreases and becomes closer to the gA length, leading to less membrane curvature. Also, ILs significantly increase lateral diffusivities of the gA dimer and lipids at 0.15M NaCl, but not at 1M NaCl because strong electrostatic interactions between salt ions and lipid head groups suppress an increase in the lateral mobility of the bilayer at high salt concentration. These findings help explain the conflicting experimental results that showed the increased ion permeability in electrophysiological experiments at 1M NaCl, but the reduced ion permeability in fluorescent experiments at 0.15M NaCl. ILs disorder lipids and make bilayers thinner, which yields less membrane curvature around the gA dimer and thus stabilizes the gA dimer, leading to the increased ion permeability. This IL effect predominantly occurs at 1M NaCl, where ILs only slightly increase the bilayer dynamics because of the strong electrostatic interactions between salt ions and lipids. In contrast, at 0.15M NaCl, ILs do not only stabilize the curved bilayer but also significantly increase the lateral mobility of gA dimers and lipids, which can reduce gA-induced pore formation, leading to the decreased ion permeability.

  20. Synergistic effect of membrane-active peptides polymyxin B and gramicidin S on multidrug-resistant strains and biofilms of Pseudomonas aeruginosa.

    PubMed

    Berditsch, Marina; Jäger, Thomas; Strempel, Nikola; Schwartz, Thomas; Overhage, Jörg; Ulrich, Anne S

    2015-09-01

    Multidrug-resistant Pseudomonas aeruginosa is a major cause of severe hospital-acquired infections. Currently, polymyxin B (PMB) is a last-resort antibiotic for the treatment of infections caused by Gram-negative bacteria, despite its undesirable side effects. The delivery of drug combinations has been shown to reduce the required therapeutic doses of antibacterial agents and thereby their toxicity if a synergistic effect is present. In this study, we investigated the synergy between two cyclic antimicrobial peptides, PMB and gramicidin S (GS), against different P. aeruginosa isolates, using a quantitative checkerboard assay with resazurin as a growth indicator. Among the 28 strains that we studied, 20 strains showed a distinct synergistic effect, represented by a fractional inhibitory concentration index (FICI) of ≤0.5. Remarkably, several clinical P. aeruginosa isolates that grew as small-colony variants revealed a nonsynergistic effect, as indicated by FICIs between >0.5 and ≤0.70. In addition to inhibiting the growth of planktonic bacteria, the peptide combinations significantly decreased static biofilm growth compared with treatment with the individual peptides. There was also a faster and more prolonged effect when the combination of PMB and GS was used compared with single-peptide treatments on the metabolic activity of pregrown biofilms. The results of the present study define a synergistic interaction between two cyclic membrane-active peptides toward 17 multidrug-resistant P. aeruginosa and biofilms of P. aeruginosa strain PAO1. Thus, the application of PMB and GS in combination is a promising option for a topical medication and in the prevention of acute and chronic infections caused by multidrug-resistant or biofilm-forming P. aeruginosa.

  1. Membrane-mediated protein-protein interactions and connection to elastic models: a coarse-grained simulation analysis of gramicidin A association.

    PubMed

    Yoo, Jejoong; Cui, Qiang

    2013-01-01

    To further foster the connection between particle based and continuum mechanics models for membrane mediated biological processes, we carried out coarse-grained (CG) simulations of gramicidin A (gA) dimer association and analyzed the results based on the combination of potential of mean force (PMF) and stress field calculations. Similar to previous studies, we observe that the association of gA dimers depends critically on the degree of hydrophobic mismatch, with the estimated binding free energy of >10 kcal/mol in a distearoylphosphatidylcholine bilayer. Qualitative trends in the computed PMF can be understood based on the stress field distributions near a single gA dimer and between a pair of gA dimers. For example, the small PMF barrier, which is ∼1 kcal/mol independent of lipid type, can be captured nearly quantitatively by considering membrane deformation energy associated with the region confined by two gA dimers. However, the PMF well depth is reproduced poorly by a simple continuum model that only considers membrane deformation energy beyond the annular lipids. Analysis of lipid orientation, configuration entropy, and stress distribution suggests that the annular lipids make a significant contribution to the association of two gA dimers. These results highlight the importance of explicitly considering contributions from annular lipids when constructing approximate models to study processes that involve a significant reorganization of lipids near proteins, such as protein-protein association and protein insertion into biomembranes. Finally, large-scale CG simulations indicate that multiple gA dimers also form clusters, although the preferred topology depends on the protein concentration. Even at high protein concentrations, every gA dimer requires contact to lipid hydrocarbons to some degree, and at most three to four proteins are in contact with each gA dimer; this observation highlights another aspect of the importance of interactions between proteins

  2. Structure-activity relationships of the antimicrobial peptide gramicidin S and its analogs: aqueous solubility, self-association, conformation, antimicrobial activity and interaction with model lipid membranes.

    PubMed

    Abraham, Thomas; Prenner, Elmar J; Lewis, Ruthven N A H; Mant, Colin T; Keller, Sandro; Hodges, Robert S; McElhaney, Ronald N

    2014-05-01

    GS10 [cyclo-(VKLdYPVKLdYP)] is a synthetic analog of the naturally occurring antimicrobial peptide gramicidin (GS) in which the two positively charged ornithine (Orn) residues are replaced by two positively charged lysine (Lys) residues and the two less polar aromatic phenylalanine (Phe) residues are replaced by the more polar tyrosine (Tyr) residues. In this study, we examine the effects of these seemingly conservative modifications to the parent GS molecule on the physical properties of the peptide, and on its interactions with lipid bilayer model and biological membranes, by a variety of biophysical techniques. We show that although GS10 retains the largely β-sheet conformation characteristic of GS, it is less structured in both water and membrane-mimetic solvents. GS10 is also more water soluble and less hydrophobic than GS, as predicted, and also exhibits a reduced tendency for self-association in aqueous solution. Surprisingly, GS10 associates more strongly with zwitterionic and anionic phospholipid bilayer model membranes than does GS, despite its greater water solubility, and the presence of anionic phospholipids and cholesterol (Chol) modestly reduces the association of both GS10 and GS to these model membranes. The strong partitioning of both peptides into lipid bilayers is driven by a large favorable entropy change opposed by a much smaller unfavorable enthalpy change. However, GS10 is also less potent than GS at inducing inverted cubic phases in phospholipid bilayer model membranes and at inhibiting the growth of the cell wall-less bacterium Acholeplasma laidlawii B. These results are discussed in terms of the comparative antibiotic and hemolytic activities of these peptides.

  3. Accumulation of 137Cs in puddle sediments within urban ecosystem.

    PubMed

    Seleznev, Andrian A; Yarmoshenko, Ilia V; Ekidin, Alexey A

    2010-08-01

    The role of puddle sediments as a final depot of (137)Cs horizontal migration within the urban landscape is studied using the example of Ekaterinburg city, Russia. Radioactive contamination in the city appeared due to fallout after atmospheric testing of nuclear weapons and nuclear accidents. Contamination density of (137)Cs in the region was assessed from archive data to be about 5.1 kBq/m(2), of which the maximum activity concentration (<30 Bq/kg) is associated with the upper 15 cm soil layer. Results of the survey reported here indicate a mean (137)Cs activity concentration in puddle sediments of 80 Bq/kg, with a maximum value of 540 Bq/kg. It is estimated that horizontal migration has led to about a fourfold concentration of (137)Cs in puddle sediments. PMID:20417997

  4. Triplet-singlet conversion in ultracold Cs{sub 2} and production of ground-state molecules

    SciTech Connect

    Bouloufa, Nadia; Aymar, Mireille; Dulieu, Olivier; Pichler, Marin

    2011-02-15

    We propose a process to convert ultracold metastable Cs{sub 2} molecules in their lowest triplet state into (singlet) ground-state molecules in their lowest vibrational levels. Molecules are first pumped into an excited triplet state, and the triplet-singlet conversion is facilitated by a two-step spontaneous decay through the coupled A {sup 1{Sigma}}{sub u}{sup +}-b {sup 3{Pi}}{sub u} states. Using spectroscopic data and accurate quantum chemistry calculations for Cs{sub 2} potential curves and transition dipole moments, we show that this process competes favorably with the single-photon decay back to the lowest triplet state. In addition, we demonstrate that this conversion process represents a loss channel for vibrational cooling of metastable triplet molecules, preventing an efficient optical pumping cycle down to low vibrational levels.

  5. Surface adsorption of Cs137 ions on quartz crystals

    USGS Publications Warehouse

    Antkiw, Stephen; Waesche, H.; Senftle, F.

    1954-01-01

    Adsorption tests were made on four large synthetic and three natural quartz crystals to see if surface defects might be detected by subsequent autoradiography techniques. The adsorbent used was radioactive Cs137 in a solution of Cs 137Cl. Natural quartz crystals adsorbed more cesium than the synthetic crystals. Certain surface defects were made evident by this method, but twinning features could not be detected.

  6. Mapping CS in starburst galaxies: Disentangling and characterising dense gas

    NASA Astrophysics Data System (ADS)

    Kelly, G.; Viti, S.; Bayet, E.; Aladro, R.; Yates, J.

    2015-06-01

    Aims: We observe the dense gas tracer CS in two nearby starburst galaxies to determine how the conditions of the dense gas varies across the circumnuclear regions in starburst galaxies. Methods: Using the IRAM-30m telescope, we mapped the distribution of the CS(2-1) and CS(3-2) lines in the circumnuclear regions of the nearby starburst galaxies NGC 3079 and NGC 6946. We also detected formaldehyde (H2CO) and methanol (CH3OH) in both galaxies. We marginally detect the isotopologue C34S. Results: We calculate column densities under LTE conditions for CS and CH3OH. Using the detections accumulated here to guide our inputs, we link a time and depth dependent chemical model with a molecular line radiative transfer model; we reproduce the observations, showing how conditions where CS is present are likely to vary away from the galactic centres. Conclusions: Using the rotational diagram method for CH3OH, we obtain a lower limit temperature of 14 K. In addition to this, by comparing the chemical and radiative transfer models to observations, we determine the properties of the dense gas as traced by CS (and CH3OH). We also estimate the quantity of the dense gas. We find that, provided there are between 105 and 106 dense cores in our beam, for both target galaxies, emission of CS from warm (T = 100-400 K), dense (n(H2) = 105-6 cm-3) cores, possibly with a high cosmic ray ionisation rate (ζ = 100ζ0) best describes conditions for our central pointing. In NGC 6946, conditions are generally cooler and/or less dense further from the centre, whereas in NGC 3079, conditions are more uniform. The inclusion of shocks allows for more efficient CS formation, which means that gas that is less dense by an order of magnitude is required to replicate observations in some cases.

  7. /sup 137/Cs radioactive dating of Lake Ontario sediment cores

    SciTech Connect

    Ward, T.E.; Breeden, J.; Komisarcik, K.; Porter, R.; Czuczwa, J.; Kaminski, R.; McVeety, B.D.

    1987-12-01

    The distribution of /sup 137/Cs in sediment cores from Lake Ontario provides estimates of the sediment accumulation rates. Geochronology with /sup 210/Pb dating and distribution of Ambrosia (ragweed) pollen compare well with /sup 137/Cs dating. These methods can determine with precision, changes in sedimentation occurring over the past 100 years or so. Typical sedimentation rates of 0.18-0.36 cm/yr were measured. 16 refs., 3 figs., 2 tabs.

  8. Microbial cleavage of organic C-S bonds

    DOEpatents

    Kilbane, II, John J.

    1994-01-01

    A microbial process for selective cleavage of organic C--S bonds which may be used for reducing the sulfur content of sulfur-containing organic carbonaceous materials, Microorganisms of Rhodococcus rhodochrous and Bacillus sphaericus have been found which have the ability of selective cleavage of organic C--S bonds. Particularly preferred microorganisms are Rhodococcus rhodochrous strain ATCC 53968 and Bacillus sphaericus strain ATCC 53969 and their derivatives.

  9. Microbial cleavage of organic C-S bonds

    DOEpatents

    Kilbane, J.J. II.

    1994-10-25

    A microbial process is described for selective cleavage of organic C-S bonds which may be used for reducing the sulfur content of sulfur-containing organic carbonaceous materials. Microorganisms of Rhodococcus rhodochrous and Bacillus sphaericus have been found which have the ability of selective cleavage of organic C-S bonds. Particularly preferred microorganisms are Rhodococcus rhodochrous strain ATCC 53968 and Bacillus sphaericus strain ATCC 53969 and their derivatives.

  10. Solvent Extraction of Tc and Cs from Alkaline Nitrate Wastes

    SciTech Connect

    Bonnesen, P.V.; Conner, C.; Delmau, L.H.; Haverlock, T.J.; Leonard, R.A.; Lumetta, G.J.; Moyer, B.A.; Sachleben, R.A.

    1999-07-11

    This paper summarizes progress at three collaborating US national laboratories on the extraction of the fission products {sup 99}Tc and {sup 137}Cs from alkaline high-level wastes (HLW). Efficient, economical processes for Tc and Cs extraction (SRTALK and alkaline-side CSEX, respectively) have been developed, and testing has progressed through batch tests on actual wastes and continuous countercurrent centrifugal-contactor tests on simulants.

  11. Equalization in redundant channels

    NASA Technical Reports Server (NTRS)

    Tulpule, Bhalchandra R. (Inventor); Collins, Robert E. (Inventor); Cominelli, Donald F. (Inventor); O'Neill, Richard D. (Inventor)

    1988-01-01

    A miscomparison between a channel's configuration data base and a voted system configuration data base in a redundant channel system having identically operating, frame synchronous channels triggers autoequalization of the channel's historical signal data bases in a hierarchical, chronological manner with that of a correctly operating channel. After equalization, symmetrization of the channel's configuration data base with that of the system permits upgrading of the previously degraded channel to full redundancy. An externally provided equalization command, e.g., manually actuated, can also trigger equalization.

  12. Integration of biological ion channels onto optically addressable micro-fluidic electrode arrays for single molecule characterization.

    SciTech Connect

    Brozik, Susan Marie; Frink, Laura J. Douglas; Bachand, George David; Keller, David J.; Patrick, Elizabeth L.; Marshall, Jason A.; Ortiz, Theodore P.; Meyer, Lauren A.; Davis, Ryan W.; Brozik, James A.; Flemming, Jeb Hunter

    2004-12-01

    The challenge of modeling the organization and function of biological membranes on a solid support has received considerable attention in recent years, primarily driven by potential applications in biosensor design. Affinity-based biosensors show great promise for extremely sensitive detection of BW agents and toxins. Receptor molecules have been successfully incorporated into phospholipid bilayers supported on sensing platforms. However, a collective body of data detailing a mechanistic understanding of membrane processes involved in receptor-substrate interactions and the competition between localized perturbations and delocalized responses resulting in reorganization of transmembrane protein structure, has yet to be produced. This report describes a systematic procedure to develop detailed correlation between (recognition-induced) protein restructuring and function of a ligand gated ion channel by combining single molecule fluorescence spectroscopy and single channel current recordings. This document is divided into three sections: (1) reported are the thermodynamics and diffusion properties of gramicidin using single molecule fluorescence imaging and (2) preliminary work on the 5HT{sub 3} serotonin receptor. Thirdly, we describe the design and fabrication of a miniaturized platform using the concepts of these two technologies (spectroscopic and single channel electrochemical techniques) for single molecule analysis, with a longer term goal of using the physical and electronic changes caused by a specific molecular recognition event as a transduction pathway in affinity based biosensors for biotoxin detection.

  13. The Transfer of Dissolved Cs-137 from Soil to Plants

    SciTech Connect

    Prorok, V.V.; Melnichenko, L.Yu.; Mason, C.F.V.; Ageyev, V.A.; Ostashko, V.V.

    2006-07-01

    Rapidly maturing plants were grown simultaneously at the same experimental sites under natural conditions at the Chernobyl Exclusion Zone. Roots of the plants were side by side in the soil. During two seasons we selected samples of the plants and of the soils several times every season. Content of Cs-137 in the plant and in the soil solution extracted from the samples of soils was measured. Results of measurements of the samples show that, for the experimental site, Cs-137 content in the plant varies with date of the sample selection. The plant:soil solution Cs-137 concentration ratio depends strongly on the date of selection and also on the type of soil. After analysis of the data we conclude that Cs-137 plant uptake is approximately proportional to the content of dissolved Cs-137 in the soil per unit of volume, and the plant:soil solution Cs-137 concentration ratio for the soil is approximately proportional to the soil moisture. (authors)

  14. Syntheses, structures, and optical properties of the indium/germanium selenides Cs{sub 4}In{sub 8}GeSe{sub 16}, CsInSe{sub 2}, and CsInGeSe{sub 4}

    SciTech Connect

    Ward, Matthew D.; Pozzi, Eric A.; Van Duyne, Richard P.; Ibers, James A.

    2014-04-01

    The three solid-state indium/germanium selenides Cs{sub 4}In{sub 8}GeSe{sub 16}, CsInSe{sub 2}, and CsInGeSe{sub 4} have been synthesized at 1173 K. The structure of Cs{sub 4}In{sub 8}GeSe{sub 16} is a three-dimensional framework whereas those of CsInSe{sub 2} and CsInGeSe{sub 4} comprise sheets separated by Cs cations. Both Cs{sub 4}In{sub 8}GeSe{sub 16} and CsInGeSe{sub 4} display In/Ge disorder. From optical absorption measurements these compounds have band gaps of 2.20 and 2.32 eV, respectively. All three compounds are charge balanced. - Graphical abstract: Structure of Cs{sub 4}In{sub 8}GeSe{sub 16}. - Highlights: • The solid-state In/Ge selenides Cs{sub 4}In{sub 8}GeSe{sub 16}, CsInSe{sub 2}, and CsInGeSe{sub 4} have been synthesized. • Both Cs{sub 4}In{sub 8}GeSe{sub 16} and CsInGeSe{sub 4} display In/Ge disorder. • Cs{sub 4}In{sub 8}GeSe{sub 16} and CsInGeSe{sub 4} have band gaps of 2.20 eV and 2.32 eV, respectively.

  15. Channel nut tool

    DOEpatents

    Olson, Marvin

    2016-01-12

    A method, system, and apparatus for installing channel nuts includes a shank, a handle formed on a first end of a shank, and an end piece with a threaded shaft configured to receive a channel nut formed on the second end of the shaft. The tool can be used to insert or remove a channel nut in a channel framing system and then removed from the channel nut.

  16. Age of Martian channels

    NASA Technical Reports Server (NTRS)

    Malin, M. C.

    1976-01-01

    The ages of large Martian channels have been studied by determining the relative abundances of craters superimposed on channels and adjacent terrains and by examining superposition relationships between channels and plains and mantle materials. The channels are extremely old, are spatially confined and temporally related to the ancient cratered terrain, and in many cases are related to the as yet poorly understood genetic processes of fretting and chaos formation. No evidence is found for recent channel activity.

  17. DEG/ENaC but not TRP channels are the major mechanoelectrical transduction channels in a C. elegans nociceptor

    PubMed Central

    Geffeney, Shana L.; Cueva, Juan G.; Glauser, Dominique A.; Doll, Joseph C.; Lee, Tim Hau-Chen; Montoya, Misty; Karania, Snetu; Garakani, Arman M.; Pruitt, Beth L.; Goodman, Miriam B.

    2011-01-01

    Summary Many nociceptors detect mechanical cues, but the ion channels responsible for mechanotransduction in these sensory neurons remain obscure. Using in vivo recordings and genetic dissection, we identified the DEG/ENaC protein, DEG-1, as the major mechanotransduction channel in ASH, a polymodal nociceptor in Caenorhabditis elegans. But, DEG-1 is not the only mechanotransduction channel in ASH: loss of deg-1 revealed a minor current whose properties differ from those expected of DEG/ENaC channels. This current was independent of two TRPV channels expressed in ASH. Although loss of these TRPV channels inhibits behavioral responses to noxious stimuli, we found that both mechanoreceptor currents and potentials were essentially wild-type in TRPV mutants. We propose that ASH nociceptors rely on two genetically-distinct mechanotransduction channels and that TRPV channels contribute to encoding and transmitting information. Because mammalian and insect nociceptors also co-express DEG/ENaCs and TRPVs, the cellular functions elaborated here for these ion channels may be conserved. PMID:21903078

  18. Properties of CsI, CsBr and GaAs thin films grown by pulsed laser deposition

    SciTech Connect

    Brendel, V M; Garnov, S V; Yagafarov, T F; Iskhakova, L D; Ermakov, R P

    2014-09-30

    CsI, CsBr and GaAs thin films have been grown by pulsed laser deposition on glass substrates. The morphology and structure of the films have been studied using X-ray diffraction and scanning electron microscopy. The CsI and CsBr films were identical in stoichiometry to the respective targets and had a polycrystalline structure. Increasing the substrate temperature led to an increase in the density of the films. All the GaAs films differed in stoichiometry from the target. An explanation was proposed for this fact. The present results demonstrate that, when the congruent transport condition is not fulfilled, films identical in stoichiometry to targets can be grown by pulsed laser deposition in the case of materials with a low melting point and thermal conductivity. (interaction of laser radiation with matter)

  19. Measurement of airborne 131I, 134)Cs and 137Cs due to the Fukushima reactor incident in Milan (Italy).

    PubMed

    Clemenza, M; Fiorini, E; Previtali, E; Sala, E

    2012-12-01

    After the earthquake and the tsunami occurred in Japan on March 2011, four of the Fukushima reactors had released in air a large amount of radioactive isotopes that diffused all over the world. The presence of airborne (131)I, (134)Cs, and (137)Cs in air particulate due to this accident were detected and measured in the Low Radioactivity Laboratory operating in the Department of Environmental Sciences of the University of Milano-Bicocca. The sensitivity of the detecting apparatus is of 0.2 uBq/m(3) of air. Concentration and time distribution of these radiocontaminations ranging from a few to 400 uBq/m(3) for the (131)I and of a few tens of uBq/m(3) for the (137)Cs and (134)Cs.

  20. The third-generation turbocharged engine for the Audi 5000 CS and 5000 CS Quattro

    SciTech Connect

    Stock, D.

    1986-01-01

    In September 1985 the new Audi 5000 CS Quattro was introduced to the American market. This luxurious high performance touring sedan has been equipped with a more advanced turbocharged engine with intercooler and electronic engine management giving improved performance, excellent torque, faster response and better fuel economy. The basic engine is the tried-and-tested Audi 5-cylinder unit. The turbocharged engine's ancillary systems, the electronic ignition control and fuel injection have all been newly developed, carefully optimized and well matched in the special demands of a turbocharged engine. The ignition system controls the engine and fuel injection and delivers analog and digital signals to the car's instrument panel display. The system also has an integrated self-diagnostic function.

  1. Update global budget of 134Cs and 137Cs derived from Fukushima NPP1 accident

    NASA Astrophysics Data System (ADS)

    Aoyama, Michio; Tsumune, Daisuke; Tsubono, Takaki; Misuimi, Kazuhiro; Inomata, Yayoi; Kumamoto, Yuichiro; Hamajima, Yasunori

    2014-05-01

    134Cs and 137Cs, hereafter radiocaesium, were released from the Fukushima NPP1 accident on March 2011 and to evaluate total amount of released radiocaesium is essential to assess impacts of released radiocaesium to iur environment. We however still have wide range of reports about total amount of released radiocaesium after the accident till today. Released radiocasium were injected in the North Pacific Ocean by two major pathways, direct discharge from the accident site and atmospheric pathway to the ocean while there was only atmospheric deposition on land. We summarize estimations of budget of radiocasium based on both observations and model studies for land deposition, deposition on the North Pacific Ocean and inventory in the North Pacific Ocean, an inventory of sediment close to the site. The major portion of released radiocaesium can be believed to be deposited mainly in the western North Pacific Ocean because deposition density at west coast of US continent was only at an order of 100 Bq m-2 while that at close to accident site was MBq m-2. An inventory in the North Pacific Ocean by some of the authors was 15-18 PBq on April-May 2011 by model-observation comparison and that in the mode water in the western North Pacific Ocean based on the observations on January- February 2012 was 6 PBq. The second portion might be on main land Japan and it is rather easy to evaluate total amount of deposited radiocaesium because there are good observation both by direct sampling of soil and aerial survey. A result of integration for land deposition was about 2.5 PBq. The third portion is in the sediment and an inventory in the sediment by observation showed 42 TBq at close to the site while the total inventory of 137Cs in sediments off the Fukushima coast is thus estimated to be the order of 0.1 PBq by a model study by some of the authors. Estimations of direct discharge from the site also still varied much from 3 PBq to 27 PBq. Two larger estimations did the backward

  2. Enterotoxigenic Escherichia coli CS6 gene products and their roles in CS6 structural protein assembly and cellular adherence.

    PubMed

    Wajima, Takeaki; Sabui, Subrata; Fukumoto, Megumi; Kano, Shigeyuki; Ramamurthy, Thandavarayan; Chatterjee, Nabendu Sekhar; Hamabata, Takashi

    2011-10-01

    Enterotoxigenic Escherichia coli (ETEC) produces a variety of colonization factors necessary for attachment to the host cell, among which CS6 is one of the most prevalent in ETEC isolates from developing countries. The CS6 operon is composed of 4 genes, cssA, cssB, cssC, and cssD. The molecular mechanism of CS6 assembly and cell surface presentation, and the contribution of each protein to the attachment of the bacterium to intestinal cells remain unclear. In the present study, a series of css gene-deletion mutants of the CS6 operon were constructed in the ETEC genetic background, and their effect on adhesion to host cells and CS6 assembly was studied. Each subunit deletion resulted in a reduction in the adhesion to intestinal cells to the same level of laboratory E. coli strains, and this effect was restored by complementary plasmids, suggesting that the 4 proteins are necessary for CS6 expression. Bacterial cell fractionation and western blotting of the mutant strains suggested that the formation of a CssA-CssB-CssC complex is necessary for recognition by CssD and transport of CssA-CssB to the outer membrane as a colonization factor. PMID:21729748

  3. Spatial and temporal variability in sedimentation rates associated with cutoff channel infill deposits: Ain River, France

    USGS Publications Warehouse

    Piegay, H.; Hupp, C.R.; Citterio, A.; Dufour, S.; Moulin, B.; Walling, D.E.

    2008-01-01

    Floodplain development is associated with lateral accretion along stable channel geometry. Along shifting rivers, the floodplain sedimentation is more complex because of changes in channel position but also cutoff channel presence, which exhibit specific overflow patterns. In this contribution, the spatial and temporal variability of sedimentation rates in cutoff channel infill deposits is related to channel changes of a shifting gravel bed river (Ain River, France). The sedimentation rates estimated from dendrogeomorphic analysis are compared between and within 14 cutoff channel infills. Detailed analyses along a single channel infill are performed to assess changes in the sedimentation rates through time by analyzing activity profiles of the fallout radionuclides 137Cs and unsupported 210Pb. Sedimentation rates are also compared within the channel infills with rates in other plots located in the adjacent floodplain. Sedimentation rates range between 0.65 and 2.4 cm a -1 over a period of 10 to 40 years. The data provide additional information on the role of distance from the bank, overbank flow frequency, and channel geometry in controlling the sedimentation rate. Channel infills, lower than adjacent floodplains, exhibit higher sedimentation rates and convey overbank sediment farther away within the floodplain. Additionally, channel degradation, aggradation, and bank erosion, which reduce or increase the distance between the main channel and the cutoff channel aquatic zone, affect local overbank flow magnitude and frequency and therefore sedimentation rates, thereby creating a complex mosaic of sedimentation zones within the floodplain and along the cutoff channel infills. Last, the dendrogeomorphic and 137Cs approaches are cross validated for estimating the sedimentation rate within a channel infill. Copyright 2008 by the American Geophysical Union.

  4. Microfluidic channel fabrication method

    DOEpatents

    Arnold, Don W.; Schoeniger, Joseph S.; Cardinale, Gregory F.

    2001-01-01

    A new channel structure for microfluidic systems and process for fabricating this structure. In contrast to the conventional practice of fabricating fluid channels as trenches or grooves in a substrate, fluid channels are fabricated as thin walled raised structures on a substrate. Microfluidic devices produced in accordance with the invention are a hybrid assembly generally consisting of three layers: 1) a substrate that can or cannot be an electrical insulator; 2) a middle layer, that is an electrically conducting material and preferably silicon, forms the channel walls whose height defines the channel height, joined to and extending from the substrate; and 3) a top layer, joined to the top of the channels, that forms a cover for the channels. The channels can be defined by photolithographic techniques and are produced by etching away the material around the channel walls.

  5. VUV fluorescence following electron-impact dissociative excitation of CS{sub 2}

    SciTech Connect

    Brotton, S. J.; McConkey, J. W.

    2011-01-15

    Electron-impact dissociation of CS{sub 2} has been studied by observation of the atomic spectral emission features in the range 115-170 nm. Absolute photoemission cross sections are presented over the complete wavelength range for an incident electron energy of 100 eV. As an example, the measured cross section of the strong C i emission at 165.7 nm, which is a prominent feature in many solar and other extraterrestrial spectra, is (1.45{+-}0.19)x10{sup -18} cm{sup 2}. Comparison with earlier cross-sectional measurements suggest that these were too high by a factor of more than three. Excitation functions of the dominant C i (156.1 nm) and S i (147.4 nm) emission lines have been measured for electron-impact energies from threshold to 360 eV. From appearance energy measurements in the near-threshold region, likely fragmentation channels are identified which involve both two-fragment breakup and total fragmentation of the parent CS{sub 2}.

  6. Low temperature phase transition and crystal structure of CsMgPO4

    NASA Astrophysics Data System (ADS)

    Orlova, Maria; Khainakov, Sergey; Michailov, Dmitriy; Perfler, Lukas; Langes, Christoph; Kahlenberg, Volker; Orlova, Albina

    2015-01-01

    CsMgPO4 doped with radioisotopes is a promising compound for usage as a radioactive medical source. However, a low temperature phase transition at temperatures close to ambient conditions (~-40 °C) was observed. Information about such kind of structural changes is important in order to understand whether it can cause any problem for medical use of this compound. The phase transition has been investigated in detail using synchrotron powder diffraction, Raman spectroscopy and DFT calculations. The structure undergoes a transformation from an orthorhombic modification, space group Pnma (RT phase) to a monoclinic polymorph, space group P21/n (LT phase). New LT modification adopts similar to RT but slightly distorted unit cell: a=9.58199(2) Å, b=8.95501(1) Å, c=5.50344(2) Å, β=90.68583(1)°, V=472.198(3) Å3. CsMgPO4 belongs to the group of framework compounds and is made up of strictly alternating MgO4- and PO4-tetrahedra sharing vertices. The cesium counter cations are located in the resulting channel-like cavities. Upon the transformation a combined tilting of the tetrahedra is observed. A comparison with other phase transitions in ABW-type framework compounds is given.

  7. New results on Efimov physics and the creation of RbCs molecules

    NASA Astrophysics Data System (ADS)

    Ferlaino, Francesca

    2011-05-01

    I will present new results from two different experiments on ultracold gases, both based on scattering length tunability near Feshbach resonances. In a first experiment, we investigate universal few-body physics with ultracold cesium atoms. We detect various Efimov states in a magnetic field range up to 1000G by measuring loss resonances in three-body recombination processes. We find in total five distinct Efimov resonances in the same spin channel, all resulting from different Feshbach resonances. Surprisingly, all Efimov resonances occur at the same value of the scattering length, which points to universality across all Feshbach resonances and rules out a significant variation of the three-body parameter. In a second experiment, we produce a degenerate or near-degenerate ultracold mixture of 87Rb and 133Cs atoms. We investigate the interspecies scattering properties by Feshbach spectroscopy and we determine the background inter-species scattering length to be unusually large and positive. Starting with magneto-associated Feshbach molecules, we perform spectroscopic measurements on the two-photon optical transition that will serve to transfer RbCs molecules to the rovibronic ground state via a STIRAP transfer scheme. This work constitutes a first step towards the production of a quantum gas of ground-state polar molecules.

  8. Ephedrine QoS: An Antidote to Slow, Congested, Bufferless NoCs

    PubMed Central

    Fang, Juan; Yao, Zhicheng; Sui, Xiufeng; Bao, Yungang

    2014-01-01

    Datacenters consolidate diverse applications to improve utilization. However when multiple applications are colocated on such platforms, contention for shared resources like networks-on-chip (NoCs) can degrade the performance of latency-critical online services (high-priority applications). Recently proposed bufferless NoCs (Nychis et al.) have the advantages of requiring less area and power, but they pose challenges in quality-of-service (QoS) support, which usually relies on buffer-based virtual channels (VCs). We propose QBLESS, a QoS-aware bufferless NoC scheme for datacenters. QBLESS consists of two components: a routing mechanism (QBLESS-R) that can substantially reduce flit deflection for high-priority applications and a congestion-control mechanism (QBLESS-CC) that guarantees performance for high-priority applications and improves overall system throughput. We use trace-driven simulation to model a 64-core system, finding that, when compared to BLESS, a previous state-of-the-art bufferless NoC design, QBLESS, improves performance of high-priority applications by an average of 33.2% and reduces network-hops by an average of 42.8%. PMID:25250386

  9. A CsI low-temperature detector for dark matter search

    NASA Astrophysics Data System (ADS)

    Angloher, G.; Dafinei, I.; Gektin, A.; Gironi, L.; Gotti, C.; Gütlein, A.; Hauff, D.; Maino, M.; Nagorny, S. S.; Nisi, S.; Pagnanini, L.; Pattavina, L.; Pessina, G.; Petricca, F.; Pirro, S.; Pröbst, F.; Reindl, F.; Schäffner, K.; Schieck, J.; Seidel, W.; Vasyukov, S.

    2016-11-01

    Cryogenic detectors have a long history of success in the field of rare event searches. In particular scintillating calorimeters are very suitable detectors for this task since two signals are induced by a particle interaction in a scintillating crystal. The thermal signal provides a precise measurement of the deposited energy while the simultaneously measured scintillation light signal yields particle discrimination as the amount of produced scintillation light depends on the nature of the interacting particle. We investigate the calorimetric properties and background rejection capabilities of two large CsI (undoped) crystals (∼122 g each) operated as scintillating calorimeters at milli-Kelvin temperatures. Furthermore, we discuss the feasibility of this detection approach towards a future background-free dark matter experiment based on alkali halide crystals, with active particle discrimination via the two-channel detection.

  10. The OH-Initiated Oxidation of CS2 in the Presence of NO: FTIR Matrix-Isolation and Theoretical Studies.

    PubMed

    Bil, A; Grzechnik, K; Sałdyka, M; Mielke, Z

    2016-09-01

    We studied the photochemistry of the carbon disulfide-nitrous acid system with the help of Fourier transform infrared (FTIR) matrix isolation spectroscopy and theoretical methods. The irradiation of the CS2···HONO complexes, isolated in solid argon, with the filtered output of the mercury lamp (λ > 345 nm) was found to produce OCS, SO2, and HNCS; HSCN was also tentatively identified. The (13)C, (15)N, and (2)H isotopic shifts as well as literature data were used for product identifications. The evolution of the measured FTIR spectra with irradiation time and the changes in the spectra after matrix annealing indicated that the identified molecules are the products of different reaction channels: OCS being a product of another reaction path than SO2 and HNCS or HSCN. The possible reaction channels between SC(OH)S/SCS(OH) radicals and NO were studied using DFT/B3LYP/aug-cc-pVTZ method. The SC(OH)S and/or SCS(OH) intermediates are formed when HONO attached to CS2 photodissociates into OH and NO. The calculations indicated that SC(OH)S radical can form with NO two stable adducts. The more stable SC(OH)S···NO structure is a reactant for a simple one-step process leading to OCS and HONS molecules. An alternative, less-stable complex formed between SC(OH)S and NO leads to formation of OCS and HSNO. The calculations predict only one stable complex between SCS(OH) radical and NO, which can dissociate along two channels leading to HNCS and SO2 or HSCN and SO2 as the end products. The identified photoproducts indicate that both SC(OH)S and SCS(OH) adducts are intermediates in the CS2 + OH + NO reaction leading to different reaction products. PMID:27491274

  11. Three new phosphates with isolated P2O7 units: noncentrosymmetric Cs2Ba3(P2O7)2 and centrosymmetric Cs2BaP2O7 and LiCsBaP2O7.

    PubMed

    Li, Lin; Han, Shujuan; Lei, Bing-Hua; Wang, Ying; Li, Hongyi; Yang, Zhihua; Pan, Shilie

    2016-03-01

    Three new phosphates, a noncentrosymmetric (NCS) Cs2Ba3(P2O7)2 and centrosymmetric (CS) Cs2BaP2O7 and LiCsBaP2O7, have been synthesized from high-temperature solutions for the first time. Analysis of the structures determined by single-crystal X-ray diffraction showed that although the three compounds contained isolated P2O7 units, they yielded different three-dimensional (3D) networks: Cs2Ba3(P2O7)2 crystallized in the NCS Orthorhombic space group P212121, Cs2BaP2O7 in the CS monoclinic space group P21/n, and LiCsBaP2O7, having an identical stoichiometry with Cs2BaP2O7, crystallized in monoclinic space group, P21/c. Structural comparisons suggested the differences between their 3D frameworks to be due to differences between the sizes and coordination environments of the cations. Characterizations including thermal and optical analyses showed Cs2Ba3(P2O7)2 and Cs2BaP2O7 to melt congruently, and Cs2Ba3(P2O7)2 to exhibit a wide transparent region with a cut-off edge below 176 nm. The NLO properties and electronic structures of these compounds were investigated using first-principles calcualtions. PMID:26831497

  12. Mapping of the putative epitope domain of Clonorchis sinensis paramyosin (CsPmy) recognized by CsPmy-specific immunoglobulin G in sera of human clonorchiasis.

    PubMed

    Kang, Jung-Mi; Ju, Hye-Lim; Lee, Jinyoung; Kim, Tae Im; Cho, Shin-Hyeong; Kim, Tong-Soo; Sohn, Woon-Mok; Na, Byoung-Kuk

    2015-05-01

    Paramyosin of Clonorchis sinensis (CsPmy) is a myofibrillar protein localized in subtegumental muscle, tegument, and the muscle layer surrounding the intestine of the parasite. Previously, we have identified that CsPmy reacted with sera of human clonorchiasis and this protein had a potential as a candidate antigen for serodiagnosis of clonorchiasis. However, we also found that CsPmy is able to bind to human immunoglobulin G (IgG) in non-specific manners, which can affect the diagnostic value of the protein. Here, we mapped CsPmy-specific IgG binding site on CsPmy to analyze the putative epitopes recognized by CsPmy-specific IgG in sera of human clonorchiasis. The fragmental expression of CsPmy followed by immunoblot analyses with sera from patients with clonorchiasis and non-specific human IgG revealed that the middle portion of CsPmy (CsPmyC: 301-600 amino acid residues) had epitopes responsible for CsPmy-specific IgG recognition. The precise CsPmy-specific IgG binding site was further narrowed down to a fragment (CsPmyC-2), which harbors 151 amino acid residues (375-525) of CsPmy. Specific antibodies for CsPmyC-2 were produced in rats after two-weeks of post-experimental infection. The CsPmyC-2 showed low levels of cross reactivity against the sera from patients with other helminth parasites. Our results suggested that CsPmyC-2 has real epitopes recognized by CsPmy-specific IgG in sera of human clonorchiasis and the fragment can be useful as a reliable serodiagnostic antigen to develop a serodiagnostic method for clonorchiasis. PMID:26099940

  13. Critical assessment of OmpF channel selectivity: merging information from different experimental protocols

    NASA Astrophysics Data System (ADS)

    López, M. L.; García-Giménez, E.; Aguilella, V. M.; Alcaraz, A.

    2010-11-01

    The ion selectivity of a channel can be quantified in several ways by using different experimental protocols. A wide, mesoscopic channel, the OmpF porin of the outer membrane of E. coli, serves as a case study for comparing and analysing several measures of the channel cation-anion permeability in chlorides of alkali metals (LiCl, NaCl, KCl, CsCl). We show how different insights can be gained and integrated to rationalize the global image of channel selectivity. To this end, reversal potential, channel conductance and bi-ionic potential (two different salts with a common anion on each side of the channel but with the same concentration) experiments are discussed in light of an electrodiffusion model based on the Poisson-Nernst-Planck formalism. Measurements and calculations based on the atomic crystal structure of the channel show that each protocol displays a particular balance between the different sources of selectivity.

  14. Channel Properties of Nax Expressed in Neurons

    PubMed Central

    Matsumoto, Masahito; Hiyama, Takeshi Y.; Kuboyama, Kazuya; Suzuki, Ryoko; Fujikawa, Akihiro; Noda, Masaharu

    2015-01-01

    Nax is a sodium-concentration ([Na+])-sensitive Na channel with a gating threshold of ~150 mM for extracellular [Na+] ([Na+]o) in vitro. We previously reported that Nax was preferentially expressed in the glial cells of sensory circumventricular organs including the subfornical organ, and was involved in [Na+] sensing for the control of salt-intake behavior. Although Nax was also suggested to be expressed in the neurons of some brain regions including the amygdala and cerebral cortex, the channel properties of Nax have not yet been adequately characterized in neurons. We herein verified that Nax was expressed in neurons in the lateral amygdala of mice using an antibody that was newly generated against mouse Nax. To investigate the channel properties of Nax expressed in neurons, we established an inducible cell line of Nax using the mouse neuroblastoma cell line, Neuro-2a, which is endogenously devoid of the expression of Nax. Functional analyses of this cell line revealed that the [Na+]-sensitivity of Nax in neuronal cells was similar to that expressed in glial cells. The cation selectivity sequence of the Nax channel in cations was revealed to be Na+ ≈ Li+ > Rb+ > Cs+ for the first time. Furthermore, we demonstrated that Nax bound to postsynaptic density protein 95 (PSD95) through its PSD95/Disc-large/ZO-1 (PDZ)-binding motif at the C-terminus in neurons. The interaction between Nax and PSD95 may be involved in promoting the surface expression of Nax channels because the depletion of endogenous PSD95 resulted in a decrease in Nax at the plasma membrane. These results indicated, for the first time, that Nax functions as a [Na+]-sensitive Na channel in neurons as well as in glial cells. PMID:25961826

  15. Cholesterol and Ion Channels

    PubMed Central

    Levitan, Irena; Fang, Yun; Rosenhouse-Dantsker, Avia; Romanenko, Victor

    2010-01-01

    A variety of ion channels, including members of all major ion channel families, have been shown to be regulated by changes in the level of membrane cholesterol and partition into cholesterol-rich membrane domains. In general, several types of cholesterol effects have been described. The most common effect is suppression of channel activity by an increase in membrane cholesterol, an effect that was described for several types of inwardly-rectifying K+ channels, voltage-gated K+ channels, Ca+2 sensitive K+ channels, voltage-gated Na+ channels, N-type voltage-gated Ca+2 channels and volume-regulated anion channels. In contrast, several types of ion channels, such as epithelial amiloride-sensitive Na+ channels and Transient Receptor Potential channels, as well as some of the types of inwardly-rectifying and voltage-gated K+ channels were shown to be inhibited by cholesterol depletion. Cholesterol was also shown to alter the kinetic properties and current-voltage dependence of several voltage-gated channels. Finally, maintaining membrane cholesterol level is required for coupling ion channels to signalling cascades. In terms of the mechanisms, three general mechanisms have been proposed: (i) specific interactions between cholesterol and the channel protein, (ii) changes in the physical properties of the membrane bilayer and (iii) maintaining the scaffolds for protein-protein interactions. The goal of this review is to describe systematically the role of cholesterol in regulation of the major types of ion channels and to discuss these effects in the context of the models proposed. PMID:20213557

  16. Fading channel simulator

    SciTech Connect

    Argo, P.E.; Fitzgerald, T.J.

    1991-12-31

    This invention relates to high frequency (HF) radio signal propagation through fading channels and, more particularly, to simulation of fading channels in order to characterize HF radio system performance in transmitting and receiving signals through such fading channels. Fading channel effects on a transmitted communication signal are simulated with both frequency and time variations using a channel scattering function to affect the transmitted signal. A conventional channel scattering function is converted to a series of channel realizations by multiplying the square root of the channel scattering function by a complex number of which the real and imaginary parts are each independent variables. The two-dimensional inverse-FFT of this complex-valued channel realization yields a matrix of channel coefficients that provide a complete frequency-time description of the channel. The transmitted radio signal is segmented to provide a series of transmitted signal and each segment is subject to FFT to generate a series of signal coefficient matrices. The channel coefficient matrices and signal coefficient matrices are then multiplied and subjected to inverse-FFT to output a signal representing the received affected radio signal. A variety of channel scattering functions can be used to characterize the response of a transmitter-receiver system to such atmospheric effects.

  17. Development of Jacketing Technologies for Iter CS and TF Conductor

    NASA Astrophysics Data System (ADS)

    Hamada, K.; Nakajima, H.; Matsui, K.; Kawano, K.; Takano, K.; Tsutsumi, F.; Okuno, K.; Teshima, O.; Soejima, K.

    2008-03-01

    The Japan Atomic Energy Agency (JAEA) has developed jacketing technologies for ITER Toroidal Field (TF) and Central Solenoid (CS) conductor. Full scale TF and CS conduits were fabricated using carbon-reduced SUS316LN and boron-added (˜40 ppm) high manganese stainless steel (0.025C -22Mn -13Cr -9Ni -0.12N: JK2LB), respectively. Welding condition was optimized so that back bead does not interfere a cable insertion. The weld joint samples were compacted by a compaction machine that was newly constructed and tested at 4.2 K. Mechanical characteristics at 4K of CS, TF conduits and CS welded joint satisfied ITER mechanical requirements. TF welded joint shows slightly lower value of 0.2% yield strength (885 MPa) than that of ITER requirement (900 MPa). The TF conduit contains nitrogen content of 0.14%, which is minimum value in ITER specification. The lower nitrogen content may be caused by the release of nitrogen from molten metal during non-filler welding resulting in a 4 K strength decrease. To satisfy the ITER requirements, minimum nitrogen contents of conduit should be increased from 0.14% to 0.15% at least. Therefore, JAEA successfully developed TF and CS conduits with welding technologies and finalized the procurement specification for ITER conductor jacketing.

  18. Photoelectron Emission Studies in CsBr at 257 nm

    SciTech Connect

    Maldonado, Juan R.; Liu, Zhi; Sun, Yun; Pianetta, Piero A.; Pease, Fabian W.; /Stanford U., Elect. Eng. Dept. /SLAC, SSRL

    2006-09-28

    CsBr/Cr photocathodes were found [1,2] to meet the requirements of a multi-electron beam lithography system operating with a light energy of 4.8 eV (257nm). The fact that photoemission was observed with a light energy below the reported 7.3 eV band gap for CsBr was not understood. This paper presents experimental results on the presence of intra-band gap absorption sites (IBAS) in CsBr thin film photo electron emitters, and presents a model based on IBAS to explain the observed photoelectron emission behavior at energies below band gap. A fluorescence band centered at 330 nm with a FWHM of about 0.34 eV was observed in CsBr/Cr samples under 257 nm laser illumination which can be attributed to IBAS and agrees well with previously obtained synchrotron photoelectron spectra[1] from the valence band of CsBr films.

  19. Electron induced surface chemistry at the Cs/sapphire interface

    SciTech Connect

    Zavadil, K.R.; Ing, J.L.

    1995-11-01

    Electron induced etching of sapphire in the presence of Cs has been studied using a variety of surface analytical techniques. We find that this process occurs on both the (0001) and (1102) orientations of sapphire. Monolayer amounts of Al and sub-oxides of Al are thermally desorbed from the surface at temperatures as low as 1000 K when the surface is irradiated with electrons in the presence of Cs. Etching is highly dependent on Cs coverage with the (0001) and (1102) surfaces requiring 2.0 {times} 10{sup 14} and 3.4 {times} 10{sup 14} atoms/cm{sup 2} to support etching, respectively. Adsorption profiles demonstrate that these coverages correspond to initial saturation of the surface with Cs. Electron damage of the surface in the absence of Cs also produces desorption of Al and sub-oxides of Al indicating a possible mechanism for etching. The impact of etching on the surface is to increase the adsorption capacity on the (0001) surface while decreasing both initial adsorption probability and capacity on the (1102) surface.

  20. In situ7Li and 133Cs nuclear magnetic resonance investigations on the role of Cs+ additive in lithium-metal deposition process

    NASA Astrophysics Data System (ADS)

    Hu, Jian Zhi; Zhao, Zhenchao; Hu, Mary Y.; Feng, Ju; Deng, Xuchu; Chen, Xilin; Xu, Wu; Liu, Jun; Zhang, Ji-Guang

    2016-02-01

    Cesium ion (Cs+) has been reported to be an effective electrolyte additive to suppress Li dendrite growth which prevents the application of lithium (Li) metal as an anode for rechargeable Li batteries. In this work, we investigated the effect of Cs+ additive on Li depositions using quantitative in situ7Li and 133Cs nuclear magnetic resonance (NMR) with planar symmetric Li cells. It's found that the addition of Cs+ can significantly enhance both the formation of well aligned Li nanorods and reversibility of the Li electrode. In situ133Cs NMR directly confirms that Cs+ migrates to Li electrode to form a positively charged electrostatic shield during the charging process. Much more electrochemical "active" Li was found in Li films deposited with Cs+ additive, while more electrochemical "dead" and thicker Li rods were identified in Li films deposited without Cs+. Combining the in situ and the previous ex-situ results, a Li deposition model has been proposed to explain these observations.

  1. Mechanosensitivity of an epithelial Na+ channel in planar lipid bilayers: release from Ca2+ block.

    PubMed Central

    Ismailov, I I; Berdiev, B K; Shlyonsky, V G; Benos, D J

    1997-01-01

    A family of novel epithelial Na+ channels (ENaCs) have recently been cloned from several different tissues. Three homologous subunits (alpha, beta, gamma-ENaCs) from the core conductive unit of Na(+)-selective, amiloride-sensitive channels that are found in epithelia. We here report the results of a study assessing the regulation of alpha,beta,gamma-rENaC by Ca2+ in planar lipid bilayers. Buffering of the bilayer bathing solutions to [Ca2+] < 1 nM increased single-channel open probability by fivefold. Further investigation of this phenomenon revealed that Ca2+ ions produced a voltage-dependent block, affecting open probability but not the unitary conductance of ENaC. Imposing a hydrostatic pressure gradient across bilayers containing alpha,beta,gamma-rENaC markedly reduced the sensitivity of these channels to inhibition by [Ca2+]. Conversely, in the nominal absence of Ca2+, the channels lost their sensitivity to mechanical stimulation. These results suggest that the previously observed mechanical activation of ENaCs reflects a release of the channels from block by Ca2+. Images FIGURE 3 FIGURE 4 PMID:9138565

  2. Optimal channels for channelized quadratic estimators.

    PubMed

    Kupinski, Meredith K; Clarkson, Eric

    2016-06-01

    We present a new method for computing optimized channels for estimation tasks that is feasible for high-dimensional image data. Maximum-likelihood (ML) parameter estimates are challenging to compute from high-dimensional likelihoods. The dimensionality reduction from M measurements to L channels is a critical advantage of channelized quadratic estimators (CQEs), since estimating likelihood moments from channelized data requires smaller sample sizes and inverting a smaller covariance matrix is easier. The channelized likelihood is then used to form ML estimates of the parameter(s). In this work we choose an imaging example in which the second-order statistics of the image data depend upon the parameter of interest: the correlation length. Correlation lengths are used to approximate background textures in many imaging applications, and in these cases an estimate of the correlation length is useful for pre-whitening. In a simulation study we compare the estimation performance, as measured by the root-mean-squared error (RMSE), of correlation length estimates from CQE and power spectral density (PSD) distribution fitting. To abide by the assumptions of the PSD method we simulate an ergodic, isotropic, stationary, and zero-mean random process. These assumptions are not part of the CQE formalism. The CQE method assumes a Gaussian channelized likelihood that can be a valid for non-Gaussian image data, since the channel outputs are formed from weighted sums of the image elements. We have shown that, for three or more channels, the RMSE of CQE estimates of correlation length is lower than conventional PSD estimates. We also show that computing CQE by using a standard nonlinear optimization method produces channels that yield RMSE within 2% of the analytic optimum. CQE estimates of anisotropic correlation length estimation are reported to demonstrate this technique on a two-parameter estimation problem. PMID:27409452

  3. Atomic sites and stability of Cs+ captured within zeolitic nanocavities.

    PubMed

    Yoshida, Kaname; Toyoura, Kazuaki; Matsunaga, Katsuyuki; Nakahira, Atsushi; Kurata, Hiroki; Ikuhara, Yumi H; Sasaki, Yukichi

    2013-01-01

    Zeolites have potential application as ion-exchangers, catalysts and molecular sieves. Zeolites are once again drawing attention in Japan as stable adsorbents and solidification materials of fission products, such as (137)Cs(+) from damaged nuclear-power plants. Although there is a long history of scientific studies on the crystal structures and ion-exchange properties of zeolites for practical application, there are still open questions, at the atomic-level, on the physical and chemical origins of selective ion-exchange abilities of different cations and detailed atomic structures of exchanged cations inside the nanoscale cavities of zeolites. Here, the precise locations of Cs(+) ions captured within A-type zeolite were analyzed using high-resolution electron microscopy. Together with theoretical calculations, the stable positions of absorbed Cs(+) ions in the nanocavities are identified, and the bonding environment within the zeolitic framework is revealed to be a key factor that influences the locations of absorbed cations.

  4. Atomic sites and stability of Cs+ captured within zeolitic nanocavities

    NASA Astrophysics Data System (ADS)

    Yoshida, Kaname; Toyoura, Kazuaki; Matsunaga, Katsuyuki; Nakahira, Atsushi; Kurata, Hiroki; Ikuhara, Yumi H.; Sasaki, Yukichi

    2013-08-01

    Zeolites have potential application as ion-exchangers, catalysts and molecular sieves. Zeolites are once again drawing attention in Japan as stable adsorbents and solidification materials of fission products, such as 137Cs+ from damaged nuclear-power plants. Although there is a long history of scientific studies on the crystal structures and ion-exchange properties of zeolites for practical application, there are still open questions, at the atomic-level, on the physical and chemical origins of selective ion-exchange abilities of different cations and detailed atomic structures of exchanged cations inside the nanoscale cavities of zeolites. Here, the precise locations of Cs+ ions captured within A-type zeolite were analyzed using high-resolution electron microscopy. Together with theoretical calculations, the stable positions of absorbed Cs+ ions in the nanocavities are identified, and the bonding environment within the zeolitic framework is revealed to be a key factor that influences the locations of absorbed cations.

  5. Spin-exchange-relaxation-free magnetometry with Cs vapor

    NASA Astrophysics Data System (ADS)

    Ledbetter, M. P.; Savukov, I. M.; Acosta, V. M.; Budker, D.; Romalis, M. V.

    2008-03-01

    We describe a Cs atomic magnetometer operating in the spin-exchange-relaxation-free (SERF) regime. With a vapor cell temperature of 103°C we achieve intrinsic magnetic resonance widths ΔB=17μG corresponding to an electron spin-relaxation rate of 300s-1 when the spin-exchange rate is ΓSE=14000s-1 . We also observe an interesting narrowing effect due to diffusion. Signal-to-noise measurements yield a sensitivity of about 400pG/Hz . Based on photon shot noise, we project a sensitivity of 40pG/Hz . A theoretical optimization of the magnetometer indicates sensitivities on the order of 2pG/Hz should be achievable in a 1cm3 volume. Because Cs has a higher saturated vapor pressure than other alkali metals, SERF magnetometers using Cs atoms are particularly attractive in applications requiring lower temperatures.

  6. CsBr/GaN Heterojunction Photoelectron Source

    SciTech Connect

    Maldonado, J.R.; Liu, Z.; Sun, Y.; Schuetter, S.; Pianetta, P.; Pease, R.F.W.; /Stanford U., Elect. Eng. Dept.

    2009-04-30

    Experimental results on a new CsBr/GaN heterojunction photocathode structure are presented. The results indicate a fourfold improvement in photoyield relative to CsBr/Cr photocathodes. A model is presented based on intraband states in CsBr and electron injection from the GaN (with 1% addition of indium) substrate to explain the observed photoyield enhancement. The photocathode lifetime at high current density (>40 A/cm{sup 2}) is limited by laser heating of the small illuminated area. Calculations are presented for sapphire and diamond substrates, indicating a factor of 20 reduction in temperature for the latter. The results are encouraging for the realization of a high photoyield photocathode operating at high current density with long lifetime.

  7. Quantum Efficiency Enhancement in CsI/Metal Photocathodes

    SciTech Connect

    Kong, Lingmei; Joly, Alan G.; Droubay, Timothy C.; Hess, Wayne P.

    2015-02-01

    High quantum efficiency enhancement is found for hybrid metal-insulator photocathodes consisting of thin films of CsI deposited on Cu(100), Ag(100), Au(111) and Au films irradiated by 266 nm laser pulses. Low work functions (near or below 2 eV) are observed following ultraviolet laser activation. Work functions are reduced by roughly 3 eV from that of clean metal surfaces. We discuss various mechanisms of quantum efficiency enhancement for alkali halide/metal photocathode systems and conclude that the large change in work function, due to Cs accumulation of Cs metal at the metal-alkali halide interface, is the dominant mechanism for quantum efficiency enhancement

  8. Quasi-freestanding graphene on Ni(111) by Cs intercalation

    NASA Astrophysics Data System (ADS)

    Alattas, M.; Schwingenschlögl, U.

    2016-05-01

    A possible approach to achieve quasi-freestanding graphene on a substrate for technological purpose is the intercalation of alkali metal atoms. Cs intercalation between graphene and Ni(111) therefore is investigated using density functional theory, incorporating van der Waals corrections. It is known that direct contact between graphene and Ni(111) perturbs the Dirac states. We find that Cs intercalation restores the linear dispersion characteristic of Dirac fermions, which agrees with experiments, but the Dirac cone is shifted to lower energy, i.e., the graphene sheet is n-doped. Cs intercalation therefore decouples the graphene sheet from the substrate except for a charge transfer. On the other hand, the spin polarization of Ni(111) does not extend through the intercalated atoms to the graphene sheet, for which we find virtually spin-degeneracy.

  9. Energetics and structural stability of Cs3C60

    SciTech Connect

    Saito, Susumu; Umemoto, Koichiro; Louie, Steven G.; Cohen, MarvinL.

    2003-12-15

    Using the ab initio pseudo potential total-energy method and the density-functional theory, we study the energetics of face-centered-cubic Cs3C60 which is a material of great interest as a possible high transition-temperature superconductor. At the optimized lattice constant the volume per C60 is found to be smaller than the most stable hexagon-coordination A15 phase, while the total energy of the fcc phase is about 0.9 eV higher than the A15 phase. These results indicate that a low-temperature and high-pressure synthesis method might be a possible way to produce the fcc Cs3C60 phase. In addition, it is also found that the A15 Cs3C60 should show a phase transformation from a hexagon-coordination phase to a pentagon-coordination phase under hydrostatic pressure.

  10. Accumulation of K+ and Cs+ in Tropical Plant Species

    NASA Astrophysics Data System (ADS)

    Velasco, H.; Anjos, R. M.; Zamboni, C. B.; Macario, K. D.; Rizzotto, M.; Cid, A. S.; Medeiros, I. M. A.; Fernández, J.; Rubio, L.; Audicio, P.; Lacerda, T.

    2010-08-01

    Concentrations of K+ and 137Cs+ in tissues of the Citrus aurantifolia were measured both by gamma spectrometry and neutron activation analysis, aiming to understand the behavior of monovalent inorganic cations in plants as well as its capability to store these elements. In contrast to K+, Cs+ ions are not essential elements to plants, what might explain the difference in bioavailability. However, our results have shown that 137Cs+ is positively correlated to 40K+ concentration within tropical plant species, suggesting that these elements might be assimilated in a similar way, and that they pass through the biological cycle together. A simple mathematical model was also proposed to describe the temporal evolution of 40K activity concentration in such tropical woody fruit species. This model exhibited close agreement with the 40K experimental results in the fruit ripening processes of lemon trees.

  11. Fundamental studies on the Cs dynamics under ion source conditions

    SciTech Connect

    Friedl, R. Fantz, U.

    2014-02-15

    The performance of surface conversion based negative hydrogen ion sources is mainly determined by the caesium dynamics. Therefore, fundamental investigations in vacuum and plasma are performed at a flexible laboratory setup with ion source parameters. Studies on the influence of Cs on the plasma parameters of H{sub 2} and D{sub 2} plasmas showed that n{sub e} and T{sub e} in the bulk plasma are not affected by relevant amounts of Cs and no isotopic differences could be observed. The coating of the vessel surfaces with Cs, however, leads to a considerable gettering of hydrogen atoms from the plasma volume and to the decrease of n{sub e} close to a sample surface due to the formation of negative ions.

  12. Ion channels and cancer.

    PubMed

    Kunzelmann, Karl

    2005-06-01

    Membrane ion channels are essential for cell proliferation and appear to have a role in the development of cancer. This has initially been demonstrated for potassium channels and is meanwhile also suggested for other cation channels and Cl- channels. For some of these channels, like voltage-gated ether à go-go and Ca2+-dependent potassium channels as well as calcium and chloride channels, a cell cycle-dependent function has been demonstrated. Along with other membrane conductances, these channels control the membrane voltage and Ca2+ signaling in proliferating cells. Homeostatic parameters, such as the intracellular ion concentration, cytosolic pH and cell volume, are also governed by the activity of ion channels. Thus it will be an essential task for future studies to unravel cell cycle-specific effects of ion channels and non-specific homeostatic functions. When studying the role of ion channels in cancer cells, it is indispensable to choose experimental conditions that come close to the in vivo situation. Thus, environmental parameters, such as low oxygen pressure, acidosis and exposure to serum proteins, have to be taken into account. In order to achieve clinical application, more studies on the original cancer tissue are required, and improved animal models. Finally, it will be essential to generate more potent and specific inhibitors of ion channels to overcome the shortcomings of some of the current approaches.

  13. Patterns and dynamics of Cs-137 soil contamination on the plot scale of the Bryansk Region (Russia): the role of processes, connectivity

    NASA Astrophysics Data System (ADS)

    Linnik, Vitaly; Sokolov, Alexander; Saveliev, Anatoly

    2014-05-01

    parameters, general additive models were used. According to results of modeling using a detailed and a generalized grid it has been found (Linnik, Saveliev et.al., 2007), that in accumulation zones (depressions) 137Cs deposit was lower when Laplace operator was positive (Laplace1>0=915 kBq/m2; Laplace2>0=921 kBq/m2) than in wash-out zones, singled out by negative values of Laplace operator (Laplace1<0=978 kBq/m2; Laplace2<0=979 kBq/m2). The inversion effect revealed in 137Cs deposit distribution could not be accounted for be processes of surface 137Cs wash-off as the chain of depressions was isolated. We found that connectivity of subsurface moving soil moisture saturation was made up by a number of small and shallow channels, covered by litter, they served as 137Cs travel paths at the period of spring wetting in April-May 1986. The total 137Cs output in soluble form from this plot calculated for the two models was 5,9% and 6,4%. References: Linnik V.G., Saveliev A.A., Govorun A.P., Ivanitsky O.M., Sokolov A.V. Spatial Variability and Topographic Factors of 137Cs Soil Contamination at a Field Scale// International Journal of Ecology & Development, 2007, Vol. 8, No.7, p.8-25.

  14. Estimation of average burnup of damaged fuels loaded in Fukushima Dai-ichi reactors by using the {sup 134}Cs/{sup 137}Cs ratio method

    SciTech Connect

    Endo, T.; Sato, S.; Yamamoto, A.

    2012-07-01

    Average burnup of damaged fuels loaded in Fukushima Dai-ichi reactors is estimated, using the {sup 134}Cs/{sup 137}Cs ratio method for measured radioactivities of {sup 134}Cs and {sup 137}Cs in contaminated soils within the range of 100 km from the Fukushima Dai-ichi nuclear power plants. As a result, the measured {sup 134}Cs/{sup 137}Cs ratio from the contaminated soil is 0.996{+-}0.07 as of March 11, 2011. Based on the {sup 134}Cs/{sup 137}Cs ratio method, the estimated burnup of damaged fuels is approximately 17.2{+-}1.5 [GWd/tHM]. It is noted that the numerical results of various calculation codes (SRAC2006/PIJ, SCALE6.0/TRITON, and MVP-BURN) are almost the same evaluation values of {sup 134}Cs/ {sup 137}Cs ratio with same evaluated nuclear data library (ENDF-B/VII.0). The void fraction effect in depletion calculation has a major impact on {sup 134}Cs/{sup 137}Cs ratio compared with the differences between JENDL-4.0 and ENDF-B/VII.0. (authors)

  15. Comparison of resonance ionization mass spectrometry systems for the determination of 135Cs/137Cs isotope ratios

    NASA Astrophysics Data System (ADS)

    Pibida, Leticia S.; McMahon, C. A.; Nortershauser, Wilfried; Bushaw, Bruce A.

    2002-03-01

    The performance of the Resonance Ionization Mass Spectrometry (RIMS) system at the National Institute of Standards and Technology (NIST) has been compared to a similar system at Pacific Northwest National Laboratory (PNNL). Efficiency and selectivity measurements were performed with both systems and compared to conventional thermal ionization mass spectrometry (TIMS). Determination of the 135Cs / 137Cs ratio was performed using single-resonance excitation 6s 2S1/2 (F equals 4) to 6p 2P3/2 (F equals 5) with an extended cavity diode laser followed by photoionization with the 488 nm line of an argon ion laser. Optical selectivity of more than 2 orders of magnitude against stable 133Cs was attained for 135Cs and 137Cs for both systems with an overall selectivity of 109 for the PNNL system and 108 for the NIST system. Overall efficiencies of 2x10-6 and 5x10-7 were measured for the PNNL and NIST systems respectively. Measurements to determine the chronological age of a nuclear burn-up sample have been performed using both RIMS systems as well as TIMS.

  16. G-protein-coupled inward rectifier potassium channels involved in corticostriatal presynaptic modulation.

    PubMed

    Meneses, David; Mateos, Verónica; Islas, Gustavo; Barral, Jaime

    2015-09-01

    Presynaptic modulation has been associated mainly with calcium channels but recent data suggests that inward rectifier potassium channels (K(IR)) also play a role. In this work we set to characterize the role of presynaptic K(IR) channels in corticostriatal synaptic transmission. We elicited synaptic potentials in striatum by stimulating cortical areas and then determined the synaptic responses of corticostriatal synapsis by using paired pulse ratio (PPR) in the presence and absence of several potassium channel blockers. Unspecific potassium channels blockers Ba(2+) and Cs(+) reduced the PPR, suggesting that these channels are presynaptically located. Further pharmacological characterization showed that application of tertiapin-Q, a specific K(IR)3 channel family blocker, also induced a reduction of PPR, suggesting that K(IR)3 channels are present at corticostriatal terminals. In contrast, exposure to Lq2, a specific K(IR)1.1 inward rectifier potassium channel, did not induce any change in PPR suggesting the absence of these channels in the presynaptic corticostriatal terminals. Our results indicate that K(IR)3 channels are functionally expressed at the corticostriatal synapses, since blockage of these channels result in PPR decrease. Our results also help to explain how synaptic activity may become sensitive to extracellular signals mediated by G-protein coupled receptors. A vast repertoire of receptors may influence neurotransmitter release in an indirect manner through regulation of K(IR)3 channels.

  17. Cs-137 concentration in reindeer and its fodder plants.

    PubMed

    Rissanen, K; Rahola, T

    1989-09-01

    Radionuclides, especially the long-lived 137Cs (physical half-life 30 years), are accumulated efficiently in the northern, subarctic, lichen-reindeer-man foodchain. Until the Chernobyl accident the fallout nuclides studied originated from nuclear weapons tests. After this accident some fresh fallout was deposited in Finnish Lapland. Lichens grow very slowly and collect nutrients very efficiently from air, rain and snow. During winter the basic fodder plants for reindeer are lichens and some winter-green plants, shrubs and dry leaves. During the bare-ground season, the reindeer eat various grasses, herbs and leaves etc. Lichens constitute 30-50 per cent of the entire vegetable mass consumed by the reindeer in a year. The highest 137Cs-concentration 2500 Bq/kg dry weight was found in lichen in the middle of the 1960s. In 1985 the concentration had decreased to about 240 Bq/kg dry weight. After the Chernobyl accident the 137Cs-concentration in lichen varied from 200 to 2000 Bq/kg dry weight in Finnish Lapland. In reindeer fodder plant samples collected in the 1980s before the Chernobyl accident the 137Cs-concentration varied from 5 to 970 Bq/kg dry weight. The highest 137Cs-concentration in reindeer meat, about 2500 Bq/kg fresh weight, was found in 1965 and thereafter decreased to about 300 Bq/kg fresh weight in the winter before the Chernobyl accident. After the accident the mean 137Cs-concentration in reindeer meat from the 1986-87 slaughtering period was 720 Bq/kg fresh weight and in 1987-88, 630 Bq/kg fresh weight. PMID:2814447

  18. Genetic effects of testicular incorporation of 137Cs in mice.

    PubMed

    Ramaiya, L K; Pomerantseva, M D; Chekhovich, A V; Lyaginskaya, A M; Kuznetsov, A S

    1994-08-01

    A comparative estimation of the frequencies of genetic disorders induced in germ cells of male mice by a single or long-term exposure to incorporated 137Cs or to external gamma-radiation has been carried out. The frequencies of dominant lethal mutations induced by a single exposure were similar with both types of radiation. In stem cell spermatogonia the frequency of reciprocal translocations was significantly lower in the case of single 137Cs administration than upon external gamma-radiation. Upon long-term administration the genetic efficiencies of both types of radiation were similar. PMID:7519738

  19. Naphthomycins L-N, ansamycin antibiotics from Streptomyces sp. CS.

    PubMed

    Yang, Yin-He; Fu, Xiao-Li; Li, Liang-Qun; Zeng, Ying; Li, Cheng-Yun; He, Yi-Neng; Zhao, Pei-Ji

    2012-07-27

    Previous analyses of the naphthomycin biosynthetic gene cluster and a comparison with known naphthomycin-type products from Streptomyces sp. CS have suggested that new products can be found from this strain. In this study, screening by LC-MS of Streptomyces sp. CS products formed under different culture conditions revealed several unknown peaks in the product spectra of extracts derived from oatmeal medium cultures. Three new naphthomycins, naphthomycins L (1), M (2), and N (3), and the known naphthomycins A (4), E (5), and D (6) were obtained. The structures were elucidated using spectroscopic data from 1D and 2D NMR and HRESIMS experiments. PMID:22742732

  20. Photoemission from Ag, Cu, and CsI

    SciTech Connect

    Srinivasan-Rao, T.; Fischer, J.; Tsang, T.

    1992-06-01

    Photoemission characteristics of three different cathodes, CsI, Ag film and Cu were investigated. CsI, upon irradiation by 213 nm, 10ps laser pulse yields a quantum efficiency of 4% at O.2[mu]J input energy. The saturation mechanism observed at higher input energies require further investigation. Ag film, upon irradiation by 630 nm, 300 fs laser emit prompt photoelectrons after absorbing 2 photons. There was no evidence of optical damage of the film up to 10[sup 11] W/cm[sup 2]. At low intensities, photoemission from Cu is a simple [nu]-e[sup [minus

  1. Collisional frequency shifts in 133Cs fountain clocks.

    PubMed

    Leo, P J; Julienne, P S; Mies, F H; Williams, C J

    2001-04-23

    We present a theoretical analysis of the density dependent frequency shift in Cs fountain clocks using the highly constrained binary collision model described by Leo et al. [Phys. Rev. Lett. 85, 2721 (2000)]. We predict a reversal in the clock shift at temperatures near 0.08 microK. Our results show that s waves dominate the collision process. However, as a consequence of the large scattering lengths in Cs the clock shift is strongly temperature dependent and does not reach a constant Wigner-law value until temperatures are less than 0.1 nK.

  2. A winning combination: the 3Cs of business continuity.

    PubMed

    Glendon, Lee

    2013-01-01

    Contingency planning is a natural part of business life and is used across identified strategic, financial and operational risks. But is it being done well and is it the right approach all of the time? This paper shows how contingency planning forms one layer of a three-line defence termed 'the 3Cs of business continuity': contingency planning; continuity capability; crisis response. Collectively, 'the 3Cs' help organisations deliver a robust response to the risks that can be seen and those that cannot. PMID:24113636

  3. A winning combination: the 3Cs of business continuity.

    PubMed

    Glendon, Lee

    2013-01-01

    Contingency planning is a natural part of business life and is used across identified strategic, financial and operational risks. But is it being done well and is it the right approach all of the time? This paper shows how contingency planning forms one layer of a three-line defence termed 'the 3Cs of business continuity': contingency planning; continuity capability; crisis response. Collectively, 'the 3Cs' help organisations deliver a robust response to the risks that can be seen and those that cannot.

  4. Rotational quenching of CS in ultracold 3He collisions

    NASA Astrophysics Data System (ADS)

    Kaur, Rajwant; Dhilip Kumar, T. J.

    2016-08-01

    Quantum mechanical scattering calculations of rotational quenching of CS (v = 0) collision with 3He are performed at ultracold temperatures and results are compared with isotopic 4He collision. Rotational quenching cross sections and rate coefficients have been calculated in the ultracold region for rotational levels up to j = 10 using the He-CS potential energy surface computed at the CCSD(T)/aug-cc-pVQZ level of theory. The quenching cross sections are found to be two orders of magnitude larger for the 3He than the 4He isotope under ultracold conditions. Wigner threshold law is found to be valid below 10-3 K temperature.

  5. Ionic Selectivity and Permeation Properties of Human PIEZO1 Channels

    PubMed Central

    Gnanasambandam, Radhakrishnan; Bae, Chilman; Gottlieb, Philip A.; Sachs, Frederick

    2015-01-01

    Members of the eukaryotic PIEZO family (the human orthologs are noted hPIEZO1 and hPIEZO2) form cation-selective mechanically-gated channels. We characterized the selectivity of human PIEZO1 (hPIEZO1) for alkali ions: K+, Na+, Cs+ and Li+; organic cations: TMA and TEA, and divalents: Ba2+, Ca2+, Mg2+ and Mn2+. All monovalent ions permeated the channel. At a membrane potential of -100 mV, Cs+, Na+ and K+ had chord conductances in the range of 35–55 pS with the exception of Li+, which had a significantly lower conductance of ~ 23 pS. The divalents decreased the single-channel permeability of K+, presumably because the divalents permeated slowly and occupied the open channel for a significant fraction of the time. In cell-attached mode, 90 mM extracellular divalents had a conductance for inward currents carried by the divalents of: 25 pS for Ba2+ and 15 pS for Ca2+ at -80 mV and 10 pS for Mg2+ at -50 mV. The organic cations, TMA and TEA, permeated slowly and attenuated K+ currents much like the divalents. As expected, the channel K+ conductance increased with K+ concentration saturating at ~ 45 pS and the KD of K+ for the channel was 32 mM. Pure divalent ion currents were of lower amplitude than those with alkali ions and the channel opening rate was lower in the presence of divalents than in the presence of monovalents. Exposing cells to the actin disrupting reagent cytochalasin D increased the frequency of openings in cell-attached patches probably by reducing mechanoprotection. PMID:25955826

  6. The preliminary results of the measurements of environmental levels of 40K and 137Cs in Venezuela

    NASA Astrophysics Data System (ADS)

    LaBrecque, J. J.; Rosales, P. A.; Carias, O.

    1992-02-01

    The study of the beta-particle radioactivity in air and 90Sr in rainwater collected at Instituto Venezolano de Investigaciones Cientificas, IVIC, that began in 1959 in conjunction with the U.S. Atomic Energy Commission was discontinued in 1963. But after the Chernobyl nuclear power plant accident in the USSR, work on the radioactive contamination in Venezuela has been renewed. Initially the measurement of 134Cs and 137Cs in imported foodstuffs, such as milk powders and meats were undertaken, and these have now been extended to measurements of environmental levels of 40K and 137Cs in soils, sea sediments and foodstuffs. The data present herein are the only readily available data on radioactive contamination in the environment in Venezuela since 1963. The measurements were performed with a simple Nal(Tl) detector and two single channel analyzers as well as with a 25% efficient hyper-pure germanium semiconductor coupled to a multichannel analyzer system. Finally, these results can be used as baseline values (environmental background) in case of a nuclear accident in the region, as well as to screen imported foodstuff which is suspected to be contaminated.

  7. Ion channels in plants

    PubMed Central

    Baluška, František; Mancuso, Stefano

    2013-01-01

    In his recent opus magnum review paper published in the October issue of Physiology Reviews, Rainer Hedrich summarized the field of plant ion channels.1 He started from the earliest electric recordings initiated by Charles Darwin of carnivorous Dionaea muscipula,1,2 known as Venus flytrap, and covered the topic extensively up to the most recent discoveries on Shaker-type potassium channels, anion channels of SLAC/SLAH families, and ligand-activated channels of glutamate receptor-like type (GLR) and cyclic nucleotide-gated channels (CNGC).1 PMID:23221742

  8. Cardiac ion channels

    PubMed Central

    Priest, Birgit T; McDermott, Jeff S

    2015-01-01

    Ion channels are critical for all aspects of cardiac function, including rhythmicity and contractility. Consequently, ion channels are key targets for therapeutics aimed at cardiac pathophysiologies such as atrial fibrillation or angina. At the same time, off-target interactions of drugs with cardiac ion channels can be the cause of unwanted side effects. This manuscript aims to review the physiology and pharmacology of key cardiac ion channels. The intent is to highlight recent developments for therapeutic development, as well as elucidate potential mechanisms for drug-induced cardiac side effects, rather than present an in-depth review of each channel subtype. PMID:26556552

  9. Intracellular potassium stabilizes human ether-à-go-go-related gene channels for export from endoplasmic reticulum.

    PubMed

    Wang, Lu; Dennis, Adrienne T; Trieu, Phan; Charron, Francois; Ethier, Natalie; Hebert, Terence E; Wan, Xiaoping; Ficker, Eckhard

    2009-04-01

    Several therapeutic compounds have been identified that prolong the QT interval on the electrocardiogram and cause torsade de pointes arrhythmias not by direct block of the cardiac potassium channel human ether-à-go-go-related gene (hERG) but via disruption of hERG trafficking to the cell surface membrane. One example of a clinically important compound class that potently inhibits hERG trafficking are cardiac glycosides. We have shown previously that inhibition of hERG trafficking by cardiac glycosides is initiated via direct block of Na(+)/K(+) pumps and not via off-target interactions with hERG or any other protein. However, it was not known how pump inhibition at the cell surface is coupled to hERG processing in the endoplasmic reticulum. Here, we show that depletion of intracellular K(+)-either indirectly after long-term exposure to cardiac glycosides or directly after exposure to gramicidin in low sodium media-is sufficient to disrupt hERG trafficking. In K(+)-depleted cells, hERG trafficking can be restored by permeating K(+) or Rb(+) ions, incubation at low temperature, exposure to the pharmacological chaperone astemizole, or specific mutations in the selectivity filter of hERG. Our data suggest a novel mechanism for drug-induced trafficking inhibition in which cardiac glycosides produce a [K(+)](i)-mediated conformational defect directly in the hERG channel protein. PMID:19139152

  10. 135Cs/137Cs isotopic composition of environmental samples across Europe: Environmental transport and source term emission applications

    SciTech Connect

    Snow, Mathew S.; Snyder, Darin C.

    2015-11-02

    135Cs/137Cs isotopic analyses represent an important tool for studying the fate and transport of radiocesium in the environment; in this work the 135Cs/137Cs isotopic composition in environmental samples taken from across Europe is reported. Surface soil and vegetation samples from western Russia, Ukraine, Austria, and Hungary show consistent aged thermal fission product 135Cs/137Cs isotope ratios of 0.58 ± 0.01 (age corrected to 1/1/15), with the exception of one sample of soil-moss from Hungary which shows an elevated 135Cs/137Cs ratio of 1.78 ± 0.12. With the exception of the outlier sample from Hungary, surface soil/vegetation data are in quantitative agreement with values previously reported for soils within the Chernobyl exclusion zone, suggesting that radiocesium at these locations is primarily composed of homogenous airborne deposition from Chernobyl. Seawater samples taken from the Irish Sea show 135Cs/137Cs isotope ratios of 1.22 ± 0.11 (age corrected to 1/1/15), suggesting aged thermal fission product Cs discharged from Sellafield. Furthermore, the differences in 135Cs/137Cs isotope ratios between Sellafield, Chernobyl, and global nuclear weapons testing fallout indicate that 135Cs/137Cs isotope ratios can be utilized to discriminate between and track radiocesium transport from different nuclear production source terms, including major emission sources in Europe.

  11. Syntheses, structures, and ion-exchange properties of the three-dimensional framework uranyl gallium phosphates, Cs4[(UO2)2(GaOH)2(PO4)4].H2O and Cs[UO2Ga(PO4)2].

    PubMed

    Shvareva, Tatiana Y; Sullens, Tyler A; Shehee, Thomas C; Albrecht-Schmitt, Thomas E

    2005-01-24

    The reaction of UO(2)(NO(3))(2).6H(2)O with Cs(2)CO(3) or CsCl, H(3)PO(4), and Ga(2)O(3) under mild hydrothermal conditions results in the formation of Cs(4)[(UO(2))(2)(GaOH)(2)(PO(4))(4)].H(2)O (UGaP-1) or Cs[UO(2)Ga(PO(4))(2)] (UGaP-2). The structure of UGaP-1 was solved from a twinned crystal revealing a three-dimensional framework structure consisting of one-dimensional (1)(infinity)[Ga(OH)(PO(4))(2)](4-) chains composed of corner-sharing GaO(6) octahedra and bridging PO(4) tetrahedra that extend along the c axis. The phosphate anions bind the UO(2)(2+) cations to form UO(7) pentagonal bipyramids. The UO(7) moieties edge-share to create dimers that link the gallium phosphate substructure into a three-dimensional (3)(infinity)[(UO(2))(2)(GaOH)(2)(PO(4))(4)](4-) anionic lattice that has intersecting channels running down the b and c axes. Cs(+) cations and water molecules occupy these channels. The structure of UGaP-2 is also three-dimensional and contains one-dimensional (1)(infinity)[Ga(PO(4))(2)](3-) gallium phosphate chains that extend down the a axis. These chains are formed from fused eight-membered rings of corner-sharing GaO(4) and PO(4) tetrahedra. The chains are in turn linked together into a three-dimensional (3)(infinity)[UO(2)Ga(PO(4))(2)](1-) framework by edge-sharing UO(7) dimers as occurs in UGaP-1. There are channels that run down the a and b axes through the framework. These channels contain the Cs(+) cations. Ion-exchange studies indicate that the Cs(+) cations in UGaP-1 and UGaP-2 can be exchanged for Ca(2+) and Ba(2+). Crystallographic data: UGaP-1, monoclinic, space group P2(1)/c, a = 18.872(1), b = 9.5105(7), c = 14.007(1) A, beta = 109.65(3)(o) , Z = 4 (T = 295 K); UGaP-2, triclinic, space group P, a = 7.7765(6), b = 8.5043(7), c = 8.9115(7) A, alpha = 66.642(1)(o), beta = 70.563(1)(o), gamma = 84.003(2)(o), Z = 2 (T = 193 K). PMID:15651876

  12. C. elegans TRP channels.

    PubMed

    Xiao, Rui; Xu, X Z Shawn

    2011-01-01

    Transient receptor potential (TRP) channels represent a superfamily of cation channels found in all eukaryotes. The C. elegans genome encodes seventeen TRP channels covering all of the seven TRP subfamilies. Genetic analyses in C. elegans have implicated TRP channels in a wide spectrum of behavioral and physiological processes, ranging from sensory transduction (e.g. chemosensation, touch sensation, proprioception and osmosensation) to fertilization, drug dependence, organelle biogenesis, apoptosis, gene expression, and neurotransmitter/hormone release. Many C. elegans TRP channels share similar activation and regulatory mechanisms with their vertebrate counterparts. Studies in C. elegans have also revealed some previously unrecognized functions and regulatory mechanisms of TRP channels. C. elegans represents an excellent genetic model organism for the study of function and regulation of TRP channels in vivo. PMID:21290304

  13. Mechanically Activated Ion Channels

    PubMed Central

    Ranade, Sanjeev S.; Syeda, Ruhma; Patapoutian, Ardem

    2015-01-01

    Mechanotransduction, the conversion of physical forces into biochemical signals, is an essential component of numerous physiological processes including not only conscious senses of touch and hearing, but also unconscious senses such as blood pressure regulation. Mechanically activated (MA) ion channels have been proposed as sensors of physical force, but the identity of these channels and an understanding of how mechanical force is transduced has remained elusive. A number of recent studies on previously known ion channels along with the identification of novel MA ion channels have greatly transformed our understanding of touch and hearing in both vertebrates and invertebrates. Here, we present an updated review of eukaryotic ion channel families that have been implicated in mechanotransduction processes and evaluate the qualifications of the candidate genes according to specified criteria. We then discuss the proposed gating models for MA ion channels and highlight recent structural studies of mechanosensitive potassium channels. PMID:26402601

  14. C. elegans TRP channels

    PubMed Central

    Xiao, Rui; Xu, X.Z. Shawn

    2010-01-01

    TRP (transient receptor potential) channels represent a superfamily of cation channels found in all eukaryotes. The C. elegans genome encodes seventeen TRP channels covering all of the seven TRP subfamilies. Genetic analyses in C. elegans have implicated TRP channels in a wide spectrum of behavioral and physiological processes, ranging from sensory transduction (e.g. chemosensation, touch sensation, proprioception and osmosensation) to fertilization, drug dependence, organelle biogenesis, apoptosis, gene expression, and neurotransmitter/hormone release. Many C. elegans TRP channels share similar activation and regulatory mechanisms with their vertebrate counterparts. Studies in C. elegans have also revealed some previously unrecognized functions and regulatory mechanisms of TRP channels. C. elegans represents an excellent genetic model organism for the study of function and regulation of TRP channels in vivo. PMID:21290304

  15. A parallel finite element simulator for ion transport through three-dimensional ion channel systems.

    PubMed

    Tu, Bin; Chen, Minxin; Xie, Yan; Zhang, Linbo; Eisenberg, Bob; Lu, Benzhuo

    2013-09-15

    A parallel finite element simulator, ichannel, is developed for ion transport through three-dimensional ion channel systems that consist of protein and membrane. The coordinates of heavy atoms of the protein are taken from the Protein Data Bank and the membrane is represented as a slab. The simulator contains two components: a parallel adaptive finite element solver for a set of Poisson-Nernst-Planck (PNP) equations that describe the electrodiffusion process of ion transport, and a mesh generation tool chain for ion channel systems, which is an essential component for the finite element computations. The finite element method has advantages in modeling irregular geometries and complex boundary conditions. We have built a tool chain to get the surface and volume mesh for ion channel systems, which consists of a set of mesh generation tools. The adaptive finite element solver in our simulator is implemented using the parallel adaptive finite element package Parallel Hierarchical Grid (PHG) developed by one of the authors, which provides the capability of doing large scale parallel computations with high parallel efficiency and the flexibility of choosing high order elements to achieve high order accuracy. The simulator is applied to a real transmembrane protein, the gramicidin A (gA) channel protein, to calculate the electrostatic potential, ion concentrations and I - V curve, with which both primitive and transformed PNP equations are studied and their numerical performances are compared. To further validate the method, we also apply the simulator to two other ion channel systems, the voltage dependent anion channel (VDAC) and α-Hemolysin (α-HL). The simulation results agree well with Brownian dynamics (BD) simulation results and experimental results. Moreover, because ionic finite size effects can be included in PNP model now, we also perform simulations using a size-modified PNP (SMPNP) model on VDAC and α-HL. It is shown that the size effects in SMPNP can

  16. Satellite bands of the RbCs molecule in the range of highly excited states

    NASA Astrophysics Data System (ADS)

    Rakić, Mario; Beuc, Robert; Bouloufa-Maafa, Nadia; Dulieu, Olivier; Vexiau, Romain; Pichler, Goran; Skenderović, Hrvoje

    2016-05-01

    We report on the observation of three RbCs satellite bands in the blue and green ranges of the visible spectrum. Absorption measurements are performed using all-sapphire cell filled with a mixture of Rb and Cs. We compare high resolution absorption spectrum of Rb-Cs vapor mixture with pure Rb and Cs vapor spectra from the literature. After detailed analysis, the new satellite bands of RbCs molecule at 418.3 nm, 468.3, and 527.5 nm are identified. The origin of these bands is discussed by direct comparison with difference potentials derived from quantum chemistry calculations of RbCs potential energy curves. These bands originate from the lower Rydberg states of the RbCs molecule. This study thus provides further insight into photoassociation of lower Rydberg molecular states, approximately between Cs(7s) + Rb(5s) and Cs(6s) + Rb(6p) asymptotes, in ultracold gases.

  17. Spatially Common Sparsity Based Adaptive Channel Estimation and Feedback for FDD Massive MIMO

    NASA Astrophysics Data System (ADS)

    Gao, Zhen; Dai, Linglong; Wang, Zhaocheng; Chen, Sheng

    2015-12-01

    This paper proposes a spatially common sparsity based adaptive channel estimation and feedback scheme for frequency division duplex based massive multi-input multi-output (MIMO) systems, which adapts training overhead and pilot design to reliably estimate and feed back the downlink channel state information (CSI) with significantly reduced overhead. Specifically, a non-orthogonal downlink pilot design is first proposed, which is very different from standard orthogonal pilots. By exploiting the spatially common sparsity of massive MIMO channels, a compressive sensing (CS) based adaptive CSI acquisition scheme is proposed, where the consumed time slot overhead only adaptively depends on the sparsity level of the channels. Additionally, a distributed sparsity adaptive matching pursuit algorithm is proposed to jointly estimate the channels of multiple subcarriers. Furthermore, by exploiting the temporal channel correlation, a closed-loop channel tracking scheme is provided, which adaptively designs the non-orthogonal pilot according to the previous channel estimation to achieve an enhanced CSI acquisition. Finally, we generalize the results of the multiple-measurement-vectors case in CS and derive the Cramer-Rao lower bound of the proposed scheme, which enlightens us to design the non-orthogonal pilot signals for the improved performance. Simulation results demonstrate that the proposed scheme outperforms its counterparts, and it is capable of approaching the performance bound.

  18. Singlet and triplet potentials of the ground-state atom pair Rb + Cs studied by Fourier-transform spectroscopy

    SciTech Connect

    Docenko, O.; Tamanis, M.; Ferber, R.; Knoeckel, H.; Tiemann, E.

    2011-05-15

    A comprehensive study of the X {sup 1}{Sigma}{sup +} and a {sup 3}{Sigma}{sup +} electronic states of the Rb and Cs atom pair in RbCs is presented. Abundant spectroscopic data for the {sup 85}Rb{sup 133}Cs and {sup 87}Rb{sup 133}Cs isotopologues were obtained from Fourier-transform spectra of laser-induced fluorescence (LIF) from the B {sup 1}{Pi} and (4){sup 1}{Sigma}{sup +} states to the a {sup 3}{Sigma}{sup +} (4549 transitions) and X {sup 1}{Sigma}{sup +} (15 709 transitions) states. The X {sup 1}{Sigma}{sup +} state data were complemented by about 15 500 transitions obtained by Fellows et al.[J. Mol. Spectrosc. 197, 19 (1999)]. LIF progressions to the a {sup 3}{Sigma}{sup +} state range from v{sub a}=0 to 37, reaching an outer turning point at 15.97 A. For the X {sup 1}{Sigma}{sup +} state, by analyzing LIF from the shelflike (4){sup 1}{Sigma}{sup +}, the data are extended from v{sub x}=119 as observed in the above-cited paper to v{sub x}=129, extending the outer turning point from 10.7 to 17.55 A. From these data, potential energy curves were constructed simultaneously for both hyperfine coupled states X {sup 1}{Sigma}{sup +} and a {sup 3}{Sigma}{sup +} in a coupled-channels fitting routine. This allowed us to accurately model the potential close to the Rb(5s) + Cs(6s) atom-pair asymptote, which is required to simulate cold collision processes and to analyze the Feshbach resonances observed for {sup 87}Rb{sup 133}Cs by Pilch et al.[Phys. Rev. A 79, 042718 (2009)]. Dissociation energies were determined for the X {sup 1}{Sigma}{sup +} state as D{sub e}=3836.373(40) cm{sup -1} and for the a {sup 3}{Sigma}{sup +} state as D{sub e}=259.341(30) cm{sup -1}.

  19. ¹³⁴Cs and ¹³⁷Cs radioactivity in soil and moss samples of Jeju Island after Fukushima nuclear reactor accident.

    PubMed

    Park, Kyung-Ho; Kang, Tae-Woo; Kim, Won-Jik; Park, Jae Woo

    2013-11-01

    Specific activities of (134)Cs and (137)Cs in surface soil and moss samples were investigated at 12 locations of Jeju Island, Korea. Specific activities of (134)Cs and (137)Cs in the surface soil vary from less than MDA to 17 Bq/kg and from 12 Bq/kg to 109 Bq/kg, respectively. Specific activities of (134)Cs and (137)Cs in moss samples lie in the range 6 Bq/kg-39 Bq/kg and 15 Bq/kg-41 Bq/kg, respectively. The activity ratios (134)Cs/(137)Cs in the soil samples are much less than the reference value of about 1.0, but they are close to 1.0 in the moss samples. Average amount of (137)Cs added to the surface soil after the Fukushima accident is estimated to be 7.8 ± 1.7 Bq/kg. The depth profile of (137)Cs specific activity has a lognormal shape with a peak between 5 cm and 7.5 cm below the ground. For the cored soil sample, (134)Cs was detected up to 3 cm below the ground.

  20. 137 Cs Activities and 135 Cs/ 137 Cs Isotopic Ratios from Soils at Idaho National Laboratory: A Case Study for Contaminant Source Attribution in the Vicinity of Nuclear Facilities

    SciTech Connect

    Snow, Mathew S.; Snyder, Darin C.; Clark, Sue B.; Kelley, Morgan; Delmore, James E.

    2015-03-03

    Radiometric and mass spectrometric analyses of Cs contamination in the environment can reveal the location of Cs emission sources, release mechanisms, modes of transport, prediction of future contamination migration, and attribution of contamination to specific generator(s) and/or process(es). The Subsurface Disposal Area (SDA) at Idaho National Laboratory (INL) represents a complicated case study for demonstrating the current capabilities and limitations to environmental Cs analyses. 137Cs distribution patterns, 135Cs/137Cs isotope ratios, known Cs chemistry at this site, and historical records enable narrowing the list of possible emission sources and release events to a single source and event, with the SDA identified as the emission source and flood transport of material from within Pit 9 and Trench 48 as the primary release event. These data combined allow refining the possible number of waste generators from dozens to a single generator, with INL on-site research and reactor programs identified as the most likely waste generator. A discussion on the ultimate limitations to the information that 135Cs/137Cs ratios alone can provide is presented and includes (1) uncertainties in the exact date of the fission event and (2) possibility of mixing between different Cs source terms (including nuclear weapons fallout and a source of interest).

  1. 137Cs activities and 135Cs/137Cs isotopic ratios from soils at Idaho National Laboratory: a case study for contaminant source attribution in the vicinity of nuclear facilities.

    PubMed

    Snow, Mathew S; Snyder, Darin C; Clark, Sue B; Kelley, Morgan; Delmore, James E

    2015-03-01

    Radiometric and mass spectrometric analyses of Cs contamination in the environment can reveal the location of Cs emission sources, release mechanisms, modes of transport, prediction of future contamination migration, and attribution of contamination to specific generator(s) and/or process(es). The Subsurface Disposal Area (SDA) at Idaho National Laboratory (INL) represents a complicated case study for demonstrating the current capabilities and limitations to environmental Cs analyses. (137)Cs distribution patterns, (135)Cs/(137)Cs isotope ratios, known Cs chemistry at this site, and historical records enable narrowing the list of possible emission sources and release events to a single source and event, with the SDA identified as the emission source and flood transport of material from within Pit 9 and Trench 48 as the primary release event. These data combined allow refining the possible number of waste generators from dozens to a single generator, with INL on-site research and reactor programs identified as the most likely waste generator. A discussion on the ultimate limitations to the information that (135)Cs/(137)Cs ratios alone can provide is presented and includes (1) uncertainties in the exact date of the fission event and (2) possibility of mixing between different Cs source terms (including nuclear weapons fallout and a source of interest). PMID:25633972

  2. 137Cs activities and 135Cs/137Cs isotopic ratios from soils at Idaho National Laboratory: a case study for contaminant source attribution in the vicinity of nuclear facilities.

    PubMed

    Snow, Mathew S; Snyder, Darin C; Clark, Sue B; Kelley, Morgan; Delmore, James E

    2015-03-01

    Radiometric and mass spectrometric analyses of Cs contamination in the environment can reveal the location of Cs emission sources, release mechanisms, modes of transport, prediction of future contamination migration, and attribution of contamination to specific generator(s) and/or process(es). The Subsurface Disposal Area (SDA) at Idaho National Laboratory (INL) represents a complicated case study for demonstrating the current capabilities and limitations to environmental Cs analyses. (137)Cs distribution patterns, (135)Cs/(137)Cs isotope ratios, known Cs chemistry at this site, and historical records enable narrowing the list of possible emission sources and release events to a single source and event, with the SDA identified as the emission source and flood transport of material from within Pit 9 and Trench 48 as the primary release event. These data combined allow refining the possible number of waste generators from dozens to a single generator, with INL on-site research and reactor programs identified as the most likely waste generator. A discussion on the ultimate limitations to the information that (135)Cs/(137)Cs ratios alone can provide is presented and includes (1) uncertainties in the exact date of the fission event and (2) possibility of mixing between different Cs source terms (including nuclear weapons fallout and a source of interest).

  3. Accumulation and distribution of 137Cs in tropical plants

    NASA Astrophysics Data System (ADS)

    Anjos, R. M.; Carvalho, C.; Mosquera, B.; Veiga, R.; Sanches, N.; Bastos, J.; Macario, K.

    2007-02-01

    The accumulation and distribution of 40K and 137Cs in several tropical plant species were studied through measurements of gamma-ray spectra, focusing on establishing the suitability of using radiocesium to trace the plant uptake of nutrients such as potassium.

  4. Polarization analysis of magnons in CsMnI3

    NASA Astrophysics Data System (ADS)

    Tun, Z.; Hsu, T. C.; Lussier, J.-G.

    1994-05-01

    A polarized neutron scattering experiment on CsMnI3 shows previously unseen magnons at wave vectors (1/3,1/3,1) and (001), and also shows that the single-ion anisotropy for this material is substantially larger than previously reported. Linear spin wave theory can adequately describe the observed magnon frequencies.

  5. CHeCS: International Space Station Medical Hardware Catalog

    NASA Technical Reports Server (NTRS)

    2008-01-01

    The purpose of this catalog is to provide a detailed description of each piece of hardware in the Crew Health Care System (CHeCS), including subpacks associated with the hardware, and to briefly describe the interfaces between the hardware and the ISS. The primary user of this document is the Space Medicine/Medical Operations ISS Biomedical Flight Controllers (ISS BMEs).

  6. Equation of state and metallization of CsI

    SciTech Connect

    Aidun, J.; Bukowinski, M.S.T.; Ross, M.

    1984-03-01

    Self-consistent, nonrelativistic augmented-plane-wave (APW) calculations for CsI were carried out to generate the band structure, the static-lattice equation of state (EOS), and the volume dependence of the electronic energy-band gap. The theoretical room-temperature isothermal compression curve agrees well with static and ultrasonic measurements at low pressure. Our calculations do not agree with two recent sets of diamond-anvil-cell measurements above 10 GPa. The calculated band gaps are too small at low pressure, but, at high pressure, are consistent with both the experimental results and the Herzfeld-model prediction. These results suggest that the insulator-to-metal transition occurs in the range 100 +- 10 GPa. A calculation of the shock compression curve of CsI shows that the thermally excited electrons cause a significant softening of the Hugoniot curve. The experimental zero-pressure band gaps of the isoelectronic compounds Xe, CsI, and BaTe are linearly correlated with ln(v/v/sub H/), where v/sub H/ is the specific volume of metallization predicted by the Herzfeld model. Based on this correlation, and on the similarity of the APW calculated EOS's of Xe and CsI, which agree closely with experimental compression measurements, we predict that BaTe will become metallic at approximately 30 GPa.

  7. Teaching a CS Introductory Course: An Active Approach

    ERIC Educational Resources Information Center

    Moura, Isabel C.; van Hattum-Janssen, Natascha

    2011-01-01

    Computer Science (CS) introductory courses that are offered by the Department of Information Systems at the University of Minho (UM), Portugal, seem to abound in non-motivated students. They are characterized by high failure and withdrawal rates and use mainly deductive teaching approaches. Deductive instruction begins with theories and progresses…

  8. Reflection of thermal Cs atoms grazing a polished glass surface

    SciTech Connect

    Anderson, A.; Haroche, S.; Hinds, E.A.; Jhe, W.; Meschede, D.; Moi, L.

    1986-10-01

    We present an experimental study which shows that a large fraction (> or =50%) of thermal Cs atoms are nearly specularly reflected by polished glass surfaces at grazing incidence. This effect is interesting in the context of projects aimed at storing cold alkali-metal atoms in boxes.

  9. Isoform Specificity of Protein Kinase Cs in Synaptic Plasticity

    ERIC Educational Resources Information Center

    Sossin, Wayne S.

    2007-01-01

    Protein kinase Cs (PKCs) are implicated in many forms of synaptic plasticity. However, the specific isoform(s) of PKC that underlie(s) these events are often not known. We have used "Aplysia" as a model system in order to investigate the isoform specificity of PKC actions due to the presence of fewer isoforms and a large number of documented…

  10. CsRAV1 induces sylleptic branching in hybrid poplar.

    PubMed

    Moreno-Cortés, Alicia; Hernández-Verdeja, Tamara; Sánchez-Jiménez, Paloma; González-Melendi, Pablo; Aragoncillo, Cipriano; Allona, Isabel

    2012-04-01

    • Sylleptic branching in trees may increase significantly branch number, leaf area and the general growth of the tree, particularly in its early years. Although this is a very important trait, so far little is known about the genes that control this process. • This article characterizes the Castanea sativa RAV1 gene, homologous to Arabidopsis TEM genes, by analyzing its circadian behavior and examining its winter expression in chestnut stems and buds. Transgenic hybrid poplars over-expressing CsRAV1 or showing RNA interference down-regulated PtaRAV1 and PtaRAV2 expression were produced and analyzed. • Over-expression of the CsRAV1 gene induces the early formation of sylleptic branches in hybrid poplar plantlets during the same growing season in which the lateral buds form. Only minor growth differences and no changes in wood anatomy are produced. • The possibility of generating trees with a greater biomass by manipulating the CsRAV1 gene makes CsRAV1 transgenic plants promising candidates for bioenergy production.

  11. Accumulation and distribution of 137Cs in tropical plants

    SciTech Connect

    Anjos, R. M.; Carvalho, C.; Mosquera, B.; Veiga, R.; Sanches, N.; Bastos, J.; Macario, K.

    2007-02-12

    The accumulation and distribution of 40K and 137Cs in several tropical plant species were studied through measurements of gamma-ray spectra, focusing on establishing the suitability of using radiocesium to trace the plant uptake of nutrients such as potassium.

  12. Fallout 137Cs in reindeer herders in Arctic Norway.

    PubMed

    Skuterud, Lavrans; Thørring, Håvard

    2015-03-01

    Reindeer herders in the Arctic were among the most heavily exposed populations to the global fallout from nuclear weapons testing in the 1950s and 1960s, due to high transfer of radionuclides in the lichens-reindeer-human food chain. Annual studies of (137)Cs in reindeer herders in Kautokeino, Norway, were initiated in 1965 to monitor radiation doses and follow environmental (137)Cs behavior. The (137)Cs concentrations declined from the peak in 1965 with effective half-times of 6-8 years, only interrupted by a temporary doubling in levels from 1986 to 1987 due to the Chernobyl fallout. During the period of 1950-2010 an average herder received an integrated effective dose from incorporated (137)Cs of about 18 mSv. This dose represents an insignificant increase in the risk for developing cancer. Health studies even show a significantly lower cancer incidence among Sámis and reindeer herders in northern Norway compared to other populations in the same area. PMID:25671344

  13. Web-CS: Infrastructure for Web-Based Competitions.

    ERIC Educational Resources Information Center

    Aerts, A. T. M.; Bierhoff, P. F. M.; De Bra, P. M. E.

    This paper presents a World Wide Web-based infrastructure for cooperation between many different parties. The infrastructure is designed for Web-based competitions involving an editorial board, designers of assignments or events, evaluators, different organizational layers, and contestants. Web-CS is entirely Web-based: all the communication…

  14. There and Back Again?: The Disappearing Pulsations of CS 1246

    NASA Astrophysics Data System (ADS)

    Vasquez Soto, Alan; Barlow, Brad

    2016-01-01

    Hot subdwarf stars were once main sequence stars, like the sun, that deviated from normal stellar evolution due to binary interactions and evolved into extreme horizontal branch stars. Several of these stars exhibit rapid pulsations driven by iron opacity instabilities. CS 1246 is a rapidly pulsating hot subdwarf discovered in 2009 that is dominated by a single 371 second pulsation. At the time of its discovery, the pulsational amplitude was one of the largest known, making CS 1246 an ideal candidate for follow up studies. Observations in 2013 implied that the pulsational amplitude had decreased significantly. Since then we have continued monitoring the star using the robotic SKYNET telescopes in Chile, in order to further characterize any changes. Our recent observations show that the pulsational amplitude has gone down by a factor of six: CS 1246 is barely a pulsator anymore. The decay in amplitude over time is reminiscent of a damped harmonic oscillator. Here we present six years of photometry for CS 1246 and discuss possible scenarios that might explain its interesting behavior.

  15. A CS1 Pedagogical Approach to Parallel Thinking

    ERIC Educational Resources Information Center

    Rague, Brian William

    2010-01-01

    Almost all collegiate programs in Computer Science offer an introductory course in programming primarily devoted to communicating the foundational principles of software design and development. The ACM designates this introduction to computer programming course for first-year students as CS1, during which methodologies for solving problems within…

  16. Fallout 137Cs in reindeer herders in Arctic Norway.

    PubMed

    Skuterud, Lavrans; Thørring, Håvard

    2015-03-01

    Reindeer herders in the Arctic were among the most heavily exposed populations to the global fallout from nuclear weapons testing in the 1950s and 1960s, due to high transfer of radionuclides in the lichens-reindeer-human food chain. Annual studies of (137)Cs in reindeer herders in Kautokeino, Norway, were initiated in 1965 to monitor radiation doses and follow environmental (137)Cs behavior. The (137)Cs concentrations declined from the peak in 1965 with effective half-times of 6-8 years, only interrupted by a temporary doubling in levels from 1986 to 1987 due to the Chernobyl fallout. During the period of 1950-2010 an average herder received an integrated effective dose from incorporated (137)Cs of about 18 mSv. This dose represents an insignificant increase in the risk for developing cancer. Health studies even show a significantly lower cancer incidence among Sámis and reindeer herders in northern Norway compared to other populations in the same area.

  17. Measurements of Cs absorption and retention in man

    SciTech Connect

    Henrichs, H.; Paretzke, H.G.; Voigt, G.; Berg, D. )

    1989-10-01

    One of the consequences of the Chernobyl reactor accident in 1986 was a comparatively high contamination of foodstuffs in Southern Federal Republic of Germany. In order to test radioecological models predicting the radiological consequences of such accidents, several thousand measurements were performed to determine Cs body burdens in members of the public. For the interpretation of these data and as a contribution to the improvement of the available database on the biokinetics of Cs isotopes in humans, we followed a small group of volunteers after their consumption of highly contaminated venison. Intakes, excretion rates and total body activities were measured during a period of more than 200 d. The data obtained were evaluated in terms of a compartment model to derive gastrointestinal uptakes, biological half-lives and dose conversion factors. The resulting uptake factors range from 65-90%, the half-lives of the long-term retention from 45 to 200 d. The majority of the resulting dose conversion factors lie below the values recommended by the ICRP, showing that the ICRP model is a reasonable and safe description of the Cs biokinetics in our study group, while the great variability of the results shows that it is not an accurate representation of the individual Cs retention.

  18. Changing CS Features Alters Evaluative Responses in Evaluative Conditioning

    ERIC Educational Resources Information Center

    Unkelbach, Christian; Stahl, Christoph; Forderer, Sabine

    2012-01-01

    Evaluative conditioning (EC) refers to changes in people's evaluative responses toward initially neutral stimuli (CSs) by mere spatial and temporal contiguity with other positive or negative stimuli (USs). We investigate whether changing CS features from conditioning to evaluation also changes people's evaluative response toward these CSs. We used…

  19. Identification and transcript analysis of two glutamate decarboxylase genes, CsGAD1 and CsGAD2, reveal the strong relationship between CsGAD1 and citrate utilization in citrus fruit.

    PubMed

    Liu, Xiao; Hu, Xiao-Mei; Jin, Long-Fei; Shi, Cai-Yun; Liu, Yong-Zhong; Peng, Shu-Ang

    2014-09-01

    Glutamate decarboxylase (GAD, EC 4.1.1.15) has been suggested to be a key, regulatory point in the biosynthesis of γ-aminobutyrate (GABA) and in the utilization of citric acid through GABA shunt pathway. In this study we discovered two GAD genes, named as CsGAD1 and CsGAD2, in citrus genome database and then successfully cloned. Both CsGAD1 and CsGAD2 have a putative pyridoxal 5-phosphate binding domain in the middle region and a putative calmodulin-binding domain at the carboxyl terminus. Gene structure analysis showed that much difference exists in the size of exons and introns or in cis-regulatory elements in promoter region between the two GAD genes. Gene expression indicated that CsGAD1 transcript was predominantly expressed in flower and CsGAD2 transcript was predominantly expressed in fruit juice sacs; in the ripening fruit, CsGAD1 transcript level was at least 2-time higher than CsGAD2 transcript level. Moreover, CsGAD1 transcript level was increased significantly along with the increase of GAD activity and accompanied by a significant decrease of titratable acid (TA), suggesting that it is CsGAD1 rather than CsGAD2 plays a role in the citric acid utilization during fruit ripening. In addition, injection of abscisic acid and foliar spray of K2SO4 significantly increased the TA content of Satsuma mandarin, and significantly decreased GAD activity as well as CsGAD1 transcript, further suggesting the important role of CsGAD1 in the citrate utilization of citrus fruit.

  20. GALAXY EVOLUTION EXPLORER OBSERVATIONS OF CS AND OH EMISSION IN COMET 9P/TEMPEL 1 DURING DEEP IMPACT

    SciTech Connect

    Feldman, Paul D.; McCandliss, Stephan R.; Morgenthaler, Jeffrey P.; Lisse, Carey M.; Weaver, Harold A.; A'Hearn, Michael F.

    2010-03-10

    Galaxy Evolution Explorer (GALEX) observations of comet 9P/Tempel 1 using the near-ultraviolet (NUV) objective grism were made before, during and after the Deep Impact event that occurred on 2005 July 4 at 05:52:03 UT when a 370 kg NASA spacecraft was maneuvered into the path of the comet. The NUV channel provides usable spectral information in a bandpass covering 2000-3400 A with a point source spectral resolving power of R {approx} 100. The primary spectral features in this range include solar continuum scattered from cometary dust and emissions from OH and CS molecular bands centered near 3085 and 2575 A, respectively. In particular, we report the only cometary CS emission detected during this event. The observations allow the evolution of these spectral features to be tracked over the period of the encounter. In general, the NUV emissions observed from Tempel 1 are much fainter than those that have been observed by GALEX from other comets. However, it is possible to derive production rates for the parent molecules of the species detected by GALEX in Tempel 1 and to determine the number of these molecules liberated by the impact. The derived quiescent production rates are Q(H{sub 2}O) = 6.4 x 10{sup 27} molecules s{sup -1} and Q(CS{sub 2}) = 6.7 x 10{sup 24} molecules s{sup -1}, while the impact produced an additional 1.6 x 10{sup 32} H{sub 2}O molecules and 1.3 x 10{sup 29} CS{sub 2} molecules, a similar ratio as in quiescent outgassing.

  1. Fading channel simulator

    DOEpatents

    Argo, Paul E.; Fitzgerald, T. Joseph

    1993-01-01

    Fading channel effects on a transmitted communication signal are simulated with both frequency and time variations using a channel scattering function to affect the transmitted signal. A conventional channel scattering function is converted to a series of channel realizations by multiplying the square root of the channel scattering function by a complex number of which the real and imaginary parts are each independent variables. The two-dimensional inverse-FFT of this complex-valued channel realization yields a matrix of channel coefficients that provide a complete frequency-time description of the channel. The transmitted radio signal is segmented to provide a series of transmitted signal and each segment is subject to FFT to generate a series of signal coefficient matrices. The channel coefficient matrices and signal coefficient matrices are then multiplied and subjected to inverse-FFT to output a signal representing the received affected radio signal. A variety of channel scattering functions can be used to characterize the response of a transmitter-receiver system to such atmospheric effects.

  2. Octupole excitations in 141,144Cs and the pronounced decrease of dipole moments with neutron number in odd- Z neutron-rich 141,143,144Cs

    NASA Astrophysics Data System (ADS)

    Luo, Y. X.; Rasmussen, J. O.; Hamilton, J. H.; Ramayya, A. V.; Liu, S. H.; Jones, E. F.; Gore, P. M.; Goodin, C.; Stone, N. J.; Zhu, S. J.; Hwang, J. K.; Li, Ke; Crowell, H. L.; Lee, I. Y.; Ter-Akopian, G. M.; Daniel, A. V.; Stoyer, M. A.; Donangelo, R.; Ma, W. C.; Cole, J. D.

    2010-07-01

    The level scheme of odd- Z neutron-rich 141Cs ( Z=55, N=86) was extended and expanded and that of 144Cs ( N=89) was identified for the first time by means of γ-γ-γ coincidence measurements of prompt γ rays in the spontaneous fission of 252Cf with Gammasphere. Spin/parity was assigned to the levels based on angular correlations and level systematics in 141,143Cs. Parity doublets characteristic of both simplex quantum number s=+i and s=-i were proposed in 141Cs. The tests by using rotational frequency ratio ω(I)/ω(I) imply octupole vibrations in 141Cs and 143Cs. B(E1)/B(E2) values and electric dipole moments D were calculated for 141Cs, and re-determined for 143Cs. It was found that B(E1)/B(E2) values of 141Cs are simplex-dependent and the average value is one order of magnitude larger than that of 143Cs, and the deduced dipole moment D of 141Cs is considerably larger than that of 143Cs, and comparable to the N=86 isotone 142Ba. For 144Cs the yrast sequence looks like a well-deformed rotational band, but no octupole band structure was identified in this nucleus. The overall variations of D in 141,143,144Cs exhibit a pronounced drop of dipole moment with increasing neutron number in this odd- Z isotopic chain, which may be analogous in nature to the quenching of D observed in even-even 146Ba ( Z=56, N=90) and 224Ra ( Z=88, N=136), and to the drop of D in the odd- Z neutron-rich 147La ( Z=57, N=90) reported by our collaboration.

  3. Prescription dose in permanent {sup 131}Cs seed prostate implants

    SciTech Connect

    Yue Ning; Heron, Dwight E.; Komanduri, Krishna; Huq, M. Saiful

    2005-08-15

    Recently, {sup 131}Cs seeds have been introduced for prostate permanent seed implants. This type of seed has a relatively short half-life of 9.7 days and has its most prominent emitted photon energy peaks in the 29-34 keV region. Traditionally, 145 and 125 Gy have been prescribed for {sup 125}I and {sup 103}Pd seed prostate implants, respectively. Since both the half-life and dosimetry characteristics of {sup 131}Cs seed are quite different from those of {sup 125}I and {sup 103}Pd, the appropriate prescription dose for {sup 131}Cs seed prostate implant may well be different. This study was designed to use a linear quadratic radiobiological model to determine an appropriate dose prescription scheme for permanent {sup 131}Cs seed prostate implants. In this model, prostate edema was taken into consideration. Calculations were also performed for tumors of different doubling times and for other related radiobiological parameters of different values. As expected, the derived prescription dose values were dependent on type of tumors and types of edema. However, for prostate cancers in which tumor cells are relatively slow growing and are reported to have a mean potential doubling time of around 40 days, the appropriate prescription dose for permanent {sup 131}Cs seed prostate implants was determined to be: 127{sub -12}{sup +5}Gy if the experiences of {sup 125}I seed implants were followed and 121{sub -3}{sup +0}Gy if the experiences of {sup 103}Pd seed implants were followed.

  4. Coherent structures near the wall in a turbulent channel flow

    NASA Astrophysics Data System (ADS)

    Jeong, J.; Hussain, F.; Schoppa, W.; Kim, J.

    1997-02-01

    Coherent structures (CS) near the wall (i.e. y+ [less-than-or-eq, slant] 60) in a numerically simulated turbulent channel flow are educed using a conditional sampling scheme which extracts the entire extent of dominant vortical structures. Such structures are detected from the instantaneous flow field using our newly developed vortex definition (Jeong & Hussain 1995) a region of negative [lambda]2, the second largest eigenvalue of the tensor SikSkj + [Omega]ik[Omega]kj which accurately captures the structure details (unlike velocity-, vorticity- or pressure-based eduction). Extensive testing has shown that [lambda]2 correctly captures vortical structures, even in the presence of the strong shear occurring near the wall of a boundary layer. We have shown that the dominant near-wall educed (i.e. ensemble averaged after proper alignment) CS are highly elongated quasi-streamwise vortices; the CS are inclined 9° in the vertical (x, y)-plane and tilted ±4° in the horizontal (x, z)-plane. The vortices of alternating sign overlap in x as a staggered array; there is no indication near the wall of hairpin vortices, not only in the educed data but also in instantaneous fields. Our model of the CS array reproduces nearly all experimentally observed events reported in the literature, such as VITA, Reynolds stress distribution, wall pressure variation, elongated low-speed streaks, spanwise shear, etc. In particular, a phase difference (in space) between streamwise and normal velocity fluctuations created by CS advection causes Q4 (‘sweep’) events to dominate Q2 (‘ejection’) and also creates counter-gradient Reynolds stresses (such as Q1 and Q3 events) above and below the CS. We also show that these effects are adequately modelled by half of a Batchelor's dipole embedded in (and decoupled from) a background shear U(y). The CS tilting (in the (x, z)-plane) is found to be responsible for sustaining CS through redistribution of streamwise turbulent kinetic energy to

  5. Jamming in Vertical Channels

    NASA Astrophysics Data System (ADS)

    Baxter, G. William; Steel, Fiona

    2011-03-01

    We study jamming of low aspect-ratio cylindrical Delrin grains in a vertical channel. Grain heights are less than their diameter so the grains resemble antacid tablets, coins, or poker chips. These grains are allowed to fall through a vertical channel with a square cross section where the channel width is greater than the diameter of a grain and constant throughout the length of the channel with no obstructions or constrictions. Grains are sometimes observed to form jams, stable structures supported by the channel walls with no support beneath them. The probability of jam occurrence and the strength or robustness of a jam is effected by grain and channel sizes. We will present experimental measurements of the jamming probability and jam strength in this system and discuss the relationship of these results to other experiments and theories. Supported by an Undergraduate Research Grant from Penn State Erie, The Behrend College.

  6. Mechanosensitive channels in microbes.

    PubMed

    Kung, Ching; Martinac, Boris; Sukharev, Sergei

    2010-01-01

    All cells, including microbes, detect and respond to mechanical forces, of which osmotic pressure is most ancient and universal. Channel proteins have evolved such that they can be directly stretched open when the membrane is under turgor pressure. Osmotic downshock, as in rain, opens bacterial mechanosensitive (MS) channels to jettison osmolytes, relieving pressure and preventing cell lysis. The ion flux through individual channel proteins can be observed directly with a patch clamp. MS channels of large and small conductance (MscL and MscS, respectively) have been cloned, crystallized, and subjected to biophysical and genetic analyses in depth. They are now models to scrutinize how membrane forces direct protein conformational changes. Eukaryotic microbes have homologs from animal sensory channels of the TRP superfamily. The MS channel in yeast is also directly sensitive to membrane stretch. This review examines the key concept that proteins embedded in the lipid bilayer can respond to the changes in the mechanical environment the lipid bilayer provides.

  7. Amyloid peptide channels.

    PubMed

    Kagan, B L; Azimov, R; Azimova, R

    2004-11-01

    At least 16 distinct clinical syndromes including Alzheimer's disease (AD), Parkinson's disease (PD), rheumatoid arthritis, type II diabetes mellitus (DM), and spongiform encephelopathies (prion diseases), are characterized by the deposition of amorphous, Congo red-staining deposits known as amyloid. These "misfolded" proteins adopt beta-sheet structures and aggregate spontaneously into similar extended fibrils despite their widely divergent primary sequences. Many, if not all, of these peptides are capable of forming ion-permeable channels in vitro and possibly in vivo. Common channel properties include irreversible, spontaneous insertion into membranes, relatively large, heterogeneous single-channel conductances, inhibition of channel formation by Congo red, and blockade of inserted channels by Zn2+. Physiologic effects of amyloid, including Ca2+ dysregulation, membrane depolarization, mitochondrial dysfunction, inhibition of long-term potentiation (LTP), and cytotoxicity, suggest that channel formation in plasma and intracellular membranes may play a key role in the pathophysiology of the amyloidoses. PMID:15702375

  8. Measurement of intrinsic radioactive backgrounds from the 137Cs and U/Th chains in CsI(Tl) crystals

    NASA Astrophysics Data System (ADS)

    Liu, Shu-Kui; Yue, Qian; Lin, Shin-Ted; Li, Yuan-Jing; Tang, Chang-Jian; Wong Tsz-King, Henry; Xing, Hao-Yang; Yang, Chao-Wen; Zhao, Wei; Zhu, Jing-Jun

    2015-04-01

    The inorganic CsI(Tl) crystal scintillator is a candidate anti-compton detector for the China Dark matter Experiment. Studying the intrinsic radiopurity of the CsI(Tl) crystal is an issue of major importance. The timing, energy and spatial correlations, as well as the capability of pulse shape discrimination provide powerful methods for the measurement of intrinsic radiopurities. The experimental design, detector performance and event-selection algorithms are described. A total of 359×3 kg-days data from three prototypes of CsI(Tl) crystals were taken at China Jinping Underground Laboratory (CJPL), which offers a good shielding environment. The contamination levels of internal isotopes from 137Cs, 232Th and 238U series, as well as the upper bounds of 235U series are reported. Identification of the whole α peaks from U/Th decay chains and derivation of those corresponding quenching factors are achieved. Supported by National Natural Science Foundation of China (11275107, 11175099)

  9. The neuronal channel NALCN contributes resting sodium permeability and is required for normal respiratory rhythm.

    PubMed

    Lu, Boxun; Su, Yanhua; Das, Sudipto; Liu, Jin; Xia, Jingsheng; Ren, Dejian

    2007-04-20

    Sodium plays a key role in determining the basal excitability of the nervous systems through the resting "leak" Na(+) permeabilities, but the molecular identities of the TTX- and Cs(+)-resistant Na(+) leak conductance are totally unknown. Here we show that this conductance is formed by the protein NALCN, a substantially uncharacterized member of the sodium/calcium channel family. Unlike any of the other 20 family members, NALCN forms a voltage-independent, nonselective cation channel. NALCN mutant mice have a severely disrupted respiratory rhythm and die within 24 hours of birth. Brain stem-spinal cord recordings reveal reduced neuronal firing. The TTX- and Cs(+)-resistant background Na(+) leak current is absent in the mutant hippocampal neurons. The resting membrane potentials of the mutant neurons are relatively insensitive to changes in extracellular Na(+) concentration. Thus, NALCN, a nonselective cation channel, forms the background Na(+) leak conductance and controls neuronal excitability. PMID:17448995

  10. HIPPI and Fibre Channel

    SciTech Connect

    Tolmie, D.E.

    1992-01-01

    The High-Performance Parallel Interface (HIPPI) and Fibre Channel are near-gigabit per second data communications interfaces being developed in ANSI standards Task Group X3T9.3. HIPPI is the current interface of choice in the high-end and supercomputer arena, and Fibre Channel is a follow-on effort. HIPPI came from a local area network background, and Fibre Channel came from a mainframe to peripheral interface background.

  11. Effect of redox potential on fixation of sup 137 Cs in lake sediment

    SciTech Connect

    Pardue, J.H.; DeLaune, R.D.; Patrick, W.H. Jr.; Whitcomb, J.H. )

    1989-11-01

    Fixation of 137Cs was determined in lake sediment suspensions under controlled redox potentials in the laboratory. The activity of previously added 137Cs on various clay sites was determined by time-series selective extractions. Monovalent cations, particularly NH+4, were much more effective at displacing 137Cs than divalent cations or Na+. Ammonium ion (NH+4) and Na+ were used to extract 137Cs from selective and non-selective 137Cs-binding sites, respectively. The activity of water-soluble 137Cs and Na-extractable 137Cs was significantly higher under anaerobic redox conditions (-200 mV), when soluble NH+4 concentrations in the anaerobic suspensions were 1000 microM or higher. Activities of 137Cs were highest (initially 40-50% of the 137Cs added) on the NH+4-extractable site. Over the long term, activities of NH+4-extractable 137Cs decreased linearly to below 10% as 137Cs was fixed on inter-lattice sites. Water-soluble 137Cs was significantly correlated with Na-extractable 137Cs in short-term experiments, suggesting the existence of an equilibrium between the different clay sites. High concentrations of NH+4 under anaerobic redox conditions could shift this equilibrium, resulting in increases in water-soluble 137Cs and increases in the activities of 137Cs bound on non-selective clay sites. Additional studies are necessary to evaluate the interaction of 137Cs with sulfides, iron oxides, and other reactive chemical species which may attenuate 137Cs in sediment.

  12. Symmetrization for redundant channels

    NASA Technical Reports Server (NTRS)

    Tulplue, Bhalchandra R. (Inventor); Collins, Robert E. (Inventor)

    1988-01-01

    A plurality of redundant channels in a system each contain a global image of all the configuration data bases in each of the channels in the system. Each global image is updated periodically from each of the other channels via cross channel data links. The global images of the local configuration data bases in each channel are separately symmetrized using a voting process to generate a system signal configuration data base which is not written into by any other routine and is available for indicating the status of the system within each channel. Equalization may be imposed on a suspect signal and a number of chances for that signal to heal itself are provided before excluding it from future votes. Reconfiguration is accomplished upon detecting a channel which is deemed invalid. A reset function is provided which permits an externally generated reset signal to permit a previously excluded channel to be reincluded within the system. The updating of global images and/or the symmetrization process may be accomplished at substantially the same time within a synchronized time frame common to all channels.

  13. Phosphoinositides regulate ion channels

    PubMed Central

    Hille, Bertil; Dickson, Eamonn J.; Kruse, Martin; Vivas, Oscar; Suh, Byung-Chang

    2014-01-01

    Phosphoinositides serve as signature motifs for different cellular membranes and often are required for the function of membrane proteins. Here, we summarize clear evidence supporting the concept that many ion channels are regulated by membrane phosphoinositides. We describe tools used to test their dependence on phosphoinositides, especially phosphatidylinositol 4,5-bisphosphate, and consider mechanisms and biological meanings of phosphoinositide regulation of ion channels. This lipid regulation can underlie changes of channel activity and electrical excitability in response to receptors. Since different intracellular membranes have different lipid compositions, the activity of ion channels still in transit towards their final destination membrane may be suppressed until they reach an optimal lipid environment. PMID:25241941

  14. IBEX channel formation

    SciTech Connect

    Jones, E.E.; Frost, C.A.; Freeman, J.R.; Jojola, J.M.

    1987-01-01

    Exploding wire experiments have been conducted to form a low-density channel for endoatmospheric channel-tracking experiments to be performed on the IBEX accelerator. Stainless steel and tungsten wires as small as six microns radius have been exploded using a 50 kJ, 200 kV fast capacitor bank designed and constructed for the purpose. Density channels have been produced. Preliminary results will be compared with a simple circuit model and hydrocode analysis. Efforts to diagnose the low-density channel are beginning. 5 refs.

  15. Measurements and Observations of 134Cs and 137Cs around a Nuclear Power Plant in Busan, South Korea

    NASA Astrophysics Data System (ADS)

    Chong, H. Y.; Park, J. N.; Kim, J. S.

    2015-12-01

    The purpose of this study is to know the change of representative species 134Cs and 137Cs of artificial radionuclides from the nuclear power station nearby for the last five years(2010~2014). The Kori Nuclear Power Plant, nuclear power facility located near Busan, is located in the south-east coast of Korea and about 21km north-east away from the Haeundae and about 25km south away from the Ulsan. An administrative district is Jangan-eup, Gijang-gun, Busan, Korea. A point was selected on the basis of the "Environmental Radiation Monitoring Plan around Nuclear Power Plants" and periodically samples were collected and analyzed. The samples were collected from the soils of the surface in the Wolnae area (NW, 1.7km). The soil samples were analyzed by gamma spectrometer with High Purity Germanium detector (HPGe) of 40% relative efficiency and were measured for 80,000 sec. As a results of soil analysis, 137Cs were detected in samples only selected in March 2013. The activity concentration of 137Cs founded in the soil sample was 0.513±0.052Bq/kg-dry. In the other hands, the concentration of 137Cs in the year 2010, 2011, 2012, and 2014 were below the minimum detectable activity (MDA). 40K, natural radionuclides which is widely present, was detected in the soil samples and other artificial radionuclides were not detected. The result of overall comparison of the environmental radioactivity survey around Kori Nuclear Power Plant for the last five years is that radioactivity levels are within average range. However, it is necessary to continue to carefully observe a fine change in regional or the monthly radiation concentration.

  16. Expression characteristics of CS-ACS1, CS-ACS2 and CS-ACS3, three members of the 1-aminocyclopropane-1-carboxylate synthase gene family in cucumber (Cucumis sativus L.) fruit under carbon dioxide stress.

    PubMed

    Mathooko, F M; Mwaniki, M W; Nakatsuka, A; Shiomi, S; Kubo, Y; Inaba, A; Nakamura, R

    1999-02-01

    We investigated the expression pattern of three 1-aminocyclopropane-1-carboxylate (ACC) synthase genes, CS-ACS1, CS-ACS2 and CS-ACS3 in cucumber (Cucumis sativus L.) fruit under CO2 stress. CO2 stress-induced ethylene production paralleled the accumulation of only CS-ACS1 transcripts which disappeared upon withdrawal of CO2. Cycloheximide inhibited the CO2 stress-induced ethylene production but superinduced the accumulation of CS-ACS1 transcript. At higher concentrations, cycloheximide also induced the accumulation of CS-ACS2 and CS-ACS3 transcripts. In the presence of CO2 and cycloheximide, the accumulation of CS-ACS2 transcript occurred within 1 h, disappeared after 3 h and increased greatly upon withdrawal of CO2. Inhibitors of protein kinase and types 1 and 2A protein phosphatases which inhibited and stimulated, respectively, CO2 stress-induced ethylene production had little effect on the expression of these genes. The results presented here identify CS-ACS1 as the main ACC synthase gene responsible for the increased ethylene biosynthesis in cucumber fruit under CO2 stress and suggest that this gene is a primary response gene and its expression is under negative control since it is expressed by treatment with cycloheximide. The results further suggest that the regulation of CO2 stress-induced ethylene biosynthesis by reversible protein phosphorylation does not result from enhanced ACC synthase transcription. PMID:10202812

  17. The Camelina aquaporin CsPIP2;1 is regulated by phosphorylation at Ser273, but not at Ser277, of the C-terminus and is involved in salt- and drought-stress responses.

    PubMed

    Jang, Ha-Young; Rhee, Jiye; Carlson, John E; Ahn, Sung-Ju

    2014-09-15

    Aquaporin (AQP) proteins are involved in water homeostasis in cells at all taxonomic levels of life. Phosphorylation of some AQPs has been proposed to regulate water permeability via gating of the channel itself. We analyzed plasma membrane intrinsic proteins (PIP) from Camelina and characterized their biological functions under both stressful and favorable conditions. A three-dimensional theoretical model of the Camelina AQP proteins was built by homology modeling which could prove useful in further functional characterization of AQPs. CsPIP2;1 was strongly and constitutively expressed in roots and leaves of Camelina, suggesting that this gene is related to maintenance of homeostasis during salt and drought stresses. CsPIP2s exhibited water channel activity in Xenopus oocytes. We then examined the roles of CsPIP2;1 phosphorylation at Ser273 and Ser277 in the regulation of water permeability using phosphorylation mutants. A single deletion strain of CsPIP2;1 was generated to serve as the primary host for testing AQP expression constructs. A Ser277 to alanine mutation (to prevent phosphorylation) did not change CsPIP2;1 water permeability while a Ser273 mutation to alanine did affect water permeability. Furthermore, a CsPIP2;1 point mutation when ectopically expressed in yeast resulted in lower growth in salt and drought conditions compared with controls, and confirmation of Ser273 as the phosphorylation site. Our results support the idea that post-translational modifications in the Ser273 regulatory domains of the C-terminus fine tune water flux through CsPIP2;1. PMID:25046761

  18. A magnesium-carboxylate framework showing luminescent sensing for CS{sub 2} and nitroaromatic compounds

    SciTech Connect

    Wu, Zhao-Feng; Tan, Bin; Feng, Mei-Ling; Du, Cheng-Feng; Huang, Xiao-Ying

    2015-03-15

    A magnesium metal-organic framework compound, namely [NH{sub 2}(CH{sub 3}){sub 2}][Mg{sub 3}(NDC){sub 2.5}(HCO{sub 2}){sub 2}(DMF){sub 0.75}(H{sub 2}O){sub 0.25}]·1.25DMF·0.75H{sub 2}O (1) (H{sub 2}NDC=1,4-naphthalene dicarboxylic acid, DMF=N,N′-dimethylformamide), has been synthesized in solvothermal conditions and structurally characterized. It features a three-dimensionally anionic framework with aligned channels parallel to the b-axis. Luminescent studies indicated that it showed significant luminescence quenching for carbon disulfide (CS{sub 2}) and nitrobenzene after being activated, at a content of only 3.0 and 0.1 vol% in DMF, respectively. In addition, the activated sample showed sensitive luminescence quenching for 1,3,5-trinitrophenol with a low concentration of 5×10{sup −5} mol/L. - Graphical abstract: Presented is a microporous 3D Mg-MOF, namely, [NH{sub 2}(CH{sub 3}){sub 2}][Mg{sub 3}(NDC){sub 2.5}(HCO{sub 2}){sub 2}(DMF){sub 0.75}(H{sub 2}O){sub 0.25}]·1.25DMF·0.75H{sub 2}O (1) (H{sub 2}NDC=1,4-naphthalene dicarboxylic acid) showing significant luminescence quenching for carbon disulfide and nitrobenzene. - Highlights: • A microporous 3D metal-organic framework based on Mg. • The compound shows significant luminescence quenching for CS{sub 2} and nitrobenzene after activated. • The compound shows sensitive luminescence quenching for 1,3,5-trinitrophenol with a low concentration of 5×10{sup −5} mol/L.

  19. Blockade of ENaCs by amiloride induces c-Fos activation of the area postrema.

    PubMed

    Miller, Rebecca L; Denny, George O; Knuepfer, Mark M; Kleyman, Thomas R; Jackson, Edwin K; Salkoff, Lawrence B; Loewy, Arthur D

    2015-03-19

    Epithelial sodium channels (ENaCs) are strongly expressed in the circumventricular organs (CVOs), and these structures may play an important role in sensing plasma sodium levels. Here, the potent ENaC blocker amiloride was injected intraperitoneally in rats and 2h later, the c-Fos activation pattern in the CVOs was studied. Amiloride elicited dose-related activation in the area postrema (AP) but only ~10% of the rats showed c-Fos activity in the organum vasculosum of the lamina terminalis (OVLT) and subfornical organ (SFO). Tyrosine hydroxylase-immunoreactive (catecholamine) AP neurons were activated, but tryptophan hydroxylase-immunoreactive (serotonin) neurons were unaffected. The AP projects to FoxP2-expressing neurons in the dorsolateral pons which include the pre-locus coeruleus nucleus and external lateral part of the parabrachial nucleus; both cell groups were c-Fos activated following systemic injections of amiloride. In contrast, another AP projection target--the aldosterone-sensitive neurons of the nucleus tractus solitarius which express the enzyme 11-β-hydroxysteriod dehydrogenase type 2 (HSD2) were not activated. As shown here, plasma concentrations of amiloride used in these experiments were near or below the IC50 level for ENaCs. Amiloride did not induce changes in blood pressure, heart rate, or regional vascular resistance, so sensory feedback from the cardiovascular system was probably not a causal factor for the c-Fos activity seen in the CVOs. In summary, amiloride may have a dual effect on sodium homeostasis causing a loss of sodium via the kidney and inhibiting sodium appetite by activating the central satiety pathway arising from the AP. PMID:25557402

  20. Worst configurations (instantons) for compressed sensing over reals: a channel coding approach

    SciTech Connect

    Chertkov, Michael; Chilappagari, Shashi K; Vasic, Bane

    2010-01-01

    We consider Linear Programming (LP) solution of a Compressed Sensing (CS) problem over reals, also known as the Basis Pursuit (BasP) algorithm. The BasP allows interpretation as a channel-coding problem, and it guarantees the error-free reconstruction over reals for properly chosen measurement matrix and sufficiently sparse error vectors. In this manuscript, we examine how the BasP performs on a given measurement matrix and develop a technique to discover sparse vectors for which the BasP fails. The resulting algorithm is a generalization of our previous results on finding the most probable error-patterns, so called instantons, degrading performance of a finite size Low-Density Parity-Check (LDPC) code in the error-floor regime. The BasP fails when its output is different from the actual error-pattern. We design CS-Instanton Search Algorithm (ISA) generating a sparse vector, called CS-instanton, such that the BasP fails on the instanton, while its action on any modification of the CS-instanton decreasing a properly defined norm is successful. We also prove that, given a sufficiently dense random input for the error-vector, the CS-ISA converges to an instanton in a small finite number of steps. Performance of the CS-ISA is tested on example of a randomly generated 512 * 120 matrix, that outputs the shortest instanton (error vector) pattern of length 11.