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Sample records for idioblasts buffer cytosolic

  1. Role of calbindin-D9k in buffering cytosolic free Ca2+ ions in pig duodenal enterocytes.

    Science.gov (United States)

    Schröder, B; Schlumbohm, C; Kaune, R; Breves, G

    1996-05-01

    1. The aim of the present study was to test whether the vitamin D-dependent Ca(2+)-binding protein calbindin-D9k could function as an important cytosolic Ca2+ buffer in duodenal enterocytes while facilitating transepithelial active transport of Ca2+ ions. For the investigations we used dual-wavelength, fluorescence ratio imaging, with fura-2 as the Ca(2+)-sensitive dye, to measure changes in cytosolic concentrations of free Ca2+ ions ([Ca2+]i) in isolated pig duodenal enterocytes affected by different cytosolic calbindin-D9k concentrations. 2. Epithelial cells were obtained from weaned piglets with normal calbindin-D9k concentrations (con-piglets), from piglets with low calbindin-D9k levels due to inherited calcitriol deficiency caused by defective renal 25-hydroxycholecalciferol D3-1 alpha-hydroxylase activity (def-piglets), and from piglets with reconstituted calbindin-D9k concentrations, i.e. def-animals treated with high doses of vitamin D3 which elevated plasma calcitriol levels by extrarenal production (def-D3-piglets). Basal levels of [Ca2+]i ranged between 170 and 205 nM and did not differ significantly between the groups. 3. After addition of 5 mM theophylline, the [Ca2+]i in enterocytes from con-piglets doubled during the 10 min incubation. This effect, however, was three times higher in enterocytes from def-piglets compared with those from con-piglets. Similar results were obtained after 4 min incubation of enterocytes from con- and def-piglets in the presence of 1 microM ionomycin. In preparations from def-D3-piglets, ionomycin-induced increases in [Ca2+]i were significantly lower compared with enterocytes from def-piglets and were not different from the control values. 4. From the results, substantial support is given for the hypothesis that one of the major functions of mucosal calbindin-D9k is the effective buffering of Ca2+ ions.

  2. Avocado fruit maturation and ripening: dynamics of aliphatic acetogenins and lipidomic profiles from mesocarp, idioblasts and seed.

    Science.gov (United States)

    Rodríguez-López, Carlos Eduardo; Hernández-Brenes, Carmen; Treviño, Víctor; Díaz de la Garza, Rocío I

    2017-09-29

    Avocado fruit contains aliphatic acetogenins (oft-acetylated, odd-chain fatty alcohols) with promising bioactivities for both medical and food industries. However, we have scarce knowledge about their metabolism. The present work aimed to study changes in acetogenin profiles from mesocarp, lipid-containing idioblasts, and seeds from 'Hass' cultivar during fruit development, germination, and three harvesting years. An untargeted LC-MS based lipidomic analysis was also conducted to profile the lipidome of avocado fruit in each tissue. The targeted analysis showed that acetogenin profiles and contents remained unchanged in avocado mesocarp during maturation and postharvest ripening, germination, and different harvesting years. However, a shift in the acetogenin profile distribution, accompanied with a sharp increase in concentration, was observed in seed during early maturation. Untargeted lipidomics showed that this shift was accompanied with remodeling of glycerolipids: TAGs and DAGs decreased during fruit growing in seed. Remarkably, the majority of the lipidome in mature seed was composed by acetogenins; we suggest that this tissue is able to synthesize them independently from mesocarp. On the other hand, lipid-containing idioblasts accumulated almost the entire acetogenin pool measured in the whole mesocarp, while only having 4% of the total fatty acids. The lipidome of this cell type changed the most when the fruit was ripening after harvesting, TAGs decreased while odd-chain DAGs increased. Notably, idioblast lipidome was more diverse than that from mesocarp. Evidence shown here suggests that idioblasts are the main site of acetogenin biosynthesis in avocado mesocarp. This work unveiled the prevalence of aliphatic acetogenins in the avocado fruit lipidome and evidenced TAGs as initial donors of the acetogenin backbones in its biosynthesis. It also sets evidence for acetogenins being included in future works aimed at characterizing the avocado seed, as they are

  3. Biosynthesis of l-Ascorbic Acid and Conversion of Carbons 1 and 2 of l-Ascorbic Acid to Oxalic Acid Occurs within Individual Calcium Oxalate Crystal Idioblasts1

    Science.gov (United States)

    Kostman, Todd A.; Tarlyn, Nathan M.; Loewus, Frank A.; Franceschi, Vincent R.

    2001-01-01

    l-Ascorbic acid (AsA) and its metabolic precursors give rise to oxalic acid (OxA) found in calcium oxalate crystals in specialized crystal idioblast cells in plants; however, it is not known if AsA and OxA are synthesized within the crystal idioblast cell or transported in from surrounding mesophyll cells. Isolated developing crystal idioblasts from Pistia stratiotes were used to study the pathway of OxA biosynthesis and to determine if idioblasts contain the entire path and are essentially independent in OxA synthesis. Idioblasts were supplied with various 14C-labeled compounds and examined by micro-autoradiography for incorporation of 14C into calcium oxalate crystals. [14C]OxA gave heavy labeling of crystals, indicating the isolated idioblasts are functional in crystal formation. Incubation with [1-14C]AsA also gave heavy labeling of crystals, whereas [6-14C]AsA gave no labeling. Labeled precursors of AsA (l-[1-14C]galactose; d-[1-14C]mannose) also resulted in crystal labeling, as did the ascorbic acid analog, d-[1-14C]erythorbic acid. Intensity of labeling of isolated idioblasts followed the pattern OxA > AsA (erythorbic acid) > l-galactose > d-mannose. Our results demonstrate that P. stratiotes crystal idioblasts synthesize the OxA used for crystal formation, the OxA is derived from the number 1 and 2 carbons of AsA, and the proposed pathway of ascorbic acid synthesis via d-mannose and l-galactose is operational in individual P. stratiotes crystal idioblasts. These results are discussed with respect to fine control of calcium oxalate precipitation and the concept of crystal idioblasts as independent physiological compartments. PMID:11161021

  4. Redox characteristics of the eukaryotic cytosol

    DEFF Research Database (Denmark)

    López-Mirabal, H Reynaldo; Winther, Jakob R

    2007-01-01

    The eukaryotic cytoplasm has long been regarded as a cellular compartment in which the reduced state of protein cysteines is largely favored. Under normal conditions, the cytosolic low-molecular weight redox buffer, comprising primarily of glutathione, is highly reducing and reactive oxygen species...... (ROS) and glutathionylated proteins are maintained at very low levels. In the present review, recent progress in the understanding of the cytosolic thiol-disulfide redox metabolism and novel analytical approaches to studying cytosolic redox properties are discussed. We will focus on the yeast model...... organism, Saccharomyces cerevisiae, where the combination of genetic and biochemical approaches has brought us furthest in understanding the mechanisms underlying cellular redox regulation. It has been shown in yeast that, in addition to the enzyme glutathione reductase, other mechanisms may exist...

  5. A membrane model for cytosolic calcium oscillations. A study using Xenopus oocytes.

    OpenAIRE

    Jafri, M S; Vajda, S; Pasik, P; Gillo, B

    1992-01-01

    Cytosolic calcium oscillations occur in a wide variety of cells and are involved in different cellular functions. We describe these calcium oscillations by a mathematical model based on the putative electrophysiological properties of the endoplasmic reticulum (ER) membrane. The salient features of our membrane model are calcium-dependent calcium channels and calcium pumps in the ER membrane, constant entry of calcium into the cytosol, calcium dependent removal from the cytosol, and buffering ...

  6. Buffers Plus

    Science.gov (United States)

    Ramette, Richard W.

    1998-11-01

    In 1989 JCE Software published The Acid-Base Package: A Collection of Useful Programs for Proton Transfer Systems (Ramette, R. W. J. Chem. Educ. Software 1989, 2B No. 2). This DOS program has been fully upgraded by the same author to the world of Windows 95. Buffers Plus takes advantage of a modern user interface and offers many new options not possible in the original version.

  7. Buffer fluid

    Energy Technology Data Exchange (ETDEWEB)

    Mirzadzhanzade, A Kh; Dedusanko, G Ya; Dinaburg, L S; Markov, Yu M; Rasizade, Ya N; Rozov, V N; Sherstnev, N M

    1979-08-30

    A drilling fluid is suggested for separating the drilling and plugging fluids which contains as the base increased solution of polyacrylamide and additive. In order to increase the viscoelastic properties of the liquid with simultaneous decrease in the periods of its fabrication, the solution contains as an additive dry bentonite clay. In cases of the use of a buffer fluid under conditions of negative temperatures, it is necessary to add to it table salt or ethylene glycol.

  8. Cytosolic glutamine synthetase in barley

    DEFF Research Database (Denmark)

    Thomsen, Hanne Cecilie

    remobilisation from ageing plant parts. Thus, GS is highly involved in determining crop yield and NUE. The major objective of this PhD project was to investigate the NUE properties of transgenic barley designed to constitutively overexpress a GS1 isogene (HvGS1.1). These transgenic lines exhibited an increased...... for N demand. Of the GS isogenes, only the transcript levels of root HvGS1.1 increased when plants were transferred from high to low N. This change coincided with an increase in total GS activity. Pronounced diurnal variation was observed for root nitrate transporter genes and GS isogenes in both root...... fertilizer requirement. The enzyme glutamine synthetase (GS) has been a major topic in plant nitrogen research for decades due to its central role in plant N metabolism. The cytosolic version of this enzyme (GS1) plays an important role in relation to primary N assimilation as well as in relation to N...

  9. Buffer mass test - Buffer materials

    International Nuclear Information System (INIS)

    Pusch, R.; Boergesson, L.

    1982-08-01

    Commercial Na bentonite (MX-80) is the clay component of the buffer material in the heater holes as well of the tunnel backfill. Important characteristics are the clay content, liquid limit, X-ray diffraction pattern, water content, and degree of granulation. The ballast material consists of quartz-rich sand and feldspar-rich filler. The preparation of highly compacted bentonite for the near-field isolation of the canister was made by using isostatic compaction technique. The resulting dense bentonite core was cut into regularly shaped blocks which were arranged around each heater and lowered as one unit - heavily instrumented - in the respective deposition holes. For three of the six holes a narrow slot was left open between the bentonite stack and the rock; for the remaining ones a wider slot was chosen with a fill of soft bentonite powder. Both arrangements are expected to yield an ultimate bulk density which is sufficiently high to fulfil the requirement of a negligible permeability and a sufficient swelling pressure as well as heat conductivity, which are the essential parameters. The tunnel backfill, which consists of a mixture of suitably graded ballast material and MX-80 powder, has a considerably lower swelling pressure and heat conductivity, and a higher permeability, all these parameters still within the requirements of the KBS 2 concept. The various zones with different bentonite/sand ratios and the technique to apply them are described in the final part of the report. (Author)

  10. Buffer regulation of calcium puff sequences

    International Nuclear Information System (INIS)

    Fraiman, Daniel; Dawson, Silvina Ponce

    2014-01-01

    Puffs are localized Ca 2+ signals that arise in oocytes in response to inositol 1,4,5-trisphosphate (IP 3 ). They are the result of the liberation of Ca 2+ from the endoplasmic reticulum through the coordinated opening of IP 3 receptor/channels clustered at a functional release site. The presence of buffers that trap Ca 2+ provides a mechanism that enriches the spatio–temporal dynamics of cytosolic calcium. The expression of different types of buffers along the cell's life provides a tool with which Ca 2+ signals and their responses can be modulated. In this paper we extend the stochastic model of a cluster of IP 3 R-Ca 2+ channels introduced previously to elucidate the effect of buffers on sequences of puffs at the same release site. We obtain analytically the probability laws of the interpuff time and of the number of channels that participate of the puffs. Furthermore, we show that under typical experimental conditions the effect of buffers can be accounted for in terms of a simple inhibiting function. Hence, by exploring different inhibiting functions we are able to study the effect of a variety of buffers on the puff size and interpuff time distributions. We find the somewhat counter-intuitive result that the addition of a fast Ca 2+ buffer can increase the average number of channels that participate of a puff. (paper)

  11. Buffer regulation of calcium puff sequences.

    Science.gov (United States)

    Fraiman, Daniel; Dawson, Silvina Ponce

    2014-02-01

    Puffs are localized Ca(2 +) signals that arise in oocytes in response to inositol 1,4,5-trisphosphate (IP3). They are the result of the liberation of Ca(2 +) from the endoplasmic reticulum through the coordinated opening of IP3 receptor/channels clustered at a functional release site. The presence of buffers that trap Ca(2 +) provides a mechanism that enriches the spatio-temporal dynamics of cytosolic calcium. The expression of different types of buffers along the cell's life provides a tool with which Ca(2 +) signals and their responses can be modulated. In this paper we extend the stochastic model of a cluster of IP3R-Ca(2 +) channels introduced previously to elucidate the effect of buffers on sequences of puffs at the same release site. We obtain analytically the probability laws of the interpuff time and of the number of channels that participate of the puffs. Furthermore, we show that under typical experimental conditions the effect of buffers can be accounted for in terms of a simple inhibiting function. Hence, by exploring different inhibiting functions we are able to study the effect of a variety of buffers on the puff size and interpuff time distributions. We find the somewhat counter-intuitive result that the addition of a fast Ca(2 +) buffer can increase the average number of channels that participate of a puff.

  12. The thermodynamic-buffer enzymes.

    Science.gov (United States)

    Stucki, J W

    1980-08-01

    Oxidative phosphorylation operates at optimal efficiency if and only if the condition of conductance matching L33/L11 = square root 1-q2 is fulfilled. In this relation L11 is the phenomenological conductance of phosphorylation, L33 the phenomenological conductance of the load, i.e. the irreversible ATP-utilizing processes in the cell, and q the degree of coupling of oxidative phosphorylation driven by respiration. Since during short time intervals L11 and q are constant whereas L33 fluctuates in the cell, oxidative phosphorylation would only rarely operate at optimal efficiency due to violation of conductance matching. This paper demonstrates that the reversible ATP-utilizing reaction catalyzed by adenylate kinase can effectively compensate deviations from conductance matching in the presence of a fluctuating L33 and hence allows oxidative phosphorylation to operate at optimal efficiency in the cell. Since the adenylate kinase reaction was found to buffer a thermodynamic potential, i.e. the phosphate potential, this finding was generalized to the concept of thermodynamic buffering. The thermodynamic buffering ability of the adenylate kinase reaction was demonstrated by experiments with incubated rat-liver mitochondria. Considerations of changes introduced in the entropy production by the adenylate kinase reaction allowed to establish the theoretical framework for thermodynamic buffering. The ability of thermodynamic buffering to compensate deviations from conductance matching in the presence of fluctuating loads was demonstrated by computer simulations. The possibility of other reversible ATP-utilizing reactions, like the ones catalyzed by creatine kinase and arginine kinase, to contribute to thermodynamic buffering is discussed. Finally, the comparison of the theoretically calculated steady-stae cytosolic adenine nucleotide concentrations with experimental data from perfused livers demonstrated that in livers from fed rats conductance matching is fulfilled on a

  13. Cytosolic cholesterol ester hydrolase in adrenal cortex

    OpenAIRE

    Tocher, Douglas R.

    1983-01-01

    Cholesterol ester hydrolase (CEH) in adrenocortical cytosol was known to be phosphorylated and activated, in response to ACTH in a cAMPdependent protein kinase mediated process. The purification of CEH from bovine adrenocortical cytosol was attempted. The use of detergents to solubilise the enzyme from lipid-rich aggregates was investigated and sodium cholate was found to be effective. A purification procedure using cholate solubilised enzyme was developed. The detergent int...

  14. Buffer Zone Fact Sheets

    Science.gov (United States)

    New requirements for buffer zones and sign posting contribute to soil fumigant mitigation and protection for workers and bystanders. The buffer provides distance between the pesticide application site and bystanders, reducing exposure risk.

  15. Redox Buffer Strength

    Science.gov (United States)

    de Levie, Robert

    1999-04-01

    The proper functioning of enzymes in bodily fluids requires that the pH be maintained within rather narrow limits. The first line of defense against large pH fluctuations in such fluids is the passive control provided by the presence of pH buffers. The ability of pH buffers to stabilize the pH is indicated by the buffer value b introduced in 1922 by van Slyke. It is equally important for many enzymes that the redox potential is kept within a narrow range. In that case, stability of the potential is most readily achieved with a redox buffer. In this communication we define the redox buffer strength by analogy with acid-base buffer strength.

  16. Organellar Calcium Buffers

    Science.gov (United States)

    Prins, Daniel; Michalak, Marek

    2011-01-01

    Ca2+ is an important intracellular messenger affecting many diverse processes. In eukaryotic cells, Ca2+ storage is achieved within specific intracellular organelles, especially the endoplasmic/sarcoplasmic reticulum, in which Ca2+ is buffered by specific proteins known as Ca2+ buffers. Ca2+ buffers are a diverse group of proteins, varying in their affinities and capacities for Ca2+, but they typically also carry out other functions within the cell. The wide range of organelles containing Ca2+ and the evidence supporting cross-talk between these organelles suggest the existence of a dynamic network of organellar Ca2+ signaling, mediated by a variety of organellar Ca2+ buffers. PMID:21421925

  17. Aeromonas caviae alters the cytosolic and mitochondrial creatine kinase activities in experimentally infected silver catfish: Impairment on renal bioenergetics.

    Science.gov (United States)

    Baldissera, Matheus D; Souza, Carine F; Júnior, Guerino B; Verdi, Camila Marina; Moreira, Karen L S; da Rocha, Maria Izabel U M; da Veiga, Marcelo L; Santos, Roberto C V; Vizzotto, Bruno S; Baldisserotto, Bernardo

    2017-09-01

    Cytosolic and mitochondrial creatine kinases (CK), through the creatine kinase-phosphocreatine (CK/PCr) system, provide a temporal and spatial energy buffer to maintain cellular energy homeostasis. However, the effects of bacterial infections on the kidney remain poorly understood and are limited only to histopathological analyses. Thus, the aim of this study was to investigate the involvement of cytosolic and mitochondrial CK activities in renal energetic homeostasis in silver catfish experimentally infected with Aeromonas caviae. Cytosolic CK activity decreased in infected animals, while mitochondrial CK activity increased compared to uninfected animals. Moreover, the activity of the sodium-potassium pump (Na + , K + -ATPase) decreased in infected animals compared to uninfected animals. Based on this evidence, it can be concluded that the inhibition of cytosolic CK activity by A. caviae causes an impairment on renal energy homeostasis through the depletion of adenosine triphosphate (ATP) levels. This contributes to the inhibition of Na + , K + -ATPase activity, although the mitochondrial CK activity acted in an attempt to restore the cytosolic ATP levels through a feedback mechanism. In summary, A. caviae infection causes a severe energetic imbalance in infected silver catfish, which may contribute to disease pathogenesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Nuclear Calcium Buffering Capacity Shapes Neuronal Architecture*

    Science.gov (United States)

    Mauceri, Daniela; Hagenston, Anna M.; Schramm, Kathrin; Weiss, Ursula; Bading, Hilmar

    2015-01-01

    Calcium-binding proteins (CaBPs) such as parvalbumin are part of the cellular calcium buffering system that determines intracellular calcium diffusion and influences the spatiotemporal dynamics of calcium signals. In neurons, CaBPs are primarily localized to the cytosol and function, for example, in nerve terminals in short-term synaptic plasticity. However, CaBPs are also expressed in the cell nucleus, suggesting that they modulate nuclear calcium signals, which are key regulators of neuronal gene expression. Here we show that the calcium buffering capacity of the cell nucleus in mouse hippocampal neurons regulates neuronal architecture by modulating the expression levels of VEGFD and the complement factor C1q-c, two nuclear calcium-regulated genes that control dendrite geometry and spine density, respectively. Increasing the levels of nuclear calcium buffers by means of expression of a nuclearly targeted form of parvalbumin fused to mCherry (PV.NLS-mC) led to a reduction in VEGFD expression and, as a result, to a decrease in total dendritic length and complexity. In contrast, mRNA levels of the synapse pruning factor C1q-c were increased in neurons expressing PV.NLS-mC, causing a reduction in the density and size of dendritic spines. Our results establish a close link between nuclear calcium buffering capacity and the transcription of genes that determine neuronal structure. They suggest that the development of cognitive deficits observed in neurological conditions associated with CaBP deregulation may reflect the loss of necessary structural features of dendrites and spines. PMID:26231212

  19. Nuclear Calcium Buffering Capacity Shapes Neuronal Architecture.

    Science.gov (United States)

    Mauceri, Daniela; Hagenston, Anna M; Schramm, Kathrin; Weiss, Ursula; Bading, Hilmar

    2015-09-18

    Calcium-binding proteins (CaBPs) such as parvalbumin are part of the cellular calcium buffering system that determines intracellular calcium diffusion and influences the spatiotemporal dynamics of calcium signals. In neurons, CaBPs are primarily localized to the cytosol and function, for example, in nerve terminals in short-term synaptic plasticity. However, CaBPs are also expressed in the cell nucleus, suggesting that they modulate nuclear calcium signals, which are key regulators of neuronal gene expression. Here we show that the calcium buffering capacity of the cell nucleus in mouse hippocampal neurons regulates neuronal architecture by modulating the expression levels of VEGFD and the complement factor C1q-c, two nuclear calcium-regulated genes that control dendrite geometry and spine density, respectively. Increasing the levels of nuclear calcium buffers by means of expression of a nuclearly targeted form of parvalbumin fused to mCherry (PV.NLS-mC) led to a reduction in VEGFD expression and, as a result, to a decrease in total dendritic length and complexity. In contrast, mRNA levels of the synapse pruning factor C1q-c were increased in neurons expressing PV.NLS-mC, causing a reduction in the density and size of dendritic spines. Our results establish a close link between nuclear calcium buffering capacity and the transcription of genes that determine neuronal structure. They suggest that the development of cognitive deficits observed in neurological conditions associated with CaBP deregulation may reflect the loss of necessary structural features of dendrites and spines. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Cytosolic delivery of materials with endosome-disrupting colloids

    Science.gov (United States)

    Helms, Brett A.; Bayles, Andrea R.

    2016-03-15

    A facile procedure to deliver nanocrystals to the cytosol of live cells that is both rapid and general. The technique employs a unique cationic core-shell polymer colloid that directs nanocrystals to the cytosol of living cells within a few hours of incubation. The present methods and compositions enable a host of advanced applications arising from efficient cytosolic delivery of nanocrystal imaging probes: from single particle tracking experiments to monitoring protein-protein interactions in live cells for extended periods.

  1. Buffer design 2012

    International Nuclear Information System (INIS)

    Juvankoski, M.

    2013-08-01

    Posiva's spent nuclear fuel disposal is based on the KBS-3V concept and on the characteristics of the Olkiluoto site. In this concept single canisters containing spent nuclear fuel surrounded by a bentonite buffer are emplaced in individual vertical boreholes drilled in the floor of deposition tunnels in bedrock at about 420 m depth below ground level. Disk type bentonite blocks are installed at the bottom of the hole and on the top of the disposal canister. Ring type bentonite blocks surround the canisters. This report describes the detailed design of the buffer for a KBS-3V repository. The report presents the design basis, the reference design, and summarises the performance analyses carried out for the design. This report addresses aspects concerning the manufacture, quality control, mechanical strength, chemical resistance, thermal dimensioning, handling of buffer components and material ageing phenomena including the effect of radiation. Interaction of buffer and other engineered barriers are included in the study. The long-term evolution of the repository and its effective drivers are considered if they have an impact on the buffer performance but operational safety aspects are also included because they may affect long-term safety. (orig.)

  2. The SVT Hit Buffer

    International Nuclear Information System (INIS)

    Belforte, S.; Dell'Orso, M.; Donati, S.

    1996-01-01

    The Hit Buffer is part of the Silicon Vertex Tracker, a trigger processor dedicated to the reconstruction of particle trajectories in the Silicon Vertex Detector and the Central Tracking Chamber of the Collider Detector at Fermilab. The Hit Buffer is a high speed data-traffic node, where thousands of words are received in arbitrary order and simultaneously organized in an internal structured data base, to be later promptly retrieved and delivered in response to specific requests. The Hit Buffer is capable of processing data at a rate of 25 MHz, thanks to the use of special fast devices like Cache-Tag RAMs and high performance Erasable Programmable Logic Devices from the XILINX XC7300 family

  3. A parallel buffer tree

    DEFF Research Database (Denmark)

    Sitchinava, Nodar; Zeh, Norbert

    2012-01-01

    We present the parallel buffer tree, a parallel external memory (PEM) data structure for batched search problems. This data structure is a non-trivial extension of Arge's sequential buffer tree to a private-cache multiprocessor environment and reduces the number of I/O operations by the number of...... in the optimal OhOf(psortN + K/PB) parallel I/O complexity, where K is the size of the output reported in the process and psortN is the parallel I/O complexity of sorting N elements using P processors....

  4. Genetically encoded pH-indicators reveal activity-dependent cytosolic acidification of Drosophila motor nerve termini in vivo

    Science.gov (United States)

    Rossano, Adam J; Chouhan, Amit K; Macleod, Gregory T

    2013-01-01

    All biochemical processes, including those underlying synaptic function and plasticity, are pH sensitive. Cytosolic pH (pHcyto) shifts are known to accompany nerve activity in situ, but technological limitations have prevented characterization of such shifts in vivo. Genetically encoded pH-indicators (GEpHIs) allow for tissue-specific in vivo measurement of pH. We expressed three different GEpHIs in the cytosol of Drosophila larval motor neurons and observed substantial presynaptic acidification in nerve termini during nerve stimulation in situ. SuperEcliptic pHluorin was the most useful GEpHI for studying pHcyto shifts in this model system. We determined the resting pH of the nerve terminal cytosol to be 7.30 ± 0.02, and observed a decrease of 0.16 ± 0.01 pH units when the axon was stimulated at 40 Hz for 4 s. Realkalinization occurred upon cessation of stimulation with a time course of 20.54 ± 1.05 s (τ). The chemical pH-indicator 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein corroborated these changes in pHcyto. Bicarbonate-derived buffering did not contribute to buffering of acid loads from short (≤4 s) trains of action potentials but did buffer slow (∼60 s) acid loads. The magnitude of cytosolic acid transients correlated with cytosolic Ca2+ increase upon stimulation, and partial inhibition of the plasma membrane Ca2+-ATPase, a Ca2+/H+ exchanger, attenuated pHcyto shifts. Repeated stimulus trains mimicking motor patterns generated greater cytosolic acidification (∼0.30 pH units). Imaging through the cuticle of intact larvae revealed spontaneous pHcyto shifts in presynaptic termini in vivo, similar to those seen in situ during fictive locomotion, indicating that presynaptic pHcyto shifts cannot be dismissed as artifacts of ex vivo preparations. PMID:23401611

  5. Cytosolic iron chaperones: Proteins delivering iron cofactors in the cytosol of mammalian cells.

    Science.gov (United States)

    Philpott, Caroline C; Ryu, Moon-Suhn; Frey, Avery; Patel, Sarju

    2017-08-04

    Eukaryotic cells contain hundreds of metalloproteins that are supported by intracellular systems coordinating the uptake and distribution of metal cofactors. Iron cofactors include heme, iron-sulfur clusters, and simple iron ions. Poly(rC)-binding proteins are multifunctional adaptors that serve as iron ion chaperones in the cytosolic/nuclear compartment, binding iron at import and delivering it to enzymes, for storage (ferritin) and export (ferroportin). Ferritin iron is mobilized by autophagy through the cargo receptor, nuclear co-activator 4. The monothiol glutaredoxin Glrx3 and BolA2 function as a [2Fe-2S] chaperone complex. These proteins form a core system of cytosolic iron cofactor chaperones in mammalian cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Buffer Zone Sign Template

    Science.gov (United States)

    The certified pesticide applicator is required to post a comparable sign, designating a buffer zone around the soil fumigant application block in order to control exposure risk. It must include the don't walk symbol, product name, and applicator contact.

  7. Buffer Zone, Nicosia

    OpenAIRE

    Sorensen, Marie Louise

    2010-01-01

    Images of the United Nations Buffer Zone or Green Line which has partitioned Cyprus since 1974 The research leading to these results has received funding from the European Community's Seventh Framework Programme [FP7/2007-2013] under grant agreement n° 217411.

  8. Influence of rapid changes in cytosolic pH on oxidative phosphorylation in skeletal muscle: theoretical studies.

    Science.gov (United States)

    Korzeniewski, Bernard; Zoladz, Jerzy A

    2002-07-01

    Cytosolic pH in skeletal muscle may vary significantly because of proton production/consumption by creatine kinase and/or proton production by anaerobic glycolysis. A computer model of oxidative phosphorylation in intact skeletal muscle developed previously was used to study the kinetic effect of these variations on the oxidative phosphorylation system. Two kinds of influence were analysed: (i) via the change in pH across the inner mitochondrial membrane and (ii) via the shift in the equilibrium of the creatine kinase-catalysed reaction. Our simulations suggest that cytosolic pH has essentially no impact on the steady-state fluxes and most metabolite concentrations. On the other hand, rapid acidification/alkalization of cytosol causes a transient decrease/increase in the respiration rate. Furthermore, changes in pH seem to affect significantly the kinetic properties of transition between resting state and active state. An increase in pH brought about by proton consumption by creatine kinase at the onset of exercise lengthens the transition time. At intensive exercise levels this pH increase could lead to loss of the stability of the system, if not compensated by glycolytic H+ production. Thus our theoretical results stress the importance of processes/mechanisms that buffer/compensate for changes in cytosolic proton concentration. In particular, we suggest that the second main role of anaerobic glycolysis, apart from additional ATP supply, may be maintaining the stability of the system at intensive exercise.

  9. Cytosolic adenylate changes during exercise in prawn muscle

    International Nuclear Information System (INIS)

    Thebault, M.T.; Raffin, J.P.; Pichon, R.

    1994-01-01

    31 P NMR and biochemical analysis were used to assess the effect of heavy exercise on cytosolic adenylate levels in Palaemon serratus abdominal muscle. At rest, the MgATP level corresponded to 85.5% of the total ATP content. The cytosolic adenylate concentrations of the prawn muscle are considerably different from that of vertebrates. The percentage of ADP bound to myofilaments was lower in the prawn muscle. Consequently, the level of free cytosolic AMP was greatly higher (thirty fold higher) than in vertebrate muscle. During vigorous work, the concentration of MgATP dropped and the cytosolic AMP accumulated, while the cytosolic adenine nucleotide pool decreased significantly. The phosphorylation potential value and the ATP/ADP ratio, calculated from the cytosolic adenylate, dropped acutely during the whole period of muscular contractions. On the contrary, the adenylate energy charge calculated from the cytosolic adenylate decreased slightly. Therefore, even in muscle displaying no AMP deamination, the adenylate charge is stabilized during exercise by the dynamic changes between cytosolic and bound adenylate species. (author). 21 refs., 2 tabs

  10. Cloning and characterization of human liver cytosolic beta-glycosidase

    NARCIS (Netherlands)

    De Graaf, M; Van Veen, IC; Van Der Meulen-Muileman, IH; Gerritsen, WR; Pinedo, HM; Haisma, HJ

    2001-01-01

    Cytosolic beta -glucosidase (EC 3.2.1.21) from mammalian liver is a member of the family 1 glycoside hydrolases and is known for its ability to hydrolyse a range of beta -D-glycosides. including beta -D-glucoside acid beta -D-galactoside. We therefore refer to this enzyme as cytosolic beta

  11. The Arabidopsis cytosolic proteome

    DEFF Research Database (Denmark)

    Ito, Jun; Parsons, Harriet Tempé; Heazlewood, Joshua L.

    2014-01-01

    compartments. However, a detailed study of enriched cytosolic fractions from Arabidopsis cell culture has been performed only recently, with over 1,000 proteins reproducibly identified by mass spectrometry. The number of proteins allocated to the cytosol nearly doubles to 1,802 if a series of targeted...

  12. Creep in buffer clay

    International Nuclear Information System (INIS)

    Pusch, R.; Adey, R.

    1999-12-01

    The study involved characterization of the microstructural arrangement and molecular forcefields in the buffer clay for getting a basis for selecting suitable creep models. It is concluded that the number of particles and wide range of the particle bond spectrum require that stochastical mechanics and thermodynamics will be considered and they are basic to the creep model proposed for predicting creep settlement of the canisters. The influence of the stress level on creep strain of MX-80 clay is not well known but for the buffer creep is approximately proportional to stress. Theoretical considerations suggest a moderate impact for temperatures up to 90 deg C and this is supported by model experiments. It is believed that the assumption of strain being proportional to temperature is conservative. The general performance of the stochastic model can be illustrated in principle by use of visco-elastic rheological models implying a time-related increase in viscosity. The shear-induced creep settlement under constant volume conditions calculated by using the proposed creep model is on the order of 1 mm in ten thousand years and up to a couple of millimeters in one million years. It is much smaller than the consolidation settlement, which is believed to be on the order of 10 mm. The general conclusion is that creep settlement of the canisters is very small and of no significance to the integrity of the buffer itself or of the canisters

  13. On the sulfation of O-desmethyltramadol by human cytosolic sulfotransferases.

    Science.gov (United States)

    Rasool, Mohammed I; Bairam, Ahsan F; Kurogi, Katsuhisa; Liu, Ming-Cheh

    2017-10-01

    Previous studies have demonstrated that sulfate conjugation is involved in the metabolism of the active metabolite of tramadol, O-desmethyltramadol (O-DMT). The current study aimed to systematically identify the human cytosolic sulfotransferases (SULTs) that are capable of mediating the sulfation of O-DMT. The sulfation of O-DMT under metabolic conditions was demonstrated using HepG2 hepatoma cells and Caco-2 human colon carcinoma cells. O-DMT-sulfating activity of thirteen known human SULTs and four human organ specimens was examined using an established sulfotransferase assay. pH-Dependency and kinetic parameters were also analyzed using, respectively, buffers at different pHs and varying O-DMT concentrations in the assays. Of the thirteen human SULTs tested, only SULT1A3 and SULT1C4 were found to display O-DMT-sulfating activity, with different pH-dependency profiles. Kinetic analysis revealed that SULT1C4 was 60 times more catalytically efficient in mediating the sulfation of O-DMT than SULT1A3 at respective optimal pH. Of the four human organ specimens tested, the cytosol prepared from the small intestine showed much higher O-DMT-sulfating activity than cytosols prepared from liver, lung, and kidney. Both cultured HepG2 and Caco-2 cells were shown to be capable of sulfating O-DMT and releasing sulfated O-DMT into cultured media. SULT1A3 and SULT1C4 were the major SULTs responsible for the sulfation of O-DMT. Collectively, the results obtained provided a molecular basis underlying the sulfation of O-DMT and contributed to a better understanding about the pharmacokinetics and pharmacodynamics of tramadol in humans. Copyright © 2017 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  14. Buffered or under Scrutiny?

    DEFF Research Database (Denmark)

    Klopf, Patricia; Nell, Phillip C.; Puck, Jonas

    – they are to a stronger extent buffered from the external environment than small firms. Thus, this paper adds to previous research by disentangling the complex effects of size on the use of political strategies – a strand of literature which has been characterized by inconsistent findings in the past.......This paper investigates political strategies of MNE subsidiaries operating in emerging markets. Our findings support previous findings of more intense political strategies in the presence of stronger institutional pressures from public and private stakeholders. Furthermore, we hypothesize...

  15. Chromosomal instability drives metastasis through a cytosolic DNA response.

    Science.gov (United States)

    Bakhoum, Samuel F; Ngo, Bryan; Laughney, Ashley M; Cavallo, Julie-Ann; Murphy, Charles J; Ly, Peter; Shah, Pragya; Sriram, Roshan K; Watkins, Thomas B K; Taunk, Neil K; Duran, Mercedes; Pauli, Chantal; Shaw, Christine; Chadalavada, Kalyani; Rajasekhar, Vinagolu K; Genovese, Giulio; Venkatesan, Subramanian; Birkbak, Nicolai J; McGranahan, Nicholas; Lundquist, Mark; LaPlant, Quincey; Healey, John H; Elemento, Olivier; Chung, Christine H; Lee, Nancy Y; Imielenski, Marcin; Nanjangud, Gouri; Pe'er, Dana; Cleveland, Don W; Powell, Simon N; Lammerding, Jan; Swanton, Charles; Cantley, Lewis C

    2018-01-25

    Chromosomal instability is a hallmark of cancer that results from ongoing errors in chromosome segregation during mitosis. Although chromosomal instability is a major driver of tumour evolution, its role in metastasis has not been established. Here we show that chromosomal instability promotes metastasis by sustaining a tumour cell-autonomous response to cytosolic DNA. Errors in chromosome segregation create a preponderance of micronuclei whose rupture spills genomic DNA into the cytosol. This leads to the activation of the cGAS-STING (cyclic GMP-AMP synthase-stimulator of interferon genes) cytosolic DNA-sensing pathway and downstream noncanonical NF-κB signalling. Genetic suppression of chromosomal instability markedly delays metastasis even in highly aneuploid tumour models, whereas continuous chromosome segregation errors promote cellular invasion and metastasis in a STING-dependent manner. By subverting lethal epithelial responses to cytosolic DNA, chromosomally unstable tumour cells co-opt chronic activation of innate immune pathways to spread to distant organs.

  16. The effect of mitochondrial dysfunction on cytosolic nucleotide metabolism

    DEFF Research Database (Denmark)

    Madsen, Claus Desler; Lykke, Anne; Rasmussen, Lene Juel

    2010-01-01

    Several enzymes of the metabolic pathways responsible for metabolism of cytosolic ribonucleotides and deoxyribonucleotides are located in mitochondria. Studies described in this paper suggest dysfunction of the mitochondria to affect these metabolic pathways and limit the available levels...

  17. Investigating the episodic buffer

    Directory of Open Access Journals (Sweden)

    Alan Baddeley

    2010-10-01

    Full Text Available A brief account is presented of the three-component working memory model proposed by Baddeley and Hitch. This is followed by an account of some of the problems it encountered in explaining how information from different subsystems with different codes could be combined, and how it was capable of communicating with long-term memory. In order to account for these, a fourth component was proposed, the episodic buffer. This was assumed to be a multidimensional store of limited capacity that can be accessed through conscious awareness. In an attempt to test and develop the concept, a series of experiments have explored the role of working memory in the binding of visual features into objects and verbal sequences into remembered sentences. The experiments use a dual task paradigm to investigate the role of the various subcomponents of working memory in binding. In contrast to our initial assumption, the episodic buffer appears to be a passive store, capable of storing bound features and making them available to conscious awareness, but not itself responsible for the process of binding.

  18. Buffer moisture protection system

    International Nuclear Information System (INIS)

    Ritola, J.; Peura, J.

    2013-11-01

    With the present knowledge, bentonite blocks have to be protected from the air relative humidity and from any moisture leakages in the environment that might cause swelling of the bentonite blocks during the 'open' installation phase before backfilling. The purpose of this work was to design the structural reference solution both for the bottom of the deposition hole and for the buffer moisture protection and dewatering system with their integrated equipment needed in the deposition hole. This report describes the Posiva's reference solution for the buffer moisture protection system and the bottom plate on basis of the demands and functional requirements set by long-term safety. The reference solution with structural details has been developed in research work made 2010-2011. The structural solution of the moisture protection system has not yet been tested in practice. On the bottom of the deposition hole a copper plate which protects the lowest bentonite block from the gathered water is installed straight to machined and even rock surface. The moisture protection sheet made of EPDM rubber is attached to the copper plate with an inflatable seal. The upper part of the moisture protection sheet is fixed to the collar structures of the lid which protects the deposition hole in the disposal tunnel. The main function of the moisture protection sheet is to protect bentonite blocks from the leaking water and from the influence of the air humidity at their installation stage. The leaking water is controlled by the dewatering and alarm system which has been integrated into the moisture protection liner. (orig.)

  19. Buffer capacity of biologics--from buffer salts to buffering by antibodies.

    Science.gov (United States)

    Karow, Anne R; Bahrenburg, Sven; Garidel, Patrick

    2013-01-01

    Controlling pH is essential for a variety of biopharmaceutical process steps. The chemical stability of biologics such as monoclonal antibodies is pH-dependent and slightly acidic conditions are favorable for stability in a number of cases. Since control of pH is widely provided by added buffer salts, the current study summarizes the buffer characteristics of acetate, citrate, histidine, succinate, and phosphate buffers. Experimentally derived values largely coincide with values calculated from a model that had been proposed in 1922 by van Slyke. As high concentrated protein formulations become more and more prevalent for biologics, the self-buffering potential of proteins becomes of relevance. The current study provides information on buffer characteristics for pH ranges down to 4.0 and up to 8.0 and shows that a monoclonal antibody at 50 mg/mL exhibits similar buffer capacity as 6 mM citrate or 14 mM histidine (pH 5.0-6.0). Buffer capacity of antibody solutions scales linearly with protein concentration up to more than 200 mg/mL. At a protein concentration of 220 mg/mL, the buffer capacity resembles the buffer capacity of 30 mM citrate or 50 mM histidine (pH 5.0-6.0). The buffer capacity of monoclonal antibodies is practically identical at the process relevant temperatures 5, 25, and 40°C. Changes in ionic strength of ΔI=0.15, in contrast, can alter the buffer capacity up to 35%. In conclusion, due to efficient self-buffering by antibodies in the pH range of favored chemical stability, conventional buffer excipients could be dispensable for pH stabilization of high concentrated protein solutions. Copyright © 2013 American Institute of Chemical Engineers.

  20. Two-Buffer Simulation Games

    Directory of Open Access Journals (Sweden)

    Milka Hutagalung

    2016-07-01

    Full Text Available We consider simulation games played between Spoiler and Duplicator on two Büchi automata in which the choices made by Spoiler can be buffered by Duplicator in two different buffers before she executes them on her structure. Previous work on such games using a single buffer has shown that they are useful to approximate language inclusion problems. We study the decidability and complexity and show that games with two buffers can be used to approximate corresponding problems on finite transducers, i.e. the inclusion problem for rational relations over infinite words.

  1. DMPD: Cytosolic DNA recognition for triggering innate immune responses. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 18280611 Cytosolic DNA recognition for triggering innate immune responses. Takaoka ...A, Taniguchi T. Adv Drug Deliv Rev. 2008 Apr 29;60(7):847-57. Epub 2007 Dec 31. (.png) (.svg) (.html) (.csml) Show Cytosol...ic DNA recognition for triggering innate immune responses. PubmedID 18280611 Title Cytosolic D

  2. Using fractional order method to generalize strengthening generating operator buffer operator and weakening buffer operator

    OpenAIRE

    Wu, L.; Liu, S.; Yang, Yingjie

    2016-01-01

    Traditional integer order buffer operator is extended to fractional order buffer operator, the corresponding relationship between the weakening buffer operator and the strengthening buffer operator is revealed. Fractional order buffer operator not only can generalize the weakening buffer operator and the strengthening buffer operator, but also realize tiny adjustment of buffer effect. The effectiveness of GM(1,1) with the fractional order buffer operator is validated by six cases.

  3. Mechanisms of buffer therapy resistance.

    Science.gov (United States)

    Bailey, Kate M; Wojtkowiak, Jonathan W; Cornnell, Heather H; Ribeiro, Maria C; Balagurunathan, Yoganand; Hashim, Arig Ibrahim; Gillies, Robert J

    2014-04-01

    Many studies have shown that the acidity of solid tumors contributes to local invasion and metastasis. Oral pH buffers can specifically neutralize the acidic pH of tumors and reduce the incidence of local invasion and metastatic formation in multiple murine models. However, this effect is not universal as we have previously observed that metastasis is not inhibited by buffers in some tumor models, regardless of buffer used. B16-F10 (murine melanoma), LL/2 (murine lung) and HCT116 (human colon) tumors are resistant to treatment with lysine buffer therapy, whereas metastasis is potently inhibited by lysine buffers in MDA-MB-231 (human breast) and PC3M (human prostate) tumors. In the current work, we confirmed that sensitive cells utilized a pH-dependent mechanism for successful metastasis supported by a highly glycolytic phenotype that acidifies the local tumor microenvironment resulting in morphological changes. In contrast, buffer-resistant cell lines exhibited a pH-independent metastatic mechanism involving constitutive secretion of matrix degrading proteases without elevated glycolysis. These results have identified two distinct mechanisms of experimental metastasis, one of which is pH-dependent (buffer therapy sensitive cells) and one which is pH-independent (buffer therapy resistant cells). Further characterization of these models has potential for therapeutic benefit. Copyright © 2014 Neoplasia Press, Inc. Published by Elsevier Inc. All rights reserved.

  4. Surface-Enhanced Raman Spectroscopy (SERS Tracking of Chelerythrine, a Na+/K+ Pump Inhibitor, into Cytosol and Plasma Membrane Fractions of Human Lens Epithelial Cell Cultures

    Directory of Open Access Journals (Sweden)

    Kevin M. Dorney

    2013-12-01

    Full Text Available Background/Aims: The quaternary benzo-phenanthridine alkaloid (QBA chelerythrine (CET is a pro-apoptotic drug and Na+/K+ pump (NKP inhibitor in human lens epithelial cells (HLECs. In order to obtain further insight into the mechanism of NKP inhibition by CET, its sub-cellular distribution was quantified in cytosolic and membrane fractions of HLEC cultures by surface-enhanced Raman spectroscopy (SERS. Methods: Silver nanoparticles (AgNPs prepared by the Creighton method were concentrated, and size-selected using a one-step tangential flow filtration approach. HLECs cultures were exposed to 50 μM CET in 300 mOsM phosphate-buffered NaCl for 30 min. A variety of cytosolic extracts, crude and purified membranes, prepared in lysing solutions in the presence and absence of a non-ionic detergent, were incubated with AgNPs and subjected to SERS analysis. Determinations of CET were based on a linear calibration plot of the integrated CET SERS intensity at its 659 cm-1 marker band as a function of CET concentration. Results: SERS detected chemically unaltered CET in both cytosol and plasma membrane fractions. Normalized for protein, the CET content was some 100 fold higher in the crude and purified plasma membrane fraction than in the soluble cytosolic extract. The total free CET concentration in the cytosol, free of membranes or containing detergent-solubilized membrane material, approached that of the incubation medium of HLECs. Conclusion: Given a negative membrane potential of HLECs the data suggest, but do not prove, that CET may traverse the plasma membrane as a positively charged monomer (CET+ accumulating near or above passive equilibrium distribution. These findings may contribute to a recently proposed hypothesis that CET binds to and inhibits the NKP through its cytosolic aspect.

  5. Surface-enhanced Raman spectroscopy (SERS) tracking of chelerythrine, a Na(+)/K(+) pump inhibitor, into cytosol and plasma membrane fractions of human lens epithelial cell cultures.

    Science.gov (United States)

    Dorney, Kevin M; Sizemore, Ioana E P; Alqahtani, Tariq; Adragna, Norma C; Lauf, Peter K

    2013-01-01

    The quaternary benzo-phenanthridine alkaloid (QBA) chelerythrine (CET) is a pro-apoptotic drug and Na(+)/K(+) pump (NKP) inhibitor in human lens epithelial cells (HLECs). In order to obtain further insight into the mechanism of NKP inhibition by CET, its sub-cellular distribution was quantified in cytosolic and membrane fractions of HLEC cultures by surface-enhanced Raman spectroscopy (SERS). Silver nanoparticles (AgNPs) prepared by the Creighton method were concentrated, and size-selected using a one-step tangential flow filtration approach. HLECs cultures were exposed to 50 μM CET in 300 mOsM phosphate-buffered NaCl for 30 min. A variety of cytosolic extracts, crude and purified membranes, prepared in lysing solutions in the presence and absence of a non-ionic detergent, were incubated with AgNPs and subjected to SERS analysis. Determinations of CET were based on a linear calibration plot of the integrated CET SERS intensity at its 659 cm(-1) marker band as a function of CET concentration. SERS detected chemically unaltered CET in both cytosol and plasma membrane fractions. Normalized for protein, the CET content was some 100 fold higher in the crude and purified plasma membrane fraction than in the soluble cytosolic extract. The total free CET concentration in the cytosol, free of membranes or containing detergent-solubilized membrane material, approached that of the incubation medium of HLECs. Given a negative membrane potential of HLECs the data suggest, but do not prove, that CET may traverse the plasma membrane as a positively charged monomer (CET(+)) accumulating near or above passive equilibrium distribution. These findings may contribute to a recently proposed hypothesis that CET binds to and inhibits the NKP through its cytosolic aspect. © 2014 S. Karger AG, Basel.

  6. Electrodialysis operation with buffer solution

    Science.gov (United States)

    Hryn, John N [Naperville, IL; Daniels, Edward J [Orland Park, IL; Krumdick, Greg K [Crete, IL

    2009-12-15

    A new method for improving the efficiency of electrodialysis (ED) cells and stacks, in particular those used in chemical synthesis. The process entails adding a buffer solution to the stack for subsequent depletion in the stack during electrolysis. The buffer solution is regenerated continuously after depletion. This buffer process serves to control the hydrogen ion or hydroxide ion concentration so as to protect the active sites of electrodialysis membranes. The process enables electrodialysis processing options for products that are sensitive to pH changes.

  7. Cytosolic antibody delivery by lipid-sensitive endosomolytic peptide

    Science.gov (United States)

    Akishiba, Misao; Takeuchi, Toshihide; Kawaguchi, Yoshimasa; Sakamoto, Kentarou; Yu, Hao-Hsin; Nakase, Ikuhiko; Takatani-Nakase, Tomoka; Madani, Fatemeh; Gräslund, Astrid; Futaki, Shiroh

    2017-08-01

    One of the major obstacles in intracellular targeting using antibodies is their limited release from endosomes into the cytosol. Here we report an approach to deliver proteins, which include antibodies, into cells by using endosomolytic peptides derived from the cationic and membrane-lytic spider venom peptide M-lycotoxin. The delivery peptides were developed by introducing one or two glutamic acid residues into the hydrophobic face. One peptide with the substitution of leucine by glutamic acid (L17E) was shown to enable a marked cytosolic liberation of antibodies (immunoglobulins G (IgGs)) from endosomes. The predominant membrane-perturbation mechanism of this peptide is the preferential disruption of negatively charged membranes (endosomal membranes) over neutral membranes (plasma membranes), and the endosomolytic peptide promotes the uptake by inducing macropinocytosis. The fidelity of this approach was confirmed through the intracellular delivery of a ribosome-inactivation protein (saporin), Cre recombinase and IgG delivery, which resulted in a specific labelling of the cytosolic proteins and subsequent suppression of the glucocorticoid receptor-mediated transcription. We also demonstrate the L17E-mediated cytosolic delivery of exosome-encapsulated proteins.

  8. Mechanistic logic underlying the axonal transport of cytosolic proteins

    Science.gov (United States)

    Scott, David A.; Das, Utpal; Tang, Yong; Roy, Subhojit

    2011-01-01

    Proteins vital to presynaptic function are synthesized in the neuronal perikarya and delivered into synapses via two modes of axonal transport. While membrane-anchoring proteins are conveyed in fast axonal transport via motor-driven vesicles, cytosolic proteins travel in slow axonal transport; via mechanisms that are poorly understood. We found that in cultured axons, populations of cytosolic proteins tagged to photoactivable-GFP (PA-GFP) move with a slow motor-dependent anterograde bias; distinct from vesicular-trafficking or diffusion of untagged PA-GFP. The overall bias is likely generated by an intricate particle-kinetics involving transient assembly and short-range vectorial spurts. In-vivo biochemical studies reveal that cytosolic proteins are organized into higher-order structures within axon-enriched fractions that are largely segregated from vesicles. Data-driven biophysical modeling best predicts a scenario where soluble molecules dynamically assemble into mobile supra-molecular structures. We propose a model where cytosolic proteins are transported by dynamically assembling into multi-protein complexes that are directly/indirectly conveyed by motors. PMID:21555071

  9. Organizers and activators: Cytosolic Nox proteins impacting on vascular function.

    Science.gov (United States)

    Schröder, Katrin; Weissmann, Norbert; Brandes, Ralf P

    2017-08-01

    NADPH oxidases of the Nox family are important enzymatic sources of reactive oxygen species (ROS) in the cardiovascular system. Of the 7 members of the Nox family, at least three depend for their activation on specific cytosolic proteins. These are p47phox and its homologue NoxO1 and p67phox and its homologue NoxA1. Also the Rho-GTPase Rac is important but as this protein has many additional functions, it will not be covered here. The Nox1 enzyme is preferentially activated by the combination of NoxO1 with NoxA1, whereas Nox2 gains highest activity with p47phox together with p67phox. As p47phox, different to NoxO1 contains an auto inhibitory region it has to be phosphorylated prior to complex formation. In the cardio-vascular system, all cytosolic Nox proteins are expressed but the evidence for their contribution to ROS production is not well established. Most data have been collected for p47phox, whereas NoxA1 has basically not yet been studied. In this article the specific aspects of cytosolic Nox proteins in the cardiovascular system with respect to Nox activation, their expression and their importance will be reviewed. Finally, it will be discussed whether cytosolic Nox proteins are suitable pharmacological targets to tamper with vascular ROS production. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Heat conductivity of buffer materials

    International Nuclear Information System (INIS)

    Boergesson, L.; Fredrikson, Anders; Johannesson, L.E.

    1994-11-01

    The report deals with the thermal conductivity of bentonite based buffer materials. An improved technique for measuring the thermal conductivity of buffer materials is described. Measurements of FLAC calculations applying this technique have led to a proposal of how standardized tests should be conducted and evaluated. The thermal conductivity of bentonite with different void ratio and degree of water saturation has been determined in the following different ways: * Theoretically according to three different investigations by other researchers. * Laboratory measurements with the proposed method. * Results from back-calculated field tests. Comparison and evaluation showed that these results agreed very well, when the buffer material was almost water saturated. However, the influence of the degree of saturation was not very well predicted with the theoretical methods. Furthermore, the field tests showed that the average thermal conductivity in situ of buffer material (compacted to blocks) with low degree of water saturation was lower than expected from laboratory tests. 12 refs, 29 figs, 11 tabs

  11. Buffers and vegetative filter strips

    Science.gov (United States)

    Matthew J. Helmers; Thomas M. Isenhart; Michael G. Dosskey; Seth M. Dabney

    2008-01-01

    This chapter describes the use of buffers and vegetative filter strips relative to water quality. In particular, we primarily discuss the herbaceous components of the following NRCS Conservation Practice Standards.

  12. Programmable pH buffers

    Science.gov (United States)

    Gough, Dara Van; Huber, Dale L.; Bunker, Bruce C.; Roberts, Mark E.

    2017-01-24

    A programmable pH buffer comprises a copolymer that changes pK.sub.a at a lower critical solution temperature (LCST) in water. The copolymer comprises a thermally programmable polymer that undergoes a hydrophobic-to-hydrophilic phase change at the LCST and an electrolytic polymer that exhibits acid-base properties that are responsive to the phase change. The programmable pH buffer can be used to sequester CO.sub.2 into water.

  13. Developmental changes in cytosolic coupling between epidermis cells as visualized by photoactivation of fluorescein

    DEFF Research Database (Denmark)

    Liu, Xiangdong; Martens, Helle; Schulz, Alexander

    Developmental changes in cytosolic coupling between epidermis cells as visualized by photoactivation of fluorescein.......Developmental changes in cytosolic coupling between epidermis cells as visualized by photoactivation of fluorescein....

  14. The chaperone BAG6 captures dislocated glycoproteins in the cytosol.

    Directory of Open Access Journals (Sweden)

    Jasper H L Claessen

    Full Text Available Secretory and membrane (glycoproteins are subject to quality control in the endoplasmic reticulum (ER to ensure that only functional proteins reach their destination. Proteins deemed terminally misfolded and hence functionally defective may be dislocated to the cytosol, where the proteasome degrades them. What we know about this process stems mostly from overexpression of tagged misfolded proteins, or from situations where viruses have hijacked the quality control machinery to their advantage. We know of only very few endogenous substrates of ER quality control, most of which are degraded as part of a signaling pathway, such as Insig-1, but such examples do not necessarily represent terminally misfolded proteins. Here we show that endogenous dislocation clients are captured specifically in association with the cytosolic chaperone BAG6, or retrieved en masse via their glycan handle.

  15. A new view of the bacterial cytosol environment.

    Directory of Open Access Journals (Sweden)

    Benjamin P Cossins

    2011-06-01

    Full Text Available The cytosol is the major environment in all bacterial cells. The true physical and dynamical nature of the cytosol solution is not fully understood and here a modeling approach is applied. Using recent and detailed data on metabolite concentrations, we have created a molecular mechanical model of the prokaryotic cytosol environment of Escherichia coli, containing proteins, metabolites and monatomic ions. We use 200 ns molecular dynamics simulations to compute diffusion rates, the extent of contact between molecules and dielectric constants. Large metabolites spend ∼80% of their time in contact with other molecules while small metabolites vary with some only spending 20% of time in contact. Large non-covalently interacting metabolite structures mediated by hydrogen-bonds, ionic and π stacking interactions are common and often associate with proteins. Mg(2+ ions were prominent in NIMS and almost absent free in solution. Κ(+ is generally not involved in NIMSs and populates the solvent fairly uniformly, hence its important role as an osmolyte. In simulations containing ubiquitin, to represent a protein component, metabolite diffusion was reduced owing to long lasting protein-metabolite interactions. Hence, it is likely that with larger proteins metabolites would diffuse even more slowly. The dielectric constant of these simulations was found to differ from that of pure water only through a large contribution from ubiquitin as metabolite and monatomic ion effects cancel. These findings suggest regions of influence specific to particular proteins affecting metabolite diffusion and electrostatics. Also some proteins may have a higher propensity for associations with metabolites owing to their larger electrostatic fields. We hope that future studies may be able to accurately predict how binding interactions differ in the cytosol relative to dilute aqueous solution.

  16. Thermophysical tests of buffer materials

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, H. [ITC, Tokyo (Japan); Taniguchi, Wataru

    1999-03-01

    Thermodynamic properties of buffer materials were measured for putting in order thermodynamic constants to be used in the near-field thermal analysis. The thermal diffusivity and thermal conductivity were measured as functions of the water content and temperature to deduce the specific heat. The thermal conductivity and specific heat varied significantly as the water content changed. Obtained values of the specific heat agreed well the expected values calculated based on the constituents of the buffer material. Temperature dependence of the thermodynamic constants was found small below 90degC. From the findings, the thermal conductivity and specific heat of the buffer material were formulated as functions of the water content. Thermodynamic study of powdery bentonite was carried out as well with a purpose of use for filling apertures in the artificial barrier. (H. Baba)

  17. ACETIC ACID AND A BUFFER

    DEFF Research Database (Denmark)

    2009-01-01

    The present invention relates to a composition comprising : a) 0.01-20% wt/wt acetic acid and b) a physiologically tolerable buffer capable of maintaining acetic acid at a pH in the range of 2-7; and use of such a composition as an antimicrobial agent.......The present invention relates to a composition comprising : a) 0.01-20% wt/wt acetic acid and b) a physiologically tolerable buffer capable of maintaining acetic acid at a pH in the range of 2-7; and use of such a composition as an antimicrobial agent....

  18. A Capital Adequacy Buffer Model

    NARCIS (Netherlands)

    D.E. Allen (David); M.J. McAleer (Michael); R.J. Powell (Robert); A.K. Singh (Abhay)

    2013-01-01

    markdownabstract__Abstract__ In this paper, we develop a new capital adequacy buffer model (CABM) which is sensitive to dynamic economic circumstances. The model, which measures additional bank capital required to compensate for fluctuating credit risk, is a novel combination of the Merton

  19. Smoothing type buffer memory device

    International Nuclear Information System (INIS)

    Podorozhnyj, D.M.; Yashin, I.V.

    1990-01-01

    The layout of the micropower 4-bit smoothing type buffer memory device allowing one to record without counting the sequence of input randomly distributed pulses in multi-channel devices with serial poll, is given. The power spent by a memory cell for one binary digit recording is not greater than 0.15 mW, the device dead time is 10 mus

  20. Formation of monofunctional cisplatin-DNA adducts in carbonate buffer.

    Science.gov (United States)

    Binter, Alexandra; Goodisman, Jerry; Dabrowiak, James C

    2006-07-01

    Carbonate in its various forms is an important component in blood and the cytosol. Since, under conditions that simulate therapy, carbonate reacts with cisplatin to form carbonato complexes, one of which is taken up and/or modified by the cell [C.R. Centerwall, J. Goodisman, D.J. Kerwood, J. Am. Chem. Soc., 127 (2005) 12768-12769], cisplatin-carbonato complexes may be important in the mechanism of action of cisplatin. In this report we study the binding of cisplatin to pBR322 DNA in two different buffers, using gel electrophoresis. In 23.8mM HEPES, N-(2-hydroxyethyl)-piperazine-N'-2-ethanesulfonic acid, 5mM NaCl, pH 7.4 buffer, cisplatin produces aquated species, which react with DNA to unwind supercoiled Form I DNA, increasing its mobility, and reducing the binding of ethidium to DNA. This behavior is consistent with the formation of the well-known intrastrand crosslink on DNA. In 23.8mM carbonate buffer, 5mM NaCl, pH 7.4, cisplatin forms carbonato species that produce DNA-adducts which do not significantly change supercoiling but enhance binding of ethidium to DNA. This behavior is consistent with the formation of a monofunctional cisplatin adduct on DNA. These results show that aquated cisplatin and carbonato complexes of cisplatin produce different types of lesions on DNA and they underscore the importance of carrying out binding studies with cisplatin and DNA using conditions that approximate those found in the cell.

  1. Targeting Cytosolic Nucleic Acid-Sensing Pathways for Cancer Immunotherapies.

    Science.gov (United States)

    Iurescia, Sandra; Fioretti, Daniela; Rinaldi, Monica

    2018-01-01

    The innate immune system provides the first line of defense against pathogen infection though also influences pathways involved in cancer immunosurveillance. The innate immune system relies on a limited set of germ line-encoded sensors termed pattern recognition receptors (PRRs), signaling proteins and immune response factors. Cytosolic receptors mediate recognition of danger damage-associated molecular patterns (DAMPs) signals. Once activated, these sensors trigger multiple signaling cascades, converging on the production of type I interferons and proinflammatory cytokines. Recent studies revealed that PRRs respond to nucleic acids (NA) released by dying, damaged, cancer cells, as danger DAMPs signals, and presence of signaling proteins across cancer types suggests that these signaling mechanisms may be involved in cancer biology. DAMPs play important roles in shaping adaptive immune responses through the activation of innate immune cells and immunological response to danger DAMPs signals is crucial for the host response to cancer and tumor rejection. Furthermore, PRRs mediate the response to NA in several vaccination strategies, including DNA immunization. As route of double-strand DNA intracellular entry, DNA immunization leads to expression of key components of cytosolic NA-sensing pathways. The involvement of NA-sensing mechanisms in the antitumor response makes these pathways attractive drug targets. Natural and synthetic agonists of NA-sensing pathways can trigger cell death in malignant cells, recruit immune cells, such as DCs, CD8 + T cells, and NK cells, into the tumor microenvironment and are being explored as promising adjuvants in cancer immunotherapies. In this minireview, we discuss how cGAS-STING and RIG-I-MAVS pathways have been targeted for cancer treatment in preclinical translational researches. In addition, we present a targeted selection of recent clinical trials employing agonists of cytosolic NA-sensing pathways showing how these pathways

  2. Protein abundance profiling of the Escherichia coli cytosol

    DEFF Research Database (Denmark)

    Ishihama, Y.; Schmidt, T.; Rappsilber, J.

    2008-01-01

    sample. Using a combination of LC-MS/MS approaches with protein and peptide fractionation steps we identified 1103 proteins from the cytosolic fraction of the Escherichia coli strain MC4100. A measure of abundance is presented for each of the identified proteins, based on the recently developed emPAI...... approach which takes into account the number of sequenced peptides per protein. The values of abundance are within a broad range and accurately reflect independently measured copy numbers per cell. As expected, the most abundant proteins were those involved in protein synthesis, most notably ribosomal...

  3. Urban Runoff: Model Ordinances for Aquatic Buffers

    Science.gov (United States)

    Aquatic Buffers serve as natural boundaries between local waterways and existing development. The model and example ordinaces below provide suggested language or technical guidance designed to create the most effective stream buffer zones possible.

  4. Buffer Zone Requirements for Soil Fumigant Applications

    Science.gov (United States)

    Updated pesticide product labels require fumigant users to establish a buffer zone around treated fields to reduce risks to bystanders. Useful information includes tarp testing guidance and a buffer zone calculator.

  5. Swelling characteristics of buffer material

    International Nuclear Information System (INIS)

    Suzuki, Hideaki; Fujita, Tomoo

    1999-12-01

    After emplacement of the engineered barrier system (EBS), it is expected that the near-field environment will be impacted by phenomena such as heat dissipation by conduction and other heat transfer mechanism, infiltration of groundwater from the surrounding rock into the EBS, generation of swelling pressure in the buffer due to water infiltration and the stress imposed by the overburden pressure. These phenomena are not all independent, but can be strongly influenced by, and coupled with, each other. Evaluating these coupled thermo-hydro-mechanical phenomena is important in order to clarify the initial transient behavior of the engineered barrier system within the near-field. This report describes the results on measurement of swelling amount and stress at boundary built up under restraint condition with water uptake. The following results are identified. (1) The swelling stress of buffer material at saturated condition tends to be independent of effects of pore water pressure and synthetic sea water, and to decrease with increasing temperature. The swelling stress can be explained by the effective dry density. (2) The strain due to swelling estimated from the results of the swelling amount of buffer material is proportional to swelling stress. (3) The swelling stress and strain under unsaturated condition increase with water uptake. (author)

  6. Glutathionylation regulates cytosolic NADP+-dependent isocitrate dehydrogenase activity.

    Science.gov (United States)

    Shin, Seoung Woo; Oh, Chang Joo; Kil, In Sup; Park, Jeen-Woo

    2009-04-01

    Cytosolic NADP+-dependent isocitrate dehydrogenase (IDPc) is susceptible to inactivation by numerous thiol-modifying reagents. This study now reports that Cys269 of IDPc is a target for S-glutathionylation and that this modification is reversed by dithiothreitol as well as enzymatically by cytosolic glutaredoxin in the presence of GSH. Glutathionylated IDPc was significantly less susceptible than native protein to peptide fragmentation by reactive oxygen species and proteolytic digestion. Glutathionylation may play a protective role in the degradation of protein through the structural alterations of IDPc. HEK293 cells treated with diamide displayed decreased IDPc activity and accumulated glutathionylated enzyme. Using immunoprecipitation with an anti-IDPc IgG and immunoblotting with an anti-GSH IgG, we purified and positively identified glutathionylated IDPc from the kidneys of mice subjected to ischemia/reperfusion injury and from the livers of ethanol-administered rats. These results suggest that IDPc activity is modulated through enzymatic glutathionylation and deglutathionylation during oxidative stress.

  7. Buffer erosion in dilute groundwater

    International Nuclear Information System (INIS)

    Schatz, T.; Kanerva, N.; Martikainen, J.; Sane, P.; Olin, M.; Seppaelae, A.; Koskinen, K.

    2013-08-01

    One scenario of interest for repository safety assessment involves the loss of bentonite buffer material in contact with dilute groundwater flowing through a transmissive fracture interface. In order to examine the extrusion/erosion behavior of bentonite buffer material under such circumstances, a series of experiments were performed in a flow-through, 1 mm aperture, artificial fracture system. These experiments covered a range of solution chemistry (salt concentration and composition), material composition (sodium montmorillonite and admixtures with calcium montmorillonite), and flow velocity conditions. No erosion was observed for sodium montmorillonite against solution compositions from 0.5 g/L to 10 g/L NaCl. No erosion was observed for 50/50 calcium/sodium montmorillonite against 0.5 g/L NaCl. Erosion was observed for both sodium montmorillonite and 50/50 calcium/sodium montmorillonite against solution compositions ≤ 0.25 g/L NaCl. The calculated erosion rates for the tests with the highest levels of measured erosion, i.e., the tests run under the most dilute conditions (ionic strength (IS) < ∼1 mM), were well-correlated to flow velocity, whereas the calculated erosion rates for the tests with lower levels of measured erosion, i.e., the tests run under somewhat less dilute conditions (∼1 mM < IS < ∼4 mM), were not similarly correlated indicating that material and solution composition can significantly affect erosion rates. In every experiment, both erosive and non-erosive, emplaced buffer material extruded into the fracture and was observed to be impermeable to water flowing in the fracture effectively forming an extended diffusive barrier around the intersecting fracture/buffer interface. Additionally, a model which was developed previously to predict the rate of erosion of bentonite buffer material in low ionic strength water in rock fracture environments was applied to three different cases: sodium montmorillonite expansion in a vertical tube, a

  8. BUFFER CAPACITY IN HETEROGENEOUS MULTICOMPONENT SYSTEMS. REVIEW

    Directory of Open Access Journals (Sweden)

    Oxana Spinu

    2015-12-01

    Full Text Available The quantitative basis of the theory of buffer properties for two-phase acid-base buffer systems and for multicomponent heterogeneous systems has been derived. The analytical equations with respect to all components for diverse multicomponent systems were deduced. It has been established, that the buffer capacities of components are mutually proportional.

  9. Doped LZO buffer layers for laminated conductors

    Science.gov (United States)

    Paranthaman, Mariappan Parans [Knoxville, TN; Schoop, Urs [Westborough, MA; Goyal, Amit [Knoxville, TN; Thieme, Cornelis Leo Hans [Westborough, MA; Verebelyi, Darren T [Oxford, MA; Rupich, Martin W [Framingham, MA

    2010-03-23

    A laminated conductor includes a metallic substrate having a surface, a biaxially textured buffer layer supported by the surface of the substrate, the biaxially textured buffer layer comprising LZO and a dopant for mitigating metal diffusion through the LZO, and a biaxially textured conductor layer supported by the biaxially textured buffer layer.

  10. RESEARCH NEEDS IN RIPARIAN BUFFER RESTORATION

    Science.gov (United States)

    Riparian buffer restorations are used as management tools to produce favorable water quality impacts; moreover, the basis for riparian buffers as an instrument of water quality restoration rests on a relatively firm foundation. However, the extent to which buffers can restore rip...

  11. Ammodytoxin, a neurotoxic secreted phospholipase A2, can act in the cytosol of the nerve cell

    International Nuclear Information System (INIS)

    Petrovic, Uros; Sribar, Jernej; Paris, Alenka; Rupnik, Marjan; Krzan, Mojca; Vardjan, Nina; Gubensek, Franc; Zorec, Robert; Krizaj, Igor

    2004-01-01

    Recent identification of intracellular proteins that bind ammodytoxin (calmodulin, 14-3-3 proteins, and R25) suggests that this snake venom presynaptically active phospholipase A 2 acts intracellularly. As these ammodytoxin acceptors are cytosolic and mitochondrial proteins, the toxin should be able to enter the cytosol of a target cell and remain stable there to interact with them. Using laser scanning confocal microscopy we show here that Alexa-labelled ammodytoxin entered the cytoplasm of the rat hippocampal neuron and subsequently also its nucleus. The transport of proteins into the nucleus proceeds via the cytosol of a cell, therefore, ammodytoxin passed the cytosol of the neuron on its way to the nucleus. Although it is not yet clear how ammodytoxin is translocated into the cytosol of the neuron, our results demonstrate that its stability in the cytosol is not in question, providing the evidence that the toxin can act in this cellular compartment

  12. The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling

    Energy Technology Data Exchange (ETDEWEB)

    Bernard, S.M.; Habash, D.Z.

    2009-07-02

    Glutamine synthetase assimilates ammonium into amino acids, thus it is a key enzyme for nitrogen metabolism. The cytosolic isoenzymes of glutamine synthetase assimilate ammonium derived from primary nitrogen uptake and from various internal nitrogen recycling pathways. In this way, cytosolic glutamine synthetase is crucial for the remobilization of protein-derived nitrogen. Cytosolic glutamine synthetase is encoded by a small family of genes that are well conserved across plant species. Members of the cytosolic glutamine synthetase gene family are regulated in response to plant nitrogen status, as well as to environmental cues, such as nitrogen availability and biotic/abiotic stresses. The complex regulation of cytosolic glutamine synthetase at the transcriptional to post-translational levels is key to the establishment of a specific physiological role for each isoenzyme. The diverse physiological roles of cytosolic glutamine synthetase isoenzymes are important in relation to current agricultural and ecological issues.

  13. Improving Water Quality With Conservation Buffers

    Science.gov (United States)

    Lowrance, R.; Dabney, S.; Schultz, R.

    2003-12-01

    Conservation buffer technologies are new approaches that need wider application. In-field buffer practices work best when used in combination with other buffer types and other conservation practices. Vegetative barriers may be used in combination with edge-of-field buffers to protect and improve their function and longevity by dispersing runoff and encouraging sediment deposition upslope of the buffer. It's important to understand how buffers can be managed to help reduce nutrient transport potential for high loading of nutrients from manure land application sites, A restored riparian wetland buffer retained or removed at least 59 percent of the nitrogen and 66 percent of the phosphorus that entered from an adjacent manure land application site. The Bear Creek National Restoration Demonstration Watershed project in Iowa has been the site of riparian forest buffers and filter strips creation; constructed wetlands to capture tile flow; stream-bank bioengineering; in-stream structures; and controlling livestock grazing. We need field studies that test various widths of buffers of different plant community compositions for their efficacy in trapping surface runoff, reducing nonpoint source pollutants in subsurface waters, and enhancing the aquatic ecosystem. Research is needed to evaluate the impact of different riparian grazing strategies on channel morphology, water quality, and the fate of livestock-associated pathogens and antibiotics. Integrating riparian buffers and other conservation buffers into these models is a key objective in future model development.

  14. Buffered Communication Analysis in Distributed Multiparty Sessions

    Science.gov (United States)

    Deniélou, Pierre-Malo; Yoshida, Nobuko

    Many communication-centred systems today rely on asynchronous messaging among distributed peers to make efficient use of parallel execution and resource access. With such asynchrony, the communication buffers can happen to grow inconsiderately over time. This paper proposes a static verification methodology based on multiparty session types which can efficiently compute the upper bounds on buffer sizes. Our analysis relies on a uniform causality audit of the entire collaboration pattern - an examination that is not always possible from each end-point type. We extend this method to design algorithms that allocate communication channels in order to optimise the memory requirements of session executions. From these analyses, we propose two refinements methods which respect buffer bounds: a global protocol refinement that automatically inserts confirmation messages to guarantee stipulated buffer sizes and a local protocol refinement to optimise asynchronous messaging without buffer overflow. Finally our work is applied to overcome a buffer overflow problem of the multi-buffering algorithm.

  15. A THEORETICAL DISCUSSION OF THE ECONOMIC EFFECTS OF BUFFER STOCKS AND BUFFER FUNDS

    OpenAIRE

    Simmons, Phil

    1988-01-01

    It has been established that the absence of risk markets justifies market intervention in principle. The form of intervention that has been discussed most widely in the literature is the buffer stock. This paper points out that other forms of intervention, specifically buffer funds, are likely to perform better. The analysis shows that buffer funds are likely to outperform buffer stocks because they address market failure more directly. A sub-theme developed in this paper is that since buffer...

  16. Extracellular but not cytosolic superoxide dismutase protects against oxidant-mediated endothelial dysfunction

    Directory of Open Access Journals (Sweden)

    Erin L. Foresman

    2013-01-01

    Full Text Available Superoxide (O2•− contributes to the development of cardiovascular disease. Generation of O2•− occurs in both the intracellular and extracellular compartments. We hypothesized that the gene transfer of cytosolic superoxide dismutase (SOD1 or extracellular SOD (SOD3 to blood vessels would differentially protect against O2•−-mediated endothelial-dependent dysfunction. Aortic ring segments from New Zealand rabbits were incubated with adenovirus (Ad containing the gene for Escherichia coli β-galactosidase, SOD1, or SOD3. Activity assays confirmed functional overexpression of both SOD3 and SOD1 isoforms in aorta 24 h following gene transfer. Histochemical staining for β-galactosidase showed gene transfer occurred in the endothelium and adventitia. Next, vessels were prepared for measurement of isometric tension in Kreb's buffer containing xanthine. After precontraction with phenylephrine, xanthine oxidase impaired relaxation to the endothelium-dependent dilator acetylcholine (ACh, max relaxation 33±4% with XO vs. 64±3% without XO, p<0.05, whereas relaxation to the endothelium-independent dilator sodium nitroprusside was unaffected. In the presence of XO, maximal relaxation to ACh was improved in vessels incubated with AdSOD3 (55±2%, p<0.05 vs. control but not AdSOD1 (34±4%. We conclude that adenoviral-mediated gene transfer of SOD3, but not SOD1, protects the aorta from xanthine/XO-mediated endothelial dysfunction. These data provide important insight into the location and enzymatic source of O2•− production in vascular disease.

  17. Structural characterization of coatomer in its cytosolic state

    Directory of Open Access Journals (Sweden)

    Shengliu Wang

    2016-07-01

    Full Text Available Abstract Studies on coat protein I (COPI have contributed to a basic understanding of how coat proteins generate vesicles to initiate intracellular transport. The core component of the COPI complex is coatomer, which is a multimeric complex that needs to be recruited from the cytosol to membrane in order to function in membrane bending and cargo sorting. Previous structural studies on the clathrin adaptors have found that membrane recruitment induces a large conformational change in promoting their role in cargo sorting. Here, pursuing negative-stain electron microscopy coupled with single-particle analyses, and also performing CXMS (chemical cross-linking coupled with mass spectrometry for validation, we have reconstructed the structure of coatomer in its soluble form. When compared to the previously elucidated structure of coatomer in its membrane-bound form we do not observe a large conformational change. Thus, the result uncovers a key difference between how COPI versus clathrin coats are regulated by membrane recruitment.

  18. Buffered Electrochemical Polishing of Niobium

    Energy Technology Data Exchange (ETDEWEB)

    Ciovati, Gianluigi [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States); Tian, Hui [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States); College of William and Mary, Williamsburg, VA (United States); Corcoran, Sean [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States)

    2011-03-01

    The standard preparation of superconducting radio-frequency (SRF) cavities made of pure niobium include the removal of a 'damaged' surface layer, by buffered chemical polishing (BCP) or electropolishing (EP), after the cavities are formed. The performance of the cavities is characterized by a sharp degradation of the quality factor when the surface magnetic field exceeds about 90 mT, a phenomenon referred to as 'Q-drop.' In cavities made of polycrystalline fine grain (ASTM 5) niobium, the Q-drop can be significantly reduced by a low-temperature (? 120 °C) 'in-situ' baking of the cavity if the chemical treatment was EP rather than BCP. As part of the effort to understand this phenomenon, we investigated the effect of introducing a polarization potential during buffered chemical polishing, creating a process which is between the standard BCP and EP. While preliminary results on the application of this process to Nb cavities have been previously reported, in this contribution we focus on the characterization of this novel electrochemical process by measuring polarization curves, etching rates, surface finish, electrochemical impedance and the effects of temperature and electrolyte composition. In particular, it is shown that the anodic potential of Nb during BCP reduces the etching rate and improves the surface finish.

  19. Characterization of the cytosolic distribution of priority pollutant metals and metalloids in the digestive gland cytosol of marine mussels: seasonal and spatial variability.

    Science.gov (United States)

    Strižak, Zeljka; Ivanković, Dušica; Pröfrock, Daniel; Helmholz, Heike; Cindrić, Ana-Marija; Erk, Marijana; Prange, Andreas

    2014-02-01

    Cytosolic profiles of several priority pollutant metals (Cu, Cd, Zn, Pb) and metalloid As were analyzed in the digestive gland of the mussel (Mytilus galloprovincialis) sampled at locations with different environmental pollution levels along the Croatian coast in the spring and summer season. Size-exclusion chromatography (SEC) connected to inductively coupled plasma mass spectrometry (ICP-MS) was used to determine selected elements bound to cytosolic biomolecules separated based on their molecular size. Copper, cadmium and zinc eluted mostly associated with high molecular weight (HMW) and medium molecular weight (MMW) biomolecules, but with a more prominent elution in the MMW peak at polluted locations which were probably associated with the 20 kDa metallothionein (MT). Elution of all three metals within this peak was also strongly correlated with cytosolic Cd as strong inducer of MT. Lead mostly eluted in HMW biomolecule range, but in elevated cytosolic Pb concentrations, significant amount eluted in low molecular weight (LMW) biomolecules. Arsenic, on the other hand eluted almost completely in LMW range, but we could not distinguish specific molecular weight biomolecules which would be predominant in detoxification mechanism. Seasonal variability in element abundance within specific peaks was present, although not in the same extent, for all elements and locations, especially for As. The results confirm the suitability of the distribution of selected metals/metalloids among different cytosolic ligands as potential indicator for metal exposure. Obtained findings can also serve as guidelines for further separation and characterization of specific cytosolic metal-binding biomolecules. © 2013.

  20. Platelet cytosolic 44-kDa protein is a substrate of cholera toxin-induced ADP-ribosylation and is not recognized by antisera against the α subunit of the stimulatory guanine nucleotide-binding regulatory protein

    International Nuclear Information System (INIS)

    Molina Y Vedia, L.M.; Reep, B.R.; Lapetina, E.G.

    1988-01-01

    ADP-ribosylation induced by cholera toxin and pertussis toxin was studied in particulate and cytosolic fractions of human platelets. Platelets were disrupted by a cycle of freezing and thawing in the presence of a hyposmotic buffer containing protease inhibitors. In both fractions, the A subunit of cholera toxin ADP-ribosylates two proteins with molecular masses of 42 and 44 kDa, whereas pertussis toxin ADP-ribosylates a 41-kDa polypeptide. Two antisera against the α subunit of the stimulatory guanine nucleotide-binding regulatory protein recognize only the 42-kDa polypeptide. Cholera toxin-induced ADP-ribosylation of the 42- and 44-kDa proteins is reduced by pretreatment of platelets with iloprost, a prostacyclin analog. The 44-kDa protein, which is substrate of cholera toxin, could be extracted completely from the membrane and recovered in the cytosolic fraction when the cells were disrupted by Dounce homogenization and the pellet was extensively washed. A 44-kDa protein can also be labeled with 8-azidoguanosine 5'-[α- 32 P]triphosphate in the cytosol and membranes. These finding indicate that cholera and pertussis toxins produced covalent modifications of proteins present in particulate and cytosolic platelet fractions. Moreover, the 44-kDa protein might be an α subunit of a guanine nucleotide-binding regulatory protein that is not recognized by available antisera

  1. Fluctuations in Cytosolic Calcium Regulate the Neuronal Malate-Aspartate NADH Shuttle

    DEFF Research Database (Denmark)

    Satrústegui, Jorgina; Bak, Lasse K

    2015-01-01

    that MAS is regulated by fluctuations in cytosolic Ca(2+) levels, and that this regulation is required to maintain a tight coupling between neuronal activity and mitochondrial respiration and oxidative phosphorylation. At cytosolic Ca(2+) fluctuations below the threshold of the mitochondrial calcium...

  2. Induction of Cytosolic Acetyl-Coenzyme A Carboxylase in Pea Leaves by Ultraviolet-B Irradiation

    OpenAIRE

    Tomokazu, Konishi; Takahiro, Kamoi; Ryuichi, Matsuno; Yukiko, Sasaki; Department of Food Science and Technology, Faculty of Agriculture, Kyoto University:(Present)Laboratory of Molecular Genetics, Biotechnology Institute, Akita Prefectural College of Agriculture; Department of Food Science and Technology, Faculty of Agriculture, Kyoto University; Department of Food Science and Technology, Faculty of Agriculture, Kyoto University; Department of Food Science and Technology, Faculty of Agriculture, Kyoto University:(Present)Laboratory of Plant Molecular Biology, School of Agricultural Sciences, Nagoya University

    1996-01-01

    Levels of subunits of two acetyl-coenzyme A carboxylases were high in small leaves of Pisum sativum, decreased with growth, and remained constant in fully expanded leaves. Irradiation of fully expanded leaves induced the cytosolic isozyme only. This result suggests a key role for the cytosolic enzyme in protection against UV-B.

  3. Different myrosinase and idioblast distribution in Arabidopsis and Brassica napus

    DEFF Research Database (Denmark)

    Andreasson, Erik; Jørgensen, Lise Bolt; Höglund, Anna-Stina

    2001-01-01

    Arabidopsis, Brassica napus, Myrosinase, Myrosinase Binding Protein, Glucosinolates, Myrosin Cell, Immunocytochemistry......Arabidopsis, Brassica napus, Myrosinase, Myrosinase Binding Protein, Glucosinolates, Myrosin Cell, Immunocytochemistry...

  4. Melatonin: Buffering the Immune System

    Directory of Open Access Journals (Sweden)

    Juan M. Guerrero

    2013-04-01

    Full Text Available Melatonin modulates a wide range of physiological functions with pleiotropic effects on the immune system. Despite the large number of reports implicating melatonin as an immunomodulatory compound, it still remains unclear how melatonin regulates immunity. While some authors argue that melatonin is an immunostimulant, many studies have also described anti-inflammatory properties. The data reviewed in this paper support the idea of melatonin as an immune buffer, acting as a stimulant under basal or immunosuppressive conditions or as an anti-inflammatory compound in the presence of exacerbated immune responses, such as acute inflammation. The clinical relevance of the multiple functions of melatonin under different immune conditions, such as infection, autoimmunity, vaccination and immunosenescence, is also reviewed.

  5. Melatonin: Buffering the Immune System

    Science.gov (United States)

    Carrillo-Vico, Antonio; Lardone, Patricia J.; Álvarez-Sánchez, Nuria; Rodríguez-Rodríguez, Ana; Guerrero, Juan M.

    2013-01-01

    Melatonin modulates a wide range of physiological functions with pleiotropic effects on the immune system. Despite the large number of reports implicating melatonin as an immunomodulatory compound, it still remains unclear how melatonin regulates immunity. While some authors argue that melatonin is an immunostimulant, many studies have also described anti-inflammatory properties. The data reviewed in this paper support the idea of melatonin as an immune buffer, acting as a stimulant under basal or immunosuppressive conditions or as an anti-inflammatory compound in the presence of exacerbated immune responses, such as acute inflammation. The clinical relevance of the multiple functions of melatonin under different immune conditions, such as infection, autoimmunity, vaccination and immunosenescence, is also reviewed. PMID:23609496

  6. Modelling of buffer material behaviour

    International Nuclear Information System (INIS)

    Boergesson, L.

    1988-12-01

    Some material models of smectite rich buffer material suited for nuclear waste isolation are accounted for in the report. The application of these models in finite element calculations of some scenarios and performance are also shown. The rock shear scenario has been closely studied with comparisons between calculated and measured results. Sensitivity analyses of the effect of changing the density of the clay and the rate of shear have been performed as well as one calculation using a hollow steel cylinder. Material models and finite element calculations of canister settlement, thermomechanical effects and swelling are also accounted for. The report shows the present state of the work to establish material models and calculation tools which can be used at the final design of the repository. (31 illustrations)

  7. Signature-based store checking buffer

    Science.gov (United States)

    Sridharan, Vilas; Gurumurthi, Sudhanva

    2015-06-02

    A system and method for optimizing redundant output verification, are provided. A hardware-based store fingerprint buffer receives multiple instances of output from multiple instances of computation. The store fingerprint buffer generates a signature from the content included in the multiple instances of output. When a barrier is reached, the store fingerprint buffer uses the signature to verify the content is error-free.

  8. SODR Memory Control Buffer Control ASIC

    Science.gov (United States)

    Hodson, Robert F.

    1994-01-01

    The Spacecraft Optical Disk Recorder (SODR) is a state of the art mass storage system for future NASA missions requiring high transmission rates and a large capacity storage system. This report covers the design and development of an SODR memory buffer control applications specific integrated circuit (ASIC). The memory buffer control ASIC has two primary functions: (1) buffering data to prevent loss of data during disk access times, (2) converting data formats from a high performance parallel interface format to a small computer systems interface format. Ten 144 p in, 50 MHz CMOS ASIC's were designed, fabricated and tested to implement the memory buffer control function.

  9. Cytosolic lipolysis and lipophagy: two sides of the same coin.

    Science.gov (United States)

    Zechner, Rudolf; Madeo, Frank; Kratky, Dagmar

    2017-11-01

    Fatty acids are the most efficient substrates for energy production in vertebrates and are essential components of the lipids that form biological membranes. Synthesis of triacylglycerols from non-esterified free fatty acids (FFAs) combined with triacylglycerol storage represents a highly efficient strategy to stockpile FFAs in cells and prevent FFA-induced lipotoxicity. Although essentially all vertebrate cells have some capacity to store and utilize triacylglycerols, white adipose tissue is by far the largest triacylglycerol depot and is uniquely able to supply FFAs to other tissues. The release of FFAs from triacylglycerols requires their enzymatic hydrolysis by a process called lipolysis. Recent discoveries thoroughly altered and extended our understanding of lipolysis. This Review discusses how cytosolic 'neutral' lipolysis and lipophagy, which utilizes 'acid' lipolysis in lysosomes, degrade cellular triacylglycerols as well as how these pathways communicate, how they affect lipid metabolism and energy homeostasis and how their dysfunction affects the pathogenesis of metabolic diseases. Answers to these questions will likely uncover novel strategies for the treatment of prevalent metabolic diseases.

  10. Surveillance for Intracellular Antibody by Cytosolic Fc Receptor TRIM21

    Directory of Open Access Journals (Sweden)

    William A. McEwan

    2016-11-01

    Full Text Available TRIM21 has emerged as an atypical Fc receptor that is broadly conserved and widely expressed in the cytoplasm of mammalian cells. Viruses that traffic surface-bound antibodies into the cell during infection recruit TRIM21 via a high affinity interaction between Fc and TRIM21 PRYSPRY domain. Following binding of intracellular antibody, TRIM21 acts as both antiviral effector and sensor for innate immune signalling. These activities serve to reduce viral replication by orders of magnitude in vitro and contribute to host survival during in vivo infection. Neutralization occurs rapidly after detection and requires the activity of the ubiquitin-proteasome system. The microbial targets of this arm of intracellular immunity are still being identified: TRIM21 activity has been reported following infection by several non-enveloped viruses and intracellular bacteria. These findings extend the sphere of influence of antibodies to the intracellular domain and have broad implications for immunity. TRIM21 has been implicated in the chronic auto-immune condition systemic lupus erythematosus and is itself an auto-antigen in Sjögren’s syndrome. This review summarises our current understanding of TRIM21’s role as a cytosolic Fc receptor and briefly discusses pathological circumstances where intracellular antibodies have been described, or are hypothesized to occur, and may benefit from further investigations of the role of TRIM21.

  11. Cytosolic Access of Intracellular Bacterial Pathogens: The Shigella Paradigm.

    Science.gov (United States)

    Mellouk, Nora; Enninga, Jost

    2016-01-01

    Shigella is a Gram-negative bacterial pathogen, which causes bacillary dysentery in humans. A crucial step of Shigella infection is its invasion of epithelial cells. Using a type III secretion system, Shigella injects several bacterial effectors ultimately leading to bacterial internalization within a vacuole. Then, Shigella escapes rapidly from the vacuole, it replicates within the cytosol and spreads from cell-to-cell. The molecular mechanism of vacuolar rupture used by Shigella has been studied in some detail during the recent years and new paradigms are emerging about the underlying molecular events. For decades, bacterial effector proteins were portrayed as main actors inducing vacuolar rupture. This includes the effector/translocators IpaB and IpaC. More recently, this has been challenged and an implication of the host cell in the process of vacuolar rupture has been put forward. This includes the bacterial subversion of host trafficking regulators, such as the Rab GTPase Rab11. The involvement of the host in determining bacterial vacuolar integrity has also been found for other bacterial pathogens, particularly for Salmonella. Here, we will discuss our current view of host factor and pathogen effector implications during Shigella vacuolar rupture and the steps leading to it.

  12. Cytosolic access of intracellular bacterial pathogens: the Shigella paradigm

    Directory of Open Access Journals (Sweden)

    Nora eMellouk

    2016-04-01

    Full Text Available Shigella is a Gram-negative bacterial pathogen, which causes bacillary dysentery in humans. A crucial step of Shigella infection is its invasion of epithelial cells. Using a type III secretion system, Shigella injects several bacterial effectors ultimately leading to bacterial internalization within a vacuole. Then, Shigella escapes rapidly from the vacuole, it replicates within the cytosol and spreads from cell-to-cell. The molecular mechanism of vacuolar rupture used by Shigella has been studied in some detail during the recent years and new paradigms are emerging about the underlying molecular events. For decades, bacterial effector proteins were portrayed as main actors inducing vacuolar rupture. This includes the effector/translocators IpaB and IpaC. More recently, this has been challenged and an implication of the host cell in the process of vacuolar rupture has been put forward. This includes the bacterial subversion of host trafficking regulators, such as the Rab GTPase Rab11. The involvement of the host in determining bacterial vacuolar integrity has also been found for other bacterial pathogens, particularly for Salmonella. Here, we will discuss our current view of host factor and pathogen effector implications during Shigella vacuolar rupture and the steps leading to it.

  13. Cytosolic sensing of immuno-stimulatory DNA, the enemy within.

    Science.gov (United States)

    Dhanwani, Rekha; Takahashi, Mariko; Sharma, Sonia

    2018-02-01

    In the cytoplasm, DNA is sensed as a universal danger signal by the innate immune system. Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor/enzyme that catalyzes formation of 2'-5'-cGAMP, an atypical cyclic di-nucleotide second messenger that binds and activates the Stimulator of Interferon Genes (STING), resulting in recruitment of Tank Binding Kinase 1 (TBK1), activation of the transcription factor Interferon Regulatory Factor 3 (IRF3), and trans-activation of innate immune response genes, including type I Interferon cytokines (IFN-I). Activation of the pro-inflammatory cGAS-STING-IRF3 response is triggered by direct recognition of the DNA genomes of bacteria and viruses, but also during RNA virus infection, neoplastic transformation, tumor immunotherapy and systemic auto-inflammatory diseases. In these circumstances, the source of immuno-stimulatory DNA has often represented a fundamental yet poorly understood aspect of the response. This review focuses on recent findings related to cGAS activation by an array of self-derived DNA substrates, including endogenous retroviral elements, mitochondrial DNA (mtDNA) and micronuclei generated as a result of genotoxic stress and DNA damage. These findings emphasize the role of the cGAS axis as a cell-intrinsic innate immune response to a wide variety of genomic insults. Copyright © 2017. Published by Elsevier Ltd.

  14. Genome-Derived Cytosolic DNA Mediates Type I Interferon-Dependent Rejection of B Cell Lymphoma Cells

    Directory of Open Access Journals (Sweden)

    Yu J. Shen

    2015-04-01

    Full Text Available The DNA damage response (DDR induces the expression of type I interferons (IFNs, but the underlying mechanisms are poorly understood. Here, we show the presence of cytosolic DNA in different mouse and human tumor cells. Treatment of cells with genotoxic agents increased the levels of cytosolic DNA in a DDR-dependent manner. Cloning of cytosolic DNA molecules from mouse lymphoma cells suggests that cytosolic DNA is derived from unique genomic loci and has the potential to form non-B DNA structures, including R-loops. Overexpression of Rnaseh1, which resolves R-loops, reduced the levels of cytosolic DNA, type I Ifn transcripts, and type I IFN-dependent rejection of lymphoma cells. Live-cell imaging showed a dynamic contact of cytosolic DNA with mitochondria, an important organelle for innate immune recognition of cytosolic nucleotides. In summary, we found that cytosolic DNA is present in many tumor cells and contributes to the immunogenicity of tumor cells.

  15. Temperature buffer test. Dismantling operation

    Energy Technology Data Exchange (ETDEWEB)

    Aakesson, Mattias [Clay Technology AB, Lund (Sweden)

    2010-12-15

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aespoe HRL. It was installed during the spring of 2003. Two heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by bentonite in the usual way, whereas the upper heater was surrounded by a ring of sand. The test was dismantled and sampled during a period from the end of October 2009 to the end of April 2010, and this report describes this operation. Different types of samples have been obtained during this operation. A large number of diameter 50 mm bentonite cores have been taken for analysis of water content and density. Large pieces, so-called big sectors, have been taken for hydro-mechanical and chemical characterizations. Finally, there has been an interest to obtain different types of interface samples in which bentonite were in contact with sand, iron or concrete. One goal has been to investigate the retrievability of the upper heater, given the possibility to remove the surrounding sand shield, and a retrieval test has therefore been performed. The sand in the shield was first removed with an industrial vacuum cleaner after loosening the material through mechanical means (with hammer drill and core machine). A front loader was subsequently used for applying a sufficient lifting force to release the heater from the bentonite underneath. The experiment has been documented in different aspects: measurements of the coordinate (height or radius) of different interfaces (between bentonite blocks and between bentonite and sand); verification of sensor positions and retrieval of sensors for subsequent

  16. Temperature buffer test. Dismantling operation

    International Nuclear Information System (INIS)

    Aakesson, Mattias

    2010-12-01

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aespoe HRL. It was installed during the spring of 2003. Two heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by bentonite in the usual way, whereas the upper heater was surrounded by a ring of sand. The test was dismantled and sampled during a period from the end of October 2009 to the end of April 2010, and this report describes this operation. Different types of samples have been obtained during this operation. A large number of diameter 50 mm bentonite cores have been taken for analysis of water content and density. Large pieces, so-called big sectors, have been taken for hydro-mechanical and chemical characterizations. Finally, there has been an interest to obtain different types of interface samples in which bentonite were in contact with sand, iron or concrete. One goal has been to investigate the retrievability of the upper heater, given the possibility to remove the surrounding sand shield, and a retrieval test has therefore been performed. The sand in the shield was first removed with an industrial vacuum cleaner after loosening the material through mechanical means (with hammer drill and core machine). A front loader was subsequently used for applying a sufficient lifting force to release the heater from the bentonite underneath. The experiment has been documented in different aspects: measurements of the coordinate (height or radius) of different interfaces (between bentonite blocks and between bentonite and sand); verification of sensor positions and retrieval of sensors for subsequent

  17. 21 CFR 520.1696a - Buffered penicillin powder, penicillin powder with buffered aqueous diluent.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Buffered penicillin powder, penicillin powder with... FORM NEW ANIMAL DRUGS § 520.1696a Buffered penicillin powder, penicillin powder with buffered aqueous diluent. (a) Specifications. When reconstituted, each milliliter contains penicillin G procaine equivalent...

  18. THE BUFFER CAPACITY OF AIRWAY EPITHELIAL SECRETIONS

    Directory of Open Access Journals (Sweden)

    Dusik eKim

    2014-06-01

    Full Text Available The pH of airway epithelial secretions influences bacterial killing and mucus properties and is reduced by acidic pollutants, gastric reflux, and respiratory diseases such as cystic fibrosis (CF. The effect of acute acid loads depends on buffer capacity, however the buffering of airway secretions has not been well characterized. In this work we develop a method for titrating micro-scale (30 µl volumes and use it to study fluid secreted by the human airway epithelial cell line Calu-3, a widely used model for submucosal gland serous cells. Microtitration curves revealed that HCO3- is the major buffer. Peak buffer capacity (β increased from 17 to 28 mM/pH during forskolin stimulation, and was reduced by >50% in fluid secreted by cystic fibrosis transmembrane conductance regulator (CFTR-deficient Calu-3 monolayers, confirming an important role of CFTR in HCO3- secretion. Back-titration with NaOH revealed non-volatile buffer capacity due to proteins synthesized and released by the epithelial cells. Lysozyme and mucin concentrations were too low to buffer Calu-3 fluid significantly, however model titrations of porcine gastric mucins at concentrations near the sol-gel transition suggest that mucins may contribute to the buffer capacity of ASL in vivo. We conclude that CFTR-dependent HCO3- secretion and epithelially-derived proteins are the predominant buffers in Calu-3 secretions.

  19. Shock buffer for nuclear control assembly

    International Nuclear Information System (INIS)

    Bevilacqua, F.

    1977-01-01

    A shock buffer is provided for the gradual deceleration of a rapidly descending control element assembly in a nuclear reactor. The interactive buffer components are associated respectively with the movable control element assembly and part of the upper guide structure independent of and spaced from the fuel assemblies of the reactor

  20. Buffer sizing for multi-hop networks

    KAUST Repository

    Shihada, Basem

    2014-01-28

    A cumulative buffer may be defined for an interference domain in a wireless mesh network and distributed among nodes in the network to maintain or improve capacity utilization of network resources in the interference domain without increasing packet queuing delay times. When an interference domain having communications links sharing resources in a network is identified, a cumulative buffer size is calculated. The cumulative buffer may be distributed among buffers in each node of the interference domain according to a simple division or according to a cost function taking into account a distance of the communications link from the source and destination. The network may be monitored and the cumulative buffer size recalculated and redistributed when the network conditions change.

  1. Optimization of protein buffer cocktails using Thermofluor.

    Science.gov (United States)

    Reinhard, Linda; Mayerhofer, Hubert; Geerlof, Arie; Mueller-Dieckmann, Jochen; Weiss, Manfred S

    2013-02-01

    The stability and homogeneity of a protein sample is strongly influenced by the composition of the buffer that the protein is in. A quick and easy approach to identify a buffer composition which increases the stability and possibly the conformational homogeneity of a protein sample is the fluorescence-based thermal-shift assay (Thermofluor). Here, a novel 96-condition screen for Thermofluor experiments is presented which consists of buffer and additive parts. The buffer screen comprises 23 different buffers and the additive screen includes small-molecule additives such as salts and nucleotide analogues. The utilization of small-molecule components which increase the thermal stability of a protein sample frequently results in a protein preparation of higher quality and quantity and ultimately also increases the chances of the protein crystallizing.

  2. DMPD: Regulation of arachidonic acid release and cytosolic phospholipase A2activation. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 10080535 Regulation of arachidonic acid release and cytosolic phospholipase A2activ...on of arachidonic acid release and cytosolic phospholipase A2activation. PubmedID 10080535 Title Regulation ...of arachidonic acid release and cytosolic phospholipase A2activation. Authors Gij

  3. Cytosolic PrP Can Participate in Prion-Mediated Toxicity

    Science.gov (United States)

    Thackray, Alana M.; Zhang, Chang; Arndt, Tina

    2014-01-01

    ABSTRACT Prion diseases are characterized by a conformational change in the normal host protein PrPC. While the majority of mature PrPC is tethered to the plasma membrane by a glycosylphosphatidylinositol anchor, topological variants of this protein can arise during its biosynthesis. Here we have generated Drosophila transgenic for cytosolic ovine PrP in order to investigate its toxic potential in flies in the absence or presence of exogenous ovine prions. While cytosolic ovine PrP expressed in Drosophila was predominantly detergent insoluble and showed resistance to low concentrations of proteinase K, it was not overtly detrimental to the flies. However, Drosophila transgenic for cytosolic PrP expression exposed to classical or atypical scrapie prion inocula showed a faster decrease in locomotor activity than similar flies exposed to scrapie-free material. The susceptibility to classical scrapie inocula could be assessed in Drosophila transgenic for panneuronal expression of cytosolic PrP, whereas susceptibility to atypical scrapie required ubiquitous PrP expression. Significantly, the toxic phenotype induced by ovine scrapie in cytosolic PrP transgenic Drosophila was transmissible to recipient PrP transgenic flies. These data show that while cytosolic PrP expression does not adversely affect Drosophila, this topological PrP variant can participate in the generation of transmissible scrapie-induced toxicity. These observations also show that PrP transgenic Drosophila are susceptible to classical and atypical scrapie prion strains and highlight the utility of this invertebrate host as a model of mammalian prion disease. IMPORTANCE During prion diseases, the host protein PrPC converts into an abnormal conformer, PrPSc, a process coupled to the generation of transmissible prions and neurotoxicity. While PrPC is principally a glycosylphosphatidylinositol-anchored membrane protein, the role of topological variants, such as cytosolic PrP, in prion-mediated toxicity and

  4. COMBINATIONS OF BUFFER-STOCKS AND BUFFER-FUNDS FOR WOOL PRICE STABILISATION IN AUSTRALIA

    OpenAIRE

    Moir, Brian; Piggott, Roley R.

    1991-01-01

    In this paper a preliminary analysis is presented of a combined buffer-fund and buffer-stock as an alternative to a pure buffer-fund or a pure buffer stock for stabilising wool prices. The alternatives analysed are designed so that each provides the same prices to producers as did the Reserve Price Scheme over the period of analysis. Least-cost combinations of policy instruments are derived. The results show that there is considerable potential for cost savings to be made by combining buffer-...

  5. Cytosolic superoxide dismutase can provide protection against Fasciola gigantica.

    Science.gov (United States)

    Jaikua, Wipaphorn; Kueakhai, Pornanan; Chaithirayanon, Kulathida; Tanomrat, Rataya; Wongwairot, Sirima; Riengrojpitak, Suda; Sobhon, Prasert; Changklungmoa, Narin

    2016-10-01

    Superoxide dismutases (SOD), antioxidant metallo-enzymes, are a part of the first line of defense in the trematode parasites which act as the chief scavengers for reactive oxygen species (ROS). A recombinant Fasciola gigantica cytosolic SOD (FgSOD) was expressed in Escherichia coli BL21 (DE3) and used for immunizing rabbits to obtain polyclonal antibodies (anti-rFgSOD). This rabbit anti-rFgSOD reacted with the native FgSOD at a molecular weight of 17.5kDa. The FgSOD protein was expressed at high level in parenchyma, caecal epithelium and egg of the parasite. The rFgSOD reacted with antisera from rabbits infected with F. gigantica metacercariae collected at 2, 5, and 7 weeks after infection, and reacted with sera of infected mice. Anti-rFgSOD exhibited cross reactivity with the other parasites' antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. A vaccination was performed in imprinting control region (ICR) mice by subcutaneous injection with 50μg of rFgSOD combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 15 metacercariae by oral route. IgG1 and IgG2a in the immune sera were determined to indicate Th2 and Th1 immune responses. It was found that the parasite burden was reduced by 45%, and both IgG1 and IgG2a levels showed correlation with the numbers of worm recoveries. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Cytosolic Calcium Coordinates Mitochondrial Energy Metabolism with Presynaptic Activity

    Science.gov (United States)

    Chouhan, Amit K.; Ivannikov, Maxim V.; Lu, Zhongmin; Sugimori, Mutsuyuki; Llinas, Rodolfo R.; Macleod, Gregory T.

    2012-01-01

    Most neurons fire in bursts, imposing episodic energy demands, but how these demands are coordinated with oxidative phosphorylation is still unknown. Here, using fluorescence imaging techniques on presynaptic termini of Drosophila motor neurons (MNs), we show that mitochondrial matrix pH (pHm), inner membrane potential (Δψm), and NAD(P)H levels ([NAD(P)H]m) increase within seconds of nerve stimulation. The elevations of pHm, Δψm, and [NAD(P)H]m indicate an increased capacity for ATP production. Elevations in pHm were blocked by manipulations which blocked mitochondrial Ca2+ uptake, including replacement of extracellular Ca2+ with Sr2+, and application of either tetraphenylphosphonium chloride or KB-R7943, indicating that it is Ca2+ that stimulates presynaptic mitochondrial energy metabolism. To place this phenomenon within the context of endogenous neuronal activity, the firing rates of a number of individually identified MNs were determined during fictive locomotion. Surprisingly, although endogenous firing rates are significantly different, there was little difference in presynaptic cytosolic Ca2+ levels ([Ca2+]c) between MNs when each fires at its endogenous rate. The average [Ca2+]c level (329±11nM) was slightly above the average Ca2+ affinity of the mitochondria (281±13nM). In summary, we show that when MNs fire at endogenous rates [Ca2+]c is driven into a range where mitochondria rapidly acquire Ca2+. As we also show that Ca2+ stimulates presynaptic mitochondrial energy metabolism, we conclude that [Ca2+]c levels play an integral role in coordinating mitochondrial energy metabolism with presynaptic activity in Drosophila MNs. PMID:22279208

  7. Buffer mass test - Site documentation

    International Nuclear Information System (INIS)

    Pusch, R.

    1983-10-01

    The purpose of this report is to compile test site data that are assumed to be of importance for the interpretation of the Buffer Mass Test. Since this test mainly concerns water uptake and migration processes in the integrated rock/backfill system and the development of temperature fields in this system, the work has been focused on the constitution and hydrology of the rock. The major constitutional rock feature of interest for the BMT is the frequency and distribution of joints and fractures. The development of models for water uptake into the highly compacted bentonite in the heater holes requires a very detailed fracture survey. The present investigation shows that two of the holes (no. 1 and 2) are located in richly fractured rock, while the others are located in fracture-poor to moderately fractured rock. The hydrological conditions of the rock in the BMT area are characterized by water pressures of as much as 100 m water head at a few meters distance from the test site. The average hydraulic conductivity of the rock that confines the BMT tunnel has been estimated at about 10 -10 m/s by Lawrence Laboratory. The actual distribution of the water that enters the tunnel has been estimated by observing the successive moistening after having switched off the ventilation, and this has offered basis of predicting the rate and uniformity of the water uptake in the tunnel backfill. As to the heater holes the detailed fracture patterns and various inflow measurements have yielded a similar basis. The report also gives major data on the rock temperature, gas conditions, mineralogy, rock mechanics, and groundwater chemistry for BMT purposes. (author)

  8. Temperature buffer test. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Aakesson, Mattias [Clay Technology AB, Lund (Sweden)

    2012-04-15

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aspo HRL. It was installed during the spring of 2003. Two steel heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by rings of compacted Wyoming bentonite only, whereas the upper heater was surrounded by a composite barrier, with a sand shield between the heater and the bentonite. The test was dismantled and sampled during the winter of 2009/2010. This report is the final report and a summary of all work performed within the TBT project. The design and the installation of the different components are summarized: the depositions hole, the heating system, the bentonite blocks with emphasis on the initial density and water content in these, the filling of slots with sand or pellets, the retaining construction with the plug, lid and nine anchor cables, the artificial saturation system, and finally the instrumentation. An overview of the operational conditions is presented: the power output from heaters, which was 1,500 W (and also 1,600 W) from each heater during the first {approx}1,700 days, and then changed to 1,000 and 2,000 W, for the upper and lower heater respectively, during the last {approx}600 days. From the start, the bentonite was hydrated with a groundwater from a nearby bore-hole, but this groundwater was replaced with de-ionized water from day {approx}1,500, due to the high flow resistance of the injections points in the filter, which implied that a high filter pressure couldn't be sustained. The sand shield around the upper heater was hydrated from day {approx}1,500 to day {approx}1

  9. Temperature buffer test. Final report

    International Nuclear Information System (INIS)

    Aakesson, Mattias

    2012-04-01

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aspo HRL. It was installed during the spring of 2003. Two steel heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by rings of compacted Wyoming bentonite only, whereas the upper heater was surrounded by a composite barrier, with a sand shield between the heater and the bentonite. The test was dismantled and sampled during the winter of 2009/2010. This report is the final report and a summary of all work performed within the TBT project. The design and the installation of the different components are summarized: the depositions hole, the heating system, the bentonite blocks with emphasis on the initial density and water content in these, the filling of slots with sand or pellets, the retaining construction with the plug, lid and nine anchor cables, the artificial saturation system, and finally the instrumentation. An overview of the operational conditions is presented: the power output from heaters, which was 1,500 W (and also 1,600 W) from each heater during the first ∼1,700 days, and then changed to 1,000 and 2,000 W, for the upper and lower heater respectively, during the last ∼600 days. From the start, the bentonite was hydrated with a groundwater from a nearby bore-hole, but this groundwater was replaced with de-ionized water from day ∼1,500, due to the high flow resistance of the injections points in the filter, which implied that a high filter pressure couldn't be sustained. The sand shield around the upper heater was hydrated from day ∼1,500 to day ∼1,800. The sensors data concerning

  10. Dynamic mechanical properties of buffer material

    International Nuclear Information System (INIS)

    Takaji, Kazuhiko; Taniguchi, Wataru

    1999-11-01

    The buffer material is expected to maintain its low water permeability, self-sealing properties, radionuclides adsorption and retardation properties, thermal conductivity, chemical buffering properties, overpack supporting properties, stress buffering properties, etc. over a long period of time. Natural clay is mentioned as a material that can relatively satisfy above. Among the kinds of natural clay, bentonite when compacted is superior because (i) it has exceptionally low water permeability and properties to control the movement of water in buffer, (ii) it fills void spaces in the buffer and fractures in the host rock as it swells upon water uptake, (iii) it has the ability to exchange cations and to adsorb cationic radioelements. In order to confirm these functions for the purpose of safety assessment, it is necessary to evaluate buffer properties through laboratory tests and engineering-scale tests, and to make assessments based on the ranges in the data obtained. This report describes the procedures, test conditions, results and examinations on the buffer material of dynamic triaxial tests, measurement of elastic wave velocity and liquefaction tests that aim at getting hold of dynamic mechanical properties. We can get hold of dependency on the shearing strain of the shearing modulus and hysteresis damping constant, the application for the mechanical model etc. by dynamic triaxial tests, the acceptability of maximum shearing modulus obtained from dynamic triaxial tests etc. by measurement of elastic wave velocity and dynamic strength caused by cyclic stress etc. by liquefaction tests. (author)

  11. Buffer-regulated biocorrosion of pure magnesium.

    Science.gov (United States)

    Kirkland, Nicholas T; Waterman, Jay; Birbilis, Nick; Dias, George; Woodfield, Tim B F; Hartshorn, Richard M; Staiger, Mark P

    2012-02-01

    Magnesium (Mg) alloys are being actively investigated as potential load-bearing orthopaedic implant materials due to their biodegradability in vivo. With Mg biomaterials at an early stage in their development, the screening of alloy compositions for their biodegradation rate, and hence biocompatibility, is reliant on cost-effective in vitro methods. The use of a buffer to control pH during in vitro biodegradation is recognised as critically important as this seeks to mimic pH control as it occurs naturally in vivo. The two different types of in vitro buffer system available are based on either (i) zwitterionic organic compounds or (ii) carbonate buffers within a partial-CO(2) atmosphere. This study investigated the influence of the buffering system itself on the in vitro corrosion of Mg. It was found that the less realistic zwitterion-based buffer did not form the same corrosion layers as the carbonate buffer, and was potentially affecting the behaviour of the hydrated oxide layer that forms on Mg in all aqueous environments. Consequently it was recommended that Mg in vitro experiments use the more biorealistic carbonate buffering system when possible.

  12. Monitoring change in refractive index of cytosol of animal cells on affinity surface under osmotic stimulus for label-free measurement of viability.

    Science.gov (United States)

    Park, Jina; Jin, Sung Il; Kim, Hyung Min; Ahn, Junhyoung; Kim, Yeon-Gu; Lee, Eun Gyo; Kim, Min-Gon; Shin, Yong-Beom

    2015-02-15

    We demonstrated that a metal-clad waveguide (MCW)-based biosensor can be applied to label-free measurements of viability of adherent animal cells with osmotic stimulation in real time. After Chinese hamster ovary (CHO) and human embryonic kidney cell 293 (HEK293) cells were attached to a Concanavalin A (Con A)-modified sensor surface, the magnitudes of cell responses to non-isotonic stimulation were compared between live and dead cells. The live cells exhibited a change in the refractive index (RI) of the cytosol caused by a redistribution of water through the cell membrane, which was induced by the osmotic stimulus, but the dead cells did not. Moreover, the normalized change in the RI measured via the MCW sensor was linearly proportional to the viability of attached cells and the resolution in monitoring cell viability was about 0.079%. Therefore, the viability of attached animal cells can be measured without labels by observing the relative differences in the RI of cytosol in isotonic and non-isotonic buffers. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Vitamin A effects on UMR 106 osteosarcoma cells are not mediated by specific cytosolic receptors.

    OpenAIRE

    Oreffo, R O; Francis, J A; Triffitt, J T

    1985-01-01

    Retinol and retinoic acid at 20 microM altered cell morphology and inhibited cell proliferation of UMR 106 osteosarcoma cells in culture. No specific cytosolic binding proteins for retinol could be detected.

  14. Buffer protection in the installation phase

    International Nuclear Information System (INIS)

    Wimelius, Hans; Pusch, Roland

    2008-12-01

    The research and development of the design and construction of the SKB's repository for final disposal of spent reactor fuel is conducted along several paths ('lines'). Issues concerning the bedrock are dealt with in the 'rock line' and those related to buffer and backfill in deposition holes and tunnels are considered in the 'buffer line' and 'backfill line', respectively. These lines also deal with sub-activities that are coupled to several other lines. One of them includes development of techniques for protecting buffer blocks from moisture and water in the installation phase. Techniques and methods for placement and removal of the 'buffer protection sheet' are dealt with in the 'buffer line'. The removal is, however, considered as being part of the backfilling sequence. Since the performance of the sheet is of fundamental importance to the placement and function of the buffer it deserves particular attention. Thus, the removal of the rubber sheet that serves to protect the buffer blocks in the installation phase may be difficult and can cause significant problems that may require retrieval of already placed canister, buffer and backfill. These matters are in focus in the present report. Arrangements for protecting already placed buffer blocks from moist air and water have been tested in earlier large-scale experiments, i.e. the Prototype Repository project at Aespoe but the experience from them has called for more effective protection of the clay blocks as described in the present report. Focus is on the construction of foundation components at the bottom of the deposition holes required for establishing a tight seal between rock and buffer blocks, and on the protection sheet and arrangements for limiting water pressure on it. Special attention is paid to the drainage of the space between rock and protection sheet that is necessary for avoiding failure of the sheet and to systems for achieving this and for providing alarm signals if the allowed pressure is

  15. Buffer protection in the installation phase

    Energy Technology Data Exchange (ETDEWEB)

    Wimelius, Hans (Swedish Nuclear Fuel and Waste Management Co., Stockholm (Sweden)); Pusch, Roland (Geodevelopment International AB, Lund (Sweden))

    2008-12-15

    The research and development of the design and construction of the SKB's repository for final disposal of spent reactor fuel is conducted along several paths ('lines'). Issues concerning the bedrock are dealt with in the 'rock line' and those related to buffer and backfill in deposition holes and tunnels are considered in the 'buffer line' and 'backfill line', respectively. These lines also deal with sub-activities that are coupled to several other lines. One of them includes development of techniques for protecting buffer blocks from moisture and water in the installation phase. Techniques and methods for placement and removal of the 'buffer protection sheet' are dealt with in the 'buffer line'. The removal is, however, considered as being part of the backfilling sequence. Since the performance of the sheet is of fundamental importance to the placement and function of the buffer it deserves particular attention. Thus, the removal of the rubber sheet that serves to protect the buffer blocks in the installation phase may be difficult and can cause significant problems that may require retrieval of already placed canister, buffer and backfill. These matters are in focus in the present report. Arrangements for protecting already placed buffer blocks from moist air and water have been tested in earlier large-scale experiments, i.e. the Prototype Repository project at Aespoe but the experience from them has called for more effective protection of the clay blocks as described in the present report. Focus is on the construction of foundation components at the bottom of the deposition holes required for establishing a tight seal between rock and buffer blocks, and on the protection sheet and arrangements for limiting water pressure on it. Special attention is paid to the drainage of the space between rock and protection sheet that is necessary for avoiding failure of the sheet and to systems for achieving

  16. Surface Water Protection by Productive Buffers

    DEFF Research Database (Denmark)

    Christen, Benjamin

    Vegetated riparian buffer zones are a widely recommended best management practice in agriculture for protecting surface and coastal waters from diffuse nutrient pollution. On the background of the EU funded research project NitroEurope (NEU; www.NitroEurope.eu), this study concentrates...... on the mitigation of nitrogen pollution in surface and groundwater, using riparian buffer zones for biomass production. The objectives are to map suitable areas for buffer implementation across the six NEU study landscapes, model tentative N-loss mitigation, calculate biomass production potential and economic...... designed for local conditions could be a way of protecting water quality attractive to many stakeholders....

  17. A possible role of rabbit heart cytosol tocopherol binding in the transfer of tocopherol into nuclei.

    OpenAIRE

    Guarnieri, C; Flamigni, F; Caldarera, C M

    1980-01-01

    An alpha-tocopherol-binding macromolecule was isolated from the heart cytosol of rabbits fed for 1 month with an alpha-tocopherol-deficient diet. The amount of [3H]-tocopherol bound to nuclear chromatin was increased when the alpha-tocopherol-deficient heart nuclei were incubated in the presence of [3H]tocopherol-cytosol complex. In this condition, large amounts of [3H]tocopherol were associated with a subnuclear fraction that contained non-histone acidic proteins.

  18. Quantitative assessment of cellular uptake and cytosolic access of antibody in living cells by an enhanced split GFP complementation assay

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ji-sun; Choi, Dong-Ki; Park, Seong-wook; Shin, Seung-Min; Bae, Jeomil [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Kim, Dong-Myung [Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Yoo, Tae Hyeon [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Kim, Yong-Sung, E-mail: kimys@ajou.ac.kr [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of)

    2015-11-27

    Considering the number of cytosolic proteins associated with many diseases, development of cytosol-penetrating molecules from outside of living cells is highly in demand. To gain access to the cytosol after cellular uptake, cell-penetrating molecules should be released from intermediate endosomes prior to the lysosomal degradation. However, it is very challenging to distinguish the pool of cytosolic-released molecules from those trapped in the endocytic vesicles. Here we describe a method to directly demonstrate the cytosolic localization and quantification of cytosolic amount of a cytosol-penetrating IgG antibody, TMab4, based on enhanced split GFP complementation system. We generated TMab4 genetically fused with one GFP fragment and separately established HeLa cells expressing the other GFP fragment in the cytosol such that the complemented GFP fluorescence is observed only when extracellular-treated TMab4 reaches the cytosol after cellular internalization. The high affinity interactions between streptavidin-binding peptide 2 and streptavidin was employed as respective fusion partners of GFP fragments to enhance the sensitivity of GFP complementation. With this method, cytosolic concentration of TMab4 was estimated to be about 170 nM after extracellular treatment of HeLa cells with 1 μM TMab4 for 6 h. We also found that after cellular internalization into living cells, nearly 1.3–4.3% of the internalized TMab4 molecules escaped into the cytosol from the endocytic vesicles. Our enhanced split GFP complementation assay provides a useful tool to directly quantify cytosolic amount of cytosol-penetrating agents and allows cell-based high-throughput screening for cytosol-penetrating agents with increased endosomal-escaping activity.

  19. Buffer sizing for multi-hop networks

    KAUST Repository

    Shihada, Basem; Jamshaid, Kamran

    2014-01-01

    domain according to a simple division or according to a cost function taking into account a distance of the communications link from the source and destination. The network may be monitored and the cumulative buffer size recalculated and redistributed

  20. Moisture Buffer Value of Building Materials

    DEFF Research Database (Denmark)

    Rode, Carsten; Peuhkuri, Ruut; Time, Berit

    2007-01-01

    When building materials are in contact with indoor air they have some effect to moderate the variations of indoor humidity in occupied buildings. But so far there has been a lack of a standardized quantity to characterize the moisture buffering capability of materials. It has been the objective o...... is a test protocol which expresses how materials should be tested for determination of their Moisture Buffer Value. Finally, the paper presents some of the results of a Round Robin Test on various typical building materials that has been carried out in the project....... of a recent Nordic project to define such a quantity, and to declare it in the form of a NORDTEST method. The Moisture Buffer Value is the figure that has been developed in the project as a way to appraise the moisture buffer effect of materials, and the value is described in the paper. Also explained...

  1. Liquid growth hormone: preservatives and buffers

    DEFF Research Database (Denmark)

    Kappelgaard, Anne-Marie; Anders, Bojesen; Skydsgaard, Karen

    2004-01-01

    injection are dependent on the preservative used in the formulation and the concentration of GH. Injection pain may also be related to the buffer substance and injection volume. A liquid formulation of GH, Norditropi SimpleXx, has been developed that dispenses with the need for reconstitution before...... solution. More pain was also reported following large volume injections and following injections with solutions containing high protein concentrations. In summary, optimization of the preservative and buffer content of a liquid GH formulation may reduce injection pain and lead to improved patient...... administration. The formulation uses phenol (3 mg/ml) as a preservative (to protect product from microbial degradation or contamination) and histidine as a buffer. Alternative preservatives used in other GH formulations include m-cresol (9 mg/ml) and benzyl alcohol (3-9 mg/ml). Buffering agents include citrate...

  2. Calculating Buffer Zones: A Guide for Applicators

    Science.gov (United States)

    Buffer zones provide distance between the application block (i.e., edge of the treated field) and bystanders, in order to control pesticide exposure risk from soil fumigants. Distance requirements may be reduced by credits such as tarps.

  3. Buffer Strips for Riparian Zone Management

    National Research Council Canada - National Science Library

    1991-01-01

    This study provides a review of technical literature concerning the width of riparian buffer strips needed to protect water quality and maintain other important values provided by riparian ecosystem...

  4. Lucas Heights buffer zone: plan of management

    International Nuclear Information System (INIS)

    1986-01-01

    This plan is being used by the Commission as a guide for its management of the Lucas Heights buffer zone, which is essentially a circular area having a 1-6 km radius around the HIFAR reactor. Aspects covered by this plan include past uses, current use, objectives for buffer zone land management, emergency evacuation, resource conservation, archaeology, fire, access, rehabilitation of disturbed areas, resource management and plan implementation

  5. Dynamic measurement of the calcium buffering properties of the sarcoplasmic reticulum in mouse skeletal muscle.

    Science.gov (United States)

    Manno, Carlo; Sztretye, Monika; Figueroa, Lourdes; Allen, Paul D; Ríos, Eduardo

    2013-01-15

    The buffering power, B, of the sarcoplasmic reticulum (SR), ratio of the changes in total and free [Ca(2+)], was determined in fast-twitch mouse muscle cells subjected to depleting membrane depolarization. Changes in total SR [Ca(2+)] were measured integrating Ca(2+) release flux, determined with a cytosolic [Ca(2+)] monitor. Free [Ca(2+)](SR) was measured using the cameleon D4cpv-Casq1. In 34 wild-type (WT) cells average B during the depolarization (ON phase) was 157 (SEM 26), implying that of 157 ions released, 156 were bound inside the SR. B was significantly greater when BAPTA, which increases release flux, was present in the cytosol. B was greater early in the pulse - when flux was greatest - than at its end, and greater in the ON than in the OFF. In 29 Casq1-null cells, B was 40 (3.6). The difference suggests that 75% of the releasable calcium is normally bound to calsequestrin. In the nulls the difference in B between ON and OFF was less than in the WT but still significant. This difference and the associated decay in B during the ON were not artifacts of a slow SR monitor, as they were also found in the WT when [Ca(2+)](SR) was tracked with the fast dye fluo-5N. The calcium buffering power, binding capacity and non-linear binding properties of the SR measured here could be accounted for by calsequestrin at the concentration present in mammalian muscle, provided that its properties were substantially different from those found in solution. Its affinity should be higher, or K(D) lower than the conventionally accepted 1 mm; its cooperativity (n in a Hill fit) should be higher and the stoichiometry of binding should be at the higher end of the values derived in solution. The reduction in B during release might reflect changes in calsequestrin conformation upon calcium loss.

  6. Low noise buffer amplifiers and buffered phase comparators for precise time and frequency measurement and distribution

    Science.gov (United States)

    Eichinger, R. A.; Dachel, P.; Miller, W. H.; Ingold, J. S.

    1982-01-01

    Extremely low noise, high performance, wideband buffer amplifiers and buffered phase comparators were developed. These buffer amplifiers are designed to distribute reference frequencies from 30 KHz to 45 MHz from a hydrogen maser without degrading the hydrogen maser's performance. The buffered phase comparators are designed to intercompare the phase of state of the art hydrogen masers without adding any significant measurement system noise. These devices have a 27 femtosecond phase stability floor and are stable to better than one picosecond for long periods of time. Their temperature coefficient is less than one picosecond per degree C, and they have shown virtually no voltage coefficients.

  7. Buffer-free therapeutic antibody preparations provide a viable alternative to conventionally buffered solutions: from protein buffer capacity prediction to bioprocess applications.

    Science.gov (United States)

    Bahrenburg, Sven; Karow, Anne R; Garidel, Patrick

    2015-04-01

    Protein therapeutics, including monoclonal antibodies (mAbs), have significant buffering capacity, particularly at concentrations>50 mg/mL. This report addresses pH-related issues critical to adoption of self-buffered monoclonal antibody formulations. We evaluated solution conditions with protein concentrations ranging from 50 to 250 mg/mL. Samples were both buffer-free and conventionally buffered with citrate. Samples were non-isotonic or adjusted for isotonicity with NaCl or trehalose. Studies included accelerated temperature stability tests, shaking stability studies, and pH changes in infusion media as protein concentrate is added. We present averaged buffering slopes of capacity that can be applied to any mAb and present a general method for calculating buffering capacity of buffer-free, highly concentrated antibody liquid formulations. In temperature stability tests, neither buffer-free nor conventionally buffered solution conditions showed significant pH changes. Conventionally buffered solutions showed significantly higher opalescence than buffer-free ones. In general, buffer-free solution conditions showed less aggregation than conventionally buffered solutions. Shaking stability tests showed no differences between buffer-free and conventionally buffered solutions. "In-use" preparation experiments showed that pH in infusion bag medium can rapidly approximate that of self-buffered protein concentrate as concentrate is added. In summary, the buffer capacity of proteins can be predicted and buffer-free therapeutic antibody preparations provide a viable alternative to conventionally buffered solutions. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Comparison of monoclonal antibodies and tritiated ligands for estrogen receptor assays in 241 breast cancer cytosols

    International Nuclear Information System (INIS)

    Goussard, J.; Lechevrel, C.; Martin, P.M.; Roussel, G.

    1986-01-01

    Estrogen receptor determinations have been performed on 241 cytosols from 160 breast cancer tumors using both radioactive ligands ([ 3 H]-estradiol, [3H]R2858) and monoclonal antibodies (Abbott ER-EIA Kit) to compare the two methods and to evaluate the clinical usefulness of the new immunological, simplified assay. Intra- and interassay reproducibility of the enzyme immunoassay (EIA) method was studied during a 6-month period on 35 standard curves with 4 different batches of monoclonal antibodies. Intraassay coefficients of variation studied on duplicates were smaller than 5% in most cases and reproducibility of the curves showed coefficients of variation lower than 10% except for standard 0 and 5 fmol/ml. Pooled cytosols used as control for the dextran coated charcoal method had interassay variation coefficients between 3.8 and 11.4%. Reproducibility has been studied on clinical specimens assayed twice at two different periods with either EIA or dextran coated charcoal methods. Slopes obtained were 1.05 and 0.96, respectively. A good stability of EIA results was obtained with protein concentrations in the range 4-0.15 mg/ml cytosol. No significant effects of dithiothreitol or monothioglycerol (1 mM) on EIA and dextran coated charcoal assay were observed. Eighty breast cancer cytosols were assayed with both EIA and Scatchard analysis. The slope of the regression curve obtained was 1.04 (r = 0.963). Cytosols were assayed by EIA and by a saturating concentration of tritiated ligand (5 nM). With 153 cytosols the EIA/5 nM slope was 1.34 (r = 0.978). This slope can be compared with the slope Scatchard/5 nM obtained with 90 cytosols: 1.29 (r = 0.985). Absence of cross-reactivity of monoclonal ER antibodies with progesterone receptor was observed

  9. Insulin Induces an Increase in Cytosolic Glucose Levels in 3T3-L1 Cells with Inhibited Glycogen Synthase Activation

    Directory of Open Access Journals (Sweden)

    Helena H. Chowdhury

    2014-10-01

    Full Text Available Glucose is an important source of energy for mammalian cells and enters the cytosol via glucose transporters. It has been thought for a long time that glucose entering the cytosol is swiftly phosphorylated in most cell types; hence the levels of free glucose are very low, beyond the detection level. However, the introduction of new fluorescence resonance energy transfer-based glucose nanosensors has made it possible to measure intracellular glucose more accurately. Here, we used the fluorescent indicator protein (FLIPglu-600µ to monitor cytosolic glucose dynamics in mouse 3T3-L1 cells in which glucose utilization for glycogen synthesis was inhibited. The results show that cells exhibit a low resting cytosolic glucose concentration. However, in cells with inhibited glycogen synthase activation, insulin induced a robust increase in cytosolic free glucose. The insulin-induced increase in cytosolic glucose in these cells is due to an imbalance between the glucose transported into the cytosol and the use of glucose in the cytosol. In untreated cells with sensitive glycogen synthase activation, insulin stimulation did not result in a change in the cytosolic glucose level. This is the first report of dynamic measurements of cytosolic glucose levels in cells devoid of the glycogen synthesis pathway.

  10. Static mechanical properties of buffer material

    International Nuclear Information System (INIS)

    Takaji, Kazuhiko; Suzuki, Hideaki

    1999-11-01

    The buffer material is expected to maintain its low water permeability, self-sealing properties, radionuclides adsorption and retardation properties, thermal conductivity, chemical buffering properties, overpack supporting properties, stress buffering properties, etc. over a long period of time. Natural clay is mentioned as a material that can relatively satisfy above. Among the kinds of natural clay, bentonite when compacted is superior because (i) it has exceptionally low water permeability and properties to control the movement of water in buffer, (ii) it fills void spaces in the buffer and fractures in the host rock as it swells upon water uptake, (iii) it has the ability to exchange cations and to adsorb cationic radioelements. In order to confirm these functions for the purpose of safety assessment, it is necessary to evaluate buffer properties through laboratory tests and engineering-scale tests, and to make assessments based on the ranges in the data obtained. This report describes the procedures, test conditions, results and examinations on the buffer material of unconfined compression tests, one-dimensional consolidation tests, consolidated-undrained triaxial compression tests and consolidated-undrained triaxial creep tests that aim at getting hold of static mechanical properties. We can get hold of the relationship between the dry density and tensile stress etc. by Brazilian tests, between the dry density and unconfined compressive strength etc. by unconfined compression tests, between the consolidation stress and void ratio etc. by one-dimensional consolidation tests, the stress pass of each effective confining pressure etc. by consolidated-undrained triaxial compression tests and the axial strain rate with time of each axial stress etc. by consolidated-undrained triaxial creep tests. (author)

  11. Temperature Buffer Test. Measurements of water content and density of the excavated buffer material

    Energy Technology Data Exchange (ETDEWEB)

    Johannesson, Lars-Erik [Clay Technology AB, Lund (Sweden)

    2010-12-15

    TBT (Temperature Buffer Test) is a joint project between SKB/ANDRA and supported by ENRESA (modeling) and DBE (instrumentation), which aims at understanding and modeling the thermo-hydromechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test was carried out at the - 420 m level in Aespoe HRL in a 8 meters deep and 1.76 m diameter deposition hole, with two heaters (3 m long, 0.6 m diameter), surrounded by a MX-80 bentonite buffer and a confining plug on top anchored with 9 rods. It was installed during spring 2003. The bentonite around upper heater was removed during the period October - December 2009 and the buffer around the lower heater was removed during January - Mars 2010. During dismantling of the buffer, samples were taken on which analyses were made. This report describes the work with the deteroemoeination of the water content and the density of the taken samples. Most of the samples were taken from the buffer by core drilling from the upper surface of each installed bentonite block. The cores had a diameter of about 50 mm and a maximum length equal to the original height of the bentonite blocks (about 500 mm). The water content of the buffer was determined by drying a sample at a temperature of 105 deg C for 24 h and the bulk density was determined by weighing a sample both in the air and immerged in paraffin oil with known density. The water content, dry density, degree of saturation and void ratio of the buffer were then plotted. The plots show that all parts of the buffer had taken up water and the degree of saturation of the buffer varied between 90 - 100%. Large variation in the dry density of the buffer was also observed.

  12. ESX-1-mediated translocation to the cytosol controls virulence of mycobacteria

    KAUST Repository

    Houben, Diane; Demangel, Caroline; Van Ingen, Jakko; Perez, Jorge; Baldeó n, Lucy R.; Abdallah, Abdallah; Caleechurn, Laxmee; Bottai, Daria; Van Zon, Maaike; De Punder, Karin; Van Der Laan, Tridia; Kant, Arie; Bossers-De Vries, Ruth; Willemsen, Peter Th J; Bitter, Wilbert M.; Van Soolingen, Dick; Brosch, Roland; Van Der Wel, Nicole N.; Peters, Peter J.

    2012-01-01

    Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium leprae, are among the most potent human bacterial pathogens. The discovery of cytosolic mycobacteria challenged the paradigm that these pathogens exclusively localize within the phagosome of host cells. As yet the biological relevance of mycobacterial translocation to the cytosol remained unclear. In this current study we used electron microscopy techniques to establish a clear link between translocation and mycobacterial virulence. Pathogenic, patient-derived mycobacteria species were found to translocate to the cytosol, while non-pathogenic species did not. We were further able to link cytosolic translocation with pathogenicity by introducing the ESX-1 (type VII) secretion system into the non-virulent, exclusively phagolysosomal Mycobacterium bovis BCG. Furthermore, we show that translocation is dependent on the C-terminus of the early-secreted antigen ESAT-6. The C-terminal truncation of ESAT-6 was shown to result in attenuation in mice, again linking translocation to virulence. Together, these data demonstrate the molecular mechanism facilitating translocation of mycobacteria. The ability to translocate from the phagolysosome to the cytosol is with this study proven to be biologically significant as it determines mycobacterial virulence. © 2012 Blackwell Publishing Ltd.

  13. Circadian Rhythm of Hepatic Cytosolic and Nuclear Estrogen and Androgen Receptors

    Science.gov (United States)

    FRANCAVILLA, ANTONIO; EAGON, PATRICIA K.; DiLEO, ALFREDO; VAN THIEL, DAVID H.; PANELLA, CARMINE; POLIMENO, LORENZO; AMORUSO, CINZIA; INGROSSO, MARCELLO; AQUILINO, A. MARIA; STARZL, THOMAS E.

    2010-01-01

    Mammalian liver is a sex steroid-responsive tissue. The effects of these hormones presumably are mediated by hepatic estrogen receptors (ER) and androgen receptors (AR). Serum levels of sex hormones display circadian rhythms. Further, estrogens and androgens are commonly administered; administration of these agents is associated frequently with liver disease. Therefore, we investigated whether the cytosolic and nuclear sex steroid receptors also display a similar circadian rhythm, and whether variations occurred in the distribution of receptors between cytosolic and nuclear compartments. Animals were killed every 4 h from midnight till the following midnight; cytosolic and nuclear levels of both ER and AR were measured. Cytosolic ER reached a maximum level at 4 AM, and a minimum at 8 PM and midnight of both days. Nuclear ER was highest at 8 AM and lowest at 4 PM and 8 PM, a pattern which parallels variations in serum estradiol levels. Cytosolic AR was highest at 8 PM and lowest at midnight and 4 AM. Nuclear AR was highest at 4 AM and lowest at 4 PM and 8 PM. The highest level of nuclear AR does not correspond to the maximum serum testosterone level, which occurred at 4 PM. The total hepatic content of both ER and AR was not constant over the 24-h period, but varied considerably with time of day. These studies suggest that both ER and AR show a distinct circadian rhythm in subcellular compartmentalization, and that total hepatic content of ER and AR varies significantly during a 24-h period. PMID:3710067

  14. ESX-1-mediated translocation to the cytosol controls virulence of mycobacteria

    KAUST Repository

    Houben, Diane

    2012-05-08

    Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium leprae, are among the most potent human bacterial pathogens. The discovery of cytosolic mycobacteria challenged the paradigm that these pathogens exclusively localize within the phagosome of host cells. As yet the biological relevance of mycobacterial translocation to the cytosol remained unclear. In this current study we used electron microscopy techniques to establish a clear link between translocation and mycobacterial virulence. Pathogenic, patient-derived mycobacteria species were found to translocate to the cytosol, while non-pathogenic species did not. We were further able to link cytosolic translocation with pathogenicity by introducing the ESX-1 (type VII) secretion system into the non-virulent, exclusively phagolysosomal Mycobacterium bovis BCG. Furthermore, we show that translocation is dependent on the C-terminus of the early-secreted antigen ESAT-6. The C-terminal truncation of ESAT-6 was shown to result in attenuation in mice, again linking translocation to virulence. Together, these data demonstrate the molecular mechanism facilitating translocation of mycobacteria. The ability to translocate from the phagolysosome to the cytosol is with this study proven to be biologically significant as it determines mycobacterial virulence. © 2012 Blackwell Publishing Ltd.

  15. Scientific Applications Performance Evaluation on Burst Buffer

    KAUST Repository

    Markomanolis, George S.

    2017-10-19

    Parallel I/O is an integral component of modern high performance computing, especially in storing and processing very large datasets, such as the case of seismic imaging, CFD, combustion and weather modeling. The storage hierarchy includes nowadays additional layers, the latest being the usage of SSD-based storage as a Burst Buffer for I/O acceleration. We present an in-depth analysis on how to use Burst Buffer for specific cases and how the internal MPI I/O aggregators operate according to the options that the user provides during his job submission. We analyze the performance of a range of I/O intensive scientific applications, at various scales on a large installation of Lustre parallel file system compared to an SSD-based Burst Buffer. Our results show a performance improvement over Lustre when using Burst Buffer. Moreover, we show results from a data hierarchy library which indicate that the standard I/O approaches are not enough to get the expected performance from this technology. The performance gain on the total execution time of the studied applications is between 1.16 and 3 times compared to Lustre. One of the test cases achieved an impressive I/O throughput of 900 GB/s on Burst Buffer.

  16. Improved indexes for targeting placement of buffers of Hortonian runoff

    Science.gov (United States)

    M.G. Dosskey; Z. Qiu; M.J. Helmers; D.E. Eisenhauer

    2011-01-01

    Targeting specific locations within agricultural watersheds for installing vegetative buffers has been advocated as a way to enhance the impact of buffers and buffer programs on stream water quality. Existing models for targeting buffers of Hortonian, or infiltration-excess, runoff are not well developed. The objective was to improve on an existing soil survey–based...

  17. A buffer overflow detection based on inequalities solution

    International Nuclear Information System (INIS)

    Xu Guoai; Zhang Miao; Yang Yixian

    2007-01-01

    A new buffer overflow detection model based on Inequalities Solution was designed, which is based on analyzing disadvantage of the old buffer overflow detection technique and successfully converting buffer overflow detection to Inequalities Solution. The new model can conquer the disadvantage of the old technique and improve efficiency of buffer overflow detection. (authors)

  18. Moisture buffer capacity of different insulation materials

    DEFF Research Database (Denmark)

    Peuhkuri, Ruut Hannele; Rode, Carsten; Hansen, Kurt Kielsgaard

    2004-01-01

    . In the isothermal tests the material samples were exposed to the same change in the relative humidity of the ambient air on both sides, while the samples were exposed to variations in relative humidity only on the cold side in the non-isothermal tests. The results of these rather different measurement principles...... lead to more durable constructions. In this paper, a large range of very different thermal insulation materials have been tested in specially constructed laboratory facilities to determine their moisture buffer capacity. Both isothermal and nonisothermal experimental set-ups have been used...... are discussed, and different ways are presented how to determine the moisture buffer capacity of the materials using partly standard material parameters and partly parameters determined from the actual measurements. The results so far show that the determination of moisture buffer capacity is very sensitive...

  19. Buffer construction technique using granular bentonite

    International Nuclear Information System (INIS)

    Masuda, Ryoichi; Asano, Hidekazu; Toguri, Satohito; Mori, Takuo; Shimura, Tomoyuki; Matsuda, Takeshi; Uyama, Masao; Noda, Masaru

    2007-01-01

    Buffer construction using bentonite pellets as filling material is a promising technology for enhancing the ease of repository operation. In this study, a test of such technology was conducted in a full-scale simulated disposal drift, using a filling system which utilizes a screw conveyor system. The simulated drift, which contained two dummy overpacks, was configured as a half-cross-section model with a height of 2.22 m and a length of 6.0 m. The average dry density of the buffer obtained in the test was 1.29 Mg/m 3 , with an angle of repose of 35 to 40 degrees. These test results indicate that buffer construction using a screw conveyor system for pellet emplacement in a waste disposal drift is a promising technology for repositories for high level radioactive wastes. (author)

  20. Labview virtual instruments for calcium buffer calculations.

    Science.gov (United States)

    Reitz, Frederick B; Pollack, Gerald H

    2003-01-01

    Labview VIs based upon the calculator programs of Fabiato and Fabiato (J. Physiol. Paris 75 (1979) 463) are presented. The VIs comprise the necessary computations for the accurate preparation of multiple-metal buffers, for the back-calculation of buffer composition given known free metal concentrations and stability constants used, for the determination of free concentrations from a given buffer composition, and for the determination of apparent stability constants from absolute constants. As implemented, the VIs can concurrently account for up to three divalent metals, two monovalent metals and four ligands thereof, and the modular design of the VIs facilitates further extension of their capacity. As Labview VIs are inherently graphical, these VIs may serve as useful templates for those wishing to adapt this software to other platforms.

  1. Developing suitable buffers to capture transport cycling behavior

    DEFF Research Database (Denmark)

    Madsen, Thomas; Schipperijn, Jasper; Christiansen, Lars Breum

    2014-01-01

    units (e.g., home-buffer-based neighborhoods) are not necessarily representative for environmental exposure. An increased understanding of appropriate neighborhoods is needed. GPS cycling tracks from 78 participants for 7 days form the basis for the development and testing of different neighborhood...... buffers for transport cycling. The percentage of GPS points per square meter was used as indicator of the effectiveness of a series of different buffer types, including home-based network buffers, shortest route to city center buffers, and city center-directed ellipse-shaped buffers. The results show...... center-directed ellipse-shaped buffers yielded better results than traditional home-based network buffer types. The ellipse-shaped buffer types could therefore be considered an alternative to more traditional buffers or administrative units in future studies of transport cycling behavior....

  2. The cytosolic exonuclease TREX1 inhibits the innate immune response to HIV-1

    Science.gov (United States)

    Yan, Nan; Regalado-Magdos, Ashton D.; Stiggelbout, Bart; Lee-Kirsch, Min Ae; Lieberman, Judy

    2010-01-01

    Viral infection triggers innate immune sensors to produce type I interferons (IFN). However, HIV infection of T cells and macrophages does not trip these alarms. How HIV avoids activating nucleic acid sensors is unknown. The cytosolic exonuclease TREX1 suppressed IFN triggered by HIV. In Trex1−/− mouse cells and human CD4+ T cells and macrophages in which TREX1 was inhibited by RNA interference, cytosolic HIV DNA accumulated, and HIV infection induced type I IFN that inhibited HIV replication and spreading. TREX1 bound to cytosolic HIV DNA and digested excess HIV DNA that would otherwise activate IFN expression via a TBK1, STING and IRF3 dependent pathway. HIV-stimulated IFN production in cells deficient in TREX1 did not involve known nucleic acid sensors. PMID:20871604

  3. Cytosol-dependent membrane fusion in ER, nuclear envelope and nuclear pore assembly: biological implications.

    Science.gov (United States)

    Rafikova, Elvira R; Melikov, Kamran; Chernomordik, Leonid V

    2010-01-01

    Endoplasmic reticulum and nuclear envelope rearrangements after mitosis are often studied in the reconstitution system based on Xenopus egg extract. In our recent work we partially replaced the membrane vesicles in the reconstitution mix with protein-free liposomes to explore the relative contributions of cytosolic and transmembrane proteins. Here we discuss our finding that cytosolic proteins mediate fusion between membranes lacking functional transmembrane proteins and the role of membrane fusion in endoplasmic reticulum and nuclear envelope reorganization. Cytosol-dependent liposome fusion has allowed us to restore, without adding transmembrane nucleoporins, functionality of nuclear pores, their spatial distribution and chromatin decondensation in nuclei formed at insufficient amounts of membrane material and characterized by only partial decondensation of chromatin and lack of nuclear transport. Both the mechanisms and the biological implications of the discovered coupling between spatial distribution of nuclear pores, chromatin decondensation and nuclear transport are discussed.

  4. Buffer of Events as a Markovian Process

    International Nuclear Information System (INIS)

    Berdugo, J.; Casaus, J.; Mana, C.

    2001-01-01

    In Particle and Asro-Particle Physics experiments, the events which get trough the detectors are read and processes on-line before they are stored for a more detailed processing and future Physics analysis. Since the events are read and, usually, processed sequentially, the time involved in these operations can lead to a significant lose of events which is, to some extent, reduced by using buffers. We present an estimate of the optimum buffer size and the fraction of events lost for a simple experimental condition which serves as an introductory example to the use of Markow Chains.(Author)

  5. Buffer of Events as a Markovian Process

    Energy Technology Data Exchange (ETDEWEB)

    Berdugo, J.; Casaus, J.; Mana, C.

    2001-07-01

    In Particle and Asro-Particle Physics experiments, the events which get trough the detectors are read and processes on-line before they are stored for a more detailed processing and future Physics analysis. Since the events are read and, usually, processed sequentially, the time involved in these operations can lead to a significant lose of events which is, to some extent, reduced by using buffers. We present an estimate of the optimum buffer size and the fraction of events lost for a simple experimental condition which serves as an introductory example to the use of Markow Chains.(Author)

  6. Emplacement of small and large buffer blocks

    International Nuclear Information System (INIS)

    Saari, H.; Nikula, M.; Suikki, M.

    2010-05-01

    The report describes emplacement of a buffer structure encircling a spent fuel canister to be deposited in a vertical hole. The report deals with installability of various size blocks and with an emplacement gear, as well as evaluates the achieved quality of emplacement and the time needed for installing the buffer. Two block assembly of unequal size were chosen for examination. A first option involved small blocks, the use of which resulted in a buffer structure consisting of small sector blocks 200 mm in height. A second option involved large blocks, resulting in a buffer structure which consists of eight blocks. In these tests, the material chosen for both block options was concrete instead of bentonite. The emplacement test was a three-phase process. A first phase included stacking a two meter high buffer structure with small blocks for ensuring the operation of test equipment and blocks. A second phase included installing buffer structures with both block options to a height matching that of a canister-encircling cylindrical component. A third phase included testing also the installability of blocks to be placed above the canister by using small blocks. In emplacement tests, special attention was paid to the installability of blocks as well as to the time required for emplacement. Lifters for both blocks worked well. Due to the mass to be lifted, the lifter for large blocks had a more heavy-duty frame structure (and other lifting gear). The employed lifters were suspended in the tests on a single steel wire rope. Stacking was managed with both block sizes at adequate precision and stacked-up towers were steady. The stacking of large blocks was considerably faster. Therefore it is probably that the overall handling of the large blocks will be more convenient at a final disposal site. From the standpoint of reliability in lifting, the small blocks were safer to install above the canister. In large blocks, there are strict shape-related requirements which are

  7. "cAMP sponge": a buffer for cyclic adenosine 3', 5'-monophosphate.

    Directory of Open Access Journals (Sweden)

    Konstantinos Lefkimmiatis

    Full Text Available BACKGROUND: While intracellular buffers are widely used to study calcium signaling, no such tool exists for the other major second messenger, cyclic AMP (cAMP. METHODS/PRINCIPAL FINDINGS: Here we describe a genetically encoded buffer for cAMP based on the high-affinity cAMP-binding carboxy-terminus of the regulatory subunit RIbeta of protein kinase A (PKA. Addition of targeting sequences permitted localization of this fragment to the extra-nuclear compartment, while tagging with mCherry allowed quantification of its expression at the single cell level. This construct (named "cAMP sponge" was shown to selectively bind cAMP in vitro. Its expression significantly suppressed agonist-induced cAMP signals and the downstream activation of PKA within the cytosol as measured by FRET-based sensors in single living cells. Point mutations in the cAMP-binding domains of the construct rendered the chimera unable to bind cAMP in vitro or in situ. Cyclic AMP sponge was fruitfully applied to examine feedback regulation of gap junction-mediated transfer of cAMP in epithelial cell couplets. CONCLUSIONS: This newest member of the cAMP toolbox has the potential to reveal unique biological functions of cAMP, including insight into the functional significance of compartmentalized signaling events.

  8. Grass buffers for playas in agricultural landscapes: An annotated bibliography

    Science.gov (United States)

    Melcher, Cynthia P.; Skagen, Susan K.

    2005-01-01

    This bibliography and associated literature synthesis (Melcher and Skagen, 2005) was developed for the Playa Lakes Joint Venture (PLJV). The PLJV sought compilation and annotation of the literature on grass buffers for protecting playas from runoff containing sediments, nutrients, pesticides, and other contaminants. In addition, PLJV sought information regarding the extent to which buffers may attenuate the precipitation runoff needed to fill playas, and avian use of buffers. We emphasize grass buffers, but we also provide information on other buffer types.

  9. Mediator-assisted Simultaneous probing of Cytosolic and Mitochondrial Redox activity in living cells

    DEFF Research Database (Denmark)

    Heiskanen, Arto; Spegel, Christer; Kostesha, Natalie

    2009-01-01

    the ferricyanide-menadione double mediator system to study the effect of dicoumarol, an inhibitor of cytosolic and mitochondrial oxidoreductases and an uncoupler of the electron transport chain. Evaluation of the role of NAD(P)H-producing pathways in mediating biological effects is facilitated by introducing...... either fructose or glucose as the carbon source, yielding either NADH or NADPH through the glycolytic or pen-rose phosphate pathway, respectively. Respiratory noncompetent cells show greater inhibition of cytosolic menadione-reducing enzymes when NADH rather than NADPH is produced. Spectrophotometric...

  10. The buffer effect in neutral electrolyte supercapacitors

    DEFF Research Database (Denmark)

    Thrane Vindt, Steffen; Skou, Eivind M.

    2016-01-01

    The observation that double-layer capacitors based on neutral aqueous electrolytes can have significantly wider usable potential windows than those based on acidic or alkaline electrolytes is studied. This effect is explained by a local pH change taking place at the electrode surfaces, leading...... potassium nitrate as the electrolyte and potassium phosphates as the buffer system....

  11. Buffer gas cooling and mixture analysis

    Science.gov (United States)

    Patterson, David S.; Doyle, John M.

    2018-03-06

    An apparatus for spectroscopy of a gas mixture is described. Such an apparatus includes a gas mixing system configured to mix a hot analyte gas that includes at least one analyte species in a gas phase into a cold buffer gas, thereby forming a supersaturated mixture to be provided for spectroscopic analysis.

  12. Buffer Overflow Period in a MAP Queue

    Directory of Open Access Journals (Sweden)

    Andrzej Chydzinski

    2007-01-01

    Full Text Available The buffer overflow period in a queue with Markovian arrival process (MAP and general service time distribution is investigated. The results include distribution of the overflow period in transient and stationary regimes and the distribution of the number of cells lost during the overflow interval. All theorems are illustrated via numerical calculations.

  13. Body Buffer Zone and Proxemics in Blocking.

    Science.gov (United States)

    Stockwell, John C.; Bahs, Clarence W.

    This paper investigates the effect of personal body buffer zones on compositional arrangements staged by novice directors. Relationships between directors' concepts of personal space and their projection of its dimensions into staging are studied through the use of a variety of proximity measures--distance, area angles of approach, and physical…

  14. Buffering action of human dentin in vitro.

    Science.gov (United States)

    Camps, J; Pashley, D H

    2000-01-01

    The purpose of this study was to determine the relative contributions of the mineral and organic phases of dentin to its total buffering capacity and to compare the buffering abilities of normal and caries-affected dentin for acids used in adhesive dentistry. Disks of normal and caries-affected human coronal dentin 0.6 mm thick were prepared. Fifty microL of various acids were applied to the surface of mineralized or completely demineralized dentin for varying lengths of time. They were collected from the surface and combined with water rinses to permit titration of the total amount of acid applied, the amount recovered, the total amount that was taken up by the dentin, and the amount that diffused across dentin into 1 mL of water. Equal volumes of acids were applied to mineralized or demineralized dentin powder or hydroxyapatite powder. About 88% to 90% of applied acid was recovered from the surface; only 10% to 12% of the acid was taken up by dentin. Of the H+ that was taken up, only 1% to 2% actually diffused across 0.6 mm of dentin. Increasing the application time of 37% phosphoric acid did not increase the amount of H+ that diffused across dentin. Increasing the concentration of phosphoric acid from 10% to 65% produced only slight increases in H+ diffusion across dentin. There was no difference in the buffering capacity of normal vs caries-affected dentin disks. Almost all of the buffering capacity of dentin is due to its mineral phase. The high buffering capacity of dentin and the high reactivity of H+ insure that little H+ diffuses through dentin more than 0.6 mm thick.

  15. A NOVEL S-ADENOSYL-L-METHIONINE: ARSENIC (III) METHYLTRANSFERASE FROM RAT LIVER CYTOSOL

    Science.gov (United States)

    A Novel S-Adenosyl-L-methionine: Arsenic(III) Methyltransferase from Rat Liver CytosolShan Lin, Qing Shi, F. Brent Nix, Miroslav Styblo, Melinda A. Beck, Karen M. Herbin-Davis, Larry L. Hall, Josef B. Simeonsson, and David J. Thomas S-adenosyl-L-methionine (AdoMet): ar...

  16. RPA and Rad51 constitute a cell intrinsic mechanism to protect the cytosol from self DNA.

    Science.gov (United States)

    Wolf, Christine; Rapp, Alexander; Berndt, Nicole; Staroske, Wolfgang; Schuster, Max; Dobrick-Mattheuer, Manuela; Kretschmer, Stefanie; König, Nadja; Kurth, Thomas; Wieczorek, Dagmar; Kast, Karin; Cardoso, M Cristina; Günther, Claudia; Lee-Kirsch, Min Ae

    2016-05-27

    Immune recognition of cytosolic DNA represents a central antiviral defence mechanism. Within the host, short single-stranded DNA (ssDNA) continuously arises during the repair of DNA damage induced by endogenous and environmental genotoxic stress. Here we show that short ssDNA traverses the nuclear membrane, but is drawn into the nucleus by binding to the DNA replication and repair factors RPA and Rad51. Knockdown of RPA and Rad51 enhances cytosolic leakage of ssDNA resulting in cGAS-dependent type I IFN activation. Mutations in the exonuclease TREX1 cause type I IFN-dependent autoinflammation and autoimmunity. We demonstrate that TREX1 is anchored within the outer nuclear membrane to ensure immediate degradation of ssDNA leaking into the cytosol. In TREX1-deficient fibroblasts, accumulating ssDNA causes exhaustion of RPA and Rad51 resulting in replication stress and activation of p53 and type I IFN. Thus, the ssDNA-binding capacity of RPA and Rad51 constitutes a cell intrinsic mechanism to protect the cytosol from self DNA.

  17. Effect of low dose radiation on thymocyte cytosol and nuclei protein synthesis in mice

    International Nuclear Information System (INIS)

    Meng Qingyong; Chen Shali; Liu Shuzheng

    2003-01-01

    Objective: To the effect of low dose radiation on thymocyte cytosol and nuclei protein synthesis in mice. Methods: The expression of proteins was analyzed by gel filtration with Sephadex G-100 and HPLC based on separation of proteins on thymocyte cytosol and nuclei after whole-body irradiation with 75 mGy X-rays and sham-irradiation, and their biological activity was examined by mouse splenocyte proliferation and chromosome aberration of human peripheral blood lymphocytes. Results: HPLC analysis showed that there was a marked increase in expression of 61.4 kD protein in the extract of thymocyte cytosol and 30.4 kD protein in the extract of thymocyte nuclei in comparison with the corresponding fractions from the sham-irradiated control mice. These protein fractions from the thymocyte cytosol and nuclei of the irradiated mice showed both stimulating effect on normal T cell proliferation and protective effect on chromosome damage induced by high dose radiation. Conclusion: These findings might have implications in study of mechanism of immunoenhancement and cytogenetic adaptive response induced by low dose radiation

  18. Nanoparticles for cytosolic delivery of important biomolecular drugs such as DNA, RNA, peptides, and proteins

    Czech Academy of Sciences Publication Activity Database

    Sedlák, M.; Koňák, Čestmír; Dybal, Jiří

    2010-01-01

    Roč. 1, č. 2010 (2010), s. 87-90 ISSN 2210-2892 Institutional research plan: CEZ:AV0Z40500505 Keywords : cytosolic delivery * nanoparticle carriers * poly(ethylacrylic acid) Subject RIV: CD - Macromolecular Chemistry http://benthamopen.com/ABSTRACT/TOPROCJ-1-87

  19. Structure and role of neutrophil cytosol factor 1 (NCF1) gene in ...

    African Journals Online (AJOL)

    Yomi

    2010-12-27

    Dec 27, 2010 ... The neutrophil cytosol factor 1 (NCF1) gene consists of 11 exons and is found in two forms; one is wild ... granulomatous disease, multiple sclerosis, arthritis and parasitic infection. ... TCR, T cell receptor; AhR, aryl hydrocarbon receptor; RA, .... During malaria, ROS production can contribute to both.

  20. The role of a cytosolic superoxide dismutase in barley-pathogen interactions

    KAUST Repository

    Lightfoot, Damien; Mcgrann, Graham R. D.; Able, Amanda J.

    2016-01-01

    susceptible background (cv. Golden Promise), when compared with wild-type plants, suggesting that cytosolic O2-HO2 contributes to the signalling required to induce a defence response to Ptt. There was no effect of HvCSD1 knockdown on infection by the hemi

  1. Live cell imaging of cytosolic NADH/NAD+ ratio in hepatocytes and liver slices.

    Science.gov (United States)

    Masia, Ricard; McCarty, William J; Lahmann, Carolina; Luther, Jay; Chung, Raymond T; Yarmush, Martin L; Yellen, Gary

    2018-01-01

    Fatty liver disease (FLD), the most common chronic liver disease in the United States, may be caused by alcohol or the metabolic syndrome. Alcohol is oxidized in the cytosol of hepatocytes by alcohol dehydrogenase (ADH), which generates NADH and increases cytosolic NADH/NAD + ratio. The increased ratio may be important for development of FLD, but our ability to examine this question is hindered by methodological limitations. To address this, we used the genetically encoded fluorescent sensor Peredox to obtain dynamic, real-time measurements of cytosolic NADH/NAD + ratio in living hepatocytes. Peredox was expressed in dissociated rat hepatocytes and HepG2 cells by transfection, and in mouse liver slices by tail-vein injection of adeno-associated virus (AAV)-encoded sensor. Under control conditions, hepatocytes and liver slices exhibit a relatively low (oxidized) cytosolic NADH/NAD + ratio as reported by Peredox. The ratio responds rapidly and reversibly to substrates of lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH). Ethanol causes a robust dose-dependent increase in cytosolic NADH/NAD + ratio, and this increase is mitigated by the presence of NAD + -generating substrates of LDH or SDH. In contrast to hepatocytes and slices, HepG2 cells exhibit a relatively high (reduced) ratio and show minimal responses to substrates of ADH and SDH. In slices, we show that comparable results are obtained with epifluorescence imaging and two-photon fluorescence lifetime imaging (2p-FLIM). Live cell imaging with Peredox is a promising new approach to investigate cytosolic NADH/NAD + ratio in hepatocytes. Imaging in liver slices is particularly attractive because it allows preservation of liver microanatomy and metabolic zonation of hepatocytes. NEW & NOTEWORTHY We describe and validate a new approach for measuring free cytosolic NADH/NAD + ratio in hepatocytes and liver slices: live cell imaging with the fluorescent biosensor Peredox. This approach yields dynamic, real

  2. Measurement of binding of adenine nucleotides and phosphate to cytosolic proteins in permeabilized rat-liver cells

    NARCIS (Netherlands)

    Gankema, H. S.; Groen, A. K.; Wanders, R. J.; Tager, J. M.

    1983-01-01

    1. A method is described for measuring the binding of metabolites to cytosolic proteins in situ in isolated rat-liver cells treated with filipin to render the plasma membrane permeable to compounds of low molecular weight. 2. There is no binding of ATP or inorganic phosphate to cytosolic proteins,

  3. Spatial modeling of the membrane-cytosolic interface in protein kinase signal transduction.

    Directory of Open Access Journals (Sweden)

    Wolfgang Giese

    2018-04-01

    Full Text Available The spatial architecture of signaling pathways and the interaction with cell size and morphology are complex, but little understood. With the advances of single cell imaging and single cell biology, it becomes crucial to understand intracellular processes in time and space. Activation of cell surface receptors often triggers a signaling cascade including the activation of membrane-attached and cytosolic signaling components, which eventually transmit the signal to the cell nucleus. Signaling proteins can form steep gradients in the cytosol, which cause strong cell size dependence. We show that the kinetics at the membrane-cytosolic interface and the ratio of cell membrane area to the enclosed cytosolic volume change the behavior of signaling cascades significantly. We suggest an estimate of average concentration for arbitrary cell shapes depending on the cell volume and cell surface area. The normalized variance, known from image analysis, is suggested as an alternative measure to quantify the deviation from the average concentration. A mathematical analysis of signal transduction in time and space is presented, providing analytical solutions for different spatial arrangements of linear signaling cascades. Quantification of signaling time scales reveals that signal propagation is faster at the membrane than at the nucleus, while this time difference decreases with the number of signaling components in the cytosol. Our investigations are complemented by numerical simulations of non-linear cascades with feedback and asymmetric cell shapes. We conclude that intracellular signal propagation is highly dependent on cell geometry and, thereby, conveys information on cell size and shape to the nucleus.

  4. Limits to anaerobic energy and cytosolic concentration in the living cell

    Science.gov (United States)

    Paglietti, A.

    2015-11-01

    For many physical systems at any given temperature, the set of all states where the system's free energy reaches its largest value can be determined from the system's constitutive equations of internal energy and entropy, once a state of that set is known. Such an approach is fraught with complications when applied to a living cell, because the cell's cytosol contains thousands of solutes, and thus thousands of state variables, which makes determination of its state impractical. We show here that, when looking for the maximum energy that the cytosol can store and release, detailed information on cytosol composition is redundant. Compatibility with cell's life requires that a single variable that represents the overall concentration of cytosol solutes must fall between defined limits, which can be determined by dehydrating and overhydrating the cell to its maximum capacity. The same limits are shown to determine, in particular, the maximum amount of free energy that a cell can supply in fast anaerobic processes, starting from any given initial state. For a typical skeletal muscle in normal physiological conditions this energy, i.e., the maximum anaerobic capacity to do work, is calculated to be about 960 J per kg of muscular mass. Such energy decreases as the overall concentration of solutes in the cytosol is increased. Similar results apply to any kind of cell. They provide an essential tool to understand and control the macroscopic response of single cells and multicellular cellular tissues alike. The applications include sport physiology, cell aging, disease produced cell damage, drug absorption capacity, to mention the most obvious ones.

  5. The effects of irradiation on the cytosol glucocorticoid receptor and concentrations of corticosterone and cyclic nucleotides in the rat liver

    International Nuclear Information System (INIS)

    Teshima, Teruki; Mori, Masaki; Honke, Yoshifumi

    1983-01-01

    The effects of irradiation on both the cytosol glucocorticoid receptor and concentrations of corticosterone and cyclic nucleotides in the rat liver were investigated. The liver concentrations of corticosterone and cyclic nucleotides were measured by radioimmunoassay before and after the irradiation of 1,000 rad/l fraction. The glucocorticoid receptor in the liver cytosol was determined by the measurement of the cytosol binding to 3 H-dexamethasone. The cytosol and nuclear corticosterone levels reached a peak 1 day after the irradiation of the rat liver and declined to the control levels after 2 days. The increase in corticosterone levels may be due to the direct stimulation of the right adrenal gland and/ or the stress induced by the irradiation. The binding capacity of the glucocorticoid receptor in rat liver cytosol decreased to the minimum 1 day after the irradiation, and the recovery occurred at 4 days. The Kd value of the glucocorticoid receptor remained unchanged from 1 hour until 4 days but was high at 4 and 7 days. The distinctly increased levels of cyclic GMP in the rat liver were found from 1 hour through 7 days after the irradiation, while cyclic AMP did not change. The inversed relationship between the cytosol glucocorticoid receptor and corticosterone levels in cytosol and the nuclei indicates that the receptor-bound corticosterone in cytosol can be transferred to a nucleus and remain there in the presence of appropriate amounts of corticosterone in cytosol, after which the receptor is released from the nucleus into cytosol. The high Kd values observed 4 -- 7 days after the irradiation may be either due to the direct effect of irradiation or to the replenishment of the receptor with a low affinity. (author)

  6. Coupled microrings data buffer using fast light

    Science.gov (United States)

    Scheuer, Jacob; Shahriar, Selim

    2013-03-01

    We present a theoretical study of a trap-door optical buffer based on a coupled microrings add/drop filter (ADF) utilizing the white light cavity (WLC). The buffer "trap-door" can be opened and closed by tuning the resonances of the microrings comprising the ADF and trap/release optical pulses. We show that the WLC based ADF yields a maximally flat filter which exhibits superior performances in terms of bandwidth and flatness compared to previous design approaches. We also present a realistic, Silicon-over-Insulator based, design and performance analysis taking into consideration the realistic properties and limitations of the materials and the fabrication process, leading to delays exceeding 850ps for 80GHz bandwidth, and a corresponding delay-bandwidth product of approximately 70.

  7. MOISTURE-BUFFERING CHARACTERISTICS OF BUILDING MATERIALS

    Directory of Open Access Journals (Sweden)

    Young Cheol Choi

    2016-05-01

    Full Text Available The humidity level of indoor air is an important factor influencing the air quality and energy consumption of buildings, as well as the durability of building components. Indoor humidity levels depend on several factors, such as moisture sources, air flow, and the adsorption/desorption properties of materials. The moisture-buffering characteristics of building materials that are in contact with indoor air may help moderate the variations of indoor humidity, especially in the summer and winter. In this study, the moisture adsorption/desorption properties of building materials were investigated experimentally and numerically. These properties can be used to characterize the ability of building materials to exchange moisture with the indoor environment. This study indicates that a building material surface resistivity was the main factor creating variations of moisture buffering.

  8. Chemical buffering capacity of clay rock

    International Nuclear Information System (INIS)

    Beaucaire, C.; Pearson, F.J.; Gautschi, A.

    2004-01-01

    The long-term performance of a nuclear waste repository is strongly dependent on the chemical properties of the host rock. The host rock establishes the chemical environment that determines such important performance attributes as radionuclide solubilities from the waste and the transport rates from the repository to the accessible environment. Clay-rich rocks are especially favourable host rocks because they provide a strong buffering capacity to resist chemical changes prompted either internally, by reactions of the waste itself and emplacement materials, or externally, by changes in the hydrologic systems surrounding the host rock. This paper will focus on three aspects of the stability of clay-rich host rocks: their ability to provide pCO 2 and redox buffering, and to resist chemical changes imposed by changes in regional hydrology and hydro-chemistry. (authors)

  9. Role of buffer gases in optoacoustic spectroscopy

    International Nuclear Information System (INIS)

    Thomas III, L.J.; Kelly, M.J.; Amer, N.M.

    1978-01-01

    The dependence of an acoustically resonant optoacoustic signal on the molecular weight and thermodynamic and transport properpties of the buffer gas is reported. Our results show that careful selection of such gases can significantly increase the sensitivity and flexibility of optoacoustic spectroscopy. We also demonstrate that such thermodynamic quantities as γ (equivalentC/sub p//C/sub v/) and sound velocity can now be measured readily and accurately. Other potential applications are suggested

  10. Microbial activity in bentonite buffers. Literature study

    Energy Technology Data Exchange (ETDEWEB)

    Ratto, M.; Itavaara, M.

    2012-07-01

    The proposed disposal concept for high-level radioactive wastes involves storing the wastes underground in copper-iron containers embedded in buffer material of compacted bentonite. Hydrogen sulphide production by sulphate-reducing prokaryotes is a potential mechanism that could cause corrosion of waste containers in repository conditions. The prevailing conditions in compacted bentonite buffer will be harsh. The swelling pressure is 7-8 MPa, the amount of free water is low and the average pore and pore throat diameters are small. This literature study aims to assess the potential of microbial activity in bentonite buffers. Literature on the environmental limits of microbial life in extreme conditions and the occurrence of sulphatereducing prokaryotes in extreme environments is reviewed briefly and the results of published studies characterizing microbes and microbial processes in repository conditions or in relevant subsurface environments are presented. The presence of bacteria, including SRBs, has been confirmed in deep groundwater and bentonite-based materials. Sulphate reducers have been detected in various high-pressure environments, and sulphate-reduction based on hydrogen as an energy source is considered a major microbial process in deep subsurface environments. In bentonite, microbial activity is strongly suppressed, mainly due to the low amount of free water and small pores, which limit the transport of microbes and nutrients. Spore-forming bacteria have been shown to survive in compacted bentonite as dormant spores, and they are able to resume a metabolically active state after decompaction. Thus, microbial sulphide production may increase in repository conditions if the dry density of the bentonite buffer is locally reduced. (orig.)

  11. Isostatic compression of buffer blocks. Middle scale

    International Nuclear Information System (INIS)

    Ritola, J.; Pyy, E.

    2012-01-01

    Manufacturing of buffer components using isostatic compression method has been studied in small scale in 2008 (Laaksonen 2010). These tests included manufacturing of buffer blocks using different bentonite materials and different compression pressures. Isostatic mould technology was also tested, along with different methods to fill the mould, such as vibration and partial vacuum, as well as a stepwise compression of the blocks. The development of manufacturing techniques has continued with small-scale (30 %) blocks (diameter 600 mm) in 2009. This was done in a separate project: Isostatic compression, manufacturing and testing of small scale (D = 600 mm) buffer blocks. The research on the isostatic compression method continued in 2010 in a project aimed to test and examine the isostatic manufacturing process of buffer blocks at 70 % scale (block diameter 1200 to 1300 mm), and the aim was to continue in 2011 with full-scale blocks (diameter 1700 mm). A total of nine bentonite blocks were manufactured at 70 % scale, of which four were ring-shaped and the rest were cylindrical. It is currently not possible to manufacture full-scale blocks, because there is no sufficiently large isostatic press available. However, such a compression unit is expected to be possible to use in the near future. The test results of bentonite blocks, produced with an isostatic pressing method at different presses and at different sizes, suggest that the technical characteristics, for example bulk density and strength values, are somewhat independent of the size of the block, and that the blocks have fairly homogenous characteristics. Water content and compression pressure are the two most important properties determining the characteristics of the compressed blocks. By adjusting these two properties it is fairly easy to produce blocks at a desired density. The commonly used compression pressure in the manufacturing of bentonite blocks is 100 MPa, which compresses bentonite to approximately

  12. Isostatic compression of buffer blocks. Middle scale

    Energy Technology Data Exchange (ETDEWEB)

    Ritola, J.; Pyy, E. [VTT Technical Research Centre of Finland, Espoo (Finland)

    2012-01-15

    Manufacturing of buffer components using isostatic compression method has been studied in small scale in 2008 (Laaksonen 2010). These tests included manufacturing of buffer blocks using different bentonite materials and different compression pressures. Isostatic mould technology was also tested, along with different methods to fill the mould, such as vibration and partial vacuum, as well as a stepwise compression of the blocks. The development of manufacturing techniques has continued with small-scale (30 %) blocks (diameter 600 mm) in 2009. This was done in a separate project: Isostatic compression, manufacturing and testing of small scale (D = 600 mm) buffer blocks. The research on the isostatic compression method continued in 2010 in a project aimed to test and examine the isostatic manufacturing process of buffer blocks at 70 % scale (block diameter 1200 to 1300 mm), and the aim was to continue in 2011 with full-scale blocks (diameter 1700 mm). A total of nine bentonite blocks were manufactured at 70 % scale, of which four were ring-shaped and the rest were cylindrical. It is currently not possible to manufacture full-scale blocks, because there is no sufficiently large isostatic press available. However, such a compression unit is expected to be possible to use in the near future. The test results of bentonite blocks, produced with an isostatic pressing method at different presses and at different sizes, suggest that the technical characteristics, for example bulk density and strength values, are somewhat independent of the size of the block, and that the blocks have fairly homogenous characteristics. Water content and compression pressure are the two most important properties determining the characteristics of the compressed blocks. By adjusting these two properties it is fairly easy to produce blocks at a desired density. The commonly used compression pressure in the manufacturing of bentonite blocks is 100 MPa, which compresses bentonite to approximately

  13. META-ANALYSIS OF NITROGEN REMOVAL IN RIPARIAN BUFFERS

    Science.gov (United States)

    Riparian buffer zones, the vegetated region adjacent to streams and wetlands, are thought to be effective at intercepting and controlling nitrogen loads entering water bodies. Riparian buffer width may be positively related to nitrogen removal effectiveness by influencing nitrog...

  14. Methyl Bromide Commodity Fumigation Buffer Zone Lookup Tables

    Science.gov (United States)

    Product labels for methyl bromide used in commodity and structural fumigation include requirements for buffer zones around treated areas. The information on this page will allow you to find the appropriate buffer zone for your planned application.

  15. Success and failure with phthalate buffers in capillary zone electrophoresis

    NARCIS (Netherlands)

    Bocek, P.; Gebauer, P.; Beckers, J.L.

    2001-01-01

    Phthalate buffers are currently used in capillary electrophoresis as robust electrolyte systems for indirect detection. This contribution demonstrates that these buffers show regularly not only successful regions of mobilities of analytes (sample window) but also regions of failure where the

  16. International comparison of observation-specific spatial buffers

    DEFF Research Database (Denmark)

    Frank, Lawrence D; Fox, Eric H; Ulmer, Jared M

    2017-01-01

    the relative differences in resulting explanatory power on self-reported physical activity outcomes. BEMs were developed in five countries using 'sausage,' 'detailed-trimmed,' and 'detailed,' network buffers at a distance of 1 km around participant household addresses (n = 5883). RESULTS: BEM values were...... significantly different (p trimmed buffer comparisons and 89% of sausage versus detailed network buffer comparisons. Results showed that BEM coefficients in physical activity models did not differ significantly across buffering methods, and in most cases BEM...

  17. Complexation of buffer constituents with neutral complexation agents: part I. Impact on common buffer properties.

    Science.gov (United States)

    Riesová, Martina; Svobodová, Jana; Tošner, Zdeněk; Beneš, Martin; Tesařová, Eva; Gaš, Bohuslav

    2013-09-17

    The complexation of buffer constituents with the complexation agent present in the solution can very significantly influence the buffer properties, such as pH, ionic strength, or conductivity. These parameters are often crucial for selection of the separation conditions in capillary electrophoresis or high-pressure liquid chromatography (HPLC) and can significantly affect results of separation, particularly for capillary electrophoresis as shown in Part II of this paper series (Beneš, M.; Riesová, M.; Svobodová, J.; Tesařová, E.; Dubský, P.; Gaš, B. Anal. Chem. 2013, DOI: 10.1021/ac401381d). In this paper, the impact of complexation of buffer constituents with a neutral complexation agent is demonstrated theoretically as well as experimentally for the model buffer system composed of benzoic acid/LiOH or common buffers (e.g., CHES/LiOH, TAPS/LiOH, Tricine/LiOH, MOPS/LiOH, MES/LiOH, and acetic acid/LiOH). Cyclodextrins as common chiral selectors were used as model complexation agents. We were not only able to demonstrate substantial changes of pH but also to predict the general complexation characteristics of selected compounds. Because of the zwitterion character of the common buffer constituents, their charged forms complex stronger with cyclodextrins than the neutral ones do. This was fully proven by NMR measurements. Additionally complexation constants of both forms of selected compounds were determined by NMR and affinity capillary electrophoresis with a very good agreement of obtained values. These data were advantageously used for the theoretical descriptions of variations in pH, depending on the composition and concentration of the buffer. Theoretical predictions were shown to be a useful tool for deriving some general rules and laws for complexing systems.

  18. Further improvements in competitive guarantees for QoS buffering

    NARCIS (Netherlands)

    Bansal, N.; Fleischer, L.K.; Kimbrel, T.; Mahdian, M.; Schieber, B.; Sviridenko, M.; Diaz, J.; Karhumäki, J.; Lepistö, A.; Sannella, D.

    2004-01-01

    We study the behavior of algorithms for buffering packets weighted by different levels of Quality of Service (QoS) guarantees in a single queue. Buffer space is limited, and packet loss occurs when the buffer overflows. We describe a modification of the previously proposed ``preemptive greedy{''}

  19. Towards Optimal Buffer Size in Wi-Fi Networks

    KAUST Repository

    Showail, Ahmad J.

    2016-01-19

    Buffer sizing is an important network configuration parameter that impacts the quality of data traffic. Falling memory cost and the fallacy that ‘more is better’ lead to over provisioning network devices with large buffers. Over-buffering or the so called ‘bufferbloat’ phenomenon creates excessive end-to-end delay in today’s networks. On the other hand, under-buffering results in frequent packet loss and subsequent under-utilization of network resources. The buffer sizing problem has been studied extensively for wired networks. However, there is little work addressing the unique challenges of wireless environment. In this dissertation, we discuss buffer sizing challenges in wireless networks, classify the state-of-the-art solutions, and propose two novel buffer sizing schemes. The first scheme targets buffer sizing in wireless multi-hop networks where the radio spectral resource is shared among a set of con- tending nodes. Hence, it sizes the buffer collectively and distributes it over a set of interfering devices. The second buffer sizing scheme is designed to cope up with recent Wi-Fi enhancements. It adapts the buffer size based on measured link characteristics and network load. Also, it enforces limits on the buffer size to maximize frame aggregation benefits. Both mechanisms are evaluated using simulation as well as testbed implementation over half-duplex and full-duplex wireless networks. Experimental evaluation shows that our proposal reduces latency by an order of magnitude.

  20. Managing Multiuser Database Buffers Using Data Mining Techniques

    NARCIS (Netherlands)

    Feng, L.; Lu, H.J.

    2004-01-01

    In this paper, we propose a data-mining-based approach to public buffer management for a multiuser database system, where database buffers are organized into two areas – public and private. While the private buffer areas contain pages to be updated by particular users, the public

  1. Heed the head: buffer benefits along headwater streams

    Science.gov (United States)

    Rhonda Mazza; Deanna (Dede) Olson

    2015-01-01

    Since the Northwest Forest Plan implemented riparian buffers along non-fish bearing streams in 1994, there have been questions about how wide those buffers need to be to protect aquatic and riparian resources from upland forest management activities. The Density Management and Riparian Buffer Study of western Oregon, also initiated in 1994, examines the effects of...

  2. Concentrated flow paths in riparian buffer zones of southern Illinois

    Science.gov (United States)

    R.C. Pankau; J.E. Schoonover; K.W.J. Willard; P.J. Edwards

    2012-01-01

    Riparian buffers in agricultural landscapes should be designed to trap pollutants in overland flow by slowing, filtering, and infiltrating surface runoff entering the buffer via sheet flow. However, observational evidence suggests that concentrated flow is prevalent from agricultural fields. Over time sediment can accumulate in riparian buffers forming berms that...

  3. Stream water responses to timber harvest: Riparian buffer width effectiveness

    Science.gov (United States)

    Barton D. Clinton

    2011-01-01

    Vegetated riparian buffers are critical for protecting aquatic and terrestrial processes and habitats in southern Appalachian ecosystems. In this case study, we examined the effect of riparian buffer width on stream water quality following upland forest management activities in four headwater catchments. Three riparian buffer widths were delineated prior to cutting; 0m...

  4. Analysis of diffusive mass transport in a cracked buffer

    International Nuclear Information System (INIS)

    Garisto, N.C.; Garisto, F.

    1989-11-01

    In the disposal vault design for the Canadian Nuclear Fuel Waste Management Program, cylindrical containers of used nuclear fuel would be placed in vertical boreholes in rock and surrounded with a bentonite-based buffer material. The buffer is expected to absorb and/or retard radionuclides leaching from the fuel after the containers fail. There is some evidence, however, that the buffer may be susceptible to cracking. In this report we investigate numerically the consequences of cracking on uranium diffusion through the buffer. The derivation of the mass-transport equations and the numerical solution method are presented for the solubility-limited diffusion of uranium in a cracked buffer system for both swept-away and semi-impermeable boundary conditions at the rock-buffer interface. The results indicate that for swept-away boundary conditions the total uranium flux through the cracked buffer system is, as expected, greater than through the uncracked buffer. The effect of the cracks is strongly dependent on the ratio D/D eff , where D and D eff are the pore-water and the effective buffer diffusion coefficient, respectively. However, although a decrease in D eff enhances the effect of cracks on the total cumulative flux (relative to the uncracked buffer), it also decreases the total cumulative flux through the cracked buffer system (relative to a cracked buffer with a larger D eff value). Finally, for semi-impermeable boundary conditions, the effect of cracks on the total radionuclide flux is relatively small

  5. Glucose acutely reduces cytosolic and mitochondrial H2O2 in rat pancreatic beta-cells.

    Science.gov (United States)

    Deglasse, Jean-Philippe; Roma, Leticia Prates; Pastor-Flores, Daniel; Gilon, Patrick; Dick, Tobias P; Jonas, Jean-Christophe

    2018-05-14

    Whether H2O2 contributes to the glucose-dependent stimulation of insulin secretion by pancreatic β-cells is highly controversial. We used two H2O2-sensitive probes, roGFP2-Orp1 and HyPer with its pH-control SypHer, to test the acute effects of glucose, monomethylsuccinate, leucine with glutamine, and α-ketoisocaproate, on β-cell cytosolic and mitochondrial H2O2 concentrations. We then tested the effects of low H2O2 and menadione concentrations on insulin secretion. RoGFP2-Orp1 was more sensitive than HyPer to H2O2 (response at 2-5 vs. 10µM) and less pH-sensitive. Under control conditions, stimulation with glucose reduced mitochondrial roGFP2-Orp1 oxidation without affecting cytosolic roGFP2-Orp1 and HyPer fluorescence ratios, except for the pH-dependent effects on HyPer. However, stimulation with glucose decreased the oxidation of both cytosolic probes by 15µM exogenous H2O2. The glucose effects were not affected by overexpression of catalase, mitochondrial catalase or superoxide dismutase 1 and 2. They followed the increase in NAD(P)H autofluorescence, were maximal at 5mM glucose in the cytosol and 10mM glucose in the mitochondria, and were partly mimicked by the other nutrients. Exogenous H2O2 (1-15µM) did not affect insulin secretion. By contrast, menadione (1-5µM) did not increase basal insulin secretion but reduced the stimulation of insulin secretion by 20mM glucose. Subcellular changes in β-cell H2O2 levels are better monitored with roGFP2-Orp1 than HyPer/SypHer. Nutrients acutely lower mitochondrial H2O2 levels in β-cells and promote degradation of exogenously supplied H2O2 in both cytosolic and mitochondrial compartments. The glucose-dependent stimulation of insulin secretion occurs independently of a detectable increase in β-cell cytosolic or mitochondrial H2O2 levels.

  6. Exchangeability of bentonite buffer and backfill materials

    Energy Technology Data Exchange (ETDEWEB)

    Savage, D. [Savage Earth Associates Ltd, Bournemouth (United Kingdom); Arthur, R. [Intera Inc, Ottawa, ON, (Canada); Luukkonen, A.

    2012-08-15

    Clay-based buffer and tunnel backfill materials are important barriers in the KBS-3 repository concept for final disposal of spent nuclear fuel in Finland. One issue that is relevant to material properties is the degree to which different bentonite compositions can be regarded as interchangeable. In Posiva's current repository design, the reference bentonite composition is MX-80, a sodium montmorillonite dominated clay. Posiva would like to be able to use bentonite with Ca-montmorillonite as the dominant clay mineral. However, at this stage, it is not clear what supporting data need to be acquired/defined to be able to place the state of knowledge of Ca-bentonite at the same level as that of Na-bentonite. In this report, the concept of bentonite exchangeability has been evaluated through consideration of how bentonite behaviour may be affected in six key performance-relevant properties, namely (1) mineralogical composition and availability of materials, (2) hydraulic conductivity, (3) mechanical and rheological properties, (4) long-term alteration, (5) colloidal properties, and (6) swelling pressure. The report evaluates implications for both buffer and backfill. Summary conclusions are drawn from these sections to suggest how bentonite exchangeability may be addressed in regulatory assessments of engineered barrier design for a future geological repository for spent fuel in Finland. Some important conclusions are: (a) There are some fundamental differences between Ca- and Na-bentonites such as colloidal behaviour, pore structure and long-term alteration that could affect the exchangeability of these materials as buffer or backfill materials and which should be further evaluated; (b) Additional experimental data are desirable for some issues such as long-term alteration, hydraulic properties and swelling behaviour, (c) The minor mineral content of bentonites is very variable, both between different bentonites and within the same bentonite type, it is not clear

  7. Cytosolic Cl- Affects the Anticancer Activity of Paclitaxel in the Gastric Cancer Cell Line, MKN28 Cell

    Directory of Open Access Journals (Sweden)

    Sachie Tanaka

    2017-05-01

    Full Text Available Background/Aims: Our previous study revealed that cytosolic Cl- affected neurite elongation promoted via assembly of microtubule in rat pheochromocytoma PC12D cells and Cl-–induced blockade of intrinsic GTPase enhanced tubulin polymerization in vitro. Paclitaxel (PTX is a microtubule-targeted chemotherapeutic drug and stabilizes microtubules resulting in mainly blockade of mitosis at the metaphase-anaphase transition and induction of apoptosis. In the present study, we tried to clarify whether the cytosolic Cl- affected PTX ability to inhibit cell growth in the gastric cancer cell line, MKN28. Methods: To clarify the cytosolic Cl- action on PTX-induced cell death and metaphase-anaphase transition in the gastric cancer cell line, MKN28 cell, and PTX-induced tubulin polymerization, we performed cell proliferation assay, cytosolic Cl- concentration measurement, immunofluorescence microscopy, and in vitro tubulin polymerization assay. Results: The decline of cytosolic Cl- weakened the cytotoxic effect of PTX on cell proliferation of MKN28 cells, which could pass through the metaphase-anaphase transition. Moreover, in vitro PTX-induced tubulin polymerization was diminished under the low Cl- condition. Conclusions: Our results strongly suggest that the upregulation of cytosolic Cl- concentration would enhance the antitumor effect of PTX, and that the cytosolic Cl- would be one of the key targets for anti-cancer therapy.

  8. Automation of metabolic stability studies in microsomes, cytosol and plasma using a 215 Gilson liquid handler.

    Science.gov (United States)

    Linget, J M; du Vignaud, P

    1999-05-01

    A 215 Gilson liquid handler was used to automate enzymatic incubations using microsomes, cytosol and plasma. The design of automated protocols are described. They were based on the use of 96 deep well plates and on HPLC-based methods for assaying the substrate. The assessment of those protocols was made with comparison between manual and automated incubations, reliability and reproducibility of automated incubations in microsomes and cytosol. Examples of the use of those programs in metabolic studies in drug research, i.e. metabolic screening in microsomes and plasma were shown. Even rapid processes (with disappearance half lives as low as 1 min) can be analysed. This work demonstrates how stability studies can be automated to save time, render experiments involving human biological media less hazardous and may be improve inter-laboratory reproducibility.

  9. Dynamin-Related Protein 1 Translocates from the Cytosol to Mitochondria during UV-Induced Apoptosis

    Science.gov (United States)

    Zhang, Zhenzhen; Wu, Shengnan; Feng, Jie

    2011-01-01

    Mitochondria are dynamic structures that frequently divide and fuse with one another to form interconnecting network. This network disintegrates into punctiform organelles during apoptosis. However, the mechanisms involved in these processes are still not well characterized. In this study, we investigate the role of dynamin-related protein 1 (Drp1), a large GTPase that mediates outer mitochondrial membrane fission, in mitochondrial dynamics in response to UV irradiation in human lung adenocarcinoma cells (ASTC-α-1) and HeLa cells. Using time-lapse fluorescent imaging, we find that Drp1 primarily distributes in cytosol under physiological conditions. After UV treatment, Drp1 translocates from cytosol to mitochondria, indicating the enhancement of Drp1 mitochondrial accumulation. Our results suggest that Drp1 is involved in the regulation of transition from an interconnecting network to a punctiform mitochondrial phenotype during UV-induced apoptosis.

  10. Nanocapsule-mediated cytosolic siRNA delivery for anti-inflammatory treatment.

    Science.gov (United States)

    Jiang, Ying; Hardie, Joseph; Liu, Yuanchang; Ray, Moumita; Luo, Xiang; Das, Riddha; Landis, Ryan F; Farkas, Michelle E; Rotello, Vincent M

    2018-06-05

    The use of nanoparticle-stabilized nanocapsules for cytosolic siRNA delivery for immunomodulation in vitro and in vivo is reported. These NPSCs deliver siRNA directly to the cytosol of macrophages in vitro with concomitant knockdown of gene expression. In vivo studies showed directed delivery of NPSCs to the spleen, enabling gene silencing of macrophages, with preliminary studies showing 70% gene knockdown at a siRNA dose of 0.28 mg/kg. Significantly, the delivery of siRNA targeting tumor necrosis factor-α efficiently silenced TNF-α expression in LPS-challenged mice, demonstrating efficacy in modulating immune response in an organ-selective manner. This research highlights the potential of the NPSC platform for targeted immunotherapy and further manipulation of the immune system. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Detecting Release of Bacterial dsDNA into the Host Cytosol Using Fluorescence Microscopy.

    Science.gov (United States)

    Dreier, Roland Felix; Santos, José Carlos; Broz, Petr

    2018-01-01

    Recognition of pathogens by the innate immune system relies on germline-encoded pattern recognition receptors (PRRs) that recognize unique microbial molecules, so-called pathogen-associated molecular patterns (PAMPs). Nucleic acids and their derivatives are one of the most important groups of PAMPs, and are recognized by a number of surface-associated as well as cytosolic PRRs. Cyclic GMP-AMP synthase (cGAS) recognizes the presence of pathogen- or host-derived dsDNA in the cytosol and initiates type-I-IFN production. Here, we describe a methodology that allows for evaluating the association of cGAS with released bacterial dsDNA during Francisella novicida infection of macrophages, by fluorescence confocal microscopy. This method can be adapted to the study of cGAS-dependent responses elicited by other intracellular bacterial pathogens and in other cell types.

  12. A Role for Cytosolic Fumarate Hydratase in Urea Cycle Metabolism and Renal Neoplasia

    Directory of Open Access Journals (Sweden)

    Julie Adam

    2013-05-01

    Full Text Available The identification of mutated metabolic enzymes in hereditary cancer syndromes has established a direct link between metabolic dysregulation and cancer. Mutations in the Krebs cycle enzyme, fumarate hydratase (FH, predispose affected individuals to leiomyomas, renal cysts, and cancers, though the respective pathogenic roles of mitochondrial and cytosolic FH isoforms remain undefined. On the basis of comprehensive metabolomic analyses, we demonstrate that FH1-deficient cells and tissues exhibit defects in the urea cycle/arginine metabolism. Remarkably, transgenic re-expression of cytosolic FH ameliorated both renal cyst development and urea cycle defects associated with renal-specific FH1 deletion in mice. Furthermore, acute arginine depletion significantly reduced the viability of FH1-deficient cells in comparison to controls. Our findings highlight the importance of extramitochondrial metabolic pathways in FH-associated oncogenesis and the urea cycle/arginine metabolism as a potential therapeutic target.

  13. Copper, Zinc Superoxide Dismutase is Primarily a Cytosolic Protein in Human Cells

    Science.gov (United States)

    Crapo, James D.; Oury, Tim; Rabouille, Catherine; Slot, Jan W.; Chang, Ling-Yi

    1992-11-01

    The intracellular localization of human copper, zinc superoxide dismutase (Cu,Zn-SOD; superoxide:superoxide oxidoreductase, EC 1.15.1.1) was evaluated by using EM immunocytochemistry and both isolated human cell lines and human tissues. Eight monoclonal antibodies raised against either native or recombinant human Cu,Zn-SOD and two polyclonal antibodies raised against either native or recombinant human Cu,Zn-SOD were used. Fixation with 2% paraformaldehyde/0.2% glutaraldehyde was found necessary to preserve normal distribution of the protein. Monoclonal antibodies were less effective than polyclonal antibodies in recognizing the antigen after adequate fixation of tissue. Cu,Zn-SOD was found widely distributed in the cell cytosol and in the cell nucleus, consistent with it being a soluble cytosolic protein. Mitochondria and secretory compartments did not label for this protein. In human cells, peroxisomes showed a labeling density slightly less than that of cytoplasm.

  14. Well-defined polypeptide-based systems as non-viral vectors for cytosolic delivery

    OpenAIRE

    Niño Pariente, Amaya

    2017-01-01

    A convenient cytosolic drug delivery constitutes a very powerful tool for the treatment and/or prevention of several relevant human diseases. Along with recent advances in therapeutic technologies based on biomacromolecules (e.g. oligonucleotides or proteins), we also require the development of technologies which improve the transport of therapeutic molecules to the cell of choice. This has led to the emergence of a variety of promising methods over the last 20 years. Despite significant prog...

  15. Role of cytosolic NADP+-dependent isocitrate dehydrogenase in ischemia-reperfusion injury in mouse kidney

    OpenAIRE

    Kim, Jinu; Kim, Ki Young; Jang, Hee-Seong; Yoshida, Takumi; Tsuchiya, Ken; Nitta, Kosaku; Park, Jeen-Woo; Bonventre, Joseph V.; Park, Kwon Moo

    2008-01-01

    Cytosolic NADP+-dependent isocitrate dehydrogenase (IDPc) synthesizes reduced NADP (NADPH), which is an essential cofactor for the generation of reduced glutathione (GSH), the most abundant and important antioxidant in mammalian cells. We investigated the role of IDPc in kidney ischemia-reperfusion (I/R) in mice. The activity and expression of IDPc were highest in the cortex, modest in the outer medulla, and lowest in the inner medulla. NADPH levels were greatest in the cortex. IDPc expressio...

  16. BAF is a cytosolic DNA sensor that leads to exogenous DNA avoiding autophagy.

    Science.gov (United States)

    Kobayashi, Shouhei; Koujin, Takako; Kojidani, Tomoko; Osakada, Hiroko; Mori, Chie; Hiraoka, Yasushi; Haraguchi, Tokuko

    2015-06-02

    Knowledge of the mechanisms by which a cell detects exogenous DNA is important for controlling pathogen infection, because most pathogens entail the presence of exogenous DNA in the cytosol, as well as for understanding the cell's response to artificially transfected DNA. The cellular response to pathogen invasion has been well studied. However, spatiotemporal information of the cellular response immediately after exogenous double-stranded DNA (dsDNA) appears in the cytosol is lacking, in part because of difficulties in monitoring when exogenous dsDNA enters the cytosol of the cell. We have recently developed a method to monitor endosome breakdown around exogenous materials using transfection reagent-coated polystyrene beads incorporated into living human cells as the objective for microscopic observations. In the present study, using dsDNA-coated polystyrene beads (DNA-beads) incorporated into living cells, we show that barrier-to-autointegration factor (BAF) bound to exogenous dsDNA immediately after its appearance in the cytosol at endosome breakdown. The BAF(+) DNA-beads then assembled a nuclear envelope (NE)-like membrane and avoided autophagy that targeted the remnants of the endosome membranes. Knockdown of BAF caused a significant decrease in the assembly of NE-like membranes and increased the formation of autophagic membranes around the DNA-beads, suggesting that BAF-mediated assembly of NE-like membranes was required for the DNA-beads to evade autophagy. Importantly, BAF-bound beads without dsDNA also assembled NE-like membranes and avoided autophagy. We propose a new role for BAF: remodeling intracellular membranes upon detection of dsDNA in mammalian cells.

  17. Effect of starvation, diabetes and insulin on the casein kinase 2 from rat liver cytosol.

    OpenAIRE

    Martos, C; Plana, M; Guasch, M D; Itarte, E

    1985-01-01

    Starvation, diabetes and insulin did not alter the concentration of casein kinases in rat liver cytosol. However, the Km for casein of casein kinase 2 from diabetic rats was about 2-fold lower than that from control animals. Administration of insulin to control rats did not alter this parameter, but increased the Km for casein of casein kinase 2 in diabetic rats. Starvation did not affect the kinetic constants of casein kinases. The effect of diabetes on casein kinase 2 persisted after partia...

  18. Unique hepatic cytosolic arginase evolved independently in ureogenic freshwater air-breathing teleost, Heteropneustes fossilis.

    Directory of Open Access Journals (Sweden)

    Shilpee Srivastava

    Full Text Available Hepatic cytosolic arginase (ARG I, an enzyme of the urea cycle operating in the liver of ureotelic animals, is reported to be present in an ammoniotelic freshwater air-breathing teleost, Heteropneustes fossilis which has ureogenic potential. Antibodies available against mammalian ARG I showed no cross reactivity with the H. fossilis ARG I. We purified unique ARG I from H. fossilis liver. Purified ARG I is a homotrimer with molecular mass 75 kDa and subunit molecular mass of 24 kDa. The pI value of the enzyme was 8.5. It showed maximum activity at pH 10.5 and 55°C. The Km of purified enzyme for L-arginine was 2.65±0.39 mM. L-ornithine and N(ω-hydroxy-L-arginine showed inhibition of the ARG I activity, with Ki values 0.52±0.02mM and 0.08±0.006mM, respectively. Antibody raised against the purified fish liver ARG I showed exclusive specificity, and has no cross reactivity against fish liver ARG II and mammalian liver ARG I and ARG II. We found another isoform of arginase bound to the outer membrane of the mitochondria which was released by 150-200 mM KCl in the extraction medium. This isoform was immunologically different from the soluble cytosolic and mitochondrial arginase. The results of present study support that hepatic cytosolic arginase evolved in this ureogenic freshwater teleost, H. fossilis. Phylogenetic analysis confirms an independent evolution event that occurred much after the evolution of the cytosolic arginase of ureotelic vertebrates.

  19. GABA transaminases from Saccharomyces cerevisiae and Arabidopsis thaliana complement function in cytosol and mitochondria.

    Science.gov (United States)

    Cao, Juxiang; Barbosa, Jose M; Singh, Narendra; Locy, Robert D

    2013-07-01

    GABA transaminase (GABA-T) catalyses the conversion of GABA to succinate semialdehyde (SSA) in the GABA shunt pathway. The GABA-T from Saccharomyces cerevisiae (ScGABA-TKG) is an α-ketoglutarate-dependent enzyme encoded by the UGA1 gene, while higher plant GABA-T is a pyruvate/glyoxylate-dependent enzyme encoded by POP2 in Arabidopsis thaliana (AtGABA-T). The GABA-T from A. thaliana is localized in mitochondria and mediated by an 18-amino acid N-terminal mitochondrial targeting peptide predicated by both web-based utilities TargetP 1.1 and PSORT. Yeast UGA1 appears to lack a mitochondrial targeting peptide and is localized in the cytosol. To verify this bioinformatic analysis and examine the significance of ScGABA-TKG and AtGABA-T compartmentation and substrate specificity on physiological function, expression vectors were constructed to modify both ScGABA-TKG and AtGABA-T, so that they express in yeast mitochondria and cytosol. Physiological function was evaluated by complementing yeast ScGABA-TKG deletion mutant Δuga1 with AtGABA-T or ScGABA-TKG targeted to the cytosol or mitochondria for the phenotypes of GABA growth defect, thermosensitivity and heat-induced production of reactive oxygen species (ROS). This study demonstrates that AtGABA-T is functionally interchangeable with ScGABA-TKG for GABA growth, thermotolerance and limiting production of ROS, regardless of location in mitochondria or cytosol of yeast cells, but AtGABA-T is about half as efficient in doing so as ScGABA-TKG. These results are consistent with the hypothesis that pyruvate/glyoxylate-limited production of NADPH mediates the effect of the GABA shunt in moderating heat stress in Saccharomyces. Copyright © 2013 John Wiley & Sons, Ltd.

  20. Cytosolic labile zinc: a marker for apoptosis in the developing rat brain.

    Science.gov (United States)

    Lee, Joo-Yong; Hwang, Jung Jin; Park, Mi-Ha; Koh, Jae-Young

    2006-01-01

    Cytosolic zinc accumulation was thought to occur specifically in neuronal death (necrosis) following acute injury. However, a recent study demonstrated that zinc accumulation also occurs in adult rat neurons undergoing apoptosis following target ablation, and in vitro experiments have shown that zinc accumulation may play a causal role in various forms of apoptosis. Here, we examined whether intraneuronal zinc accumulation occurs in central neurons undergoing apoptosis during development. Embryonic and newborn Sprague-Dawley rat brains were double-stained for terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) detection of apoptosis and immunohistochemical detection of stage-specific neuronal markers, such as nestin, proliferating cell nuclear antigen (PCNA), TuJ1 and neuronal nuclear specific protein (NeuN). The results revealed that apoptotic cell death occurred in neurons of diverse stages (neural stem cells, and dividing, young and adult neurons) throughout the brain during the embryonic and early postnatal periods. Further staining of brain sections with acid fuchsin or zinc-specific fluorescent dyes showed that all of the apoptotic neurons were acidophilic and contained labile zinc in their cell bodies. Cytosolic zinc accumulation was also observed in cultured cortical neurons undergoing staurosporine- or sodium nitroprusside (SNP)-induced apoptosis. In contrast, zinc chelation with CaEDTA or N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) reduced SNP-induced apoptosis but not staurosporine-induced apoptosis, indicating that cytosolic zinc accumulation does not play a causal role in all forms of apoptosis. Finally, the specific cytosolic zinc accumulation may have a practical application as a relatively simple marker for neurons undergoing developmental apoptosis.

  1. A cytosolic copper storage protein provides a second level of copper tolerance in Streptomyces lividans.

    Science.gov (United States)

    Straw, Megan L; Chaplin, Amanda K; Hough, Michael A; Paps, Jordi; Bavro, Vassiliy N; Wilson, Michael T; Vijgenboom, Erik; Worrall, Jonathan A R

    2018-01-24

    Streptomyces lividans has a distinct dependence on the bioavailability of copper for its morphological development. A cytosolic copper resistance system is operative in S. lividans that serves to preclude deleterious copper levels. This system comprises of several CopZ-like copper chaperones and P 1 -type ATPases, predominantly under the transcriptional control of a metalloregulator from the copper sensitive operon repressor (CsoR) family. In the present study, we discover a new layer of cytosolic copper resistance in S. lividans that involves a protein belonging to the newly discovered family of copper storage proteins, which we have named Ccsp (cytosolic copper storage protein). From an evolutionary perspective, we find Ccsp homologues to be widespread in Bacteria and extend through into Archaea and Eukaryota. Under copper stress Ccsp is upregulated and consists of a homotetramer assembly capable of binding up to 80 cuprous ions (20 per protomer). X-ray crystallography reveals 18 cysteines, 3 histidines and 1 aspartate are involved in cuprous ion coordination. Loading of cuprous ions to Ccsp is a cooperative process with a Hill coefficient of 1.9 and a CopZ-like copper chaperone can transfer copper to Ccsp. A Δccsp mutant strain indicates that Ccsp is not required under initial copper stress in S. lividans, but as the CsoR/CopZ/ATPase efflux system becomes saturated, Ccsp facilitates a second level of copper tolerance.

  2. The Hepatitis B Virus X Protein Elevates Cytosolic Calcium Signals by Modulating Mitochondrial Calcium Uptake

    Science.gov (United States)

    Yang, Bei

    2012-01-01

    Chronic hepatitis B virus (HBV) infections are associated with the development of hepatocellular carcinoma (HCC). The HBV X protein (HBx) is thought to play an important role in the development of HBV-associated HCC. One fundamental HBx function is elevation of cytosolic calcium signals; this HBx activity has been linked to HBx stimulation of cell proliferation and transcription pathways, as well as HBV replication. Exactly how HBx elevates cytosolic calcium signals is not clear. The studies described here show that HBx stimulates calcium entry into cells, resulting in an increased plateau level of inositol 1,4,5-triphosphate (IP3)-linked calcium signals. This increased calcium plateau can be inhibited by blocking mitochondrial calcium uptake and store-operated calcium entry (SOCE). Blocking SOCE also reduced HBV replication. Finally, these studies also demonstrate that there is increased mitochondrial calcium uptake in HBx-expressing cells. Cumulatively, these studies suggest that HBx can increase mitochondrial calcium uptake and promote increased SOCE to sustain higher cytosolic calcium and stimulate HBV replication. PMID:22031934

  3. TRIM56-mediated monoubiquitination of cGAS for cytosolic DNA sensing.

    Science.gov (United States)

    Seo, Gil Ju; Kim, Charlotte; Shin, Woo-Jin; Sklan, Ella H; Eoh, Hyungjin; Jung, Jae U

    2018-02-09

    Intracellular nucleic acid sensors often undergo sophisticated modifications that are critical for the regulation of antimicrobial responses. Upon recognition of DNA, the cytosolic sensor cyclic GMP-AMP (cGAMP) synthase (cGAS) produces the second messenger cGAMP, which subsequently initiates downstream signaling to induce interferon-αβ (IFNαβ) production. Here we report that TRIM56 E3 ligase-induced monoubiquitination of cGAS is important for cytosolic DNA sensing and IFNαβ production to induce anti-DNA viral immunity. TRIM56 induces the Lys335 monoubiquitination of cGAS, resulting in a marked increase of its dimerization, DNA-binding activity, and cGAMP production. Consequently, TRIM56-deficient cells are defective in cGAS-mediated IFNαβ production upon herpes simplex virus-1 (HSV-1) infection. Furthermore, TRIM56-deficient mice show impaired IFNαβ production and high susceptibility to lethal HSV-1 infection but not to influenza A virus infection. This adds TRIM56 as a crucial component of the cytosolic DNA sensing pathway that induces anti-DNA viral innate immunity.

  4. The RAB2B-GARIL5 Complex Promotes Cytosolic DNA-Induced Innate Immune Responses.

    Science.gov (United States)

    Takahama, Michihiro; Fukuda, Mitsunori; Ohbayashi, Norihiko; Kozaki, Tatsuya; Misawa, Takuma; Okamoto, Toru; Matsuura, Yoshiharu; Akira, Shizuo; Saitoh, Tatsuya

    2017-09-19

    Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that induces the IFN antiviral response. However, the regulatory mechanisms that mediate cGAS-triggered signaling have not been fully explored. Here, we show the involvement of a small GTPase, RAB2B, and its effector protein, Golgi-associated RAB2B interactor-like 5 (GARIL5), in the cGAS-mediated IFN response. RAB2B-deficiency affects the IFN response induced by cytosolic DNA. Consistent with this, RAB2B deficiency enhances replication of vaccinia virus, a DNA virus. After DNA stimulation, RAB2B colocalizes with stimulator of interferon genes (STING), the downstream signal mediator of cGAS, on the Golgi apparatus. The GTP-binding activity of RAB2B is required for its localization on the Golgi apparatus and for recruitment of GARIL5. GARIL5 deficiency also affects the IFN response induced by cytosolic DNA and enhances replication of vaccinia virus. These findings indicate that the RAB2B-GARIL5 complex promotes IFN responses against DNA viruses by regulating the cGAS-STING signaling axis. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  5. The RAB2B-GARIL5 Complex Promotes Cytosolic DNA-Induced Innate Immune Responses

    Directory of Open Access Journals (Sweden)

    Michihiro Takahama

    2017-09-01

    Full Text Available Cyclic GMP-AMP synthase (cGAS is a cytosolic DNA sensor that induces the IFN antiviral response. However, the regulatory mechanisms that mediate cGAS-triggered signaling have not been fully explored. Here, we show the involvement of a small GTPase, RAB2B, and its effector protein, Golgi-associated RAB2B interactor-like 5 (GARIL5, in the cGAS-mediated IFN response. RAB2B-deficiency affects the IFN response induced by cytosolic DNA. Consistent with this, RAB2B deficiency enhances replication of vaccinia virus, a DNA virus. After DNA stimulation, RAB2B colocalizes with stimulator of interferon genes (STING, the downstream signal mediator of cGAS, on the Golgi apparatus. The GTP-binding activity of RAB2B is required for its localization on the Golgi apparatus and for recruitment of GARIL5. GARIL5 deficiency also affects the IFN response induced by cytosolic DNA and enhances replication of vaccinia virus. These findings indicate that the RAB2B-GARIL5 complex promotes IFN responses against DNA viruses by regulating the cGAS-STING signaling axis.

  6. Cytosolic calcium homeostasis in fungi: Roles of plasma membrane transport and intracellular sequestration of calcium

    International Nuclear Information System (INIS)

    Miller, A.J.; Vogg, G.; Sanders, D.

    1990-01-01

    Cytosolic free calcium ([Ca 2+ ] c ) has been measured in the mycelial fungus Neurospora crassa with Ca 2+ - selective microelectrodes. The mean value of [Ca 2+ ] c is 92 ± 15 nM and it is insensitive to external pH values between 5.8 and 8.4. Simultaneous measurement of membrane potential enables the electrochemical potential difference for Ca 2+ across the plasma membrane to be estimated as about -60 kJmol -1 - a value that cannot be sustained either by a simple Ca 2+ - ATPase, or, in alkaline conditions, by straightforward H + /Ca 2+ exchange with a stoichiometric ratio of + /Ca 2+ . The authors propose that the most likely alternative mechanism of Ca 2+ efflux is ATP-driven H + /Ca 2+ exchange, with a stoichiometric ratio of at least 2 H + /Ca 2+ . The increase in [Ca 2+ ] c in the presence of CN - at pH 8.4 is compared with 45 Ca 2+ influx under the same conditions. The proportion of entering Ca 2+ remaining free in the cytosol is only 8 x 10 -5 , and since the concentration of available chelation sites on Ca 2+ binding proteins is unlikely to exceed 100 μM, a major role for the fungal vacuole in short-term Ca 2+ homeostasis is indicated. This notion is supported by the observation that cytosolic Ca 2+ homeostasis is disrupted by a protonophore, which rapidly abolishes the driving force for Ca 2+ uptake into fungal vacuoles

  7. Organization of cytoskeleton controls the changes in cytosolic calcium of cold-shocked Nicotiana plumbaginifolia protoplasts.

    Science.gov (United States)

    Mazars, C; Thion, L; Thuleau, P; Graziana, A; Knight, M R; Moreau, M; Ranjeva, R

    1997-11-01

    Using Nicotiana plumbaginifolia constitutively expressing the recombinant bioluminescent calcium indicator, aequorin, it has been previously demonstrated that plant cells react to cold-shock by an immediate rise in cytosolic calcium. Such an opportune system has been exploited to address the regulatory pathway involved in the calcium response. For this purpose, we have used protoplasts derived from N. plumbaginifolia leaves that behave as the whole plant but with a better reproducibility. By both immunodetecting cytoskeletal components on membrane ghosts and measuring the relative change in cytosolic calcium, we demonstrate that the organization of the cytoskeleton has profound influences on the calcium response. The disruption of the microtubule meshwork by various active drugs, such as colchicin, oryzalin and vinblastin, leads to an important increase in the cytosolic calcium (up to 400 nM) in cold-shocked protoplasts over control. beta-Lumicolchicin, an inactive analogue of colchicin, is ineffective either on cytoplasmic calcium increase or on microtubule organization. A microfilament disrupting drug, cytochalasin D, exerts a slight stimulatory effect, whereas the simultaneous disruption of microtubule and microfilament meshworks results in a dramatic increase in the calcium response to cold-shock. The results described in the present paper illustrate the role of the intracellular organization and, more specifically, the role of cytoskeleton in controlling the intensity of calcium response to an extracellular stimulus.

  8. AM fungal exudates activate MAP kinases in plant cells in dependence from cytosolic Ca(2+) increase.

    Science.gov (United States)

    Francia, Doriana; Chiltz, Annick; Lo Schiavo, Fiorella; Pugin, Alain; Bonfante, Paola; Cardinale, Francesca

    2011-09-01

    The molecular dialogue occurring prior to direct contact between the fungal and plant partners of arbuscular-mycorrhizal (AM) symbioses begins with the release of fungal elicitors, so far only partially identified chemically, which can activate specific signaling pathways in the host plant. We show here that the activation of MAPK is also induced by exudates of germinating spores of Gigaspora margarita in cultured cells of the non-leguminous species tobacco (Nicotiana tabacum), as well as in those of the model legume Lotus japonicus. MAPK activity peaked about 15 min after the exposure of the host cells to the fungal exudates (FE). FE were also responsible for a rapid and transient increase in free cytosolic Ca(2+) in Nicotiana plumbaginifolia and tobacco cells, and pre-treatment with a Ca(2+)-channel blocker (La(3+)) showed that in these cells, MAPK activation was dependent on the cytosolic Ca(2+) increase. A partial dependence of MAPK activity on the common Sym pathway could be demonstrated for a cell line of L. japonicus defective for LjSym4 and hence unable to establish an AM symbiosis. Our results show that MAPK activation is triggered by an FE-induced cytosolic Ca(2+) transient, and that a Sym genetic determinant acts to modulate the intensity and duration of this activity. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  9. Chloride concentrations in human hepatic cytosol and mitochondria are a function of age.

    Science.gov (United States)

    Jahn, Stephan C; Rowland-Faux, Laura; Stacpoole, Peter W; James, Margaret O

    2015-04-10

    We recently reported that, in a concentration-dependent manner, chloride protects hepatic glutathione transferase zeta 1 from inactivation by dichloroacetate, an investigational drug used in treating various acquired and congenital metabolic diseases. Despite the importance of chloride ions in normal physiology, and decades of study of chloride transport across membranes, the literature lacks information on chloride concentrations in animal tissues other than blood. In this study we measured chloride concentrations in human liver samples from male and female donors aged 1 day to 84 years (n = 97). Because glutathione transferase zeta 1 is present in cytosol and, to a lesser extent, in mitochondria, we measured chloride in these fractions by high-performance liquid chromatography analysis following conversion of the free chloride to pentafluorobenzylchloride. We found that chloride concentration decreased with age in hepatic cytosol but increased in liver mitochondria. In addition, chloride concentrations in cytosol, (105.2 ± 62.4 mM; range: 24.7-365.7 mM) were strikingly higher than those in mitochondria (4.2 ± 3.8 mM; range 0.9-22.2 mM). These results suggest a possible explanation for clinical observations seen in patients treated with dichloroacetate, whereby children metabolize the drug more rapidly than adults following repeated doses, and also provide information that may influence our understanding of normal liver physiology. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. A Buffer Management Issue in Designing SSDs for LFSs

    Science.gov (United States)

    Kim, Jaegeuk; Seol, Jinho; Maeng, Seungryoul

    This letter introduces a buffer management issue in designing SSDs for log-structured file systems (LFSs). We implemented a novel trace-driven SSD simulator in SystemC language, and simulated several SSD architectures with the NILFS2 trace. From the results, we give two major considerations related to the buffer management as follows. (1) The write buffer is used as a buffer not a cache, since all write requests are sequential in NILFS2. (2) For better performance, the main architectural factor is the bus bandwidth, but 332MHz is enough. Instead, the read buffer makes a key role in performance improvement while caching data. To enhance SSDs, accordingly, it is an effective way to make efficient read buffer management policies, and one of the examples is tracking the valid data zone in NILFS2, which can increase the data hit ratio in read buffers significantly.

  11. Magnetorheological elastomer and its application on impact buffer

    International Nuclear Information System (INIS)

    Fu, J; Yu, M; Zhu, L X; Dong, X M

    2013-01-01

    In this study, a new magnetorheological elastomer (MRE) based buffer is proposed and its vibration isolation performance is investigated. The MRE buffer with a compact structure is first designed in order to accomplish the maximization of the variable stiffness range. The working characteristics of the MRE buffer are then measured and the model of MRE is established. On the basis of the experimental data, the control model of the MRE buffer is also formulated. A two-degree-of-freedom dynamic model with an MRE buffer is then developed. An intelligent control strategy, human simulated intelligent control (HSIC), is proposed to reduce the impact during the drop crash. Finally, the proposed MRE buffer and controller are validated numerically and experimentally. The results show that the proposed MRE buffer and the control strategy can reduce the impact acceleration effectively.

  12. Buffer layers and articles for electronic devices

    Science.gov (United States)

    Paranthaman, Mariappan P.; Aytug, Tolga; Christen, David K.; Feenstra, Roeland; Goyal, Amit

    2004-07-20

    Materials for depositing buffer layers on biaxially textured and untextured metallic and metal oxide substrates for use in the manufacture of superconducting and other electronic articles comprise RMnO.sub.3, R.sub.1-x A.sub.x MnO.sub.3, and combinations thereof; wherein R includes an element selected from the group consisting of La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, and Y, and A includes an element selected from the group consisting of Be, Mg, Ca, Sr, Ba, and Ra.

  13. Shock buffer for nuclear control element assembly

    International Nuclear Information System (INIS)

    Bevilacqua, F.

    1977-01-01

    A shock buffer for a control element assembly in a nuclear reactor is described, comprising a piston and a cylinder. The piston is affixed to and extends upward from the control rod guide structure; the cylinder is supported by the upper portion of the control element assembly and is vertically oriented with open end downward for receiving the piston. Coolant liquid normally has free access to the cylinder. The piston displaces liquid from the cylinder when inserted, thereby decelerating the control element assembly near its lower extent of travel. (LL)

  14. Prognostic significance of cytosolic pS2 content in ovarian tumors

    International Nuclear Information System (INIS)

    Raigoso, P.; Allende, T.; Zeidan, N.; Llana, B.; Bernardo, L.; Roiz, C.; Tejuca, S.; Vazquez, J.; Lamelas, M.L.

    2002-01-01

    Aim: pS2 is an estrogen regulated peptide which has been associated with a good prognosis an with a more favorable response to treatment in breast cancer patients. In ovarian tumors, the expression of pS2 was demonstrated at both mRNA and protein levels. In addition, it has been showed significant association of pS2 with mucinous differentiation or well differentiation grade of the tumors. However, it is little know about the prognostic significance of the pS2 content in ovarian carcinomas. The aims of the present work were to analyze the cytosolic pS2 content in benign and malignant ovarian tumors, its relationship with clinico-pathologic parameters, steroid receptor status, and prognostic significance. Material and Methods: We analysed the cytosolic concentrations of pS2 in 91 specimen ovarian tissues by an immunoradiometric assay (ELSA-pS2, CIS, France). The tissues were 8 normal ovaries, 43 benign tumors and 40 malignant ovarian tumors. The same ovarian tissues processed to pS2 were analyzed to Estrogen (ER) and Progesterone (PgR) Receptor status. These steroid receptors were quantified biochemically following commercial ELISA method (ABBOTT Diagnostics, Germany). The relationship between cytosolic content and clinico-pathologic factors was examined by the Mann-Whitney or Kruskall-Wallis test. Correlation between steroid receptors and pS2 content was calculated with the Spearman test. Survival curves were calculated using the Kaplan-Meier method and compared by the log-rank test. Differences were considered significant at 5% probability level. Results: pS2 could be detected in 30 cases (32.9%) with values ranged from 0.04 to 89 ng/mg prt. Only one normal ovary showed detectable levels of pS2 and there were not differences in cytosolic content between benign and malignant ovarian tumors. The pS2 levels were only associated to mucinous differentiation in both benign and malignant ovarian tumors (p=0.029 and p=0.015, respectively). Significantly higher

  15. Clad buffer rod sensors for liquid metals

    International Nuclear Information System (INIS)

    Jen, C.-K.; Ihara, I.

    1999-01-01

    Clad buffer rods, consisting of a core and a cladding, have been developed for ultrasonic monitoring of liquid metal processing. The cores of these rods are made of low ultrasonic-loss materials and the claddings are fabricated by thermal spray techniques. The clad geometry ensures proper ultrasonic guidance. The lengths of these rods ranges from tens of centimeters to 1m. On-line ultrasonic level measurements in liquid metals such as magnesium at 700 deg C and aluminum at 960 deg C are presented to demonstrate their operation at high temperature and their high ultrasonic performance. A spherical concave lens is machined at the rod end for improving the spatial resolution. High quality ultrasonic images have been obtained in the liquid zinc at 600 deg C. High spatial resolution is needed for the detection of inclusions in liquid metals during processing. We also show that the elastic properties such as density, longitudinal and shear wave velocities of liquid metals can be measured using a transducer which generates and receives both longitudinal and shear waves and is mounted at the end of a clad buffer rod. (author)

  16. Unsaturated hydraulic property of buffer material

    International Nuclear Information System (INIS)

    Suzuki, Hideaki; Fujita, Tomoo

    1999-09-01

    After emplacement of the engineered barrier system (EBS), it is expected that the near-field environment will be impacted by phenomena such as heat dissipation by conduction and other heat transfer mechanism, infiltration of groundwater from the surrounding rock into the EBS, generation of swelling pressure in the buffer due to water infiltration and the stress imposed by the overburden pressure. These phenomena are not all independent, but can be strongly influenced by, and coupled with, each other. Evaluating these coupled thermo-hydro-mechanical phenomena is important in order to clarify the initial transient behavior of the engineered barrier system within the near-field. This report describes the results on measurement of chemical potential, water diffusivity, and thermal water diffusivity of bentonite that is considered as a candidate material of buffer and on comparison between measurements and theoretical studies for these properties. The following results are identified; (l) The hysteresis of chemical potential in wet and dry conditions for compacted bentonite is not shown clearly. The chemical potential depends on temperature and amount of montmorillonite. When chemical potential of compacted bentonite is zero, the specimen is saturated. The van Genuchten model is applicable to the measured chemical potential of compacted bentonite. (2) The Darcy's law and Philip and de Vries model are applicable to the measured water diffusivity and thermal water diffusivity of compacted bentonite. (author)

  17. Phylogenetic Analysis of Nucleus-Encoded Acetyl-CoA Carboxylases Targeted at the Cytosol and Plastid of Algae.

    KAUST Repository

    Huerlimann, Roger; Zenger, Kyall R; Jerry, Dean R; Heimann, Kirsten

    2015-01-01

    as Chromalveolata, forming the red lineage. However, recent genetic evidence groups the Stramenopiles, Alveolata and green plastid containing Rhizaria as SAR, excluding Haptophyta and Cryptophyta. Sequences coding for plastid and cytosol targeted homomeric ACCases

  18. The use of 0.01M phosphate buffered saline as detection buffer for ...

    African Journals Online (AJOL)

    Insufficient supply of manufacture's buffers/diluents in relation to the number of strips per kit has been found to have negative impact on patients' results. Some laboratories personnel tend to use diluents from other rapid tests manufacturers such as Bioline, Unigold as well as malaria rapid diagnostic test (MRDT). This study ...

  19. Vegetative buffer strips for reducing herbicide transport in runoff: effects of buffer width, vegetation, and season

    Science.gov (United States)

    The effect of vegetative buffer strip (VBS) width, vegetation, and season of the year on herbicide transport in runoff has not been well documented for runoff prone soils. A multi-year replicated plot-scale study was conducted on an eroded claypan soil with the following objectives: 1) assess the ef...

  20. Buffer Sizing in 802.11 Wireless Mesh Networks

    KAUST Repository

    Jamshaid, Kamran; Shihada, Basem; Xia, Li; Levis, Philip

    2011-01-01

    We analyze the problem of buffer sizing for TCP flows in 802.11-based Wireless Mesh Networks. Our objective is to maintain high network utilization while providing low queueing delays. The problem is complicated by the time-varying capacity of the wireless channel as well as the random access mechanism of 802.11 MAC protocol. While arbitrarily large buffers can maintain high network utilization, this results in large queueing delays. Such delays may affect TCP stability characteristics, and also increase queueing delays for other flows (including real-time flows) sharing the buffer. In this paper we propose sizing link buffers collectively for a set of nodes within mutual interference range called the 'collision domain'. We aim to provide a buffer just large enough to saturate the available capacity of the bottleneck collision domain that limits the carrying capacity of the network. This neighborhood buffer is distributed over multiple nodes that constitute the network bottleneck; a transmission by any of these nodes fully utilizes the available spectral resource for the duration of the transmission. We show that sizing routing buffers collectively for this bottleneck allows us to have small buffers (as low as 2 - 3 packets) at individual nodes without any significant loss in network utilization. We propose heuristics to determine these buffer sizes in WMNs. Our results show that we can reduce the end-to-end delays by 6× to 10× at the cost of losing roughly 5% of the network capacity achievable with large buffers.

  1. Buffer Sizing in 802.11 Wireless Mesh Networks

    KAUST Repository

    Jamshaid, Kamran

    2011-10-01

    We analyze the problem of buffer sizing for TCP flows in 802.11-based Wireless Mesh Networks. Our objective is to maintain high network utilization while providing low queueing delays. The problem is complicated by the time-varying capacity of the wireless channel as well as the random access mechanism of 802.11 MAC protocol. While arbitrarily large buffers can maintain high network utilization, this results in large queueing delays. Such delays may affect TCP stability characteristics, and also increase queueing delays for other flows (including real-time flows) sharing the buffer. In this paper we propose sizing link buffers collectively for a set of nodes within mutual interference range called the \\'collision domain\\'. We aim to provide a buffer just large enough to saturate the available capacity of the bottleneck collision domain that limits the carrying capacity of the network. This neighborhood buffer is distributed over multiple nodes that constitute the network bottleneck; a transmission by any of these nodes fully utilizes the available spectral resource for the duration of the transmission. We show that sizing routing buffers collectively for this bottleneck allows us to have small buffers (as low as 2 - 3 packets) at individual nodes without any significant loss in network utilization. We propose heuristics to determine these buffer sizes in WMNs. Our results show that we can reduce the end-to-end delays by 6× to 10× at the cost of losing roughly 5% of the network capacity achievable with large buffers.

  2. Studies of the activity of cytosol on the mixed disulfide bond formed by proteins and radioprotector mercaptoethylguanidine

    Energy Technology Data Exchange (ETDEWEB)

    Horvath, M [National Inst. of Oncology, Budapest (Hungary); Holland, J [Orszagos Onkologiai Intezet, Budapest (Hungary)

    1979-01-01

    The cytoplasm of normal and tumorous rat liver cells contains a heat-resistant compound with reducing ability to break the mixed disulfide bond of albumin-/sup 14/C-mercaptoethylguanidine. The reducing activity of cytosol is destoryed by 1000 krd /sup 60/Co-gamma-ray doses in diluted solution. In vivo supralethal of rats does not affect the activity of cytosol prepared from liver cells.

  3. Pressurized liquid extraction-assisted mussel cytosol preparation for the determination of metals bound to metallothionein-like proteins

    International Nuclear Information System (INIS)

    Santiago-Rivas, Sandra; Moreda-Pineiro, Antonio; Bermejo-Barrera, Pilar; Moreda-Pineiro, Jorge; Alonso-Rodriguez, Elia; Muniategui-Lorenzo, Soledad; Lopez-Mahia, Purificacion; Prada-Rodriguez, Dario

    2007-01-01

    The possibilities of pressurized liquid extraction (PLE) have been novelty tested to assist the cytosol preparation from wet mussel soft tissue before the determination of metals bound to metallothionein-like proteins (MLPs). Results obtained after PLE were compared with those obtained after a classical blending procedure for mussel cytosolic preparation. Isoforms MLP-1 (retention time of 4.1 min) and MLP-2 (retention time of 7.4 min) were separated by anion exchange high-performance liquid chromatography (HPLC) and the concentrations of Ba, Cu, Mn, Sr and Zn bound to MLP isoforms were directly measured by inductively coupled plasma-atomic emission spectrometry (ICP-OES) as a multi-element detector. The optimized PLE-assisted mussel cytosol preparation has consisted of one extraction cycle at room temperature and 1500 psi for 2 min. Since separation between the solid mussel residue and the extract (cytosol) is performed by the PLE system, the cytosol preparation method is faster than conventional cytosol preparation methods by cutting/blending using Ultraturrax or Stomacher devices

  4. Alterations in cytosol free calcium in horseradish roots simultaneously exposed to lanthanum(III) and acid rain.

    Science.gov (United States)

    Zhang, Xuanbo; Wang, Lihong; Zhou, Anhua; Zhou, Qing; Huang, Xiaohua

    2016-04-01

    The extensive use of rare earth elements (REEs) has increased their environmental levels. REE pollution concomitant with acid rain in many agricultural regions can affect crop growth. Cytosol free calcium ions (Ca(2+)) play an important role in almost all cellular activities. However, no data have been reported regarding the role of cytosol free Ca(2+) in plant roots simultaneously exposed to REE and acid rain. In this study, the effects of exposures to lanthanum(III) and acid rain, independently and in combination, on cytosol free Ca(2+) levels, root activity, metal contents, biomass, cytosol pH and La contents in horseradish roots were investigated. The simultaneous exposures to La(III) and acid rain increased or decreased the cytosol free Ca(2+) levels, depending on the concentration of La(III), and these effects were more evident than independent exposure to La(III) or acid rain. In combined exposures, cytosol free Ca(2+) played an important role in the regulation of root activity, metal contents and biomass. These roles were closely related to La(III) dose, acid rain strength and treatment mode (independent exposure or simultaneous exposure). A low concentration of La(III) (20 mg L(-1)) could alleviate the adverse effects on the roots caused by acid rain, and the combined exposures at higher concentrations of La(III) and acid rain had synergic effects on the roots. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. A Genetic Screen Reveals that Synthesis of 1,4-Dihydroxy-2-Naphthoate (DHNA), but Not Full-Length Menaquinone, Is Required for Listeria monocytogenes Cytosolic Survival.

    Science.gov (United States)

    Chen, Grischa Y; McDougal, Courtney E; D'Antonio, Marc A; Portman, Jonathan L; Sauer, John-Demian

    2017-03-21

    Through unknown mechanisms, the host cytosol restricts bacterial colonization; therefore, only professional cytosolic pathogens are adapted to colonize this host environment. Listeria monocytogenes is a Gram-positive intracellular pathogen that is highly adapted to colonize the cytosol of both phagocytic and nonphagocytic cells. To identify L. monocytogenes determinants of cytosolic survival, we designed and executed a novel screen to isolate L. monocytogenes mutants with cytosolic survival defects. Multiple mutants identified in the screen were defective for synthesis of menaquinone (MK), an essential molecule in the electron transport chain. Analysis of an extensive set of MK biosynthesis and respiratory chain mutants revealed that cellular respiration was not required for cytosolic survival of L. monocytogenes but that, instead, synthesis of 1,4-dihydroxy-2-naphthoate (DHNA), an MK biosynthesis intermediate, was essential. Recent discoveries showed that modulation of the central metabolism of both host and pathogen can influence the outcome of host-pathogen interactions. Our results identify a potentially novel function of the MK biosynthetic intermediate DHNA and specifically highlight how L. monocytogenes metabolic adaptations promote cytosolic survival and evasion of host immunity. IMPORTANCE Cytosolic bacterial pathogens, such as Listeria monocytogenes and Francisella tularensis , are exquisitely evolved to colonize the host cytosol in a variety of cell types. Establishing an intracellular niche shields these pathogens from effectors of humoral immunity, grants access to host nutrients, and is essential for pathogenesis. Through yet-to-be-defined mechanisms, the host cytosol restricts replication of non-cytosol-adapted bacteria, likely through a combination of cell autonomous defenses (CADs) and nutritional immunity. Utilizing a novel genetic screen, we identified determinants of L. monocytogenes cytosolic survival and virulence and identified a role

  6. Experiments on thermal conductivity in buffer materials for geologic repository

    International Nuclear Information System (INIS)

    Kanno, T.; Yano, T.; Wakamatsu, H.; Matsushima, E.

    1989-01-01

    Engineered barriers for geologic disposal for HLW are planned to consist of canister, overpack and buffer elements. One of important physical characteristics of buffer materials is determining temperature profiles within the near field in a repository. Buffer materials require high thermal conductivity to disperse radiogenic heat away to the host rock. As the buffer materials, compacted blocks of the mixture of sodium bentonite and sand have been the most promising candidate in some countries, e.g. Sweden, Switzerland and Japan. The authors have been carrying out a series of thermal dispersion experiments to evaluate thermal conductivity of bentonite/quartz sand blocks. In this study, the following two factors considered to affect thermal properties of the near field were examined: effective thermal conductivities of buffer materials, and heat transfer characteristics of the gap between overpack and buffer materials

  7. Moisture buffering capacity of highly absorbing materials

    Energy Technology Data Exchange (ETDEWEB)

    Cerolini, S.; D' Orazio, M.; Stazi, A. [Department of Architecture, Construction and Structures (DACS), Faculty of Engineering, Polytechnic University of Marche, Via Brecce Bianche, 60100 Ancona (Italy); Di Perna, C. [Department of Energetics, Faculty of Engineering, Polytechnic University of Marche, Via Brecce Bianche, 60100 Ancona (Italy)

    2009-02-15

    This research investigates the possibility to use highly absorbing materials to dampen indoor RH% variations. The practical MBV of sodium polyacrylate, cellulose-based material, perlite and gypsum is evaluated for a daily cyclic exposure that alternates high (75%) and low (33%) RH% levels for 8 h and 16 h, respectively. The adjustment velocity to RH% variations and the presence of hysteretic phenomena are also presented. The cellulose-based material proves to be the most suitable for moisture buffering applications. Starting from this material's properties, the effect of thickness, vapour resistance factor ({mu}) and mass surface exchange coefficient (Z{sub v}) on sorption capacity is evaluated by the use of a numerical model. (author)

  8. A high capacity FASTBUS multiple event buffer

    International Nuclear Information System (INIS)

    Appel, J.A.; Farr, W.D.; Kaplan, D.M.; Levit, L.B.; Napier, T.M.

    1985-01-01

    We have developed a front-end data acquisition and event buffering memory. This single-width FASTBUS module has a capacity of 256K X 32 bits plus parity. The module is dual ported, and its front panel ECLport accepts data at up to 20 MB/sec. It may also be written to and read from as a standard FASTBUS Slave. The module records events as variable length records. Each record is accepted or rejected via front panel control signal. Circuitry to automate FASTBUS record readout and record skip is provided. In its ''linear'' mode, the module may be used as a single pass list. Alternatively, in the ''circular'' mode, the module's internal read pointer can follow its write pointer continuously around the memory. Circular mode is well suited to handling of a continuous data stream. Modules may be linked for larger memory capacity

  9. Buffer mass test - Heater design and operation

    International Nuclear Information System (INIS)

    Nilsson, J.; Ramqvist, G.; Pusch, R.

    1984-06-01

    The nuclear waste is assumed to be contained in cylindrical metal canisters which will be inserted in deposition holes. Heat is generated as a result of the continuing decay of the radioactive waste and in the Buffer Mass Test (BMT) the heat flux expected from such canisters was simulated by the use of six electric heaters. the heaters were constructed partly of aluminium and partly of stainless steel. They are 1520 mm in length and 380 mm in diameter, and give a maximum power output of 3000 W. The heater power can be monitored by panel meters coupled to a computer-based data acquisition system. Both the heater and the control system were manufactured with a high degree of redundancy in case of component failure. This report describes the design, construction, testing, installation and necessary tools for heater installation and dismantling operation. (author)

  10. A study on manufacturing and construction method of buffer

    International Nuclear Information System (INIS)

    Chijimatsu, Masakazu; Sugita, Yutaka; Amemiya, Kiyoshi

    1999-09-01

    As an engineered barrier system in the geological disposal of high-level waste, multibarrier system is considered. Multibarrier system consists of the vitrified waste, the overpack and the buffer. Bentonite is one of the potential material as the buffer because of its low water permeability, self-sealing properties, radionuclides adsorption and retardation properties, thermal conductivity, chemical buffering properties, overpack supporting properties, stress buffering properties, etc. In order to evaluate the functions of buffer, a lot of experiments has been conducted. The evaluations of these functions are based on the assumption that the buffer is emplaced or constructed in the disposal tunnel (or disposal pit) properly. Therefore, it is necessary to study on the manufacturing / construction method of buffer. As the manufacturing / construction technology of the buffer, the block installation method and in-situ compaction method, etc, are being investigated. The block installation method is to emplace the buffer blocks manufactured in advance at the ground facility, and construction processes of the block installation method at the underground will be simplified compared with the in-situ compaction method. On the other hand, the in-situ compaction method is to introduce the buffer material with specified water content into the disposal tunnel and to make the buffer with high density at the site using a compaction machine. In regard to the in-situ compaction method, it is necessary to investigate the optimum finished thickness of one layer because it is impossible to construct the buffer at one time. This report describes the results of compaction property test and the summary of the past investigation results in connection with the manufacturing / construction method. Then this report shows the construction method that will be feasible in the actual disposal site. (J.P.N.)

  11. Moisture Buffer Effect and its Impact on Indoor Environment

    DEFF Research Database (Denmark)

    Zhang, Mingjie; Qin, Menghao; Chen, Zhi

    2017-01-01

    The moisture buffer effect of building materials may have great influence on indoor hygrothermal environment. In order to characterize the moisture buffering ability of materials, the basic concept of moisture buffer value (MBV) is adopted. Firstly, a theoretical correction factor is introduced...... in this paper. The moisture uptake/release by hygroscopic materials can be calculated with the factor and the basic MBV. Furthermore, the validation of the correction factor is carried out. The impact of moisture buffering on indoor environment is assessed by using numerical simulations. The results show...

  12. Effects of node buffer and capacity on network traffic

    International Nuclear Information System (INIS)

    Ling Xiang; Ding Jian-Xun; Hu Mao-Bin

    2012-01-01

    In this paper, we study the optimization of network traffic by considering the effects of node buffer ability and capacity. Two node buffer settings are considered. The node capacity is considered to be proportional to its buffer ability. The node effects on network traffic systems are studied with the shortest path protocol and an extension of the optimal routing [Phys. Rev. E 74 046106 (2006)]. In the diagrams of flux—density relationships, it is shown that a nodes buffer ability and capacity have profound effects on the network traffic

  13. Back contact buffer layer for thin-film solar cells

    Science.gov (United States)

    Compaan, Alvin D.; Plotnikov, Victor V.

    2014-09-09

    A photovoltaic cell structure is disclosed that includes a buffer/passivation layer at a CdTe/Back contact interface. The buffer/passivation layer is formed from the same material that forms the n-type semiconductor active layer. In one embodiment, the buffer layer and the n-type semiconductor active layer are formed from cadmium sulfide (CdS). A method of forming a photovoltaic cell includes the step of forming the semiconductor active layers and the buffer/passivation layer within the same deposition chamber and using the same material source.

  14. Solubilization of proteins: the importance of lysis buffer choice.

    Science.gov (United States)

    Peach, Mandy; Marsh, Noelle; Miskiewicz, Ewa I; MacPhee, Daniel J

    2015-01-01

    The efficient extraction of proteins of interest from cells and tissues is not always straightforward. Here we demonstrate the differences in extraction of the focal adhesion protein Kindlin-2 from choriocarcinoma cells using NP-40 and RIPA lysis buffer. Furthermore, we demonstrate the use of a more denaturing urea/thiourea lysis buffer for solubilization, by comparing its effectiveness for solubilization of small heat-shock proteins from smooth muscle with the often utilized RIPA lysis buffer. Overall, the results demonstrate the importance of establishing the optimal lysis buffer for specific protein solubilization within the experimental workflow.

  15. How Do Stream Buffers Reduce the Offsite Impact of Pollution?

    OpenAIRE

    Easton, Zachary M.

    2012-01-01

    Summarizes the importance of buffers to reduce pollution and specifically examines where they can be used, how they work, their limitations, required maintenance, performance levels, and expected cost.

  16. Effect of Buffer Bow Structure in Ship-Ship Collision

    DEFF Research Database (Denmark)

    Yamada, Yasuhira; Endo, Hisayoshi; Pedersen, Preben Terndrup

    2008-01-01

    tankers, the introduction of buffer bulbous bows has been proposed. Relatively soft buffer bows absorb part of the kinetic energy of the striking ship before penetrating the inner hull of the struck vessel. The purpose of the present paper is to verify the effectiveness of a prototype buffer bulbous bow......) and the forward velocity of the struck ship on the collapse mode of the bow of the striking vessel are investigated. Collapse modes, contact forces and energy absorption capabilities of the buffer bows are compared with those of conventional bows....

  17. Role of cytosolic NADP+-dependent isocitrate dehydrogenase in ischemia-reperfusion injury in mouse kidney.

    Science.gov (United States)

    Kim, Jinu; Kim, Ki Young; Jang, Hee-Seong; Yoshida, Takumi; Tsuchiya, Ken; Nitta, Kosaku; Park, Jeen-Woo; Bonventre, Joseph V; Park, Kwon Moo

    2009-03-01

    Cytosolic NADP+-dependent isocitrate dehydrogenase (IDPc) synthesizes reduced NADP (NADPH), which is an essential cofactor for the generation of reduced glutathione (GSH), the most abundant and important antioxidant in mammalian cells. We investigated the role of IDPc in kidney ischemia-reperfusion (I/R) in mice. The activity and expression of IDPc were highest in the cortex, modest in the outer medulla, and lowest in the inner medulla. NADPH levels were greatest in the cortex. IDPc expression in the S1 and S2 segments of proximal tubules was higher than in the S3 segment, which is much more susceptible to I/R. IDPc protein was also highly expressed in the mitochondrion-rich intercalated cells of the collecting duct. IDPc activity was 10- to 30-fold higher than the activity of glucose-6-phosphate dehydrogenase, another producer of cytosolic NADPH, in various kidney regions. This study identifies that IDPc may be the primary source of NADPH in the kidney. I/R significantly reduced IDPc expression and activity and NADPH production and increased the ratio of oxidized glutathione to total glutathione [GSSG/(GSH+GSSG)], resulting in kidney dysfunction, tubular cell damage, and lipid peroxidation. In LLC-PK(1) cells, upregulation of IDPc by IDPc gene transfer protected the cells against hydrogen peroxide, enhancing NADPH production, inhibiting the increase of GSSG/(GSH+GSSG), and reducing lipid peroxidation. IDPc downregulation by small interference RNA treatment presented results contrasting with the upregulation. In conclusion, these results demonstrate that IDPc is expressed differentially along tubules in patterns that may contribute to differences in susceptibility to injury, is a major enzyme in cytosolic NADPH generation in kidney, and is downregulated with I/R.

  18. A cytosolic juxtamembrane interface modulates plexin A3 oligomerization and signal transduction.

    Directory of Open Access Journals (Sweden)

    Rachael Barton

    Full Text Available Plexins (plxns are transmembrane (TM receptors involved in the guidance of vascular, lymphatic vessel, and neuron growth as well as cancer metastasis. Plxn signaling results in cytosolic GTPase-activating protein activity, and previous research implicates dimerization as important for activation of plxn signaling. Purified, soluble plxn extracellular and cytosolic domains exhibit only weak homomeric interactions, suggesting a role for the plxn TM and juxtamembrane regions in homooligomerization. In this study, we consider a heptad repeat in the Danio rerio PlxnA3 cytosolic juxtamembrane domain (JM for its ability to influence PlxnA3 homooligomerization in TM-domain containing constructs. Site-directed mutagenesis in conjunction with the AraTM assay and bioluminescent energy transfer (BRET² suggest an interface involving a JM heptad repeat, in particular residue M1281, regulates PlxnA3 homomeric interactions when examined in constructs containing an ectodomain, TM and JM domain. In the presence of a neuropilin-2a co-receptor and semaphorin 3F ligand, disruption to PlxnA3 homodimerization caused by an M1281F mutation is eliminated, suggesting destabilization of the PlxnA3 homodimer in the JM is not sufficient to disrupt co-receptor complex formation. In contrast, enhanced homodimerization of PlxnA3 caused by mutation M1281L remains even in the presence of ligand semaphorin 3F and co-receptor neuropilin-2a. Consistent with this pattern of PlxnA3 dimerization in the presence of ligand and co-receptor, destabilizing mutations to PlxnA3 homodimerization (M1281F are able to rescue motor patterning defects in sidetracked zebrafish embryos, whereas mutations that enhance PlxnA3 homodimerization (M1281L are not. Collectively, our results indicate the JM heptad repeat, in particular residue M1281, forms a switchable interface that modulates both PlxnA3 homomeric interactions and signal transduction.

  19. The role of a cytosolic superoxide dismutase in barley-pathogen interactions

    KAUST Repository

    Lightfoot, Damien

    2016-03-19

    Reactive oxygen species (ROS), including superoxide (O2-HO2) and hydrogen peroxide (H2O2), are differentially produced during resistance responses to biotrophic pathogens and during susceptible responses to necrotrophic and hemi-biotrophic pathogens. Superoxide dismutase (SOD) is responsible for the catalysis of the dismutation of O2-HO2 to H2O2, regulating the redox status of plant cells. Increased SOD activity has been correlated previously with resistance in barley to the hemi-biotrophic pathogen Pyrenophora teres f. teres (Ptt, the causal agent of the net form of net blotch disease), but the role of individual isoforms of SOD has not been studied. A cytosolic CuZnSOD, HvCSD1, was isolated from barley and characterized as being expressed in tissue from different developmental stages. HvCSD1 was up-regulated during the interaction with Ptt and to a greater extent during the resistance response. Net blotch disease symptoms and fungal growth were not as pronounced in transgenic HvCSD1 knockdown lines in a susceptible background (cv. Golden Promise), when compared with wild-type plants, suggesting that cytosolic O2-HO2 contributes to the signalling required to induce a defence response to Ptt. There was no effect of HvCSD1 knockdown on infection by the hemi-biotrophic rice blast pathogen Magnaporthe oryzae or the biotrophic powdery mildew pathogen Blumeria graminis f. sp. hordei, but HvCSD1 also played a role in the regulation of lesion development by methyl viologen. Together, these results suggest that HvCSD1 could be important in the maintenance of the cytosolic redox status and in the differential regulation of responses to pathogens with different lifestyles.

  20. Fatty acid translocase promoted hepatitis B virus replication by upregulating the levels of hepatic cytosolic calcium.

    Science.gov (United States)

    Huang, Jian; Zhao, Lei; Yang, Ping; Chen, Zhen; Ruan, Xiong Z; Huang, Ailong; Tang, Ni; Chen, Yaxi

    2017-09-15

    Hepatitis B virus (HBV) is designated a "metabolovirus" due to the intimate connection between the virus and host metabolism. The nutrition state of the host plays a relevant role in the severity of HBV infection. Metabolic syndrome (MS) is prone to increasing HBV DNA loads and accelerating the progression of liver disease in patients with chronic hepatitis B (CHB). Cluster of differentiation 36 (CD36), also named fatty acid translocase, is known to facilitate long-chain fatty acid uptake and contribute to the development of MS. We recently found that CD36 overexpression enhanced HBV replication. In this study, we further explored the mechanism by which CD36 overexpression promotes HBV replication. Our data showed that CD36 overexpression increased HBV replication, and CD36 knockdown inhibited HBV replication. RNA sequencing found some of the differentially expressed genes were involved in calcium ion homeostasis. CD36 overexpression elevated the cytosolic calcium level, and CD36 knockdown decreased the cytosolic calcium level. Calcium chelator BAPTA-AM could override the HBV replication increased by CD36 overexpression, and the calcium activator thapsigargin could improve the HBV replication reduced by CD36 knockdown. We further found that CD36 overexpression activated Src kinase, which plays an important role in the regulation of the store-operated Ca 2+ channel. An inhibitor of Src kinase (SU6656) significantly reduced the CD36-induced HBV replication. We identified a novel link between CD36 and HBV replication, which is associated with cytosolic calcium and the Src kinase pathway. CD36 may represent a potential therapeutic target for the treatment of CHB patients with MS. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Cytosolic nucleotides block and regulate the Arabidopsis vacuolar anion channel AtALMT9.

    Science.gov (United States)

    Zhang, Jingbo; Martinoia, Enrico; De Angeli, Alexis

    2014-09-12

    The aluminum-activated malate transporters (ALMTs) form a membrane protein family exhibiting different physiological roles in plants, varying from conferring tolerance to environmental Al(3+) to the regulation of stomatal movement. The regulation of the anion channels of the ALMT family is largely unknown. Identifying intracellular modulators of the activity of anion channels is fundamental to understanding their physiological functions. In this study we investigated the role of cytosolic nucleotides in regulating the activity of the vacuolar anion channel AtALMT9. We found that cytosolic nucleotides modulate the transport activity of AtALMT9. This modulation was based on a direct block of the pore of the channel at negative membrane potentials (open channel block) by the nucleotide and not by a phosphorylation mechanism. The block by nucleotides of AtALMT9-mediated currents was voltage dependent. The blocking efficiency of intracellular nucleotides increased with the number of phosphate groups and ATP was the most effective cellular blocker. Interestingly, the ATP block induced a marked modification of the current-voltage characteristic of AtALMT9. In addition, increased concentrations of vacuolar anions were able to shift the ATP block threshold to a more negative membrane potential. The block of AtALMT9-mediated anion currents by ATP at negative membrane potentials acts as a gate of the channel and vacuolar anion tune this gating mechanism. Our results suggest that anion transport across the vacuolar membrane in plant cells is controlled by cytosolic nucleotides and the energetic status of the cell. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Cytosolic Nucleotides Block and Regulate the Arabidopsis Vacuolar Anion Channel AtALMT9*

    Science.gov (United States)

    Zhang, Jingbo; Martinoia, Enrico; De Angeli, Alexis

    2014-01-01

    The aluminum-activated malate transporters (ALMTs) form a membrane protein family exhibiting different physiological roles in plants, varying from conferring tolerance to environmental Al3+ to the regulation of stomatal movement. The regulation of the anion channels of the ALMT family is largely unknown. Identifying intracellular modulators of the activity of anion channels is fundamental to understanding their physiological functions. In this study we investigated the role of cytosolic nucleotides in regulating the activity of the vacuolar anion channel AtALMT9. We found that cytosolic nucleotides modulate the transport activity of AtALMT9. This modulation was based on a direct block of the pore of the channel at negative membrane potentials (open channel block) by the nucleotide and not by a phosphorylation mechanism. The block by nucleotides of AtALMT9-mediated currents was voltage dependent. The blocking efficiency of intracellular nucleotides increased with the number of phosphate groups and ATP was the most effective cellular blocker. Interestingly, the ATP block induced a marked modification of the current-voltage characteristic of AtALMT9. In addition, increased concentrations of vacuolar anions were able to shift the ATP block threshold to a more negative membrane potential. The block of AtALMT9-mediated anion currents by ATP at negative membrane potentials acts as a gate of the channel and vacuolar anion tune this gating mechanism. Our results suggest that anion transport across the vacuolar membrane in plant cells is controlled by cytosolic nucleotides and the energetic status of the cell. PMID:25028514

  3. From Proteomics to Structural Studies of Cytosolic/Mitochondrial-Type Thioredoxin Systems in Barley Seeds

    DEFF Research Database (Denmark)

    Shahpiri, Azar; Svensson, Birte; Finnie, Christine

    2009-01-01

    Thioredoxins (Trx) are ubiquitous proteins that participate in thiol disulfide reactions via two active site cysteine residues, allowing Trx to reduce disulfide bonds in target proteins. Recent progress in proteome analysis has resulted in identification of a wide range of potential target proteins...... for Trx, indicating that Trx plays a key role in several aspects of cell metabolism. In contrast to other organisms, plants contain multiple forms of Trx that are classified based on their primary structures and sub-cellular localization. The reduction of cytosolic and mitochondrial types of Trx...

  4. Purification and characterization of a novel cytosolic NADP(H)-dependent retinol oxidoreductase from rabbit liver.

    Science.gov (United States)

    Huang, D Y; Ichikawa, Y

    1997-03-07

    Rabbit liver cytosol exhibits very high retinol dehydrogenase activity. At least two retinol dehydrogenases were demonstrated to exist in rabbit liver cytosol, and the major one, a cytosolic NADP(H)-dependent retinol dehydrogenase (systematic name: retinol oxidoreductase) was purified about 1795-fold to electrophoretic and column chromatographic homogeneity by a procedure involving column chromatography on AF-Red Toyopearl twice and then hydroxyapatite. Its molecular mass was estimated to be 34 kDa by SDS-PAGE, and 144 kDa by HPLC gel filtration, suggesting that it is a homo-tetramer. The enzyme uses free retinol and retinal, and their complexes with CRBP as substrates in vitro. The optimum pH values for retinol oxidation of free retinol and CRBP-retinol were 8.8-9.2 and 8.0-9.0, respectively, and those for retinal reduction of free retinal and retinal-CRBP were the same, 7.0-7.6. Km for free retinol and Vmax for retinal formation were 2.8 microM and 2893 nmol/min per mg protein at 37 degrees C (pH 9.0) and the corresponding values with retinol-CRBP as a substrate were 2.5 microM and 2428 nmol/min per mg protein at 37 degrees C (pH 8.6); Km for free retinal and Vmax for retinol formation were 6.5 microM and 4108 nmol/min per mg protein, and the corresponding values with retinal-CRBP as a substrate were 5.1 microM and 3067 nmol/min per mg protein at 37 degrees C, pH 7.4. NAD(H) was not effective as a cofactor. 4-Methylpyrazole was a weak inhibitor (IC50 = 28 mM) of the enzyme, and ethanol was neither a substrate nor an inhibitor of the enzyme. This enzyme exhibits relatively broad aldehyde reductase activity and some ketone reductase activity, the activity for aromatic substitutive aldehydes being especially high and effective. Whereas, except in the case of retinol, oxidative activity toward the corresponding alcohols was not detected. This novel cytosolic enzyme may play an important role in vivo in maintaining the homeostasis of retinal, the substrate of retinoic

  5. Transmembrane proteoglycans control stretch-activated channels to set cytosolic calcium levels

    DEFF Research Database (Denmark)

    Gopal, Sandeep; Søgaard, Pernille; Multhaupt, Hinke A B

    2015-01-01

    show that syndecans regulate transient receptor potential canonical (TRPCs) channels to control cytosolic calcium equilibria and consequent cell behavior. In fibroblasts, ligand interactions with heparan sulfate of syndecan-4 recruit cytoplasmic protein kinase C to target serine714 of TRPC7...... with subsequent control of the cytoskeleton and the myofibroblast phenotype. In epidermal keratinocytes a syndecan-TRPC4 complex controls adhesion, adherens junction composition, and early differentiation in vivo and in vitro. In Caenorhabditis elegans, the TRPC orthologues TRP-1 and -2 genetically complement...

  6. Temperature buffer test. Installation of buffer, heaters and instruments in the deposition hole

    Energy Technology Data Exchange (ETDEWEB)

    Johannesson, Lars-Erik; Sanden, Torbjoern; Aakesson, Mattias [Clay Technology AB, Lund (Sweden); Barcena, Ignacio; Garcia-Sineriz, Jose Luis [Aitemin, Madrid (Spain)

    2010-12-15

    During 2003 the Temperature Buffer Test was installed in Aespoe Hard Rock Laboratory. Temperature, water pressure, relative humidity, total pressure and displacements etc. are measured in numerous points in the test. Most of the cables from the transducers are led in the deposition hole through slots in the rock surface of the deposition hole in watertight tubes to the data collection system in a container placed in the tunnel close to the deposition hole. This report describes the work with the installations of the buffer, heaters, and instruments and yields a description of the final location of all instruments. The report also contains a description of the materials that were installed and the densities yielded after placement.

  7. Temperature buffer test. Installation of buffer, heaters and instruments in the deposition hole

    International Nuclear Information System (INIS)

    Johannesson, Lars-Erik; Sanden, Torbjoern; Aakesson, Mattias; Barcena, Ignacio; Garcia-Sineriz, Jose Luis

    2010-12-01

    During 2003 the Temperature Buffer Test was installed in Aespoe Hard Rock Laboratory. Temperature, water pressure, relative humidity, total pressure and displacements etc. are measured in numerous points in the test. Most of the cables from the transducers are led in the deposition hole through slots in the rock surface of the deposition hole in watertight tubes to the data collection system in a container placed in the tunnel close to the deposition hole. This report describes the work with the installations of the buffer, heaters, and instruments and yields a description of the final location of all instruments. The report also contains a description of the materials that were installed and the densities yielded after placement

  8. The buffer/container experiment: results, synthesis, issues

    International Nuclear Information System (INIS)

    Graham, J.; Chandler, N.A.; Dixon, D.A.; Roach, P.J.; To, T.; Wan, A.W.L.

    1997-12-01

    A large in-ground experiment has examined how heat affects the performance of the dense sand bentonite 'buffer' that has been proposed for use in the Canadian Nuclear Fuel Waste Management Program. The experiment was performed by Atomic Energy of Canada Limited at its Underground Research Laboratory, Lac du Bonnet, Manitoba between 1991 and 1994. The experiment placed a full-size heater representing a container of nuclear fuel waste in a 1.24-m diameter borehole filled with buffer below the floor of a room excavated at 240-m depth in granitic rock of the Canadian Shield. The buffer and surrounding rock were extensively instrumented for temperatures, total pressures, water pressures, suctions, and rock displacements. Power was provided to the heater for almost 900 days. The experiment showed that good rock conditions can be pre-selected, a borehole can be drilled, and buffer can be placed at controlled densities and water contents. The instrumentation generally worked well, and an extensive data base was successfully organized. Drying was observed in buffer close to the heater. This caused some desiccation cracking. However the cracks only extended approximately one third of the distance to the buffer-rock interface and did not form an advective pathway. Following sampling at the time of decommissioning, cracked samples of buffer were transported to the laboratory and given access to water. The hydraulic conductivities and swelling pressures of these resaturated samples were very similar to those of uncracked buffer. A good balance was achieved between the mass of water flowing into the experiment from the surrounding rock and the increased mass of water in the buffer. A good understanding was developed of the relationships between suctions, water contents, and total pressures in buffer near the buffer-rock interface. Comparisons between measurements and predictions of measured parameters show that a good understanding has been developed of the processes operating

  9. The buffer/container experiment: results, synthesis, issues

    Energy Technology Data Exchange (ETDEWEB)

    Graham, J. [Univ. of Manitoba, Dept. of Civil Engineering, Winnipeg, MB (Canada); Chandler, N.A.; Dixon, D.A.; Roach, P.J.; To, T.; Wan, A.W.L

    1997-12-01

    A large in-ground experiment has examined how heat affects the performance of the dense sand bentonite 'buffer' that has been proposed for use in the Canadian Nuclear Fuel Waste Management Program. The experiment was performed by Atomic Energy of Canada Limited at its Underground Research Laboratory, Lac du Bonnet, Manitoba between 1991 and 1994. The experiment placed a full-size heater representing a container of nuclear fuel waste in a 1.24-m diameter borehole filled with buffer below the floor of a room excavated at 240-m depth in granitic rock of the Canadian Shield. The buffer and surrounding rock were extensively instrumented for temperatures, total pressures, water pressures, suctions, and rock displacements. Power was provided to the heater for almost 900 days. The experiment showed that good rock conditions can be pre-selected, a borehole can be drilled, and buffer can be placed at controlled densities and water contents. The instrumentation generally worked well, and an extensive data base was successfully organized. Drying was observed in buffer close to the heater. This caused some desiccation cracking. However the cracks only extended approximately one third of the distance to the buffer-rock interface and did not form an advective pathway. Following sampling at the time of decommissioning, cracked samples of buffer were transported to the laboratory and given access to water. The hydraulic conductivities and swelling pressures of these resaturated samples were very similar to those of uncracked buffer. A good balance was achieved between the mass of water flowing into the experiment from the surrounding rock and the increased mass of water in the buffer. A good understanding was developed of the relationships between suctions, water contents, and total pressures in buffer near the buffer-rock interface. Comparisons between measurements and predictions of measured parameters show that a good understanding has been developed of the processes

  10. Cytosolic monoterpene biosynthesis is supported by plastid-generated geranyl diphosphate substrate in transgenic tomato fruits.

    Science.gov (United States)

    Gutensohn, Michael; Orlova, Irina; Nguyen, Thuong T H; Davidovich-Rikanati, Rachel; Ferruzzi, Mario G; Sitrit, Yaron; Lewinsohn, Efraim; Pichersky, Eran; Dudareva, Natalia

    2013-08-01

    Geranyl diphosphate (GPP), the precursor of most monoterpenes, is synthesized in plastids from dimethylallyl diphosphate and isopentenyl diphosphate by GPP synthases (GPPSs). In heterodimeric GPPSs, a non-catalytic small subunit (GPPS-SSU) interacts with a catalytic large subunit, such as geranylgeranyl diphosphate synthase, and determines its product specificity. Here, snapdragon (Antirrhinum majus) GPPS-SSU was over-expressed in tomato fruits under the control of the fruit ripening-specific polygalacturonase promoter to divert the metabolic flux from carotenoid formation towards GPP and monoterpene biosynthesis. Transgenic tomato fruits produced monoterpenes, including geraniol, geranial, neral, citronellol and citronellal, while exhibiting reduced carotenoid content. Co-expression of the Ocimum basilicum geraniol synthase (GES) gene with snapdragon GPPS-SSU led to a more than threefold increase in monoterpene formation in tomato fruits relative to the parental GES line, indicating that the produced GPP can be used by plastidic monoterpene synthases. Co-expression of snapdragon GPPS-SSU with the O. basilicum α-zingiberene synthase (ZIS) gene encoding a cytosolic terpene synthase that has been shown to possess both sesqui- and monoterpene synthase activities resulted in increased levels of ZIS-derived monoterpene products compared to fruits expressing ZIS alone. These results suggest that re-direction of the metabolic flux towards GPP in plastids also increases the cytosolic pool of GPP available for monoterpene synthesis in this compartment via GPP export from plastids. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

  11. Endosomolytic Nano-Polyplex Platform Technology for Cytosolic Peptide Delivery To Inhibit Pathological Vasoconstriction.

    Science.gov (United States)

    Evans, Brian C; Hocking, Kyle M; Kilchrist, Kameron V; Wise, Eric S; Brophy, Colleen M; Duvall, Craig L

    2015-06-23

    A platform technology has been developed and tested for delivery of intracellular-acting peptides through electrostatically complexed nanoparticles, or nano-polyplexes, formulated from an anionic endosomolytic polymer and cationic therapeutic peptides. This delivery platform has been initially tested and optimized for delivery of two unique vasoactive peptides, a phosphomimetic of heat shock protein 20 and an inhibitor of MAPKAP kinase II, to prevent pathological vasoconstriction (i.e., vasospasm) in human vascular tissue. These peptides inhibit vasoconstriction and promote vasorelaxation by modulating actin dynamics in vascular smooth muscle cells. Formulating these peptides into nano-polyplexes significantly enhances peptide uptake and retention, facilitates cytosolic delivery through a pH-dependent endosomal escape mechanism, and enhances peptide bioactivity in vitro as measured by inhibition of F-actin stress fiber formation. In comparison to treatment with the free peptides, which were endowed with cell-penetrating sequences, the nano-polyplexes significantly increased vasorelaxation, inhibited vasoconstriction, and decreased F-actin formation in the human saphenous vein ex vivo. These results suggest that these formulations have significant potential for treatment of conditions such as cerebral vasospasm following subarachnoid hemorrhage. Furthermore, because many therapeutic peptides include cationic cell-penetrating segments, this simple and modular platform technology may have broad applicability as a cost-effective approach for enhancing the efficacy of cytosolically active peptides.

  12. A Plant Phytosulfokine Peptide Initiates Auxin-Dependent Immunity through Cytosolic Ca2+ Signaling in Tomato.

    Science.gov (United States)

    Zhang, Huan; Hu, Zhangjian; Lei, Cui; Zheng, Chenfei; Wang, Jiao; Shao, Shujun; Li, Xin; Xia, Xiaojian; Cai, Xinzhong; Zhou, Jie; Zhou, Yanhong; Yu, Jingquan; Foyer, Christine H; Shi, Kai

    2018-03-01

    Phytosulfokine (PSK) is a disulfated pentapeptide that is an important signaling molecule. Although it has recently been implicated in plant defenses to pathogen infection, the mechanisms involved remain poorly understood. Using surface plasmon resonance and gene silencing approaches, we showed that the tomato ( Solanum lycopersicum ) PSK receptor PSKR1, rather than PSKR2, functioned as the major PSK receptor in immune responses. Silencing of PSK signaling genes rendered tomato more susceptible to infection by the economically important necrotrophic pathogen Botrytis cinerea Analysis of tomato mutants defective in either defense hormone biosynthesis or signaling demonstrated that PSK-induced immunity required auxin biosynthesis and associated defense pathways. Here, using aequorin-expressing tomato plants, we provide evidence that PSK perception by tomato PSKR1 elevated cytosolic [Ca 2+ ], leading to auxin-dependent immune responses via enhanced binding activity between calmodulins and the auxin biosynthetic YUCs. Thus, our data demonstrate that PSK acts as a damage-associated molecular pattern and is perceived mainly by PSKR1, which increases cytosolic [Ca 2+ ] and activates auxin-mediated pathways that enhance immunity of tomato plants to B. cinerea . © 2018 American Society of Plant Biologists. All rights reserved.

  13. Nitrate Activation of Cytosolic Protein Kinases Diverts Photosynthetic Carbon from Sucrose to Amino Acid Biosynthesis

    Science.gov (United States)

    Champigny, Marie-Louise; Foyer, Christine

    1992-01-01

    The regulation of carbon partitioning between carbohydrates (principally sucrose) and amino acids has been only poorly characterized in higher plants. The hypothesis that the pathway of sucrose and amino acid biosynthesis compete for carbon skeletons and energy is widely accepted. In this review, we suggest a mechanism involving the regulation of cytosolic protein kinases whereby the flow of carbon is regulated at the level of partitioning between the pathways of carbohydrate and nitrogen metabolism via the covalent modulation of component enzymes. The addition of nitrate to wheat seedlings (Triticum aestivum) grown in the absence of exogenous nitrogen has a dramatic, if transient, impact on sucrose formation and on the activities of sucrose phosphate synthase (which is inactivated) and phosphoenolpyruvate carboxylase (which is activated). The activities of these two enzymes are modulated by protein phosphorylation in response to the addition of nitrate, but they respond in an inverse fashion. Sucrose phosphate synthase in inactivated and phosphoenolpyruvate carboxylase is activated. Nitrate functions as a signal metabolite activating the cytosolic protein kinase, thereby modulating the activities of at least two of the key enzymes in assimilate partitioning and redirecting the flow of carbon away from sucrose biosynthesis toward amino acid synthesis. PMID:16653003

  14. Aspartic cathepsin D degrades the cytosolic cysteine cathepsin inhibitor stefin B in the cells.

    Science.gov (United States)

    Železnik, Tajana Zajc; Kadin, Andrey; Turk, Vito; Dolenc, Iztok

    2015-09-18

    Stefin B is the major general cytosolic protein inhibitor of cysteine cathepsins. Its main function is to protect the organism against the activity of endogenous potentially hazardous proteases accidentally released from lysosomes. In this study, we investigated the possible effect of endosomal/lysosomal aspartic cathepsins D and E on stefin B after membrane permeabilization. Loss of membrane integrity of lysosomes and endosomes was induced by a lysosomotropic agent L-Leucyl-L-leucine methyl ester (Leu-Leu-OMe). The rat thyroid cell line FRTL-5 was selected as a model cell line owing to its high levels of proteases, including cathepsin D and E. Permeabilization of acid vesicles from FRTL-5 cells induced degradation of stefin B. The process was inhibited by pepstatin A, a potent inhibitor of aspartic proteases. However, degradation of stefin B was prevented by siRNA-mediated silencing of cathepsin D expression. In contrast, cathepsin E silencing had no effect on stefin B degradation. These results showed that cathepsin D and not cathepsin E degrades stefin B. It can be concluded that the presence of cathepsin D in the cytosol affects the inhibitory potency of stefin B, thus preventing the regulation of cysteine cathepsin activities in various biological processes. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Upregulation of cytosolic NADP+-dependent isocitrate dehydrogenase by hyperglycemia protects renal cells against oxidative stress.

    Science.gov (United States)

    Lee, Soh-Hyun; Ha, Sun-Ok; Koh, Ho-Jin; Kim, KilSoo; Jeon, Seon-Min; Choi, Myung-Sook; Kwon, Oh-Shin; Huh, Tae-Lin

    2010-02-28

    Hyperglycemia-induced oxidative stress is widely recognized as a key mediator in the pathogenesis of diabetic nephropathy, a complication of diabetes. We found that both expression and enzymatic activity of cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) were upregulated in the renal cortexes of diabetic rats and mice. Similarly, IDPc was induced in murine renal proximal tubular OK cells by high hyperglycemia, while it was abrogated by co-treatment with the antioxidant N-Acetyl-Cysteine (NAC). In OK cells, increased expression of IDPc by stable transfection prevented hyperglycemia-mediated reactive oxygen species (ROS) production, subsequent cellular oxidative stress and extracellular matrix accumulation, whereas these processes were all stimulated by decreased IDPc expression. In addition, production of NADPH and GSH in the cytosol was positively correlated with the expression level of IDPc in OK cells. These results together indicate that upregulation of IDPc in response to hyperglycemia might play an essential role in preventing the progression of diabetic nephropathy, which is accompanied by ROS-induced cellular damage and fibrosis, by providing NADPH, the reducing equivalent needed for recycling reduced glutathione and low molecular weight antioxidant thiol proteins.

  16. Cytosolic NADP(+)-dependent isocitrate dehydrogenase regulates cadmium-induced apoptosis.

    Science.gov (United States)

    Shin, Seoung Woo; Kil, In Sup; Park, Jeen-Woo

    2010-04-01

    Cadmium ions have a high affinity for thiol groups. Therefore, they may disturb many cellular functions. We recently reported that cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) functions as an antioxidant enzyme to supply NADPH, a major source of reducing equivalents to the cytosol. Cadmium decreased the activity of IDPc both as a purified enzyme and in cultured cells. In the present study, we demonstrate that the knockdown of IDPc expression in HEK293 cells greatly enhances apoptosis induced by cadmium. Transfection of HEK293 cells with an IDPc small interfering RNA significantly decreased the activity of IDPc and enhanced cellular susceptibility to cadmium-induced apoptosis as indicated by the morphological evidence of apoptosis, DNA fragmentation and condensation, cellular redox status, mitochondria redox status and function, and the modulation of apoptotic marker proteins. Taken together, our results suggest that suppressing the expression of IDPc enhances cadmium-induced apoptosis of HEK293 cells by increasing disruption of the cellular redox status. Copyright 2009 Elsevier Inc. All rights reserved.

  17. AIM2-Like Receptors Positively and Negatively Regulate the Interferon Response Induced by Cytosolic DNA

    Directory of Open Access Journals (Sweden)

    Yuki Nakaya

    2017-07-01

    Full Text Available Cytosolic DNAs derived from retrotransposons serve as pathogen-associated molecular patterns for pattern recognition receptors (PRRs that stimulate the induction of interferons (IFNs and other cytokines, leading to autoimmune disease. Cyclic GMP-AMP synthase is one PRR that senses retrotransposon DNA, activating type I IFN responses through the stimulator of IFN genes (STING. Absent in melanoma 2 (AIM2-like receptors (ALRs have also been implicated in these pathways. Here we show that the mouse ALR IFI205 senses cytosolic retrotransposon DNA independently of cyclic GMP-AMP production. AIM2 antagonizes IFI205-mediated IFN induction activity by sequestering it from STING. We also found that the complement of genes located in the ALR locus in C57BL/6 and AIM2 knockout mice are different and unique, which has implications for interpretation of the sensing of pathogens in different mouse strains. Our data suggest that members of the ALR family are critical to the host IFN response to endogenous DNA.

  18. Human cytosolic thymidine kinase: purification and physical characterization of the enzyme from HeLa cells

    International Nuclear Information System (INIS)

    Sherley, J.L.; Kelly, T.J.

    1988-01-01

    The mammalian cytosolic thymidine kinase is one of a number of enzymes involved in DNA replication whose activities increase dramatically during S phase of the cell cycle. As a first step in defining the mechanisms that control the S phase induction of thymidine kinase activity, the authors have purified the human enzyme from HeLa cells and raised a specific immune serum against the purified protein. The enzyme was isolated from cells arrested in S phase by treatment with methotrexate and purified to near homogeneity by ion-exchange and affinity chromatography. Stabilization of the purified enzyme was achieved by the addition of digitonin. An electrophoretic R/sub m/ of 0.2 in nondenaturing gels characterizes the purified enzyme activity as cytosolic thymidine kinase. The enzyme has a Stoke's radius of 40 A determined by gel filtration and a sedimentation coefficient of 5.5 S determined by glycerol gradient sedimentation. Based on these hydrodynamic values, a native molecular weight of 96,000 was calculated for the purified enzyme. When electrophoresed in denaturing sodium dodecyl sulfate-polyacrylamide gels under reducing conditions, the most purified enzyme fraction was found to contain one predominant polypeptide of M/sub r/ = 24,000. Several lines of evidence indicate that this polypeptide is responsible for thymidine kinase enzymatic activity

  19. Cytosolic malate dehydrogenase regulates RANKL-mediated osteoclastogenesis via AMPK/c-Fos/NFATc1 signaling

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Se Jeong [Department of Oral Microbiology and Immunology, College of Dentistry, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Gu, Dong Ryun [Department of Oral Microbiology and Immunology, College of Dentistry, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Center for Metabolic Function Regulation (CMFR), School of Medicine, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Jin, Su Hyun [Center for Metabolic Function Regulation (CMFR), School of Medicine, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Park, Keun Ha [Department of Oral Microbiology and Immunology, College of Dentistry, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Center for Metabolic Function Regulation (CMFR), School of Medicine, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Lee, Seoung Hoon, E-mail: leesh2@wku.ac.kr [Department of Oral Microbiology and Immunology, College of Dentistry, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Center for Metabolic Function Regulation (CMFR), School of Medicine, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of); Wonkwang Institute of Biomaterials and Implant, Wonkwang University, Iksan, Jeonbuk 54538 (Korea, Republic of)

    2016-06-17

    Cytosolic malate dehydrogenase (malate dehydrogenase 1, MDH1) plays pivotal roles in the malate/aspartate shuttle that might modulate metabolism between the cytosol and mitochondria. In this study, we investigated the role of MDH1 in osteoclast differentiation and formation. MDH1 expression was induced by receptor activator of nuclear factor kappa-B ligand (RANKL) treatment. Knockdown of MDH1 by infection with retrovirus containing MDH1-specific shRNA (shMDH1) reduced mature osteoclast formation and bone resorption activity. Moreover, the expression of marker genes associated with osteoclast differentiation was downregulated by shMDH1 treatment, suggesting a role of MDH1 in osteoclast differentiation. In addition, intracellular ATP production was reduced following the activation of adenosine 5′ monophosphate-activated protein kinase (AMPK), a cellular energy sensor and negative regulator of RANKL-induced osteoclast differentiation, in shMDH1-infected osteoclasts compared to control cells. In addition, the expression of c-Fos and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), a critical transcription factor of osteoclastogenesis, was decreased with MDH1 knockdown during RANKL-mediated osteoclast differentiation. These findings provide strong evidence that MDH1 plays a critical role in osteoclast differentiation and function via modulation of the intracellular energy status, which might affect AMPK activity and NFATc1 expression.

  20. Cytosolic malate dehydrogenase regulates RANKL-mediated osteoclastogenesis via AMPK/c-Fos/NFATc1 signaling

    International Nuclear Information System (INIS)

    Oh, Se Jeong; Gu, Dong Ryun; Jin, Su Hyun; Park, Keun Ha; Lee, Seoung Hoon

    2016-01-01

    Cytosolic malate dehydrogenase (malate dehydrogenase 1, MDH1) plays pivotal roles in the malate/aspartate shuttle that might modulate metabolism between the cytosol and mitochondria. In this study, we investigated the role of MDH1 in osteoclast differentiation and formation. MDH1 expression was induced by receptor activator of nuclear factor kappa-B ligand (RANKL) treatment. Knockdown of MDH1 by infection with retrovirus containing MDH1-specific shRNA (shMDH1) reduced mature osteoclast formation and bone resorption activity. Moreover, the expression of marker genes associated with osteoclast differentiation was downregulated by shMDH1 treatment, suggesting a role of MDH1 in osteoclast differentiation. In addition, intracellular ATP production was reduced following the activation of adenosine 5′ monophosphate-activated protein kinase (AMPK), a cellular energy sensor and negative regulator of RANKL-induced osteoclast differentiation, in shMDH1-infected osteoclasts compared to control cells. In addition, the expression of c-Fos and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), a critical transcription factor of osteoclastogenesis, was decreased with MDH1 knockdown during RANKL-mediated osteoclast differentiation. These findings provide strong evidence that MDH1 plays a critical role in osteoclast differentiation and function via modulation of the intracellular energy status, which might affect AMPK activity and NFATc1 expression.

  1. Enhanced fluorescence imaging of live cells by effective cytosolic delivery of probes.

    Directory of Open Access Journals (Sweden)

    Marzia Massignani

    Full Text Available BACKGROUND: Microscopic techniques enable real-space imaging of complex biological events and processes. They have become an essential tool to confirm and complement hypotheses made by biomedical scientists and also allow the re-examination of existing models, hence influencing future investigations. Particularly imaging live cells is crucial for an improved understanding of dynamic biological processes, however hitherto live cell imaging has been limited by the necessity to introduce probes within a cell without altering its physiological and structural integrity. We demonstrate herein that this hurdle can be overcome by effective cytosolic delivery. PRINCIPAL FINDINGS: We show the delivery within several types of mammalian cells using nanometre-sized biomimetic polymer vesicles (a.k.a. polymersomes that offer both highly efficient cellular uptake and endolysomal escape capability without any effect on the cellular metabolic activity. Such biocompatible polymersomes can encapsulate various types of probes including cell membrane probes and nucleic acid probes as well as labelled nucleic acids, antibodies and quantum dots. SIGNIFICANCE: We show the delivery of sufficient quantities of probes to the cytosol, allowing sustained functional imaging of live cells over time periods of days to weeks. Finally the combination of such effective staining with three-dimensional imaging by confocal laser scanning microscopy allows cell imaging in complex three-dimensional environments under both mono-culture and co-culture conditions. Thus cell migration and proliferation can be studied in models that are much closer to the in vivo situation.

  2. Cloning and Expression of a Cytosolic HSP90 Gene in Chlorella vulgaris

    Directory of Open Access Journals (Sweden)

    Zhengyi Liu

    2014-01-01

    Full Text Available Heat shock protein 90 (HSP90, a highly conserved molecular chaperone, plays essential roles in folding, keeping structural integrity, and regulating the subset of cytosolic proteins. We cloned the cDNA of Chlorella vulgaris HSP90 (named CvHSP90 by combining homology cloning with rapid amplification of cDNA ends (RACE. Sequence analysis indicated that CvHSP90 is a cytosolic member of the HSP90 family. Quantitative RT-PCR was applied to determine the expression level of messenger RNA (mRNA in CvHSP90 under different stress conditions. C. vulgaris was kept in different temperatures (5–45°C for 1 h. The mRNA expression level of CvHSP90 increased with temperature from 5 to 10°C, went further from 35 to 40°C, and reached the maximum at 40°C. On the other hand, for C. vulgaris kept at 35°C for different durations, the mRNA expression level of CvHSP90 increased gradually and reached the peak at 7 h and then declined progressively. In addition, the expression level of CvHSP90 at 40 or 45 in salinity (‰ was almost fourfold of that at 25 in salinity (‰ for 2 h. Therefore, CvHSP90 may be a potential biomarker to monitor environment changes.

  3. Subcellular Distribution of NAD+ between Cytosol and Mitochondria Determines the Metabolic Profile of Human Cells*

    Science.gov (United States)

    VanLinden, Magali R.; Dölle, Christian; Pettersen, Ina K. N.; Kulikova, Veronika A.; Niere, Marc; Agrimi, Gennaro; Dyrstad, Sissel E.; Palmieri, Ferdinando; Nikiforov, Andrey A.; Tronstad, Karl Johan; Ziegler, Mathias

    2015-01-01

    The mitochondrial NAD pool is particularly important for the maintenance of vital cellular functions. Although at least in some fungi and plants, mitochondrial NAD is imported from the cytosol by carrier proteins, in mammals, the mechanism of how this organellar pool is generated has remained obscure. A transporter mediating NAD import into mammalian mitochondria has not been identified. In contrast, human recombinant NMNAT3 localizes to the mitochondrial matrix and is able to catalyze NAD+ biosynthesis in vitro. However, whether the endogenous NMNAT3 protein is functionally effective at generating NAD+ in mitochondria of intact human cells still remains to be demonstrated. To modulate mitochondrial NAD+ content, we have expressed plant and yeast mitochondrial NAD+ carriers in human cells and observed a profound increase in mitochondrial NAD+. None of the closest human homologs of these carriers had any detectable effect on mitochondrial NAD+ content. Surprisingly, constitutive redistribution of NAD+ from the cytosol to the mitochondria by stable expression of the Arabidopsis thaliana mitochondrial NAD+ transporter NDT2 in HEK293 cells resulted in dramatic growth retardation and a metabolic shift from oxidative phosphorylation to glycolysis, despite the elevated mitochondrial NAD+ levels. These results suggest that a mitochondrial NAD+ transporter, similar to the known one from A. thaliana, is likely absent and could even be harmful in human cells. We provide further support for the alternative possibility, namely intramitochondrial NAD+ synthesis, by demonstrating the presence of endogenous NMNAT3 in the mitochondria of human cells. PMID:26432643

  4. Subcellular Distribution of NAD+ between Cytosol and Mitochondria Determines the Metabolic Profile of Human Cells.

    Science.gov (United States)

    VanLinden, Magali R; Dölle, Christian; Pettersen, Ina K N; Kulikova, Veronika A; Niere, Marc; Agrimi, Gennaro; Dyrstad, Sissel E; Palmieri, Ferdinando; Nikiforov, Andrey A; Tronstad, Karl Johan; Ziegler, Mathias

    2015-11-13

    The mitochondrial NAD pool is particularly important for the maintenance of vital cellular functions. Although at least in some fungi and plants, mitochondrial NAD is imported from the cytosol by carrier proteins, in mammals, the mechanism of how this organellar pool is generated has remained obscure. A transporter mediating NAD import into mammalian mitochondria has not been identified. In contrast, human recombinant NMNAT3 localizes to the mitochondrial matrix and is able to catalyze NAD(+) biosynthesis in vitro. However, whether the endogenous NMNAT3 protein is functionally effective at generating NAD(+) in mitochondria of intact human cells still remains to be demonstrated. To modulate mitochondrial NAD(+) content, we have expressed plant and yeast mitochondrial NAD(+) carriers in human cells and observed a profound increase in mitochondrial NAD(+). None of the closest human homologs of these carriers had any detectable effect on mitochondrial NAD(+) content. Surprisingly, constitutive redistribution of NAD(+) from the cytosol to the mitochondria by stable expression of the Arabidopsis thaliana mitochondrial NAD(+) transporter NDT2 in HEK293 cells resulted in dramatic growth retardation and a metabolic shift from oxidative phosphorylation to glycolysis, despite the elevated mitochondrial NAD(+) levels. These results suggest that a mitochondrial NAD(+) transporter, similar to the known one from A. thaliana, is likely absent and could even be harmful in human cells. We provide further support for the alternative possibility, namely intramitochondrial NAD(+) synthesis, by demonstrating the presence of endogenous NMNAT3 in the mitochondria of human cells. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Phosphocitrate inhibits mitochondrial and cytosolic accumulation of calcium in kidney cells in vivo.

    Science.gov (United States)

    Tew, W P; Malis, C D; Howard, J E; Lehninger, A L

    1981-01-01

    Synthetic 3-phosphocitrate, an extremely potent inhibitor of calcium phosphate crystallization as determined in a nonbiological physical-chemical assay, has many similarities to a mitochondrial factor that inhibits crystallization of nondiffracting amorphous calcium phosphate. In order to determine whether phosphocitrate can prevent uptake and crystallization of calcium phosphate in mitochondria in vivo, it was administered intraperitoneally to animals given large daily doses of calcium gluconate or parathyroid hormone, a regimen that causes massive accumulation and crystallization of calcium phosphate in the mitochondria and cytosol of renal tubule cells in vivo. Administration of phosphocitrate greatly reduced the net uptake of Ca2+ by the kidneys and prevented the appearance of apatite-like crystalline structures within the mitochondrial matrix and cytosol of renal tubule cells. Phosphocitrate, which is a poor chelator of Ca2+, did not reduce the hypercalcemia induced by either agent. These in vivo observations therefore indicate that phosphocitrate acts primarily at the cellular level to prevent the extensive accumulation of calcium phosphate in kidney cells by inhibiting the mitochondrial accumulation or crystallization of calcium phosphate. Images PMID:6946490

  6. The tobacco carcinogen NNK is stereoselectively reduced by human pancreatic microsomes and cytosols.

    Science.gov (United States)

    Trushin, Neil; Leder, Gerhard; El-Bayoumy, Karam; Hoffmann, Dietrich; Beger, Hans G; Henne-Bruns, Doris; Ramadani, Marco; Prokopczyk, Bogdan

    2008-07-01

    Cigarette smoking increases the risk of cancer of the pancreas. The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is the only known environmental compound that induces pancreatic cancer in laboratory animals. Concentrations of NNK are significantly higher in the pancreatic juice of smokers than in that of nonsmokers. The chiral NNK metabolite, (R,S)-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is itself a potent pancreatic carcinogen in rats. The carcinogenicity of NNAL is related to its stereochemistry; (S)-NNAL is a more potent lung tumorigen in the A/J mouse than is (R)-NNAL. In this study, we determined the potential of the human pancreas to convert NNK into NNAL. Human pancreatic microsomes and cytosols were incubated with [5-(3)H]NNK, and the metabolic products were determined by high-performance liquid chromatography (HPLC). (S)-NNAL was the predominant isomer formed in all cytosolic incubations. In ten microsomal samples, NNAL was formed at an average rate of 3.8 +/- 1.6 pmol/mg/min; (R)-NNAL was the predominant isomer in this group. The average rate of NNAL formation in 18 other microsomal samples was significantly lower, 0.13 +/- 0.12 pmol/mg/min (p < 0.001); (S)-NNAL was the predominant isomer formed in this group. In human pancreatic tissues, there is intraindividual variability regarding the capacity for, and stereoselectivity of, carbonyl reduction of NNK.

  7. Denatured protein-coated docetaxel nanoparticles: Alterable drug state and cytosolic delivery.

    Science.gov (United States)

    Zhang, Li; Xiao, Qingqing; Wang, Yiran; Zhang, Chenshuang; He, Wei; Yin, Lifang

    2017-05-15

    Many lead compounds have a low solubility in water, which substantially hinders their clinical application. Nanosuspensions have been considered a promising strategy for the delivery of water-insoluble drugs. Here, denatured soy protein isolate (SPI)-coated docetaxel nanosuspensions (DTX-NS) were developed using an anti-solvent precipitation-ultrasonication method to improve the water-solubility of DTX, thus improving its intracellular delivery. DTX-NS, with a diameter of 150-250nm and drug-loading up to 18.18%, were successfully prepared by coating drug particles with SPI. Interestingly, the drug state of DTX-NS was alterable. Amorphous drug nanoparticles were obtained at low drug-loading, whereas at a high drug-loading, the DTX-NS drug was mainly present in the crystalline state. Moreover, DTX-NS could be internalized at high levels by cancer cells and enter the cytosol by lysosomal escape, enhancing cell cytotoxicity and apoptosis compared with free DTX. Taken together, denatured SPI has a strong stabilization effect on nanosuspensions, and the drug state in SPI-coated nanosuspensions is alterable by changing the drug-loading. Moreover, DTX-NS could achieve cytosolic delivery, generating enhanced cell cytotoxicity against cancer cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Retargeting the Clostridium botulinum C2 toxin to the neuronal cytosol.

    Science.gov (United States)

    Pavlik, Benjamin J; Hruska, Elizabeth J; Van Cott, Kevin E; Blum, Paul H

    2016-03-30

    Many biological toxins are known to attack specific cell types, delivering their enzymatic payloads to the cytosol. This process can be manipulated by molecular engineering of chimeric toxins. Using toxins with naturally unlinked components as a starting point is advantageous because it allows for the development of payloads separately from the binding/translocation components. Here the Clostridium botulinum C2 binding/translocation domain was retargeted to neural cell populations by deleting its non-specific binding domain and replacing it with a C. botulinum neurotoxin binding domain. This fusion protein was used to deliver fluorescently labeled payloads to Neuro-2a cells. Intracellular delivery was quantified by flow cytometry and found to be dependent on artificial enrichment of cells with the polysialoganglioside receptor GT1b. Visualization by confocal microscopy showed a dissociation of payloads from the early endosome indicating translocation of the chimeric toxin. The natural Clostridium botulinum C2 toxin was then delivered to human glioblastoma A172 and synchronized HeLa cells. In the presence of the fusion protein, native cytosolic enzymatic activity of the enzyme was observed and found to be GT1b-dependent. This retargeted toxin may enable delivery of therapeutics to peripheral neurons and be of use in addressing experimental questions about neural physiology.

  9. Riparian ecosystems and buffers - multiscale structure, function, and management: introduction

    Science.gov (United States)

    Kathleen A. Dwire; Richard R. Lowrance

    2006-01-01

    Given the importance of issues related to improved understanding and management of riparian ecosystems and buffers, the American Water Resources Association (AWRA) sponsored a Summer Specialty Conference in June 2004 at Olympic Valley, California, entitled 'Riparian Ecosystems and Buffers: Multiscale Structure, Function, and Management.' The primary objective...

  10. Universal buffers for use in biochemistry and biophysical experiments

    Directory of Open Access Journals (Sweden)

    Dewey Brooke

    2015-08-01

    Full Text Available The use of buffers that mimic biological solutions is a foundation of biochemical and biophysical studies. However, buffering agents have both specific and nonspecific interactions with proteins. Buffer molecules can induce changes in conformational equilibria, dynamic behavior, and catalytic properties merely by their presence in solution. This effect is of concern because many of the standard experiments used to investigate protein structure and function involve changing solution conditions such as pH and/or temperature. In experiments in which pH is varied, it is common practice to switch buffering agents so that the pH is within the working range of the weak acid and conjugate base. If multiple buffers are used, it is not always possible to decouple buffer induced change from pH or temperature induced change. We have developed a series of mixed biological buffers for protein analysis that can be used across a broad pH range, are compatible with biologically relevant metal ions, and avoid complications that may arise from changing the small molecule composition of buffers when pH is used as an experimental variable.

  11. Social Buffering of Stress in Development: A Career Perspective

    Science.gov (United States)

    Gunnar, Megan R.

    2016-01-01

    This review provides a broad overview of my research group's work on social buffering in human development in the context of the field. Much of the focus is on social buffering of the hypothalamic-pituitary-adrenocortical (HPA) system, one of the two major arms of the mammalian stress system. This focus reflects the centrality of the HPA system in research on social buffering in the fields of developmental psychobiology and developmental science. However, buffering of the cardiovascular and autonomic nervous system is also discussed. The central developmental question in this area derives from attachment theory which argues that the infant's experience of stress and arousal regulation in the context of her early attachment relationships is not an immature form of social buffering experienced in adulthood, but rather the foundation out of which individual differences in the capacity to gain stress relief from social partners emerge. The emergence of social buffering in infancy, changes in social buffering throughout childhood and adolescence, the influence of early experience on later individual differences in social buffering, and critical gaps in our knowledge are described. PMID:28544861

  12. Dynamic Buffer Capacity in Acid-Base Systems.

    Science.gov (United States)

    Michałowska-Kaczmarczyk, Anna M; Michałowski, Tadeusz

    The generalized concept of 'dynamic' buffer capacity β V is related to electrolytic systems of different complexity where acid-base equilibria are involved. The resulting formulas are presented in a uniform and consistent form. The detailed calculations are related to two Britton-Robinson buffers, taken as examples.

  13. Molecular Buffers Permit Sensitivity Tuning and Inversion of Riboswitch Signals

    DEFF Research Database (Denmark)

    Rugbjerg, Peter; Genee, Hans Jasper; Jensen, Kristian

    2016-01-01

    transcription factor, while interacting DNA-binding domains mediate the transduction of signal and form an interacting molecular buffer. The molecular buffer system enables modular signal inversion through integration with repressor modules. Further, tuning of input sensitivity was achieved through perturbation...

  14. Reduction of buffering requirements: Another advantage of cooperative transmission

    KAUST Repository

    Bader, Ahmed

    2015-04-01

    Yet another advent of cooperative transmission is exposed in this letter. It is shown that cooperation lends itself to the reduction of buffer sizes of wireless sensor nodes. It is less likely to find the channel busy when cooperative transmission is employed in the network. Otherwise, in the lack of cooperation, the probability of build up of packet queues in transmission buffers increases.

  15. Dynamic Buffer Capacity in Acid?Base Systems

    OpenAIRE

    Micha?owska-Kaczmarczyk, Anna M.; Micha?owski, Tadeusz

    2015-01-01

    The generalized concept of ?dynamic? buffer capacity ? V is related to electrolytic systems of different complexity where acid?base equilibria are involved. The resulting formulas are presented in a uniform and consistent form. The detailed calculations are related to two Britton?Robinson buffers, taken as examples.

  16. Methods for improved growth of group III nitride buffer layers

    Science.gov (United States)

    Melnik, Yurity; Chen, Lu; Kojiri, Hidehiro

    2014-07-15

    Methods are disclosed for growing high crystal quality group III-nitride epitaxial layers with advanced multiple buffer layer techniques. In an embodiment, a method includes forming group III-nitride buffer layers that contain aluminum on suitable substrate in a processing chamber of a hydride vapor phase epitaxy processing system. A hydrogen halide or halogen gas is flowing into the growth zone during deposition of buffer layers to suppress homogeneous particle formation. Some combinations of low temperature buffers that contain aluminum (e.g., AlN, AlGaN) and high temperature buffers that contain aluminum (e.g., AlN, AlGaN) may be used to improve crystal quality and morphology of subsequently grown group III-nitride epitaxial layers. The buffer may be deposited on the substrate, or on the surface of another buffer. The additional buffer layers may be added as interlayers in group III-nitride layers (e.g., GaN, AlGaN, AlN).

  17. Sampling phase lock loop (PLL) with low power clock buffer

    NARCIS (Netherlands)

    Gao, X.; Bahai, A.; Bohsali, M.; Djabbari, A.; Klumperink, Eric A.M.; Nauta, Bram; Socci, G.

    2013-01-01

    A sampling phase locked loop (PLL) circuit includes a pull-up/down buffer configured to convert an oscillator reference clock into a square wave sampling control signal input to a sampling phase detector. The buffer circuit is configured to reduce power by controlling the switching of the pull-up

  18. Bus Implementation Using New Low Power PFSCL Tristate Buffers

    Directory of Open Access Journals (Sweden)

    Neeta Pandey

    2016-01-01

    Full Text Available This paper proposes new positive feedback source coupled logic (PFSCL tristate buffers suited to bus applications. The proposed buffers use switch to attain high impedance state and modify the load or the current source section. An interesting consequence of this is overall reduction in the power consumption. The proposed tristate buffers consume half the power compared to the available switch based counterpart. The issues with available PFSCL tristate buffers based bus implementation are identified and benefits of employing the proposed tristate buffer topologies are put forward. SPICE simulation results using TSMC 180 nm CMOS technology parameters are included to support the theoretical formulations. The performance of proposed tristate buffer topologies is examined on the basis of propagation delay, output enable time, and power consumption. It is found that one of the proposed tristate buffer topology outperforms the others in terms of all the performance parameters. An examination of behavior of available and the proposed PFSCL tristate buffer topologies under parameter variations and mismatch shows a maximum variation of 14%.

  19. Effects of riparian buffers on hydrology of northern seasonal ponds

    Science.gov (United States)

    Randall K. Kolka; Brian J. Palik; Daniel P. Tersteeg; James C. Bell

    2011-01-01

    Although seasonal ponds are common in northern, glaciated, forested landscapes, forest management guidelines are generally lacking for these systems. The objective of this study was to determine the effect of riparian buffer type on seasonal pond hydrology following harvest of the adjacent upland forest. A replicated block design consisting of four buffer treatments...

  20. Glutathione redox potential in the mitochondrial intermembrane space is linked to the cytosol and impacts the Mia40 redox state

    Science.gov (United States)

    Kojer, Kerstin; Bien, Melanie; Gangel, Heike; Morgan, Bruce; Dick, Tobias P; Riemer, Jan

    2012-01-01

    Glutathione is an important mediator and regulator of cellular redox processes. Detailed knowledge of local glutathione redox potential (EGSH) dynamics is critical to understand the network of redox processes and their influence on cellular function. Using dynamic oxidant recovery assays together with EGSH-specific fluorescent reporters, we investigate the glutathione pools of the cytosol, mitochondrial matrix and intermembrane space (IMS). We demonstrate that the glutathione pools of IMS and cytosol are dynamically interconnected via porins. In contrast, no appreciable communication was observed between the glutathione pools of the IMS and matrix. By modulating redox pathways in the cytosol and IMS, we find that the cytosolic glutathione reductase system is the major determinant of EGSH in the IMS, thus explaining a steady-state EGSH in the IMS which is similar to the cytosol. Moreover, we show that the local EGSH contributes to the partially reduced redox state of the IMS oxidoreductase Mia40 in vivo. Taken together, we provide a comprehensive mechanistic picture of the IMS redox milieu and define the redox influences on Mia40 in living cells. PMID:22705944

  1. Smooth Nb surfaces fabricated by buffered electropolishing

    International Nuclear Information System (INIS)

    Wu, Andy T.; Mammosser, John; Phillips, Larry; Delayen, Jean; Reece, Charles; Wilkerson, Amy; Smith, David; Ike, Robert

    2007-01-01

    It was demonstrated that smooth Nb surfaces could be obtained through buffered electropolishing (BEP) employing an electrolyte consisting of lactic, sulfuric, and hydrofluoric acids. Parameters that control the polishing process were optimized to achieve a smooth surface finish. The polishing rate of BEP was determined to be 0.646 μm/min which was much higher than 0.381 μm/min achieved by the conventional electropolishing (EP) process widely used in the superconducting radio frequency (SRF) community. Root mean square measurements using a 3D profilometer revealed that Nb surfaces treated by BEP were an order of magnitude smoother than those treated by the optimized EP process. The chemical composition of the Nb surfaces after BEP was analyzed by static and dynamic secondary ion mass spectrometry (SIMS) systems. SIMS results implied that the surface oxide structure of Nb might be more complicated than what usually believed and could be inhomogeneous. Preliminary results of BEP on Nb SRF single cell cavities and half-cells were reported. It was shown that smooth and bright surfaces could be obtained in 1800 s when the electric field inside a SRF cavity was uniform during a BEP process. This study showed that BEP is a promising technique for surface treatment on Nb SRF cavities to be used in particle accelerators

  2. Buffers for biomass production in temperate European agriculture

    DEFF Research Database (Denmark)

    Christen, Benjamin; Dalgaard, Tommy

    2013-01-01

    , environmental pressures from intensive agriculture and policy developments. Use of conservation buffers by farmers outside of designated schemes is limited to date, but the increasing demand for bioenergy and the combination of agricultural production with conservation calls for a much wider implementation....... This paper reviews the biophysical knowledge on buffer functioning and associated ecosystem services. It describes how a three-zone buffer design, with arable fields buffered in combination by grassland, short rotation forestry (SRF) or coppice (SRC) and undisturbed vegetation along water courses, can...... be incorporated into farming landscapes as productive conservation elements and reflects on the potential for successful implementation. Land use plays a much greater role in determining catchment hydrology than soil type: shelterbelts or buffer strips have markedly higher infiltration capacity than arable...

  3. Evaluation on elution feature of bentonite buffer materials

    Energy Technology Data Exchange (ETDEWEB)

    Ishikawa, Hirohisa; Kanno, Takeshi; Matsumoto, Kazuhiro

    1997-09-01

    In order to evaluate long term physical stability of artificial barrier in land disposal of high level radioactive wastes, it is necessary to know quantitatively elution behavior of buffering materials from disposal road (or cavity) to circumferential rock crack. When elution of the buffer material occurs on large scale, amount of bentonite in the disposal road (or cavity) reduces and reduction of various functions expected to the buffer materials is presumed. According to specification examples of road transverse arrangement and disposal vertical arrangement systems, evaluation on elution amount of the buffer materials at disposal environment was conducted. Opening width of rock crack in the disposal environment was supposed to be 0.5 mm. As a result, obtained mass elution ratios of the buffer materials due to extrusion phenomenon were 0.04 to 0.2% after 10,000 year and 2 to 12% after 1,000,000 years. (G.K.)

  4. Thin film photovoltaic devices with a minimally conductive buffer layer

    Science.gov (United States)

    Barnes, Teresa M.; Burst, James

    2016-11-15

    A thin film photovoltaic device (100) with a tunable, minimally conductive buffer (128) layer is provided. The photovoltaic device (100) may include a back contact (150), a transparent front contact stack (120), and an absorber (140) positioned between the front contact stack (120) and the back contact (150). The front contact stack (120) may include a low resistivity transparent conductive oxide (TCO) layer (124) and a buffer layer (128) that is proximate to the absorber layer (140). The photovoltaic device (100) may also include a window layer (130) between the buffer layer (128) and the absorber (140). In some cases, the buffer layer (128) is minimally conductive, with its resistivity being tunable, and the buffer layer (128) may be formed as an alloy from a host oxide and a high-permittivity oxide. The high-permittivity oxide may further be chosen to have a bandgap greater than the host oxide.

  5. Multi-Buffer Simulations for Trace Language Inclusion

    Directory of Open Access Journals (Sweden)

    Milka Hutagalung

    2016-09-01

    Full Text Available We consider simulation games played between Spoiler and Duplicator on two Büchi automata in which the choices made by Spoiler can be buffered by Duplicator in several buffers before she executes them on her structure. We show that the simulation games are useful to approximate the inclusion of trace closures of languages accepted by finite-state automata, which is known to be undecidable. We study the decidability and complexity and show that the game with bounded buffers can be decided in polynomial time, whereas the game with one unbounded and one bounded buffer is highly undecidable. We also show some sufficient conditions on the automata for Duplicator to win the game (with unbounded buffers.

  6. Replenishing data descriptors in a DMA injection FIFO buffer

    Science.gov (United States)

    Archer, Charles J [Rochester, MN; Blocksome, Michael A [Rochester, MN; Cernohous, Bob R [Rochester, MN; Heidelberger, Philip [Cortlandt Manor, NY; Kumar, Sameer [White Plains, NY; Parker, Jeffrey J [Rochester, MN

    2011-10-11

    Methods, apparatus, and products are disclosed for replenishing data descriptors in a Direct Memory Access (`DMA`) injection first-in-first-out (`FIFO`) buffer that include: determining, by a messaging module on an origin compute node, whether a number of data descriptors in a DMA injection FIFO buffer exceeds a predetermined threshold, each data descriptor specifying an application message for transmission to a target compute node; queuing, by the messaging module, a plurality of new data descriptors in a pending descriptor queue if the number of the data descriptors in the DMA injection FIFO buffer exceeds the predetermined threshold; establishing, by the messaging module, interrupt criteria that specify when to replenish the injection FIFO buffer with the plurality of new data descriptors in the pending descriptor queue; and injecting, by the messaging module, the plurality of new data descriptors into the injection FIFO buffer in dependence upon the interrupt criteria.

  7. Buffer erosion: An overview of concepts and potential safety consequences

    International Nuclear Information System (INIS)

    Apted, Michael J.; Arthur, Randy; Bennett, David; Savage, David; Saellfors, Goeran; Wennerstroem, Haakan

    2010-11-01

    In its safety analysis SR-Can, SKB reported preliminary results and conclusions on the mechanisms of bentonite colloid formation and stability, with a rough estimate of the consequences of loss of bentonite buffer by erosion. With the review of SR-Can the authorities (SKI and SSI) commented that erosion of the buffer had the greatest safety significance, that the understanding of the mechanisms of buffer erosion was inadequate, and that more work would be required to arrive at robust estimates of the extent and impacts of buffer erosion. After the SR-Can report, SKB started a two-year research project on buffer erosion. The results from this two-year project have been reported in several SKB technical reports. SSM started this project to build up its own competence in the related scientific areas by a preliminary evaluation of SKB's research results

  8. Buffer erosion: An overview of concepts and potential safety consequences

    Energy Technology Data Exchange (ETDEWEB)

    Apted, Michael J.; Arthur, Randy (INTERA Incorporated, Denver, CO (United States)); Bennett, David (TerraSalus Limited, Rutland (United Kingdom)); Savage, David (Savage Earth Associates Limited, Bournemouth (United Kingdom)); Saellfors, Goeran (GeoForce AB, Billdal (Sweden)); Wennerstroem, Haakan (Dept. of Chemistry, Lund Univ., Lund (Sweden))

    2010-11-15

    In its safety analysis SR-Can, SKB reported preliminary results and conclusions on the mechanisms of bentonite colloid formation and stability, with a rough estimate of the consequences of loss of bentonite buffer by erosion. With the review of SR-Can the authorities (SKI and SSI) commented that erosion of the buffer had the greatest safety significance, that the understanding of the mechanisms of buffer erosion was inadequate, and that more work would be required to arrive at robust estimates of the extent and impacts of buffer erosion. After the SR-Can report, SKB started a two-year research project on buffer erosion. The results from this two-year project have been reported in several SKB technical reports. SSM started this project to build up its own competence in the related scientific areas by a preliminary evaluation of SKB's research results

  9. Phosphorus retention in riparian buffers: review of their efficiency.

    Science.gov (United States)

    Hoffmann, Carl Christian; Kjaergaard, Charlotte; Uusi-Kämppä, Jaana; Hansen, Hans Christian Bruun; Kronvang, Brian

    2009-01-01

    Ground water and surface water interactions are of fundamental importance for the biogeochemical processes governing phosphorus (P) dynamics in riparian buffers. The four most important conceptual hydrological pathways for P losses from and P retention in riparian buffers are reviewed in this paper: (i) The diffuse flow path with ground water flow through the riparian aquifer, (ii) the overland flow path across the riparian buffer with water coming from adjacent agricultural fields, (iii) irrigation of the riparian buffer with tile drainage water from agricultural fields where disconnected tile drains irrigate the riparian buffer, and (iv) inundation of the riparian buffer (floodplain) with river water during short or longer periods. We have examined how the different flow paths in the riparian buffer influence P retention mechanisms theoretically and from empirical evidence. The different hydrological flow paths determine where and how water-borne P compounds meet and interact with iron and aluminum oxides or other minerals in the geochemical cycling of P in the complex and dynamic environment that constitutes a riparian buffer. The main physical process in the riparian buffer-sedimentation-is active along several flow paths and may account for P retention rates of up to 128 kg P ha(-1) yr(-1), while plant uptake may temporarily immobilize up to 15 kg P ha(-1) yr(-1). Retention of dissolved P in riparian buffers is not as pronounced as retention of particulate P and is often below 0.5 kg P ha(-1) yr(-1). Several studies show significant release of dissolved P (i.e., up to 8 kg P ha(-1) yr(-1)).

  10. Visualization of Buffer Capacity with 3-D "Topo" Surfaces: Buffer Ridges, Equivalence Point Canyons and Dilution Ramps

    Science.gov (United States)

    Smith, Garon C.; Hossain, Md Mainul

    2016-01-01

    BufCap TOPOS is free software that generates 3-D topographical surfaces ("topos") for acid-base equilibrium studies. It portrays pH and buffer capacity behavior during titration and dilution procedures. Topo surfaces are created by plotting computed pH and buffer capacity values above a composition grid with volume of NaOH as the x axis…

  11. BENTO buffer development program in Finland - Key issues

    International Nuclear Information System (INIS)

    Autio, J.; Korkiala-Tanttu, L.; Vaehaenen, M.; Koskinen, K.; Korkeakoski, P.; Haapala, K.

    2010-01-01

    Document available in extended abstract form only. Posiva launched a programme, BENTO, to develop technology of using bentonite in spent nuclear fuel repositories. The main purpose of the BENTO programme is to produce buffer designs and verify that they fulfil the requirements, especially safety requirements. To achieve this objective, resources and the level of expertise and know-how has to be increased. There are several uncertainties related to the functioning of the buffer components at present. An issue is defined as being significant if there is sufficient uncertainty that the buffer system might not fulfil the requirements because of the issue. These significant issues need to be resolved in order to develop a proper design and to verify the fulfilment of the requirements. The list of significant issues may change with time. Therefore it is crucial to develop adequate expertise, know-how and laboratory facilities to manage the changes. Moreover, there is confidence that by solving the open issues a defendable construction license application can be submitted in 2012. The basic nature of the programme is a combination of material and process research with the design and manufacturing of buffer components to produce feasible buffer design with proven long-term functional properties. The development work carried out under BENTO-programme has been initially divided into four different projects. During the course of work the number of projects and their content can be adjusted. The four BENTO projects are: 1. Manufacturing (MANU); 2. Design (DESI); 3. Modelling (MODE); 4. Material and Process Research (MARE). BENTO programme aims at producing feasible buffer designs which fulfil the requirements specified in Posiva's requirement management system. The designs are produced in DESI-project by following the design development scheme which starts from specification of design basis and ends in documented detailed designs and therefore DESIgn is specified as one

  12. Two-Phase Acto-Cytosolic Fluid Flow in a Moving Keratocyte: A 2D Continuum Model.

    Science.gov (United States)

    Nikmaneshi, M R; Firoozabadi, B; Saidi, M S

    2015-09-01

    The F-actin network and cytosol in the lamellipodia of crawling cells flow in a centripetal pattern and spout-like form, respectively. We have numerically studied this two-phase flow in the realistic geometry of a moving keratocyte. Cytosol has been treated as a low viscosity Newtonian fluid flowing through the high viscosity porous medium of F-actin network. Other involved phenomena including myosin activity, adhesion friction, and interphase interaction are also discussed to provide an overall view of this problem. Adopting a two-phase coupled model by myosin concentration, we have found new accurate perspectives of acto-cytosolic flow and pressure fields, myosin distribution, as well as the distribution of effective forces across the lamellipodia of a keratocyte with stationary shape. The order of magnitude method is also used to determine the contribution of forces in the internal dynamics of lamellipodia.

  13. Erosion of buffer caused by groundwater leakages

    International Nuclear Information System (INIS)

    Autio, J.; Hanana, K.; Punkkinen, O.; Koskinen, K.; Olin, M.

    2010-01-01

    Document available in extended abstract form only. In the Finnish HLW disposal concept the most important properties of the bentonite clay being considered for these isolation purposes are its thermal behaviour, low hydraulic conductivity, diffusion limited transport, rheology, plasticity, sufficient swelling potential, and exchange capacity. All of these properties depend critically on bentonite density; therefore, any potential mass loss or redistribution events must be well characterized. One such event or process is the erosion of bentonite by flowing groundwater and the groundwater flowing in newly formed channels, in special. Mechanical erosion during the operational phase, due to high groundwater pressure gradients in open excavations, has been identified as a critical issue in TKS-2006 and SR-Can. This work addresses the mechanical erosion of bentonite by fluid shear. In order for buffer erosion to occur three processes must take place: detachment, entrainment, and transport. These processes are followed by the settling of the material and redistribution of buffer mass. Erosion begins with the detachment of a particle from surrounding material, which requires the application of shear forces greater than the attractive force between the particle and parent structure. Entrainment is the process by which the eroding medium lifts the detached particle into the flow. The most important aspect in entrainment is transfer of fluid's inertial forces via surface friction to particles' inertial forces, which, in turn, must overcome the frictional resistance between the particle and its surroundings. Factors influencing frictional resistance include gravity, particle mass, saturation degree of parent structure, composition of water present in parent structure, particle size, and surface roughness. Recent erosion tests, whereby water flow was directed over compacted bentonite blocks or through a system of bentonite pellets, have indicated that bentonite erodes

  14. MANU. Instrumentation of Buffer Demo. Preliminary Study

    International Nuclear Information System (INIS)

    Laaksonen, R.

    2010-01-01

    The purpose of this work is to describe feasible measuring and monitoring alternatives which can be used, if needed, in medium to full scale nuclear waste repository deposition hole mock-up tests. The focus of the work was to determine what variables can actually be measured, how to achieve the measurements and what kind of demands comes from the modelling, scientific, and technical points of view. This project includes a review of the previous waste repository mock-up tests carried out in several European countries such as Belgium, Czech Republic, Spain and Sweden. Also information was gathered by interviewing domestic and foreign scientists specialized in the fields of measurement instrumentation and related in-situ and laboratory work. On the basis of this review, recommendations were developed for the necessary actions needed to be done from the instrumentation point of view for future tests. It is possible to measure and monitor the processes going on in a deposition hole in-situ conditions. The data received during a test in real repository conditions enables to follow the processes and to verify the hypothesis made on the behaviour of various components of the repository: buffer, canister, rock and backfill. Because full scale testing is expensive, the objectives and hypothesis must be carefully set and the test itself with its instrumentation must serve very specific objectives. The main purpose of mock-up tests is to verify that the conditions surrounding the canister are according to the design requirements. A whole mock-up test and demonstration process requires a lot of time and effort. The instrumentation part of the work must also start at early stages to ensure that the instrumentation itself will not become bottlenecked nor suffer from low quality solutions. The planning of the instrumentation work could be done in collaboration with foreign scientists which have participated to previous instrumentation projects. (orig.)

  15. Temperature Buffer Test. Final THM modelling

    International Nuclear Information System (INIS)

    Aakesson, Mattias; Malmberg, Daniel; Boergesson, Lennart; Hernelind, Jan; Ledesma, Alberto; Jacinto, Abel

    2012-01-01

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aespoe HRL. It was installed during the spring of 2003. Two heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by bentonite only, whereas the upper heater was surrounded by a composite barrier, with a sand shield between the heater and the bentonite. The test was dismantled and sampled during the winter of 2009/2010. This report presents the final THM modelling which was resumed subsequent to the dismantling operation. The main part of this work has been numerical modelling of the field test. Three different modelling teams have presented several model cases for different geometries and different degree of process complexity. Two different numerical codes, Code B right and Abaqus, have been used. The modelling performed by UPC-Cimne using Code B right, has been divided in three subtasks: i) analysis of the response observed in the lower part of the test, by inclusion of a number of considerations: (a) the use of the Barcelona Expansive Model for MX-80 bentonite; (b) updated parameters in the vapour diffusive flow term; (c) the use of a non-conventional water retention curve for MX-80 at high temperature; ii) assessment of a possible relation between the cracks observed in the bentonite blocks in the upper part of TBT, and the cycles of suction and stresses registered in that zone at the start of the experiment; and iii) analysis of the performance, observations and interpretation of the entire test. It was however not possible to carry out a full THM analysis until the end of the test due to

  16. Temperature Buffer Test. Final THM modelling

    Energy Technology Data Exchange (ETDEWEB)

    Aakesson, Mattias; Malmberg, Daniel; Boergesson, Lennart; Hernelind, Jan [Clay Technology AB, Lund (Sweden); Ledesma, Alberto; Jacinto, Abel [UPC, Universitat Politecnica de Catalunya, Barcelona (Spain)

    2012-01-15

    The Temperature Buffer Test (TBT) is a joint project between SKB/ANDRA and supported by ENRESA (modelling) and DBE (instrumentation), which aims at improving the understanding and to model the thermo-hydro-mechanical behavior of buffers made of swelling clay submitted to high temperatures (over 100 deg C) during the water saturation process. The test has been carried out in a KBS-3 deposition hole at Aespoe HRL. It was installed during the spring of 2003. Two heaters (3 m long, 0.6 m diameter) and two buffer arrangements have been investigated: the lower heater was surrounded by bentonite only, whereas the upper heater was surrounded by a composite barrier, with a sand shield between the heater and the bentonite. The test was dismantled and sampled during the winter of 2009/2010. This report presents the final THM modelling which was resumed subsequent to the dismantling operation. The main part of this work has been numerical modelling of the field test. Three different modelling teams have presented several model cases for different geometries and different degree of process complexity. Two different numerical codes, Code{sub B}right and Abaqus, have been used. The modelling performed by UPC-Cimne using Code{sub B}right, has been divided in three subtasks: i) analysis of the response observed in the lower part of the test, by inclusion of a number of considerations: (a) the use of the Barcelona Expansive Model for MX-80 bentonite; (b) updated parameters in the vapour diffusive flow term; (c) the use of a non-conventional water retention curve for MX-80 at high temperature; ii) assessment of a possible relation between the cracks observed in the bentonite blocks in the upper part of TBT, and the cycles of suction and stresses registered in that zone at the start of the experiment; and iii) analysis of the performance, observations and interpretation of the entire test. It was however not possible to carry out a full THM analysis until the end of the test due to

  17. Liver/kidney microsomal antibody type 1 and liver cytosol antibody type 1 concentrations in type 2 autoimmune hepatitis

    OpenAIRE

    Muratori, L; Cataleta, M; Muratori, P; Lenzi, M; Bianchi, F

    1998-01-01

    Background—Liver/kidney microsomal antibody type 1 (LKM1) and liver cytosol antibody type 1 (LC1) are the serological markers of type 2 autoimmune hepatitis (AIH). 
Aims—Since LKM1 and LC1 react against two distinct liver specific autoantigens (cytochrome P450IID6 (CYP2D6) and a 58 kDa cytosolic polypeptide respectively), the aim was to see whether LKM1 and LC1 concentrations correlate with liver disease activity. 
Patients—Twenty one patients with type 2 AIH were studied. 
Methods—A...

  18. The Interplay between Feedback and Buffering in Cellular Homeostasis.

    Science.gov (United States)

    Hancock, Edward J; Ang, Jordan; Papachristodoulou, Antonis; Stan, Guy-Bart

    2017-11-22

    Buffering, the use of reservoirs of molecules to maintain concentrations of key molecular species, and negative feedback are the primary known mechanisms for robust homeostatic regulation. To our knowledge, however, the fundamental principles behind their combined effect have not been elucidated. Here, we study the interplay between buffering and negative feedback in the context of cellular homeostasis. We show that negative feedback counteracts slow-changing disturbances, whereas buffering counteracts fast-changing disturbances. Furthermore, feedback and buffering have limitations that create trade-offs for regulation: instability in the case of feedback and molecular noise in the case of buffering. However, because buffering stabilizes feedback and feedback attenuates noise from slower-acting buffering, their combined effect on homeostasis can be synergistic. These effects can be explained within a traditional control theory framework and are consistent with experimental observations of both ATP homeostasis and pH regulation in vivo. These principles are critical for studying robustness and homeostasis in biology and biotechnology. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  19. Buffer lining manufacturing method for radioactive waste container

    International Nuclear Information System (INIS)

    Kawakami, Susumu; Sugino, Hiroyuki

    1998-01-01

    A recessed portion is formed on an upper surface of a filler layer made of a buffer powder filled into a container main body, the upper portion of the vessel main body is closed by a shrinkable liquid tight film. It is placed in a pressurizing container and pressed to mold a buffer lining base material integrated with the vessel main body. A flat upper surface and a containing space are formed by shaving to form a buffer lining. A disposing vessel containing radioactive wastes is inserted into the containing space, and the containing space is closed by a buffer block. The upper surface is sealed by a lid. With such a constitution, since a buffer lining integrated with the vessel main body can be formed easily inside the vessel main body, the disposing vessel can be contained in the containing vessel in a state surrounded by the buffer easily and stably without laying or piling over a large quantity of buffer blocks. (T.M.)

  20. Accelerating Science with the NERSC Burst Buffer Early User Program

    Energy Technology Data Exchange (ETDEWEB)

    Bhimji, Wahid [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Bard, Debbie [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Romanus, Melissa [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Rutgers Univ., New Brunswick, NJ (United States); Paul, David [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Ovsyannikov, Andrey [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Friesen, Brian [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Bryson, Matt [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Correa, Joaquin [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Lockwood, Glenn K. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Tsulaia, Vakho [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Byna, Suren [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Farrell, Steve [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Gursoy, Doga [Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS); Daley, Chris [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Beckner, Vince [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Van Straalen, Brian [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Trebotich, David [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Tull, Craig [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Weber, Gunther H. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Wright, Nicholas J. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Antypas, Katie [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Prabhat, none [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

    2016-01-01

    NVRAM-based Burst Buffers are an important part of the emerging HPC storage landscape. The National Energy Research Scientific Computing Center (NERSC) at Lawrence Berkeley National Laboratory recently installed one of the first Burst Buffer systems as part of its new Cori supercomputer, collaborating with Cray on the development of the DataWarp software. NERSC has a diverse user base comprised of over 6500 users in 700 different projects spanning a wide variety of scientific computing applications. The use-cases of the Burst Buffer at NERSC are therefore also considerable and diverse. We describe here performance measurements and lessons learned from the Burst Buffer Early User Program at NERSC, which selected a number of research projects to gain early access to the Burst Buffer and exercise its capability to enable new scientific advancements. To the best of our knowledge this is the first time a Burst Buffer has been stressed at scale by diverse, real user workloads and therefore these lessons will be of considerable benefit to shaping the developing use of Burst Buffers at HPC centers.

  1. pH variations during diafiltration due to buffer nonidealities.

    Science.gov (United States)

    Baek, Youngbin; Yang, Deyu; Singh, Nripen; Arunkumar, Abhiram; Ghose, Sanchayita; Li, Zheng Jian; Zydney, Andrew L

    2017-11-01

    Diafiltration is used for final formulation of essentially all biotherapeutics. Several studies have demonstrated that buffer/excipient concentrations in the final diafiltered product can be different than that in the diafiltration buffer due to interactions between buffer species and the protein product. However, recent work in our lab has shown variations in solution pH that are largely independent of the protein concentration during the first few diavolumes. Our hypothesis is that these pH variations are due to nonidealities in the acid-base equilibrium coefficient. A model was developed for the diafiltration process accounting for the ionic strength dependence of the pK a . Experimental results obtained using phosphate and histidine buffers were in excellent agreement with model predictions. A decrease in ionic strength leads to an increase in the pK a for the phosphate buffer, causing a shift in the solution pH, even under conditions where the initial feed and the diafiltration buffer are at the same pH. This effect could be eliminated by matching the ionic strength of the feed and diafiltration buffer. The experimental data and model provide new insights into the factors controlling the pH profile during diafiltration processes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1555-1560, 2017. © 2017 American Institute of Chemical Engineers.

  2. Protein buffering in model systems and in whole human saliva.

    Directory of Open Access Journals (Sweden)

    Andreas Lamanda

    Full Text Available The aim of this study was to quantify the buffer attributes (value, power, range and optimum of two model systems for whole human resting saliva, the purified proteins from whole human resting saliva and single proteins. Two model systems, the first containing amyloglucosidase and lysozyme, and the second containing amyloglucosidase and alpha-amylase, were shown to provide, in combination with hydrogencarbonate and di-hydrogenphosphate, almost identical buffer attributes as whole human resting saliva. It was further demonstrated that changes in the protein concentration as small as 0.1% may change the buffer value of a buffer solution up to 15 times. Additionally, it was shown that there was a protein concentration change in the same range (0.16% between saliva samples collected at the time periods of 13:00 and others collected at 9:00 am and 17:00. The mode of the protein expression changed between these samples corresponded to the change in basic buffer power and the change of the buffer value at pH 6.7. Finally, SDS Page and Ruthenium II tris (bathophenantroline disulfonate staining unveiled a constant protein expression in all samples except for one 50 kDa protein band. As the change in the expression pattern of that 50 kDa protein band corresponded to the change in basic buffer power and the buffer value at pH 6.7, it was reasonable to conclude that this 50 kDa protein band may contain the protein(s belonging to the protein buffer system of human saliva.

  3. The stochastic nuclide transport model for buffer/backfill materials

    International Nuclear Information System (INIS)

    Ma Liping; Han Yongguo

    2014-01-01

    Currently, study on nuclide migration law in geological disposal repository of high level waste is assumed buffer/backfill layer to be continuous medium, utilized the continuity equation, equation of state, the equations of motion, etc, formed a set of theory and method to estimate nuclide concentration distribution in buffer/backfill layer, and provided an important basis for nuclide migration rules of repository. However, it is necessary to study the buffer/backfill layer microstructure and subtly describe the pore structure and fracture system of the buffer/backfill layer, and reflect the changes in connectivity and in different directions of the buffer/backfill layer. Through using random field theory, the nuclide transport for the buffer/backfill layer in geological disposal repository of nuclear waste is described in the paper. This paper mainly includes that, t represents the time, ξ t ⊂ Z d = d represents the integer lattice, Z represents collectivity integers, d = l, 2, 3, for instance, d = 2, Z d = {(m, n) : m, n ∈ Z} the state point of ξ t is typically considered to be occupied by the nuclide concentration values of the buffer/backfill layer, ξ t also represents random set in the diagram of two dimensional integer lattice, namely, t ∈ [0, T], {ξ t ,0 ≤ t ≤ ⊂ T} Consequently, according to the stochastic process obtained above, the changes of the nuclide concentration values of the buffer/backfill layer or the buffer/backfill laboratory materials in the repository with the time can be known. (authors)

  4. Gene deletion of cytosolic ATP: citrate lyase leads to altered organic acid production in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; Nielsen, Michael Lynge; Olsson, Lisbeth

    2009-01-01

    With the availability of the genome sequence of the filamentous fungus Aspergillus niger, the use of targeted genetic modifications has become feasible. This, together with the fact that A. niger is well established industrially, makes this fungus an attractive micro-organism for creating a cell...... factory platform for production of chemicals. Using molecular biology techniques, this study focused on metabolic engineering of A. niger to manipulate its organic acid production in the direction of succinic acid. The gene target for complete gene deletion was cytosolic ATP: citrate lyase (acl), which...... the acl gene. Additionally, the total amount of organic acids produced in the deletion strain was significantly increased. Genome-scale stoichiometric metabolic model predictions can be used for identifying gene targets. Deletion of the acl led to increased succinic acid production by A. niger....

  5. Knockdown of cytosolic NADP(+) -dependent isocitrate dehydrogenase enhances MPP(+) -induced oxidative injury in PC12 cells.

    Science.gov (United States)

    Yang, Eun Sun; Park, Jeen-Woo

    2011-05-01

    1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its toxic metabolite 1-methyl-4-phenylpyridium ion (MPP(+)) have been shown to induce Parkinson's disease-like symptoms as well as neurotoxicity in humans and animal species. Recently, we reported that maintenance of redox balance and cellular defense against oxidative damage are primary functions of the novel antioxidant enzyme cytosolic NADP(+) -dependent isocitrate dehydrogenase (IDPc). In this study, we examined the role of IDPc in cellular defense against MPP(+) -induced oxidative injury using PC12 cells transfected with IDPc small interfering RNA (siRNA). Our results demonstrate that MPP(+) -mediated disruption of cellular redox status, oxidative damage to cells, and apoptotic cell death were significantly enhanced by knockdown of IDPc.

  6. Decrease in the cytosolic NADP+-dependent isocitrate dehydrogenase activity through porcine sperm capacitation.

    Science.gov (United States)

    Katoh, Yuki; Tamba, Michiko; Matsuda, Manabu; Kikuchi, Kazuhiro; Okamura, Naomichi

    2018-02-26

    In order to understand the molecular mechanisms involved in the sperm capacitation, we have identified the proteins tyrosine-phosphorylated during the capacitation especially in conjunction with the regulation of the levels of reactive oxygen species (ROS) in sperm. In the present study, the effects of the tyrosine phosphorylation of cytosolic NADP + -dependent isocitrate dehydrogenase (IDPc) on its catalytic activity and on the levels of ROS in sperm have been studied. The tyrosine phosphorylated IDPc showed a significantly lowered enzymatic activity. The immunocytochemical analyses using the highly specific antisera against IDPc revealed that IDPc was mainly localized to the principal piece of the porcine sperm flagellum. As IDPc is one of the major NADPH regenerating enzymes in porcine sperm, it is strongly suggested that the decrease in IDPc activity is involved in the increased levels of ROS, which results in the induction of hyperactivated flagellar movement and capacitation. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Regulation of singlet oxygen-induced apoptosis by cytosolic NADP+-dependent isocitrate dehydrogenase.

    Science.gov (United States)

    Kim, Sun Yee; Lee, Su Min; Tak, Jean Kyoung; Choi, Kyeong Sook; Kwon, Taeg Kyu; Park, Jeen-Woo

    2007-08-01

    Singlet oxygen is a highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules and it also promotes deleterious processes such as cell death. Recently, we demonstrated that the control of redox balance and the cellular defense against oxidative damage are the primary functions of cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) through supplying NADPH for antioxidant systems. In this report, we demonstrate that modulation of IDPc activity in HL-60 cells regulates singlet oxygen-induced apoptosis. When we examined the protective role of IDPc against singlet oxygen-induced apoptosis with HL-60 cells transfected with the cDNA for mouse IDPc in sense and antisense orientations, a clear inverse relationship was observed between the amount of IDPc expressed in target cells and their susceptibility to apoptosis. The results suggest that IDPc plays an important protective role in apoptosis of HL-60 cells induced by singlet oxygen.

  8. Regulation and function of the cGAS-STING pathway of cytosolic DNA sensing.

    Science.gov (United States)

    Chen, Qi; Sun, Lijun; Chen, Zhijian J

    2016-09-20

    The recognition of microbial nucleic acids is a major mechanism by which the immune system detects pathogens. Cyclic GMP-AMP (cGAMP) synthase (cGAS) is a cytosolic DNA sensor that activates innate immune responses through production of the second messenger cGAMP, which activates the adaptor STING. The cGAS-STING pathway not only mediates protective immune defense against infection by a large variety of DNA-containing pathogens but also detects tumor-derived DNA and generates intrinsic antitumor immunity. However, aberrant activation of the cGAS pathway by self DNA can also lead to autoimmune and inflammatory disease. Thus, the cGAS pathway must be properly regulated. Here we review the recent advances in understanding of the cGAS-STING pathway, focusing on the regulatory mechanisms and roles of this pathway in heath and disease.

  9. Cytosolic phosphoenolpyruvate carboxykinase is a response gene involved in porcine adipocyte adaptation to heat stress.

    Science.gov (United States)

    Qu, Huan; Ajuwon, Kolapo M

    2018-05-04

    Heat stress (HS) leads to increased lipid storage and expression of cytosolic phosphoenolpyruvate carboxykinase (PCK1) in pig adipocytes. However, the importance of PCK1 activation and lipid storage in the adaptive response to HS is unknown. Therefore, in vitro experiments were conducted to investigate the effect of PCK1 inhibition with 3-mercaptopicolinic acid (3MPA) on lipid storage and adipocyte response during HS. In vitro culture of adipocytes under HS (41.0 °C) increased (P cultured adipocytes were less able to induce adaptive responses such as upregulation of HSP70 and triglycerides, and this exacerbated ER stress during HS. Thus, PCK1 may function to alleviate ER stress that occurs during HS.

  10. Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport.

    Science.gov (United States)

    Makiuchi, Takashi; Mi-ichi, Fumika; Nakada-Tsukui, Kumiko; Nozaki, Tomoyoshi

    2013-01-01

    Under anaerobic environments, the mitochondria have undergone remarkable reduction and transformation into highly reduced structures, referred as mitochondrion-related organelles (MROs), which include mitosomes and hydrogenosomes. In agreement with the concept of reductive evolution, mitosomes of Entamoeba histolytica lack most of the components of the TOM (translocase of the outer mitochondrial membrane) complex, which is required for the targeting and membrane translocation of preproteins into the canonical aerobic mitochondria. Here we showed, in E. histolytica mitosomes, the presence of a 600-kDa TOM complex composed of Tom40, a conserved pore-forming subunit, and Tom60, a novel lineage-specific receptor protein. Tom60, containing multiple tetratricopeptide repeats, is localized to the mitosomal outer membrane and the cytosol, and serves as a receptor of both mitosomal matrix and membrane preproteins. Our data indicate that Entamoeba has invented a novel lineage-specific shuttle receptor of the TOM complex as a consequence of adaptation to an anaerobic environment.

  11. Cytosolic fatty acid-binding proteins: subjects and tools in metabolic research

    Energy Technology Data Exchange (ETDEWEB)

    Binas, B. [Max Delbrueck Center for Molecular Medicine, Berlin-Buch (Germany)

    1998-12-31

    Fatty acid-binding proteins (FABPs) are major targets for specific binding of fatty acids in vivo. They constitute a widely expressed family of genetically related, small cytosolic proteins which very likely mediate intracellular transport of free long chain fatty acids. Genetic inhibition of FABP expression in vivo should therefore provide a useful tool to investigate and engineer fatty acid metabolism. (orig.) [Deutsch] Fettsaeurebindungsproteine (FABPs) sind wichtige Bindungsstellen fuer Fettsaeuren in vivo; sie bilden eine breit exprimierte Familie genetisch verwandter kleiner Zytosoleiweisse, die sehr wahrscheinlich den intrazellulaeren Transport unveresterter langkettiger Fettsaeuren vermitteln. Die genetische Hemmung der FABP-Expanssion in vivo bietet sich deshalb als Werkzeug zur Erforschung und gezielten Veraenderung des Fettsaeurestoffwechsels an. (orig.)

  12. On Buffer-Aided Multiple-Access Relay Channel

    DEFF Research Database (Denmark)

    Liu, Rongkuan; Popovski, Petar; Wang, Gang

    2016-01-01

    This letter treats uplink scenario where M user equipments (UEs) send to a base station (BS), possibly via a common relay station (RS) that is equipped with a buffer. This is a multiple-access relay channel aided by a buffer. We devise a protocol in which the transmission mode is selected...... entirely at the BS, while simultaneously a number of UEs sends new messages to the BS. The results show that the adaptive selection of direct and buffer-aided relay transmissions leads to significant average throughput gains....

  13. Lean buffering in serial production lines with Bernoulli machines

    Directory of Open Access Journals (Sweden)

    A. B. Hu

    2006-01-01

    Full Text Available Lean buffering is the smallest buffer capacity necessary to ensure the desired production rate of a manufacturing system. In this paper, analytical methods for selecting lean buffering in serial production lines are developed under the assumption that the machines obey the Bernoulli reliability model. Both closed-form expressions and recursive approaches are investigated. The cases of identical and nonidentical machines are analyzed. Results obtained can be useful for production line designers and production managers to maintain the required production rate with the smallest possible inventories.

  14. Buffer-gas-induced absorption resonances in Rb vapor

    International Nuclear Information System (INIS)

    Mikhailov, Eugeniy E.; Novikova, Irina; Rostovtsev, Yuri V.; Welch, George R.

    2004-01-01

    We observe transformation of the electromagnetically induced transparency (EIT) resonance into an absorption resonance in a Λ interaction configuration in a cell filled with 87 Rb and a buffer gas. This transformation occurs as one-photon detuning of the coupling fields is varied from the atomic transition. No such absorption resonance is found in the absence of a buffer gas. The width of the absorption resonance is several times smaller than the width of the EIT resonance, and the changes of absorption near these resonances are about the same. Similar absorption resonances are detected in the Hanle configuration in a buffered cell

  15. Buffer layers for REBCO films for use in superconducting devices

    Science.gov (United States)

    Goyal, Amit; Wee, Sung-Hun

    2014-06-10

    A superconducting article includes a substrate having a biaxially textured surface. A biaxially textured buffer layer, which can be a cap layer, is supported by the substrate. The buffer layer includes a double perovskite of the formula A.sub.2B'B''O.sub.6, where A is rare earth or alkaline earth metal and B' and B'' are different transition metal cations. A biaxially textured superconductor layer is deposited so as to be supported by the buffer layer. A method of making a superconducting article is also disclosed.

  16. Stacking by electroinjection with discontinuous buffers in capillary zone electrophoresis.

    Science.gov (United States)

    Shihabi, Zak K

    2002-08-01

    The work presented here demonstrates that electroinjection can be performed using discontinuous buffers, which can result in better stacking than that obtained by hydrodynamic injection. The sample can be concentrated at the tip of the capillary leaving practically the whole capillary for sample separation. This results in several advantages, such as better sample concentration, higher plate number and shorter time of stacking. However, sample introduction by electromigration is suited for samples free or low in salt content. Samples, which are high in salt content, are better introduced by the hydrodynamic injection for stacking by the discontinuous buffers. Different simple methods to introduce the discontinuity in the buffer for electroinjection are discussed.

  17. Localization of age-related macular degeneration-associated ARMS2 in cytosol, not mitochondria

    Science.gov (United States)

    Wang, Gaofeng; Spencer, Kylee L.; Court, Brenda L.; Olson, Lana M.; Scott, William K.; Haines, Jonathan L.; Pericak-Vance, Margaret A.

    2010-01-01

    PURPOSE To analyze the relationship between ARMS2 and HTRA1 in the association with age-related macular degeneration (AMD) in an independent case-control dataset, and to investigate the subcellular localization of the ARMS2 protein in an in vitro system. METHOD Two SNPs in ARMS2 and HTRA1 were genotyped in 685 cases and 269 controls by Taqman Assay. Allelic association was tested by a χ2 test. A likelihood ratio test (LRT) of full vs. reduced models was utilized to analyze the interaction between ARMS2 and smoking and HTRA1 and smoking, after adjusting for CFH and age. Immunofluorescence and immunoblot were applied to localize ARMS2 in retinal epithelial ARPE-19 cells and COS7 cell transfected by ARMS2 constructs. RESULT Both significantly associated SNP rs10490924 and rs11200638 (P<0.0001) are in strong linkage disequilibrium (LD) (D′=0.97, r2=0.93) that generates virtually identical association test and odds ratios. In separate logistic regression models the interaction effect for both smoking with ARMS2 and with HTRA1 was not statistically significant. Immunofluorescence and immunoblot show that both endogenous and exogenous ARMS2 are mainly distributed in the cytosol, not the mitochondria. Comparing to wild type, ARMS2 A69S is more likely to be associated with cytoskeleton in COS7 cells. CONCLUSIONS The significant associations in ARMS2 and HTRA1 are with polymorphisms in strong LD that confer virtually identical risks, preventing differentiation at the statistical level. We found that ARMS2 was mainly distributed in the cytosol, not in mitochondrial outer membrane as previously reported, suggesting that ARMS2 may not confer risk to AMD through the mitochondrial pathway. PMID:19255159

  18. Dose enhancement effects to the nucleus and mitochondria from gold nanoparticles in the cytosol

    Science.gov (United States)

    McNamara, AL; Kam, WW-Y; Scales, N; McMahon, SJ; Bennett, JW; Byrne, HL; Schuemann, J; Paganetti, H; Banati, R; Kuncic, Z

    2016-01-01

    Gold nanoparticles (GNPs) have shown potential as dose enhancers for radiation therapy. Since damage to the genome affects the viability of a cell, it is generally assumed that GNPs have to localise within the cell nucleus. In practice, however, GNPs tend to localise in the cytoplasm yet still appear to have a dose enhancing effect on the cell. Whether this effect can be attributed to stress-induced biological mechanisms or to physical damage to extra-nuclear cellular targets is still unclear. There is however growing evidence to suggest that the cellular response to radiation can also be influenced by indirect processes induced when the nucleus is not directly targeted by radiation. The mitochondrion in particular may be an effective extra-nuclear radiation target given its many important functional roles in the cell. To more accurately predict the physical effect of radiation within different cell organelles, we measured the full chemical composition of a whole human lymphocytic JURKAT cell as well as two separate organelles; the cell nucleus and the mitochondrion. The experimental measurements found that all three biological materials had similar ionisation energies ~ 70 eV, substantially lower than that of liquid water ~ 78 eV. Monte Carlo simulations for 10 – 50 keV incident photons showed higher energy deposition and ionisation numbers in the cell and organelle materials compared to liquid water. Adding a 1% mass fraction of gold to each material increased the energy deposition by a factor of ~ 1.8 when averaged over all incident photon energies. Simulations of a realistic compartmentalised cell show that the presence of gold in the cytosol increases the energy deposition in the mitochondrial volume more than within the nuclear volume. We find this is due to sub-micron delocalisation of energy by photoelectrons, making the mitochondria a potentially viable indirect radiation target for GNPs that localise to the cytosol. PMID:27435339

  19. Expression of cytosolic NADP(+)-dependent isocitrate dehydrogenase in melanocytes and its role as an antioxidant.

    Science.gov (United States)

    Kim, Ji Young; Shin, Jae Yong; Kim, Miri; Hann, Seung-Kyung; Oh, Sang Ho

    2012-02-01

    Cytosolic NADP(+)-dependent ICDH (IDPc) has an antioxidant effect as a supplier of NADPH to the cytosol, which is needed for the production of glutathione. To evaluate the expression of IDPc in melanocytes and to elucidate its role as an antioxidant. The knock-down of IDPc expression in immortalized mouse melanocyte cell lines (melan-a) was performed using the short interfering RNA (siRNA)-targeted gene silencing method. After confirming the silencing of IDPc expression with mRNA and protein levels, viability, apoptosis and necrosis, as well as ROS production in IDPc-silenced melanocytes were monitored under conditions of oxidative stress and non-stress. Also, the ratio of oxidized glutathione to total glutathione was examined, and whether the addition of glutathione recovered cell viability, decreased by oxidant stress, was checked. The expression of IDPc in both primary human melanocytes and melan-a cells was confirmed by Western blot and RT-PCR. The silencing of IDPc expression by transfecting IDPc siRNA in melan-a cells was observed by Western blotting and real-time RT-PCR. IDPc knock-down cells showed significantly decreased cell viability and an increased number of cells under apoptosis and necrosis. IDPc siRNA-treated melanocytes demonstrated a higher intensity of DCFDA after the addition of H(2)O(2) compared with scrambled siRNA-treated melanocytes, and a lower ratio of reduced glutathione to oxidized glutathione were observed in IDPc siRNA transfected melanocytes. In addition, the addition of glutathione recovered cell viability, which was previously decreased after incubation with H(2)O(2). This study suggests that decreased IDPc expression renders melanocytes more vulnerable to oxidative stress, and IDPc plays an important antioxidant function in melanocytes. Copyright © 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

  20. AIM2-Like Receptors Positively and Negatively Regulate the Interferon Response Induced by Cytosolic DNA.

    Science.gov (United States)

    Nakaya, Yuki; Lilue, Jingtao; Stavrou, Spyridon; Moran, Eileen A; Ross, Susan R

    2017-07-05

    Cytosolic DNAs derived from retrotransposons serve as pathogen-associated molecular patterns for pattern recognition receptors (PRRs) that stimulate the induction of interferons (IFNs) and other cytokines, leading to autoimmune disease. Cyclic GMP-AMP synthase is one PRR that senses retrotransposon DNA, activating type I IFN responses through the stimulator of IFN genes (STING). Absent in melanoma 2 (AIM2)-like receptors (ALRs) have also been implicated in these pathways. Here we show that the mouse ALR IFI205 senses cytosolic retrotransposon DNA independently of cyclic GMP-AMP production. AIM2 antagonizes IFI205-mediated IFN induction activity by sequestering it from STING. We also found that the complement of genes located in the ALR locus in C57BL/6 and AIM2 knockout mice are different and unique, which has implications for interpretation of the sensing of pathogens in different mouse strains. Our data suggest that members of the ALR family are critical to the host IFN response to endogenous DNA. IMPORTANCE Autoimmune diseases like Aicardi-Goutières syndrome and lupus erythematosus arise when cells of the immune system become activated and attack host cells and tissues. We found that DNA generated by endogenous retroviruses and retroelements in inbred mice and mouse cells is recognized by several host proteins found in macrophages that are members of the ALR family and that these proteins both suppress and activate the pathways leading to the generation of cytokines and IFNs. We also show that there is great genetic diversity between different inbred mouse strains in the ALR genes, which might contribute to differential susceptibility to autoimmunity. Understanding how immune cells become activated is important to the control of disease. Copyright © 2017 Nakaya et al.

  1. Stereoselective sulfate conjugation of racemic 4-hydroxypropranolol by human and rat liver cytosol

    Energy Technology Data Exchange (ETDEWEB)

    Walle, T.; Walle, U.K. (Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston (USA))

    1991-03-01

    The objective of this study was to determine the stereochemistry of sulfoconjugation of a chiral phenolic amine drug, 4-hydroxypropranolol (HOP), by the human liver. The reaction was catalyzed by the 100,000 g cytosol as the phenolsulfotransferase (PST) enzyme source with PAP35S as the co-substrate. The enantiomers of the intact sulfate conjugate formed, (+)-HOP35S and (-)-HOP35S, were separated by HPLC and measured by liquid scintillation spectrometry. Complex velocity vs. substrate concentration curves were obtained with two peaks of activity, one at 3 microM (high affinity) and one at 500 microM (low affinity). The high-affinity reaction demonstrated a high degree of stereoselectivity. Whereas the affinity of the enantiomers for this reaction was identical, with a very low apparent KM value of 0.59 microM, the apparent Vmax value for (+)-HOPS formation was 4.6-fold higher than for (-)-HOPS. In sharp contrast, the low-affinity reaction, with an apparent KM of 65 microM, was not stereoselective. Inhibition of the high-affinity reaction by elevated temperature, but not by dichloronitrophenol, indicated that this activity was due to a monoamine form of PST. Inhibition of the low-affinity reaction by dichloronitrophenol, but not by elevated temperature, indicated that this activity was due to a phenol form of PST. As a comparison, experiments with the rat liver cytosol demonstrated only one activity, with apparent KM values of 50 microM for both enantiomers and opposite stereoselectivity in maximum velocity compared to humans, {plus minus}-HOPS ratio 0.72. The results of this study demonstrate stereoselectivity in human hepatic sulfation of a chiral phenolic amine, with clear differences between PST isoenzymes.

  2. Cytosolic calcium rises and related events in ergosterol-treated Nicotiana cells.

    Science.gov (United States)

    Vatsa, Parul; Chiltz, Annick; Luini, Estelle; Vandelle, Elodie; Pugin, Alain; Roblin, Gabriel

    2011-07-01

    The typical fungal membrane component ergosterol was previously shown to trigger defence responses and protect plants against pathogens. Most of the elicitors mobilize the second messenger calcium, to trigger plant defences. We checked the involvement of calcium in response to ergosterol using Nicotiana plumbaginifolia and Nicotiana tabacum cv Xanthi cells expressing apoaequorin in the cytosol. First, it was verified if ergosterol was efficient in these cells inducing modifications of proton fluxes and increased expression of defence-related genes. Then, it was shown that ergosterol induced a rapid and transient biphasic increase of free [Ca²⁺](cyt) which intensity depends on ergosterol concentration in the range 0.002-10 μM. Among sterols, this calcium mobilization was specific for ergosterol and, ergosterol-induced pH and [Ca²⁺](cyt) changes were specifically desensitized after two subsequent applications of ergosterol. Specific modulators allowed elucidating some events in the signalling pathway triggered by ergosterol. The action of BAPTA, LaCl₃, nifedipine, verapamil, neomycin, U73122 and ruthenium red suggested that the first phase was linked to calcium influx from external medium which subsequently triggered the second phase linked to calcium release from internal stores. The calcium influx and the [Ca²⁺](cyt) increase depended on upstream protein phosphorylation. The extracellular alkalinization and ROS production depended on calcium influx but, the ergosterol-induced MAPK activation was calcium-independent. ROS were not involved in cytosolic calcium rise as described in other models, indicating that ROS do not systematically participate in the amplification of calcium signalling. Interestingly, ergosterol-induced ROS production is not linked to cell death and ergosterol does not induce any calcium elevation in the nucleus. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  3. Cytosolic 5'-nucleotidase II interacts with the leucin rich repeat of NLR family member Ipaf.

    Directory of Open Access Journals (Sweden)

    Federico Cividini

    Full Text Available IMP/GMP preferring cytosolic 5'-nucleotidase II (cN-II is a bifunctional enzyme whose activities and expression play crucial roles in nucleotide pool maintenance, nucleotide-dependent pathways and programmed cell death. Alignment of primary amino acid sequences of cN-II from human and other organisms show a strong conservation throughout the entire vertebrata taxon suggesting a fundamental role in eukaryotic cells. With the aim to investigate the potential role of this homology in protein-protein interactions, a two hybrid system screening of cN-II interactors was performed in S. cerevisiae. Among the X positive hits, the Leucin Rich Repeat (LRR domain of Ipaf was found to interact with cN-II. Recombinant Ipaf isoform B (lacking the Nucleotide Binding Domain was used in an in vitro affinity chromatography assay confirming the interaction obtained in the screening. Moreover, co-immunoprecipitation with proteins from wild type Human Embryonic Kidney 293 T cells demonstrated that endogenous cN-II co-immunoprecipitated both with wild type Ipaf and its LRR domain after transfection with corresponding expression vectors, but not with Ipaf lacking the LRR domain. These results suggest that the interaction takes place through the LRR domain of Ipaf. In addition, a proximity ligation assay was performed in A549 lung carcinoma cells and in MDA-MB-231 breast cancer cells and showed a positive cytosolic signal, confirming that this interaction occurs in human cells. This is the first report of a protein-protein interaction involving cN-II, suggesting either novel functions or an additional level of regulation of this complex enzyme.

  4. SAXS analysis of a soluble cytosolic NgBR construct including extracellular and transmembrane domains.

    Directory of Open Access Journals (Sweden)

    Joshua Holcomb

    Full Text Available The Nogo-B receptor (NgBR is involved in oncogenic Ras signaling through directly binding to farnesylated Ras. It recruits farnesylated Ras to the non-lipid-raft membrane for interaction with downstream effectors. However, the cytosolic domain of NgBR itself is only partially folded. The lack of several conserved secondary structural elements makes this domain unlikely to form a complete farnesyl binding pocket. We find that inclusion of the extracellular and transmembrane domains that contain additional conserved residues to the cytosolic region results in a well folded protein with a similar size and shape to the E.coli cis-isoprenyl transferase (UPPs. Small Angle X-ray Scattering (SAXS analysis reveals the radius of gyration (Rg of our NgBR construct to be 18.2 Å with a maximum particle dimension (Dmax of 61.0 Å. Ab initio shape modeling returns a globular molecular envelope with an estimated molecular weight of 23.0 kD closely correlated with the calculated molecular weight. Both Kratky plot and pair distribution function of NgBR scattering reveal a bell shaped peak which is characteristic of a single globularly folded protein. In addition, circular dichroism (CD analysis reveals that our construct has the secondary structure contents similar to the UPPs. However, this result does not agree with the currently accepted topological orientation of NgBR which might partition this construct into three separate domains. This discrepancy suggests another possible NgBR topology and lends insight into a potential molecular basis of how NgBR facilitates farnesylated Ras recruitment.

  5. Towards Optimal Buffer Size in Wi-Fi Networks

    KAUST Repository

    Showail, Ahmad

    2016-01-01

    extensively for wired networks. However, there is little work addressing the unique challenges of wireless environment. In this dissertation, we discuss buffer sizing challenges in wireless networks, classify the state-of-the-art solutions, and propose two

  6. Studies of Latent Acidity and Neutral Buffered Chloroaluminate Ionic Liquids

    National Research Council Canada - National Science Library

    Osteryoung, Robert

    2000-01-01

    Studies on ionic liquids composed of aluminum chloride and 1-ethyl-3-methylimidazolium chloride were carried out, with emphasis on understanding and explaining acidity and latent acidity in "neutral buffered" melts...

  7. EnviroAtlas - Phoenix, AZ - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  8. EnviroAtlas - Tampa, FL - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  9. Full scale tests of moisture buffer capacity of wall materials

    DEFF Research Database (Denmark)

    Mortensen, Lone Hedegaard; Rode, Carsten; Peuhkuri, Ruut Hannele

    2005-01-01

    that are harmful such as growth of house dust mites, surface condensation and mould growth. Therefore a series of experiments has been carried out in a full scale test facility to determine the moisture buffer effect of interior walls of cellular concrete and plaster board constructions. For the cellular concrete......Moisture buffer capacity of hygroscopic materials can be used to moderate peaks in the relative humidity (RH) of indoor air as well as moisture content variations in building materials and furnishing. This can help to ensure healthier indoor environments by preventing many processes...... of the changes of moisture content in specimens of the wall composites exposed to the same environment. It was found that the finishes had a big impact on the buffer performance of the underlying materials. Even though the untreated cellular concrete had a very high buffer capacity, the effect was strongly...

  10. EnviroAtlas - Woodbine, IA - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  11. Buffer management in wireless full-duplex systems

    KAUST Repository

    Bouacida, Nader; Showail, Ahmad; Shihada, Basem

    2015-01-01

    , we address the problem of buffer management in full-duplex networks by using Wireless Queue Management (WQM), which is an active queue management technique for wireless networks. Our solution is based on Relay Full-Duplex MAC (RFD

  12. Design and analysis of hybrid optical and electronic buffer based ...

    Indian Academy of Sciences (India)

    ARUNENDRA SINGH

    2018-03-10

    Mar 10, 2018 ... need and requirement of internet traffic is increasing in a very rapid pace. ... infrastructure of telecommunication and data networking. One of the significant .... packet loss rate, large buffer space would be required, and.

  13. EnviroAtlas - Paterson, NJ - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  14. Swelling of the buffer of KBS-3V deposition hole

    International Nuclear Information System (INIS)

    Lempinen, A.

    2006-12-01

    At the time of the installation of spent nuclear fuel canister in the KBS-3V deposition hole, empty space is left around bentonite buffer for technical reasons. The gap between the buffer and the canister is about 10 mm, and the gap between the buffer and the rock is 30 to 35 mm. In this study, the swelling of the buffer to fill the gaps was simulated, when the gaps are initially filled with water and no external water is available. The model used here is a thermodynamical model for swelling clay, with parameters determined for bentonite. The simulations presented here were performed with Freefem++ software, which is a finite element application for partial differential equations. These equations come from the material model. The simulation results show that the swelling fills the outer gaps in few years, but no significant swelling pressure is generated. For swelling pressure, external water supply is required. (orig.)

  15. EnviroAtlas - New York, NY - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  16. EnviroAtlas - Durham, NC - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  17. EnviroAtlas - Austin, TX - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  18. EnviroAtlas - Green Bay, WI - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  19. Stacking and discontinuous buffers in capillary zone electrophoresis.

    Science.gov (United States)

    Shihabi, Z K

    2000-08-01

    Discontinuous buffers for capillary zone electrophoresis (CZE) can be used under less rigid conditions compared to those for isotachophoresis for stacking. They can be prepared simply by modifying the sample itself, either by addition of small inorganic ions, low conductivity diluents, or both, and also by adjusting its pH, meanwhile injecting a large volume on the capillary. Zwitterionic and organic-based buffers such as triethanolamine and tris(hydroxymethyl)aminomethane (Tris) are well suited for stacking due to their low conductivity, provided the buffer is discontinuous as demonstrated here. A simple mechanism based on discontinuous buffers is described to explain many of the observed stacking types in CZE, pointing out the many similarities to transient isotachophoresis.

  20. Optimizing the availability of a buffered industrial process

    Science.gov (United States)

    Martz, Jr., Harry F.; Hamada, Michael S.; Koehler, Arthur J.; Berg, Eric C.

    2004-08-24

    A computer-implemented process determines optimum configuration parameters for a buffered industrial process. A population size is initialized by randomly selecting a first set of design and operation values associated with subsystems and buffers of the buffered industrial process to form a set of operating parameters for each member of the population. An availability discrete event simulation (ADES) is performed on each member of the population to determine the product-based availability of each member. A new population is formed having members with a second set of design and operation values related to the first set of design and operation values through a genetic algorithm and the product-based availability determined by the ADES. Subsequent population members are then determined by iterating the genetic algorithm with product-based availability determined by ADES to form improved design and operation values from which the configuration parameters are selected for the buffered industrial process.

  1. RESEARCH SHOWS IMPORTANCE OF RIPARIAN BUFFERS FOR AQUATIC HEALTH

    Science.gov (United States)

    Issue: Excess nitrogen from fertilizer, septic tanks, animal feedlots, and runoff from pavement can threaten aquatic ecosystem health. Riparian buffers -- the vegetated region adjacent to streams and wetlands -- are thought to be effective at intercepting and controlling excess ...

  2. EnviroAtlas - Des Moines, IA - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  3. EnviroAtlas - Portland, ME - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  4. EnviroAtlas - Portland, OR - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  5. EnviroAtlas - Cleveland, OH - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  6. EnviroAtlas - Memphis, TN - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  7. EnviroAtlas - Fresno, CA - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  8. EnviroAtlas - New Bedford, MA - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  9. EnviroAtlas - Pittsburgh, PA - Near Road Tree Buffer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This EnviroAtlas dataset addresses the tree buffer along heavily traveled roads. The roads are interstates, arterials, and collectors within the EnviroAtlas...

  10. The buffer/container experiment design and construction report

    Energy Technology Data Exchange (ETDEWEB)

    Chandler, N.A.; Wan, A.W.L.; Roach, P.J

    1998-03-01

    The Buffer/Container Experiment was a full-scale in situ experiment, installed at a depth of 240 m in granitic rock at AECL's Underground Research Laboratory (URL). The experiment was designed to examine the performance of a compacted sand-bentonite buffer material under the influences of elevated temperature and in situ moisture conditions. Buffer material was compacted in situ into a 5-m-deep, 1.24-m-diameter borehole drilled into the floor of an excavation. A 2.3-m long heater, representative of a nuclear fuel waste container, was placed within the buffer, and instrumentation was installed to monitor changes in buffer moisture conditions, temperature and stress. The experiment was sealed at the top of the borehole and restrained against vertical displacement. Instrumentation in the rock monitored pore pressures, temperatures and rock displacement. The heater was operated at a constant power of 1200 W, which provided a heater skin temperature of approximately 85 degrees C. Experiment construction and installation required two years, followed by two and a half years of heater operation and two years of monitoring the rock conditions during cooling. The construction phase of the experiment included the design, construction and testing of a segmental heater and controller, geological and hydrogeological characterization of the rock, excavation of the experiment room, drilling of the emplacement borehole using high pressure water, mixing and in situ compaction of buffer material, installation of instrumentation in the rock, buffer and on the heater, and the construction of concrete curb and steel vertical restraint system at the top of emplacement borehole. Upon completion of the experiment, decommissioning sampling equipment was designed and constructed and sampling methods were developed which allowed approximately 2000 samples of buffer material to be taken over a 12-day period. Quality assurance procedures were developed for all aspects of experiment

  11. Design of a flux buffer based on the flux shuttle

    International Nuclear Information System (INIS)

    Gershenson, M.

    1991-01-01

    This paper discusses the design considerations for a flux buffer based on the flux-shuttle concept. Particular attention is given to the issues of flux popping, stability of operation and saturation levels for a large input. Modulation techniques used in order to minimize 1/f noise, in addition to offsets are also analyzed. Advantages over conventional approaches using a SQUID for a flux buffer are discussed. Results of computer simulations are presented

  12. Current status of mechanical erosion studies of bentonite buffer

    International Nuclear Information System (INIS)

    Sane, P.; Olin, M.; Koskinen, K.

    2013-08-01

    The performance of the bentonite buffer in KBS-3-type nuclear waste repository concept relies to a great extent on the buffer surrounding the canister having sufficient dry density. Loss of buffer material caused by erosion remains as the most significant process reducing the density of the buffer. The mechanical erosion, or pre-saturation erosion, is the process where flowing groundwater transports buffer material away from the deposition hole towards the deposition tunnel. This process reduces the overall buffer density and potentially creates localized regions of low density. In the worst case the process is assumed to last as long as the free volume between the pellets in the pellets filled regions is filled with groundwater. With fixed environmental and material parameters a set of experiments was performed, testing the erosive properties of different buffer and backfill materials (MX-80 and Friedland Clay) in different groundwater conditions. The method used was a pinhole erosion test using two sizescales; 100 mm and 400 mm of cell length. The purpose of the pinhole tests was to test the scenario where piping channel is formed in the buffer and water flows through a single channel. The erosion data was produced with two methods, firstly the time-related erosion rates measured in-situ during the measurement and secondly the overall mass loss in the sample cell measured after dismantling of the test. It was observed that erosion in piping channels decreases rapidly (∼24 h) and irreversibly to a level that is an order of magnitude lower than the peak values. (orig.)

  13. Implementing the countercyclical capital buffer in South Africa: Practical considerations

    OpenAIRE

    Pravin Burra; Pieter Juriaan de Jongh; Helgard Raubenheimer; Gary van Vuuren; Henco Wiid

    2015-01-01

    The Basel II regulatory framework significantly increased the resilience of the banking system, but proved ineffective in preventing the 2008/9 financial crisis. The subsequent introduction of Basel III aimed, inter alia, to supplement bank capital using buffers. The countercyclical buffer boosts existing minimum capital requirements when systemic risk surges are detected. Bolstering capital in favourable economic conditions cushions losses in unfavourable conditions, thereby addressing capit...

  14. The buffer/container experiment design and construction report

    International Nuclear Information System (INIS)

    Chandler, N.A.; Wan, A.W.L.; Roach, P.J.

    1998-03-01

    The Buffer/Container Experiment was a full-scale in situ experiment, installed at a depth of 240 m in granitic rock at AECL's Underground Research Laboratory (URL). The experiment was designed to examine the performance of a compacted sand-bentonite buffer material under the influences of elevated temperature and in situ moisture conditions. Buffer material was compacted in situ into a 5-m-deep, 1.24-m-diameter borehole drilled into the floor of an excavation. A 2.3-m long heater, representative of a nuclear fuel waste container, was placed within the buffer, and instrumentation was installed to monitor changes in buffer moisture conditions, temperature and stress. The experiment was sealed at the top of the borehole and restrained against vertical displacement. Instrumentation in the rock monitored pore pressures, temperatures and rock displacement. The heater was operated at a constant power of 1200 W, which provided a heater skin temperature of approximately 85 degrees C. Experiment construction and installation required two years, followed by two and a half years of heater operation and two years of monitoring the rock conditions during cooling. The construction phase of the experiment included the design, construction and testing of a segmental heater and controller, geological and hydrogeological characterization of the rock, excavation of the experiment room, drilling of the emplacement borehole using high pressure water, mixing and in situ compaction of buffer material, installation of instrumentation in the rock, buffer and on the heater, and the construction of concrete curb and steel vertical restraint system at the top of emplacement borehole. Upon completion of the experiment, decommissioning sampling equipment was designed and constructed and sampling methods were developed which allowed approximately 2000 samples of buffer material to be taken over a 12-day period. Quality assurance procedures were developed for all aspects of experiment construction

  15. Traveling waves in the discrete fast buffered bistable system.

    Science.gov (United States)

    Tsai, Je-Chiang; Sneyd, James

    2007-11-01

    We study the existence and uniqueness of traveling wave solutions of the discrete buffered bistable equation. Buffered excitable systems are used to model, among other things, the propagation of waves of increased calcium concentration, and discrete models are often used to describe the propagation of such waves across multiple cells. We derive necessary conditions for the existence of waves, and, under some restrictive technical assumptions, we derive sufficient conditions. When the wave exists it is unique and stable.

  16. The Arabidopsis thalianaK+-uptake permease 7 (AtKUP7) contains a functional cytosolic adenylate cyclase catalytic centre

    KAUST Repository

    Al-Younis, Inas; Wong, Aloysius Tze; Gehring, Christoph A

    2015-01-01

    KUP7) as one of several candidate ACs. Firstly, we show that a recombinant N-terminal, cytosolic domain of AtKUP71-100 is able to complement the AC-deficient mutant cyaA in Escherichia coli and thus restoring the fermentation of lactose, and secondly

  17. Ca2+-mobilizing agonists increase mitochondrial ATP production to accelerate cytosolic Ca2+ removal: aberrations in human complex I deficiency.

    NARCIS (Netherlands)

    Visch, H.J.; Koopman, W.J.H.; Zeegers, D.; Emst-de Vries, S.E. van; Kuppeveld, F.J.M. van; Heuvel, L.W. van den; Smeitink, J.A.M.; Willems, P.H.G.M.

    2006-01-01

    Previously, we reported that both the bradykinin (Bk)-induced increase in mitochondrial ATP concentration ([ATP]M) and the rate of cytosolic Ca2+ removal are significantly decreased in skin fibroblasts from a patient with an isolated complex I deficiency. Here we demonstrate that the mitochondrial

  18. Ach1 is involved in shuttling mitochondrial acetyl units for cytosolic C2 provision in Saccharomyces cerevisiae lacking pyruvate decarboxylase

    DEFF Research Database (Denmark)

    Chen, Yun; Zhang, Yiming; Siewers, Verena

    2015-01-01

    Saccharomyces cerevisiae, acetyl-CoA is compartmentalized in the cytosol, mitochondrion, peroxisome and nucleus, and cannot be directly transported between these compartments. With the acetyl-carnitine or glyoxylate shuttle, acetyl-CoA produced in peroxisomes or the cytoplasm can be transported...

  19. Effect of C-terminal of human cytosolic thymidine kinase (TK1) on in vitro stability and enzymatic properties

    DEFF Research Database (Denmark)

    Munch-Petersen, Birgitte; Munch-Petersen, Sune; Berenstein, Dvora

    2006-01-01

    Thymidine kinase (TK1) is a key enzyme in the salvage pathway of nucleotide metabolism and catalyzes the first rate-limiting step in the synthesis of dTTP, transfer of a gamma-phosphate group from a nucleoside triphosphate to the 5′-hydroxyl group of thymidine, thus forming dTMP. TK1 is cytosolic...

  20. The chlamydial periplasmic stress response serine protease cHtrA is secreted into host cell cytosol

    Directory of Open Access Journals (Sweden)

    Flores Rhonda

    2011-04-01

    Full Text Available Abstract Background The periplasmic High Temperature Requirement protein A (HtrA plays important roles in bacterial protein folding and stress responses. However, the role of chlamydial HtrA (cHtrA in chlamydial pathogenesis is not clear. Results The cHtrA was detected both inside and outside the chlamydial inclusions. The detection was specific since both polyclonal and monoclonal anti-cHtrA antibodies revealed similar intracellular labeling patterns that were only removed by absorption with cHtrA but not control fusion proteins. In a Western blot assay, the anti-cHtrA antibodies detected the endogenous cHtrA in Chlamydia-infected cells without cross-reacting with any other chlamydial or host cell antigens. Fractionation of the infected cells revealed cHtrA in the host cell cytosol fraction. The periplasmic cHtrA protein appeared to be actively secreted into host cell cytosol since no other chlamydial periplasmic proteins were detected in the host cell cytoplasm. Most chlamydial species secreted cHtrA into host cell cytosol and the secretion was not inhibitable by a type III secretion inhibitor. Conclusion Since it is hypothesized that chlamydial organisms possess a proteolysis strategy to manipulate host cell signaling pathways, secretion of the serine protease cHtrA into host cell cytosol suggests that the periplasmic cHtrA may also play an important role in chlamydial interactions with host cells.

  1. THE CYTOSOLIC AND GLYCOSOMAL GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE FROM TRYPANOSOMA-BRUCEI - KINETIC-PROPERTIES AND COMPARISON WITH HOMOLOGOUS ENZYMES

    NARCIS (Netherlands)

    LAMBEIR, AM; LOISEAU, AM; KUNTZ, DA; VELLIEUX, FM; MICHELS, PAM; OPPERDOES, FR

    1991-01-01

    The protozoan haemoflagellate Trypanosoma brucei has two NAD-dependent glyceraldehyde-3-phosphate dehydrogenase isoenzymes, each with a different localization within the cell. One isoenzyme is found in the cytosol, as in other eukaryotes, while the other is found in the glycosome, a microbody-like

  2. Cutting edge: HLA-B27 acquires many N-terminal dibasic peptides: coupling cytosolic peptide stability to antigen presentation

    NARCIS (Netherlands)

    Herberts, Carla A.; Neijssen, Joost J.; de Haan, Jolanda; Janssen, Lennert; Drijfhout, Jan Wouter; Reits, Eric A.; Neefjes, Jacques J.

    2006-01-01

    Ag presentation by MHC class I is a highly inefficient process because cytosolic peptidases destroy most peptides after proteasomal generation. Various mechanisms shape the MHC class I peptidome. We define a new one: intracellular peptide stability. Peptides with two N-terminal basic amino acids are

  3. Deflating link buffers in a wireless mesh network

    KAUST Repository

    Jamshaid, Kamran; Shihada, Basem; Showail, Ahmad; Levis, Philip

    2014-01-01

    We analyze the problem of buffer sizing for backlogged TCP flows in 802.11-based wireless mesh networks. Our objective is to maintain high network utilization while providing low queueing delays. Unlike wired networks where a single link buffer feeds a bottleneck link, the radio spectral resource in a mesh network is shared among a set of contending mesh routers. We account for this by formulating the buffer size problem as sizing a collective buffer distributed over a set of interfering nodes. In this paper we propose mechanisms for sizing and distributing this collective buffer among the mesh nodes constituting the network bottleneck. Our mechanism factors in the network topology and wireless link rates, improving on pre-set buffer allocations that cannot optimally work across the range of configurations achievable with 802.11 radios. We evaluate our mechanisms using simulations as well as experiments on a testbed. Our results show that we can reduce the RTT of a flow by 6× or more, at the cost of less than 10% drop in end-to-end flow throughput.

  4. Deflating link buffers in a wireless mesh network

    KAUST Repository

    Jamshaid, Kamran

    2014-05-01

    We analyze the problem of buffer sizing for backlogged TCP flows in 802.11-based wireless mesh networks. Our objective is to maintain high network utilization while providing low queueing delays. Unlike wired networks where a single link buffer feeds a bottleneck link, the radio spectral resource in a mesh network is shared among a set of contending mesh routers. We account for this by formulating the buffer size problem as sizing a collective buffer distributed over a set of interfering nodes. In this paper we propose mechanisms for sizing and distributing this collective buffer among the mesh nodes constituting the network bottleneck. Our mechanism factors in the network topology and wireless link rates, improving on pre-set buffer allocations that cannot optimally work across the range of configurations achievable with 802.11 radios. We evaluate our mechanisms using simulations as well as experiments on a testbed. Our results show that we can reduce the RTT of a flow by 6× or more, at the cost of less than 10% drop in end-to-end flow throughput.

  5. Buffering capacity: its relevance in soil and water pollution

    Energy Technology Data Exchange (ETDEWEB)

    Stigliani, W.M. [University of Northern Iowa, Cedar Falls (United States)

    1996-02-01

    Buffering capacities in soils are essential for neutralizing or immobilizing inputs of acids and heavy metals. There are four major buffering regimes, each of which operates in a defined pH range. When the rate of pollutant inputs exceeds the rate of natural replenishment of the buffer in a specific regime, the supply of buffer will be exhausted over time, and the soil may switch to a regime at lower pH. As the pH of the soil declines, so does its capacity to immobilize heavy metals. If the soil initially has a moderately high buffering capacity, the time scale before exhaustion is on the order of decades to a century. Over this time there may be no observable environmental effects. When the effects do become obvious, it may be too late to reverse the damage. The importance of buffering capacity as a valued environmental resource must be recognized, and its preservation must be a major component of long-term soil protection policies. (Author). 17 refs., 6 figs., 1 tab.

  6. Mechanisms of social buffering of fear in zebrafish.

    Science.gov (United States)

    Faustino, Ana I; Tacão-Monteiro, André; Oliveira, Rui F

    2017-03-31

    Some humans thrive whereas others resign when exposed to threatening situations throughout life. Social support has been identified as an important modulator of these discrepancies in human behaviour, and other social animals also exhibit phenomena in which individuals recover better from aversive events when conspecifics are present - aka social buffering. Here we studied social buffering in zebrafish, by exposing focal fish to an aversive stimulus (alarm substance - AS) either in the absence or presence of conspecific cues. When exposed to AS in the presence of both olfactory (shoal water) and visual (sight of shoal) conspecific cues, focal fish exhibited a lower fear response than when tested alone, demonstrating social buffering in zebrafish. When separately testing each cue's effectiveness, we verified that the visual cue was more effective than the olfactory in reducing freezing in a persistent threat scenario. Finally, we verified that social buffering was independent of shoal size and coincided with a distinct pattern of co-activation of brain regions known to be involved in mammalian social buffering. Thus, this study suggests a shared evolutionary origin for social buffering in vertebrates, bringing new evidence on the behavioural, sensory and neural mechanisms underlying this phenomenon.

  7. Design, production and initial state of the buffer

    Energy Technology Data Exchange (ETDEWEB)

    Boerjesson, Lennart; Gunnarsson, David; Johannesson, Lars-Erik; Jonsson, Esther

    2010-12-15

    The report is included in a set of Production reports, presenting how the KBS-3 repository is designed, produced and inspected. The set of reports is included in the safety report for the KBS-3 repository and repository facility. The report provides input on the initial state of the buffer for the assessment of the long-term safety, SR-Site. The initial state refers to the properties of the engineered barriers once they have been finally placed in the KBS-3 repository and will not be further handled within the repository facility. In addition, the report provides input to the operational safety report, SR-Operation, on how the buffer shall be handled and installed. The report presents the design premises and reference design of the buffer and verifies the conformity of the reference design to the design premises. It also describes the production of the buffer, from excavation and delivery of buffer material to installation in the deposition hole. Finally, the initial state of the buffer and its conformity to the reference design and design premises is presented

  8. Effects of buffer thickness on ATW blanket performances

    International Nuclear Information System (INIS)

    Yang, Won Sik

    2001-01-01

    This paper presents the preliminary results of target and buffer design studies for a lead-bismuth eutectic (LBE) cooled accelerator transmutation of waste (ATW) system, aimed at maximizing the source importance while simultaneously reducing the irradiation damage to fuel. Using an 840 MWt LBE cooled ATW design, the effects of buffer thickness on the blanket performances have been studied. Varying the buffer thickness for a given blanket configuration, system performances have been estimated by a series of calculations using MCNPX and REBUS-3 codes. The effects of source importance change are studied by investigating the low-energy (< 20 MeV) neutron source distribution and the equilibrium cycle blanket performance parameters such as fuel inventory, discharge burnup, burnup reactivity loss, and peak fast fluence. As the irradiation damage to fuel, the displacements per atom (dpa), hydrogen production, and helium production rates are evaluated at the buffer and blanket interface where the peak fast fluence occurs. The results show that the damage rates and the source importance increase monotonically as the buffer thickness decreases. Based on a compromise between the competing objectives of increasing the source importance and reducing the damage rates, a buffer thickness of around 20 cm appears to be reasonable

  9. Effects of buffer ionization in protein transition volumes.

    Science.gov (United States)

    Lee, Soyoung; Heerklotz, Heiko; Chalikian, Tigran V

    2010-05-01

    Protein denaturation events are generally associated with a change in the state of ionization of abnormally titrating groups and, therefore, are coupled with changes in buffer ionization/neutralization equilibria. Consequently, buffer ionization should influence the measured change in volume accompanying protein denaturation. Changes in volume accompanying protein denaturation reflect the differential packing and hydration of polypeptide chains in their native and denatured conformations while also describing the pressure stability of proteins. A characteristic feature of conformational transitions of globular proteins is a near zero change in volume that is comparable in magnitude with the volume of ionization of biologically relevant buffers. Thus, the impact of buffer ionization on the volume of protein denaturation could be very significant with the potential to affect not only its magnitude but also its sign. To investigate this point quantitatively, we performed pressure perturbation calorimetric (PPC) studies of lysozyme and ribonuclease A at pH 3.0 in four buffers differing in their ionization volumes. Our results identify buffer ionization as an important determinant of protein transition volume that needs to be carefully taken into account. We emphasize that the importance of our results is not limited to PPC measurements but is more general and applies to all volumetric investigations, in particular, extending to the derivation of the pressure-temperature phase diagram of protein stability.

  10. Phylogenetic Analysis of Nucleus-Encoded Acetyl-CoA Carboxylases Targeted at the Cytosol and Plastid of Algae.

    KAUST Repository

    Huerlimann, Roger

    2015-07-01

    The understanding of algal phylogeny is being impeded by an unknown number of events of horizontal gene transfer (HGT), and primary and secondary/tertiary endosymbiosis. Through these events, previously heterotrophic eukaryotes developed photosynthesis and acquired new biochemical pathways. Acetyl-CoA carboxylase (ACCase) is a key enzyme in the fatty acid synthesis and elongation pathways in algae, where ACCase exists in two locations (cytosol and plastid) and in two forms (homomeric and heteromeric). All algae contain nucleus-encoded homomeric ACCase in the cytosol, independent of the origin of the plastid. Nucleus-encoded homomeric ACCase is also found in plastids of algae that arose from a secondary/tertiary endosymbiotic event. In contrast, plastids of algae that arose from a primary endosymbiotic event contain heteromeric ACCase, which consists of three nucleus-encoded and one plastid-encoded subunits. These properties of ACCase provide the potential to inform on the phylogenetic relationships of hosts and their plastids, allowing different hypothesis of endosymbiotic events to be tested. Alveolata (Dinoflagellata and Apicomplexa) and Chromista (Stramenopiles, Haptophyta and Cryptophyta) have traditionally been grouped together as Chromalveolata, forming the red lineage. However, recent genetic evidence groups the Stramenopiles, Alveolata and green plastid containing Rhizaria as SAR, excluding Haptophyta and Cryptophyta. Sequences coding for plastid and cytosol targeted homomeric ACCases were isolated from Isochrysis aff. galbana (TISO), Chromera velia and Nannochloropsis oculata, representing three taxonomic groups for which sequences were lacking. Phylogenetic analyses show that cytosolic ACCase strongly supports the SAR grouping. Conversely, plastidial ACCase groups the SAR with the Haptophyta, Cryptophyta and Prasinophyceae (Chlorophyta). These two ACCase based, phylogenetic relationships suggest that the plastidial homomeric ACCase was acquired by the

  11. Faster and stronger manifestation of mitochondrial diseases in skeletal muscle than in heart related to cytosolic inorganic phosphate (Pi) accumulation.

    Science.gov (United States)

    Korzeniewski, Bernard

    2016-08-01

    A model of the cell bioenergetic system was used to compare the effect of oxidative phosphorylation (OXPHOS) deficiencies in a broad range of moderate ATP demand in skeletal muscle and heart. Computer simulations revealed that kinetic properties of the system are similar in both cases despite the much higher mitochondria content and "basic" OXPHOS activity in heart than in skeletal muscle, because of a much higher each-step activation (ESA) of OXPHOS in skeletal muscle than in heart. Large OXPHOS deficiencies lead in both tissues to a significant decrease in oxygen consumption (V̇o2) and phosphocreatine (PCr) and increase in cytosolic ADP, Pi, and H(+) The main difference between skeletal muscle and heart is a much higher cytosolic Pi concentration in healthy tissue and much higher cytosolic Pi accumulation (level) at low OXPHOS activities in the former, caused by a higher PCr level in healthy tissue (and higher total phosphate pool) and smaller Pi redistribution between cytosol and mitochondria at OXPHOS deficiency. This difference does not depend on ATP demand in a broad range. A much greater Pi increase and PCr decrease during rest-to-moderate work transition in skeletal muscle at OXPHOS deficiencies than at normal OXPHOS activity significantly slows down the V̇o2 on-kinetics. Because high cytosolic Pi concentrations cause fatigue in skeletal muscle and can compromise force generation in skeletal muscle and heart, this system property can contribute to the faster and stronger manifestation of mitochondrial diseases in skeletal muscle than in heart. Shortly, skeletal muscle with large OXPHOS deficiencies becomes fatigued already during low/moderate exercise. Copyright © 2016 the American Physiological Society.

  12. Ca2+ dependence of gluconeogenesis stimulation by glucagon at different cytosolic NAD+-NADH redox potentials

    Directory of Open Access Journals (Sweden)

    Marques-da-Silva A.C.

    1997-01-01

    Full Text Available The influence of Ca2+ on hepatic gluconeogenesis was measured in the isolated perfused rat liver at different cytosolic NAD+-NADH potentials. Lactate and pyruvate were the gluconeogenic substrates and the cytosolic NAD+-NADH potentials were changed by varying the lactate to pyruvate ratios from 0.01 to 100. The following results were obtained: a gluconeogenesis from lactate plus pyruvate was not affected by Ca2+-free perfusion (no Ca2+ in the perfusion fluid combined with previous depletion of the intracellular pools; gluconeogenesis was also poorly dependent on the lactate to pyruvate ratios in the range of 0.1 to 100; only for a ratio equal to 0.01 was a significantly smaller gluconeogenic activity observed in comparison to the other ratios. b In the presence of Ca2+, the increase in oxygen uptake caused by the infusion of lactate plus pyruvate at a ratio equal to 10 was the most pronounced one; in Ca2+-free perfusion the increase in oxygen uptake caused by lactate plus pyruvate infusion tended to be higher for all lactate to pyruvate ratios; the most pronounced difference was observed for a lactate/pyruvate ratio equal to 1. c In the presence of Ca2+ the effects of glucagon on gluconeogenesis showed a positive correlation with the lactate to pyruvate ratios; for a ratio equal to 0.01 no stimulation occurred, but in the 0.1 to 100 range stimulation increased progressively, producing a clear parabolic dependence between the effects of glucagon and the lactate to pyruvate ratio. d In the absence of Ca2+ the relationship between the changes caused by glucagon in gluconeogenesis and the lactate to pyruvate ratio was substantially changed; the dependence curve was no longer parabolic but sigmoidal in shape with a plateau beginning at a lactate/pyruvate ratio equal to 1; there was inhibition at the lactate to pyruvate ratios of 0.01 and 0.1 and a constant stimulation starting with a ratio equal to 1; for the lactate to pyruvate ratios of 10 and 100

  13. Agarose electrophoresis of DNA in discontinuous buffers, using a horizontal slab apparatus and a buffer system with improved properties.

    Science.gov (United States)

    Zsolnai, A; Orbán, L; Chrambach, A

    1993-03-01

    Using a horizontal slab apparatus with a buffer in the reservoirs at the level of the gel ("sea-level electrophoresis"), the retrograde discontinuous buffer system reported by Wiltfang et al. for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins was applied to DNA electrophoresis. This application yielded the advantages of an increased displacement rate of the moving boundary front and a decrease in the concentration of the counterion base in the resolving phase, which yielded reduced relative mobility values at equivalent gel concentrations and practicable low buffer concentrations. The change of relative mobilities (Rf) with a variation of field strength is decreased compared to that of the migration rate in the continuous Tris-boric-acid-EDTA (TBE) buffer and thus the robustness of the system is improved, as well as the efficiency of separation. The system of Wiltfang et al. has in common with previously described discontinuous DNA system, that it is able to stack DNA from dilute samples and is insensitive to sample components with lower net mobilities than DNA, such as acetate. However, the variance of Rf at constant current density in the discontinuous buffer system is not improved over that of the migration rate at constant field strength in the continuous TBE buffer.

  14. Interplay of Mg2+, ADP, and ATP in the cytosol and mitochondria: unravelling the role of Mg2+ in cell respiration.

    Science.gov (United States)

    Gout, Elisabeth; Rébeillé, Fabrice; Douce, Roland; Bligny, Richard

    2014-10-28

    In animal and plant cells, the ATP/ADP ratio and/or energy charge are generally considered key parameters regulating metabolism and respiration. The major alternative issue of whether the cytosolic and mitochondrial concentrations of ADP and ATP directly mediate cell respiration remains unclear, however. In addition, because only free nucleotides are exchanged by the mitochondrial ADP/ATP carrier, whereas MgADP is the substrate of ATP synthase (EC 3.6.3.14), the cytosolic and mitochondrial Mg(2+) concentrations must be considered as well. Here we developed in vivo/in vitro techniques using (31)P-NMR spectroscopy to simultaneously measure these key components in subcellular compartments. We show that heterotrophic sycamore (Acer pseudoplatanus L.) cells incubated in various nutrient media contain low, stable cytosolic ADP and Mg(2+) concentrations, unlike ATP. ADP is mainly free in the cytosol, but complexed by Mg(2+) in the mitochondrial matrix, where [Mg(2+)] is tenfold higher. In contrast, owing to a much higher affinity for Mg(2+), ATP is mostly complexed by Mg(2+) in both compartments. Mg(2+) starvation used to alter cytosolic and mitochondrial [Mg(2+)] reversibly increases free nucleotide concentration in the cytosol and matrix, enhances ADP at the expense of ATP, decreases coupled respiration, and stops cell growth. We conclude that the cytosolic ADP concentration, and not ATP, ATP/ADP ratio, or energy charge, controls the respiration of plant cells. The Mg(2+) concentration, remarkably constant and low in the cytosol and tenfold higher in the matrix, mediates ADP/ATP exchange between the cytosol and matrix, [MgADP]-dependent mitochondrial ATP synthase activity, and cytosolic free ADP homeostasis.

  15. Inherited human group IVA cytosolic phospholipase A2 deficiency abolishes platelet, endothelial, and leucocyte eicosanoid generation

    Science.gov (United States)

    Kirkby, Nicholas S.; Reed, Daniel M.; Edin, Matthew L.; Rauzi, Francesca; Mataragka, Stefania; Vojnovic, Ivana; Bishop-Bailey, David; Milne, Ginger L.; Longhurst, Hilary; Zeldin, Darryl C.; Mitchell, Jane A.; Warner, Timothy D.

    2016-01-01

    Eicosanoids are important vascular regulators, but the phospholipase A2 (PLA2) isoforms supporting their production within the cardiovascular system are not fully understood. To address this, we have studied platelets, endothelial cells, and leukocytes from 2 siblings with a homozygous loss-of-function mutation in group IVA cytosolic phospholipase A2 (cPLA2α). Chromatography/mass spectrometry was used to determine levels of a broad range of eicosanoids produced by isolated vascular cells, and in plasma and urine. Eicosanoid release data were paired with studies of cellular function. Absence of cPLA2α almost abolished eicosanoid synthesis in platelets (e.g., thromboxane A2, control 20.5 ± 1.4 ng/ml vs. patient 0.1 ng/ml) and leukocytes [e.g., prostaglandin E2 (PGE2), control 21.9 ± 7.4 ng/ml vs. patient 1.9 ng/ml], and this was associated with impaired platelet activation and enhanced inflammatory responses. cPLA2α-deficient endothelial cells showed reduced, but not absent, formation of prostaglandin I2 (prostacyclin; control 956 ± 422 pg/ml vs. patient 196 pg/ml) and were primed for inflammation. In the urine, prostaglandin metabolites were selectively influenced by cPLA2α deficiency. For example, prostacyclin metabolites were strongly reduced (18.4% of control) in patients lacking cPLA2α, whereas PGE2 metabolites (77.8% of control) were similar to healthy volunteer levels. These studies constitute a definitive account, demonstrating the fundamental role of cPLA2α to eicosanoid formation and cellular responses within the human circulation.—Kirkby, N. S., Reed, D. M., Edin, M. L., Rauzi, F., Mataragka, S., Vojnovic, I., Bishop-Bailey, D., Milne, G. L., Longhurst, H., Zeldin, D. C., Mitchell, J. A., Warner, T. D. Inherited human group IVA cytosolic phospholipase A2 deficiency abolishes platelet, endothelial, and leucocyte eicosanoid generation. PMID:26183771

  16. Overexpression of human virus surface glycoprotein precursors induces cytosolic unfolded protein response in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Sasnauskas Kęstutis

    2011-05-01

    Full Text Available Abstract Background The expression of human virus surface proteins, as well as other mammalian glycoproteins, is much more efficient in cells of higher eukaryotes rather than yeasts. The limitations to high-level expression of active viral surface glycoproteins in yeast are not well understood. To identify possible bottlenecks we performed a detailed study on overexpression of recombinant mumps hemagglutinin-neuraminidase (MuHN and measles hemagglutinin (MeH in yeast Saccharomyces cerevisiae, combining the analysis of recombinant proteins with a proteomic approach. Results Overexpressed recombinant MuHN and MeH proteins were present in large aggregates, were inactive and totally insoluble under native conditions. Moreover, the majority of recombinant protein was found in immature form of non-glycosylated precursors. Fractionation of yeast lysates revealed that the core of viral surface protein aggregates consists of MuHN or MeH disulfide-linked multimers involving eukaryotic translation elongation factor 1A (eEF1A and is closely associated with small heat shock proteins (sHsps that can be removed only under denaturing conditions. Complexes of large Hsps seem to be bound to aggregate core peripherally as they can be easily removed at high salt concentrations. Proteomic analysis revealed that the accumulation of unglycosylated viral protein precursors results in specific cytosolic unfolded protein response (UPR-Cyto in yeast cells, characterized by different action and regulation of small Hsps versus large chaperones of Hsp70, Hsp90 and Hsp110 families. In contrast to most environmental stresses, in the response to synthesis of recombinant MuHN and MeH, only the large Hsps were upregulated whereas sHsps were not. Interestingly, the amount of eEF1A was also increased during this stress response. Conclusions Inefficient translocation of MuHN and MeH precursors through ER membrane is a bottleneck for high-level expression in yeast. Overexpression of

  17. Cigarette smoke toxicants as substrates and inhibitors for human cytosolic SULTs

    International Nuclear Information System (INIS)

    Yasuda, Shin; Idell, Steven; Fu Jian; Carter, Glendora; Snow, Rhodora; Liu, M.-C.

    2007-01-01

    The current study was designed to examine the role of sulfation in the metabolism of cigarette smoke toxicants and clarify whether these toxicants, by serving as substrates for the cytosolic sulfotransferases (SULTs), may interfere with the sulfation of key endogenous compounds. By metabolic labeling, [ 35 S]sulfated species were found to be generated and released into the media of HepG2 human hepatoma cells and primary human lung endothelial cells labeled with [ 35 S]sulfate in the presence of cigarette smoke extract (CSE). Concomitantly, several [ 35 S]sulfated metabolites observed in the medium in the absence of CSE either decreased or disappeared. Eleven previously prepared human cytosolic SULTs were tested for sulfating activity with CSE and known cigarette smoke toxicants as substrates. Activity data revealed SULT1A1, SULT1A2, SULT1A3, and SULT1C2 as major enzymes responsible for their sulfation. To examine their inhibitory effects on the sulfation of 17β-estradiol by SULT1A1, enzymatic assays were performed in the presence of three representative toxicant compounds, namely N-hydroxy-4-aminobiphenyl (N-OH-4-ABP), 4-aminobiphenyl (4-ABP) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). IC 50 values determined for the sulfation of 17β-estradiol by SULT1A1 were 11.8 μM, 28.2 μM, and 500 μM, respectively, for N-OH-4-ABP, 4-ABP and PhIP. Kinetic analyses indicated that the mechanism underlying the inhibition of 17β-estradiol sulfation by these cigarette smoke toxicants is of a mixed competitive-noncompetitive type. Metabolic labeling experiments clearly showed inhibition of the production of [ 35 S]sulfated 17β-estradiol by N-OH-4-ABP in a concentration-dependent manner in HepG2 cells. Taken together, these results suggest that sulfation plays a significant role in the metabolism of cigarette smoke compounds. By serving as substrates for SULTs, cigarette smoke toxicants may interfere with the metabolism of 17β-estradiol and other endogenous

  18. Effects of deuterated water upon specific activity of some marker enzymes for cytosol and plasmatic membrane

    International Nuclear Information System (INIS)

    Buzgariu, Wanda; Coroiu, Viorica; Moldovan, Lucia; Titescu, G.; Stefanescu, I.

    2004-01-01

    Recently, numerous studies were devoted to the effects of an increased environmental deuterium concentration on physiological characteristics of various biological systems, from monocellular organisms up to mammals. Within these preoccupations the experiments on enzyme activity and parameters are of special interest since they throw light upon the mechanisms in metabolic biochemical reactions (glycolysis, photosynthesis, transport across membranes, etc). The present work concerns the effects of heavy water upon the activity of some enzymes (dehydrogenase-LDH lactate and 5' nucleotidase) implied in different metabolic pathways, serving as functional indicators for some cellular compartments such as the cytosols and cellular membranes. Enzyme activity was determined by growing for 6 days the cells (Hep 2, CHO, fibroblasts) in deuterated culture media at different concentration levels (20%, 40%, 65% si 90%), as well as in a reaction medium deuterated at 99.96%. In case of the first experimental run the LDH activity was monitored for the three cellular lines (Hep 2, CHO, fibroblasts) for different time intervals (1 d, 3 d and 6 d). After the first 24 h of cells' exposure the activity values were similar regardless of the heavy water concentration in the medium. Exposing the cells for longer time (6 days) led to modifications of LDH activity. In contrast to the case of media with relatively moderate D 2 O content, cell growing in conditions of intense deuteration 65% and 90 % D 2 O) led to an increase of cytosolic enzyme activity of about 50%. In case of 5' nucleotidase after 6 days of cell cultivation in deuteration conditions the activity decreased to 50% and 70% from the value corresponding to normal conditions for cell growth. This diminution of the activity was characteristic for the media with 65% and 90% D 2 O. In the second experimental run the activities of dehydrogenase lactate and 5' nucleotidase from the cellular homogenate obtained from cells grown in

  19. Cytosolic and nuclear caspase-8 have opposite impact on survival after liver resection for hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Koschny, Ronald; Schemmer, Peter; Schirmacher, Peter; Ganten, Tom M; Brost, Sylvia; Hinz, Ulf; Sykora, Jaromir; Batke, Emanuela M; Singer, Stephan; Breuhahn, Kai; Stremmel, Wolfgang; Walczak, Henning

    2013-01-01

    An imbalance between proliferation and apoptosis is one of the main features of carcinogenesis. TRAIL (TNF-related apoptosis-inducing ligand) induces apoptosis upon binding to the TRAIL death receptors, TRAIL receptor 1 (TRAIL-R1) and TRAIL-R2, whereas binding to TRAIL-R3 and TRAIL-R4 might promote cell survival and proliferation. The anti-tumor activity of TRAIL-R1 and TRAIL-R2 agonists is currently investigated in clinical trials. To gain further insight into the regulation of apoptosis in hepatocellular carcinoma (HCC), we investigated the TRAIL pathway and the regulators of apoptosis caspase-8, Bcl-xL and Mcl-1 in patients with HCC regarding patient survival. We analyzed 157 hepatocellular carcinoma patients who underwent partial liver resection or orthotopic liver transplantation and healthy control liver tissue using immunohistochemistry on tissue microarrays for the expression of TRAIL-R1 to TRAIL-R4, caspase-8, Bcl-xL and Mcl-1. Immunohistochemical data were evaluated for potential associations with clinico-pathological parameters and survival. Whereas TRAIL-R1 was downregulated in HCC in comparison to normal liver tissue, TRAIL-R2 and –R4 were upregulated in HCC, especially in G2 and G3 tumors. TRAIL-R1 downregulation and upregulation of TRAIL-R2 and TRAIL-R4 correlated with tumor dedifferentiation (G2/G3). TRAIL-R3, Bcl-xL and Mcl-1 showed no differential expression in tumor tissue compared to normal tissue. The expression levels of TRAIL receptors did not correlate with patient survival after partial hepatectomy. Interestingly, in tumor tissue, but not in normal hepatocytes, caspase-8 showed a strong nuclear staining. Low cytosolic and high nuclear staining intensity of caspase-8 significantly correlated with impaired survival after partial hepatectomy, which, for cytosolic caspase-8, was independent from tumor grade. Assessment of TRAIL-receptor expression patterns may have therapeutic implications for the use of TRAIL receptor agonists in HCC therapy

  20. Effects of buffer thickness on ATW blanket performance

    International Nuclear Information System (INIS)

    Yang, W. S.; Mercatali, L.; Taiwo, T. A.; Hill, R. N.

    2001-01-01

    This paper presents preliminary results of target and buffer design studies for liquid metal cooled accelerator transmutation of waste (ATW) systems, aimed at maximizing the source importance while simultaneously reducing the irradiation damage to fuel. Using 840 MWt liquid metal cooled ATW designs, the effects of buffer thickness on the blanket performance have been studied. Varying the buffer thickness for a given blanket configuration, system performance parameters have been estimated by a series of calculations using the MCNPX and REBUS-3 codes. The effects of source importance variation are studied by investigating the low-energy ( and lt; 20 MeV) neutron source distribution and the equilibrium cycle blanket performance parameters such as fuel inventory, discharge burnup, burnup reactivity loss, and peak fast fluence. For investigating irradiation damage to fuel, the displacements per atom (dpa), hydrogen production, and helium production rates are evaluated at the buffer and blanket interface where the peak fast fluence occurs. Results for the liquid-metal-cooled designs show that the damage rates and the source importance increase monotonically as the buffer thickness decreases. Based on a compromise between the competing objectives of increasing the source importance and reducing the damage rates, a buffer thickness of around 20 cm appears to be reasonable. Investigation of the impact of the proton beam energy on the target and buffer design shows that for a given blanket power level, a lower beam energy (0.6 GeV versus 1 GeV) results in a higher irradiation damage to the beam window. This trend occurs because of the increase in the beam intensity required to maintain the power level

  1. Effects of Buffer Thickness on ATW Blanket Performance

    International Nuclear Information System (INIS)

    Yang, W.S.; Mercatali, L.; Taiwo, T.A.; Hill, R.N.

    2002-01-01

    This paper presents preliminary results of target and buffer design studies for liquid metal cooled accelerator transmutation of waste (ATW) systems, aimed at maximizing the source importance while simultaneously reducing the irradiation damage to fuel. Using 840 MWt liquid metal cooled ATW designs, the effects of buffer thickness on the blanket performance have been studied. Varying the buffer thickness for a given blanket configuration, system performance parameters have been estimated by a series of calculations using the MCNPX and REBUS-3 codes. The effects of source importance variation are studied by investigating the low-energy (< 20 MeV) neutron source distribution and the equilibrium cycle blanket performance parameters such as fuel inventory, discharge burnup, burnup reactivity loss, and peak fast fluence. For investigating irradiation damage to fuel, the displacements per atom (dpa), hydrogen production, and helium production rates are evaluated at the buffer and blanket interface where the peak fast fluence occurs. Results for the liquid-metal-cooled designs show that the damage rates and the source importance increase monotonically as the buffer thickness decreases. Based on a compromise between the competing objectives of increasing the source importance and reducing the damage rates, a buffer thickness of around 20 cm appears to be reasonable. Investigation of the impact of the proton beam energy on the target and buffer design shows that for a given blanket power level, a lower beam energy (0.6 GeV versus 1 GeV) results in a higher irradiation damage to the beam window. This trend occurs because of the increase in the beam intensity required to maintain the power level. (authors)

  2. Flow and transport through a damaged buffer - exploration of the impact of a cemented and an eroded buffer

    Energy Technology Data Exchange (ETDEWEB)

    Neretnieks, Ivars [Dept. of Chemical Engineering and Technology, Royal Institute of Technology, Stockholm (Sweden)

    2006-12-15

    The compacted bentonite buffer that surrounds the canister with spent fuel is designed not to allow water to flow through it in any appreciable quantities. Should the buffer be severely damaged water could flow to the canister and bring with it corrosive agent such as sulphide. This could speed up the rate of copper corrosion. Should the canister have a damage or be corroded so that the spent fuel inside the canister comes in contact with the water radionuclides could dissolve in the water and be carried away. Two modes of canister damage and their consequences for solute transport are explored in this report. In one case the buffer can potentially be eroded by the water seeping in the fractures in the surrounding rock and attain a high hydraulic conductivity allowing water to flow through it, at least locally. In another case it is conjectured that the buffer is cemented and becomes brittle. Then a crack could conceivably form in the cemented buffer allowing water to flow through it directly to the canister surface. Both cases are hypothetical although in the erosion case the erosion rate could possibly be quantified. The cases studied are to be seen as exploratory calculations for 'what if' cases. Some worst case assumptions have been used to explore bounding scenarios. Thus for the erosion case it is assumed that the hydraulic conductivity is so high that the buffer exerts negligible resistance to water flow. This implies that all the water that can flow through the fractures intersecting the deposition hole can reach the canister. Similarly for the cemented buffer case it is assumed that the crack in the buffer directly connects the rock fracture to the canister surface. For these circumstances it is found that the flowrate that can enter the deposition hole will be approximately two times as large as the water flow that would pass a projected area equal to that of the deposition hole. Thus if the flux in the rock some distance from the deposition

  3. Flow and transport through a damaged buffer - exploration of the impact of a cemented and an eroded buffer

    International Nuclear Information System (INIS)

    Neretnieks, Ivars

    2006-12-01

    The compacted bentonite buffer that surrounds the canister with spent fuel is designed not to allow water to flow through it in any appreciable quantities. Should the buffer be severely damaged water could flow to the canister and bring with it corrosive agent such as sulphide. This could speed up the rate of copper corrosion. Should the canister have a damage or be corroded so that the spent fuel inside the canister comes in contact with the water radionuclides could dissolve in the water and be carried away. Two modes of canister damage and their consequences for solute transport are explored in this report. In one case the buffer can potentially be eroded by the water seeping in the fractures in the surrounding rock and attain a high hydraulic conductivity allowing water to flow through it, at least locally. In another case it is conjectured that the buffer is cemented and becomes brittle. Then a crack could conceivably form in the cemented buffer allowing water to flow through it directly to the canister surface. Both cases are hypothetical although in the erosion case the erosion rate could possibly be quantified. The cases studied are to be seen as exploratory calculations for 'what if' cases. Some worst case assumptions have been used to explore bounding scenarios. Thus for the erosion case it is assumed that the hydraulic conductivity is so high that the buffer exerts negligible resistance to water flow. This implies that all the water that can flow through the fractures intersecting the deposition hole can reach the canister. Similarly for the cemented buffer case it is assumed that the crack in the buffer directly connects the rock fracture to the canister surface. For these circumstances it is found that the flowrate that can enter the deposition hole will be approximately two times as large as the water flow that would pass a projected area equal to that of the deposition hole. Thus if the flux in the rock some distance from the deposition hole is 1

  4. Enzymatic sulfation of tocopherols and tocopherol metabolites by human cytosolic sulfotransferases.

    Science.gov (United States)

    Hashiguchi, Takuyu; Kurogi, Katsuhisa; Sakakibara, Yoichi; Yamasaki, Masao; Nishiyama, Kazuo; Yasuda, Shin; Liu, Ming-Cheh; Suiko, Masahito

    2011-01-01

    Tocopherols are essential micronutrients for mammals widely known as potent lipid-soluble antioxidants that are present in cell membranes. Recent studies have demonstrated that most of the carboxychromanol (CEHC), a tocopherol metabolite, in the plasma exists primarily in sulfate- and glucuronide-conjugated forms. To gain insight into the enzymatic sulfation of tocopherols and their metabolites, a systematic investigation was performed using all 14 known human cytosolic sulfotransferases (SULTs). The results showed that the members of the SULT1 family displayed stronger sulfating activities toward tocopherols and their metabolites. These enzymes showed a substrate preference for γ-tocopherol over α-tocopherol and for γ-CEHC over other CEHCs. Using A549 human lung epithelial cells in a metabolic labeling study, a similar trend in the sulfation of tocopherols and CEHCs was observed. Collectively, the results obtained indicate that SULT-mediated enzymatic sulfation of tocopherols and their metabolites is a significant pathway for regulation of the homeostasis and physiological functions of these important compounds.

  5. Multifunctional polymersomes for cytosolic delivery of gemcitabine and doxorubicin to cancer cells.

    Science.gov (United States)

    Nahire, Rahul; Haldar, Manas K; Paul, Shirshendu; Ambre, Avinash H; Meghnani, Varsha; Layek, Buddhadev; Katti, Kalpana S; Gange, Kara N; Singh, Jagdish; Sarkar, Kausik; Mallik, Sanku

    2014-08-01

    Although liposomes are widely used as carriers of drugs and imaging agents, they suffer from a lack of stability and the slow release of the encapsulated contents at the targeted site. Polymersomes (vesicles of amphiphilic polymers) are considerably more stable compared to liposomes; however, they also demonstrate a slow release for the encapsulated contents, limiting their efficacy as a drug-delivery tool. As a solution, we prepared and characterized echogenic polymersomes, which are programmed to release the encapsulated drugs rapidly when incubated with cytosolic concentrations of glutathione. These vesicles encapsulated air bubbles inside and efficiently reflected diagnostic-frequency ultrasound. Folate-targeted polymersomes showed an enhanced uptake by breast and pancreatic-cancer cells in a monolayer as well as in three-dimensional spheroid cultures. Polymersomes encapsulated with the anticancer drugs gemcitabine and doxorubicin showed significant cytotoxicity to these cells. With further improvements, these vesicles hold the promise to serve as multifunctional nanocarriers, offering a triggered release as well as diagnostic ultrasound imaging. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. New insights into the posttranslational regulation of human cytosolic thioredoxin by S-palmitoylation

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Zhiyu; Zhong, Liangwei, E-mail: liazho@ucas.ac.cn

    2015-05-15

    High level of palmitate is associated with metabolic disorders. We recently showed that enhanced level of S-palmitoylated cytosolic thioredoxin (Trx1) in mouse liver was new characteristic feature of insulin resistance. However, our understanding of the effect of S-palmitoylation on Trx1 is limited, and the tissue specificity of Trx1 S-palmitoylation is unclear. Here we show that S-palmitoylation also occurs at Cys73 of Trx1 in living endothelial cells, and the level of S-palmitoylated Trx1 undergoes regulation by insulin signaling. Trx1 prefers thiol-thioester exchange with palmitoyl-CoA to acetyl-CoA. S-palmitoylation alters conformation or secondary structure of Trx1, as well as decreases the ability of Trx1 to transfer electrons from thioredoxin reductase to S-nitrosylated protein–tyrosine phosphatase 1B and S-nitroso-glutathione. Our results demonstrate that S-palmitoylation is an important post-translational modification of human Trx1. - Highlights: • S-palmitoylation occurs at Cys73 of Trx1 in living endothelial cells. • Insulin signaling may regulate level of S-palmitoylated Trx1 in the cells. • S-palmitoylation plays significant effects on Trx1 structure and functions.

  7. Amino acid starvation has opposite effects on mitochondrial and cytosolic protein synthesis.

    Directory of Open Access Journals (Sweden)

    Mark A Johnson

    Full Text Available Amino acids are essential for cell growth and proliferation for they can serve as precursors of protein synthesis, be remodelled for nucleotide and fat biosynthesis, or be burnt as fuel. Mitochondria are energy producing organelles that additionally play a central role in amino acid homeostasis. One might expect mitochondrial metabolism to be geared towards the production and preservation of amino acids when cells are deprived of an exogenous supply. On the contrary, we find that human cells respond to amino acid starvation by upregulating the amino acid-consuming processes of respiration, protein synthesis, and amino acid catabolism in the mitochondria. The increased utilization of these nutrients in the organelle is not driven primarily by energy demand, as it occurs when glucose is plentiful. Instead it is proposed that the changes in the mitochondrial metabolism complement the repression of cytosolic protein synthesis to restrict cell growth and proliferation when amino acids are limiting. Therefore, stimulating mitochondrial function might offer a means of inhibiting nutrient-demanding anabolism that drives cellular proliferation.

  8. Cellular defense against UVB-induced phototoxicity by cytosolic NADP+-dependent isocitrate dehydrogenase

    International Nuclear Information System (INIS)

    Jo, Seung-Hee; Lee, So-Hyun; Suk Chun, Hang; Min Lee, Su; Koh, Ho-Jin; Lee, Sung-Eun; Chun, Jang-Soo; Park, Jeen-Woo; Huh, Tae-Lin

    2002-01-01

    Ultraviolet (UV) radiation is known as a major cause of skin photoaging and photocarcinogenesis. Many harmful effects of UV radiation are associated with the generation of reactive oxygen species. Recently, we have shown that NADP + -dependent isocitrate dehydrogenase is involved in the supply of NADPH needed for GSH production against cellular oxidative damage. In this study we investigated the role of cytosolic form of NADP + -dependent isocitrate dehydrogenase (IDPc) against UV radiation-induced cytotoxicity by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 2.3-fold higher and 39% lower, respectively, than that in the parental cells carrying the vector alone. Upon exposure to UVB (312 nm), the cells with low levels of IDPc became more sensitive to cell killing. Lipid peroxidation, protein oxidation, oxidative DNA damage, and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly overexpressed IDPc exhibited enhanced resistance against UV radiation, compared to the control cells. The data indicate that IDPc plays an important role in cellular defense against UV radiation-induced oxidative injury

  9. Cellular defense against UVB-induced phototoxicity by cytosolic NADP(+)-dependent isocitrate dehydrogenase.

    Science.gov (United States)

    Jo, Seung-Hee; Lee, So-Hyun; Chun, Hang Suk; Lee, Su Min; Koh, Ho-Jin; Lee, Sung-Eun; Chun, Jang-Soo; Park, Jeen-Woo; Huh, Tae-Lin

    2002-03-29

    Ultraviolet (UV) radiation is known as a major cause of skin photoaging and photocarcinogenesis. Many harmful effects of UV radiation are associated with the generation of reactive oxygen species. Recently, we have shown that NADP(+)-dependent isocitrate dehydrogenase is involved in the supply of NADPH needed for GSH production against cellular oxidative damage. In this study we investigated the role of cytosolic form of NADP(+)-dependent isocitrate dehydrogenase (IDPc) against UV radiation-induced cytotoxicity by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 2.3-fold higher and 39% lower, respectively, than that in the parental cells carrying the vector alone. Upon exposure to UVB (312 nm), the cells with low levels of IDPc became more sensitive to cell killing. Lipid peroxidation, protein oxidation, oxidative DNA damage, and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly overexpressed IDPc exhibited enhanced resistance against UV radiation, compared to the control cells. The data indicate that IDPc plays an important role in cellular defense against UV radiation-induced oxidative injury. (c)2002 Elsevier Science (USA).

  10. Cellular defense against singlet oxygen-induced oxidative damage by cytosolic NADP+-dependent isocitrate dehydrogenase.

    Science.gov (United States)

    Kim, Sun Yee; Park, Jeen-Woo

    2003-03-01

    Singlet oxygen (1O2) is a highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules. Recently, we have shown that NADP+-dependent isocitrate dehydrogenase is involved in the supply of NADPH needed for GSH production against cellular oxidative damage. In this study, we investigated the role of cytosolic form of NADP+-dependent isocitrate dehydrogenase (IDPc) against singlet oxygen-induced cytotoxicity by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 2.3-fold higher and 39% lower, respectively, than that in the parental cells carrying the vector alone. Upon exposure to singlet oxygen generated from photoactivated dye, the cells with low levels of IDPc became more sensitive to cell killing. Lipid peroxidation, protein oxidation, oxidative DNA damage and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly over-expressed IDPc exhibited enhanced resistance against singlet oxygen, compared to the control cells. The data indicate that IDPc plays an important role in cellular defense against singlet oxygen-induced oxidative injury.

  11. Cytosolically expressed PrP GPI-signal peptide interacts with mitochondria.

    Science.gov (United States)

    Guizzunti, Gianni; Zurzolo, Chiara

    2015-01-01

    We previously reported that PrP GPI-anchor signal peptide (GPI-SP) is specifically degraded by the proteasome. Additionally, we showed that the point mutation P238S, responsible for a genetic form of prion diseases, while not affecting the GPI-anchoring process, results in the accumulation of PrP GPI-SP, suggesting the possibility that PrP GPI-anchor signal peptide could play a role in neurodegenerative prion diseases. We now show that PrP GPI-SP, when expressed as a cytosolic peptide, is able to localize to the mitochondria and to induce mitochondrial fragmentation and vacuolarization, followed by loss in mitochondrial membrane potential, ultimately resulting in apoptosis. Our results identify the GPI-SP of PrP as a novel candidate responsible for the impairment in mitochondrial function involved in the synaptic pathology observed in prion diseases, establishing a link between PrP GPI-SP accumulation and neuronal death.

  12. Ecto- and cytosolic 5'-nucleotidases in normal and AMP deaminase-deficient human skeletal muscle

    DEFF Research Database (Denmark)

    Hanisch, Frank; Hellsten, Ylva; Zierz, Stephan

    2006-01-01

    homogenate 5'-nucleotidase and ectoN, or in cN-I expression on Western blots. No correlation for age, fibre type distribution and AMPD1 genotype was found for whole homogenate nucleotidase, total cN and cN-I using multiple linear regression analysis. There was no gender-specific difference in the activities...... with a homozygous C34T mutation, cN-I might be a more important pathway for AMP removal. We determined activities of AMP deaminase, cN-I, total cytosolic 5'-nucleotidase (total cN), ecto-5'-nucleotidase (ectoN) and whole homogenate 5'-nucleotidase activity in skeletal muscle biopsies from patients with different...... AMPD1 genotypes [homozygotes for C34T mutation (TT); heterozygotes for C34T mutation (CT); and homozygotes for wild type (CC): diseased controls CC; and normal controls CC]. AMP deaminase activity showed genotype-dependent differences. Total cN activity in normal controls accounted for 57...

  13. Cytosolic Phospholipase A2 Protein as a Novel Therapeutic Target for Spinal Cord Injury

    Science.gov (United States)

    Liu, Nai-Kui; Deng, Ling-Xiao; Zhang, Yi Ping; Lu, Qing-Bo; Wang, Xiao-Fei; Hu, Jian-Guo; Oakes, Eddie; Bonventre, Joseph V; Shields, Christopher B; Xu, Xiao-Ming

    2014-01-01

    Objective The objective of this study was to investigate whether cytosolic phospholipase A2 (cPLA2), an important isoform of PLA2 that mediates the release of arachidonic acid, plays a role in the pathogenesis of spinal cord injury (SCI). Methods A combination of molecular, histological, immunohistochemical, and behavioral assessments were used to test whether blocking cPLA2 activation pharmacologically or genetically reduced cell death, protected spinal cord tissue, and improved behavioral recovery after a contusive SCI performed at the 10th thoracic level in adult mice. Results SCI significantly increased cPLA2 expression and activation. Activated cPLA2 was localized mainly in neurons and oligodendrocytes. Notably, the SCI-induced cPLA2 activation was mediated by the extracellular signal-regulated kinase signaling pathway. In vitro, activation of cPLA2 by ceramide-1-phosphate or A23187 induced spinal neuronal death, which was substantially reversed by arachidonyl trifluoromethyl ketone, a cPLA2 inhibitor. Remarkably, blocking cPLA2 pharmacologically at 30 minutes postinjury or genetically deleting cPLA2 in mice ameliorated motor deficits, and reduced cell loss and tissue damage after SCI. Interpretation cPLA2 may play a key role in the pathogenesis of SCI, at least in the C57BL/6 mouse, and as such could be an attractive therapeutic target for ameliorating secondary tissue damage and promoting recovery of function after SCI. PMID:24623140

  14. Prefoldin Promotes Proteasomal Degradation of Cytosolic Proteins with Missense Mutations by Maintaining Substrate Solubility.

    Directory of Open Access Journals (Sweden)

    Sophie A Comyn

    2016-07-01

    Full Text Available Misfolded proteins challenge the ability of cells to maintain protein homeostasis and can accumulate into toxic protein aggregates. As a consequence, cells have adopted a number of protein quality control pathways to prevent protein aggregation, promote protein folding, and target terminally misfolded proteins for degradation. In this study, we employed a thermosensitive allele of the yeast Guk1 guanylate kinase as a model misfolded protein to investigate degradative protein quality control pathways. We performed a flow cytometry based screen to identify factors that promote proteasomal degradation of proteins misfolded as the result of missense mutations. In addition to the E3 ubiquitin ligase Ubr1, we identified the prefoldin chaperone subunit Gim3 as an important quality control factor. Whereas the absence of GIM3 did not impair proteasomal function or the ubiquitination of the model substrate, it led to the accumulation of the poorly soluble model substrate in cellular inclusions that was accompanied by delayed degradation. We found that Gim3 interacted with the Guk1 mutant allele and propose that prefoldin promotes the degradation of the unstable model substrate by maintaining the solubility of the misfolded protein. We also demonstrated that in addition to the Guk1 mutant, prefoldin can stabilize other misfolded cytosolic proteins containing missense mutations.

  15. Lipid-Based Liquid Crystalline Nanoparticles Facilitate Cytosolic Delivery of siRNA via Structural Transformation.

    Science.gov (United States)

    He, Shufang; Fan, Weiwei; Wu, Na; Zhu, Jingjing; Miao, Yunqiu; Miao, Xiaran; Li, Feifei; Zhang, Xinxin; Gan, Yong

    2018-04-11

    RNA interference (RNAi) technology has shown great promise for the treatment of cancer and other genetic disorders. Despite the efforts to increase the target tissue distribution, the safe and effective delivery of siRNA to the diseased cells with sufficient cytosolic transport is another critical factor for successful RNAi clinical application. Here, the constructed lipid-based liquid crystalline nanoparticles, called nano-Transformers, can transform thestructure in the intracellular acidic environment and perform high-efficient siRNA delivery for cancer treatment. The developed nano-Transformers have satisfactory siRNA loading efficiency and low cytotoxicity. Different from the traditional cationic nanocarriers, the endosomal membrane fusion induced by the conformational transition of lipids contributes to the easy dissociation of siRNA from nanocarriers and direct release of free siRNA into cytoplasm. We show that transfection with cyclin-dependent kinase 1 (CDK1)-siRNA-loaded nano-Transformers causes up to 95% reduction of relevant mRNA in vitro and greatly inhibits the tumor growth without causing any immunogenic response in vivo. This work highlights that the lipid-based nano-Transformers may become the next generation of siRNA delivery system with higher efficacy and improved safety profiles.

  16. Sorbitol dehydrogenase is a cytosolic protein required for sorbitol metabolism in Arabidopsis thaliana.

    Science.gov (United States)

    Aguayo, María Francisca; Ampuero, Diego; Mandujano, Patricio; Parada, Roberto; Muñoz, Rodrigo; Gallart, Marta; Altabella, Teresa; Cabrera, Ricardo; Stange, Claudia; Handford, Michael

    2013-05-01

    Sorbitol is converted to fructose in Rosaceae species by SORBITOL DEHYDROGENASE (SDH, EC 1.1.1.14), especially in sink organs. SDH has also been found in non-Rosaceae species and here we show that the protein encoded by At5g51970 in Arabidopsis thaliana (L.) Heynh. possesses the molecular characteristics of an SDH. Using a green fluorescent protein-tagged version and anti-SDH antisera, we determined that SDH is cytosolically localized, consistent with bioinformatic predictions. We also show that SDH is widely expressed, and that SDH protein accumulates in both source and sink organs. In the presence of NAD+, recombinant SDH exhibited greatest oxidative activity with sorbitol, ribitol and xylitol as substrates; other sugar alcohols were oxidized to a lesser extent. Under standard growth conditions, three independent sdh- mutants developed as wild-type. Nevertheless, all three exhibited reduced dry weight and primary root length compared to wild-type when grown in the presence of sorbitol. Additionally, under short-day conditions, the mutants were more resistant to dehydration stress, as shown by a reduced loss of leaf water content when watering was withheld, and a greater survival rate on re-watering. This evidence suggests that limitations in the metabolism of sugar alcohols alter the growth of Arabidopsis and its response to drought. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  17. Predominant Role of Cytosolic Phospholipase A2α in Dioxin-induced Neonatal Hydronephrosis in Mice

    Science.gov (United States)

    Yoshioka, Wataru; Kawaguchi, Tatsuya; Fujisawa, Nozomi; Aida-Yasuoka, Keiko; Shimizu, Takao; Matsumura, Fumio; Tohyama, Chiharu

    2014-01-01

    Hydronephrosis is a common disease characterized by dilation of the renal pelvis and calices, resulting in loss of kidney function in the most severe cases. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces nonobstructive hydronephrosis in mouse neonates through upregulation of prostaglandin E2 (PGE2) synthesis pathway consisting of cyclooxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) by a yet unknown mechanism. We here studied possible involvement of cytosolic phospholipase A2α (cPLA2α) in this mechanism. To this end, we used a cPLA2α-null mouse model and found that cPLA2α has a significant role in the upregulation of the PGE2 synthesis pathway through a noncanonical pathway of aryl hydrocarbon receptor. This study is the first to demonstrate the predominant role of cPLA2α in hydronephrosis. Elucidation of the pathway leading to the onset of hydronephrosis using the TCDD-exposed mouse model will deepen our understanding of the molecular basis of nonobstructive hydronephrosis in humans. PMID:24509627

  18. OAS proteins and cGAS: unifying concepts in sensing and responding to cytosolic nucleic acids.

    Science.gov (United States)

    Hornung, Veit; Hartmann, Rune; Ablasser, Andrea; Hopfner, Karl-Peter

    2014-08-01

    Recent discoveries in the field of innate immunity have highlighted the existence of a family of nucleic acid-sensing proteins that have similar structural and functional properties. These include the well-known oligoadenylate synthase (OAS) family proteins and the recently identified OAS homologue cyclic GMP-AMP (cGAMP) synthase (cGAS). The OAS proteins and cGAS are template-independent nucleotidyltransferases that, once activated by double-stranded nucleic acids in the cytosol, produce unique classes of 2'-5'-linked second messenger molecules, which - through distinct mechanisms - have crucial antiviral functions. 2'-5'-linked oligoadenylates limit viral propagation through the activation of the enzyme RNase L, which degrades host and viral RNA, and 2'-5'-linked cGAMP activates downstream signalling pathways to induce de novo antiviral gene expression. In this Progress article, we describe the striking functional and structural similarities between OAS proteins and cGAS, and highlight their roles in antiviral immunity.

  19. Structure of Human GIVD Cytosolic Phospholipase A2 Reveals Insights into Substrate Recognition

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hui; Klein, Michael G.; Snell, Gyorgy; Lane, Weston; Zou, Hua; Levin, Irena; Li, Ke; Sang, Bi-Ching (Takeda Cali)

    2016-07-01

    Cytosolic phospholipases A2 (cPLA2s) consist of a family of calcium-sensitive enzymes that function to generate lipid second messengers through hydrolysis of membrane-associated glycerophospholipids. The GIVD cPLA2 (cPLA2δ) is a potential drug target for developing a selective therapeutic agent for the treatment of psoriasis. Here, we present two X-ray structures of human cPLA2δ, capturing an apo state, and in complex with a substrate-like inhibitor. Comparison of the apo and inhibitor-bound structures reveals conformational changes in a flexible cap that allows the substrate to access the relatively buried active site, providing new insight into the mechanism for substrate recognition. The cPLA2δ structure reveals an unexpected second C2 domain that was previously unrecognized from sequence alignments, placing cPLA2δ into the class of membrane-associated proteins that contain a tandem pair of C2 domains. Furthermore, our structures elucidate novel inter-domain interactions and define three potential calcium-binding sites that are likely important for regulation and activation of enzymatic activity. These findings provide novel insights into the molecular mechanisms governing cPLA2's function in signal transduction.

  20. Analysis and effects of cytosolic free calcium increases in response to elicitors in Nicotiana plumbaginifolia cells.

    Science.gov (United States)

    Lecourieux, David; Mazars, Christian; Pauly, Nicolas; Ranjeva, Raoul; Pugin, Alain

    2002-10-01

    Cell suspensions obtained from Nicotiana plumbaginifolia plants stably expressing the apoaequorin gene were used to analyze changes in cytosolic free calcium concentrations ([Ca(2+)](cyt)) in response to elicitors of plant defenses, particularly cryptogein and oligogalacturonides. The calcium signatures differ in lag time, peak time, intensity, and duration. The intensities of both signatures depend on elicitor concentration and extracellular calcium concentration. Cryptogein signature is characterized by a long-sustained [Ca(2+)](cyt) increase that should be responsible for sustained mitogen-activated protein kinase activation, microtubule depolymerization, defense gene activation, and cell death. The [Ca(2+)](cyt) increase in elicitor-treated cells first results from a calcium influx, which in turns leads to calcium release from internal stores and additional Ca(2+) influx. H(2)O(2) resulting from the calcium-dependent activation of the NADPH oxidase also participates in [Ca(2+)](cyt) increase and may activate calcium channels from the plasma membrane. Competition assays with different elicitins demonstrate that [Ca(2+)](cyt) increase is mediated by cryptogein-receptor interaction.

  1. Properties of purified cytosolic isoenzyme I of Cu,Zn-superoxide dismutase from Nicotiana plumbaginifolia leaves.

    Science.gov (United States)

    Ragusa, S; Cambria, M T; Scarpa, M; Di Paolo, M L; Falconi, M; Rigo, A; Cambria, A

    2001-11-01

    The isoenzyme I of cytosolic Cu,Zn-superoxide dismutase (SOD) from Nicotiana plumbaginifolia (tobacco) leaves has been purified to apparent homogeneity. The relative molecular mass of the native isoenzyme, determined by gel filtration chromatography, is about 33.2 kDa. SDS-polyacrylamide gel electrophoresis shows that the enzyme is composed of two equal subunits of 16.6 kDa The isolectric point, assayed by isoelectric focusing, in the pH range of 3.5-6.5, is 4.3. The enzyme stability was tested at different temperatures, pH, and concentration of inhibitors (KCN and H(2)O(2)). The catalytic constant (k(cat)) was 1.17 +/- 0.14 x 10(9) M(-1) s(-1) at pH 9.9 and 0.1 M ionic strength. The activation energy of the thermal denaturation process is 263 kJ mol(-1). The electrostatic surface potential of the modeled tobacco Cu,Zn-SOD I was calculated showing that the functional spatial network of charges on the protein surface has been maintained, independently of the amino acid substitution around the active sites. Copyright 2001 Academic Press.

  2. Holocarboxylase Synthetase: A Moonlighting Transcriptional Coregulator of Gene Expression and a Cytosolic Regulator of Biotin Utilization.

    Science.gov (United States)

    León-Del-Río, Alfonso; Valadez-Graham, Viviana; Gravel, Roy A

    2017-08-21

    The vitamin biotin is an essential nutrient for the metabolism and survival of all organisms owing to its function as a cofactor of enzymes collectively known as biotin-dependent carboxylases. These enzymes use covalently attached biotin as a vector to transfer a carboxyl group between donor and acceptor molecules during carboxylation reactions. In human cells, biotin-dependent carboxylases catalyze key reactions in gluconeogenesis, fatty acid synthesis, and amino acid catabolism. Biotin is attached to apocarboxylases by a biotin ligase: holocarboxylase synthetase (HCS) in mammalian cells and BirA in microbes. Despite their evolutionary distance, these proteins share structural and sequence similarities, underscoring their importance across all life forms. However, beyond its role in metabolism, HCS participates in the regulation of biotin utilization and acts as a nuclear transcriptional coregulator of gene expression. In this review, we discuss the function of HCS and biotin in metabolism and human disease, a putative role for the enzyme in histone biotinylation, and its participation as a nuclear factor in chromatin dynamics. We suggest that HCS be classified as a moonlighting protein, with two biotin-dependent cytosolic metabolic roles and a distinct biotin-independent nuclear coregulatory function.

  3. Parathyroid hormone depresses cytosolic pH and DNA synthesis in osteoblast-like cells

    International Nuclear Information System (INIS)

    Reid, I.R.; Civitelli, R.; Avioli, L.V.; Hruska, K.A.

    1988-01-01

    It has recently become apparent that a number of hormones and growth factors modulate cytosolic pH (pH i ) and there is some evidence that this in turn may influence cell growth. The authors have examined the effects of parathyroid hormone (PTH) on both these parameters in an osteoblast-like cell line, UMR 106. Preliminary studies, using the pH-sensitive fluorescent probe 2',7'-bis(2-carboxyethyl)-5,(6)-carboxyfluorescein indicated that these cells regulate pH i by means of an amiloride-inhibitable Na + -H + exchanger. Rat PTH-(1-34) (rPTH) caused a progressive dose-related decrease in pH i with a half-maximal effect at 10 -11 M. The diacylglycerol analogue, phorbol 12-myristate 13-acetate, increased both pH i and [ 3 H]thymidine incorporation, and amiloride reduced both indexes. However, rPTH remained a potent inhibitor of [ 3 H]thymidine incorporation in the presence of amiloride, even though it did not affect pH i in these circumstances. It is concluded that PTH decreases pH i and growth in UMR 106 cells but that these changes can be dissociated. Depression of pH i may have other important effects on bone metabolism, such as reducing cell-cell communication, and may be associated with alkalinization of the bone fluid compartment

  4. Thermo-hydro-mechanical modelling of buffer, synthesis report

    International Nuclear Information System (INIS)

    Toprak, E.; Mokni, N.; Olivella, S.; Pintado, X.

    2013-08-01

    This study addresses analyses of coupled thermo-hydro-mechanical (THM) processes in a scheme considered for the spent nuclear fuel repository in Olkiluoto (Finland). The finite element code CODE B RIGHT is used to perform modelling calculations. The objective of the THM modelling was to study some fundamental design parameters. The time required to reach full saturation, the maximum temperature reached in the canister, the deformations in the buffer-backfill interface, the stress-deformation balance between the buffer and the backfill, the swelling pressure developed and the homogenization process development are critical variables. Because of the complexity of the THM processes developed, only a single deposition hole has been modelled with realistic boundary conditions which take into account the entire repository. A thermal calculation has been performed to adopt appropriate boundary conditions for a reduced domain. The modelling has been done under axisymmetric conditions. As a material model for the buffer bentonite and backfill soil, the Barcelona Basic Model (BBM) has been used. Simulation of laboratory tests conducted at B and Tech under supervision of Posiva has been carried out in order to determine the fundamental mechanical parameters for modelling the behaviour of MX-80 bentonite using the BBM model. The modelling process of the buffer-backfill interface is an essential part of tunnel backfill design. The calculations will aim to determine deformations in this intersection, the behaviour of which is important for the buffer swelling. The homogenization process is a key issue as well. Porosity evolution during the saturation process is evaluated in order to check if the final saturated density accomplishes the homogenization requirements. This report also describes the effect of the existence of an air-filled gap located between the canister and the bentonite block rings in thermo-hydro-mechanical behaviour of the future spent nuclear fuel repository in

  5. Thermo-hydro-mechanical modelling of buffer, synthesis report

    Energy Technology Data Exchange (ETDEWEB)

    Toprak, E.; Mokni, N.; Olivella, S. [Universitat Politecnica de Catalunya, Barcelona (Spain); Pintado, X. [B and Tech Oy, Helsinki (Finland)

    2013-08-15

    This study addresses analyses of coupled thermo-hydro-mechanical (THM) processes in a scheme considered for the spent nuclear fuel repository in Olkiluoto (Finland). The finite element code CODE{sub B}RIGHT is used to perform modelling calculations. The objective of the THM modelling was to study some fundamental design parameters. The time required to reach full saturation, the maximum temperature reached in the canister, the deformations in the buffer-backfill interface, the stress-deformation balance between the buffer and the backfill, the swelling pressure developed and the homogenization process development are critical variables. Because of the complexity of the THM processes developed, only a single deposition hole has been modelled with realistic boundary conditions which take into account the entire repository. A thermal calculation has been performed to adopt appropriate boundary conditions for a reduced domain. The modelling has been done under axisymmetric conditions. As a material model for the buffer bentonite and backfill soil, the Barcelona Basic Model (BBM) has been used. Simulation of laboratory tests conducted at B and Tech under supervision of Posiva has been carried out in order to determine the fundamental mechanical parameters for modelling the behaviour of MX-80 bentonite using the BBM model. The modelling process of the buffer-backfill interface is an essential part of tunnel backfill design. The calculations will aim to determine deformations in this intersection, the behaviour of which is important for the buffer swelling. The homogenization process is a key issue as well. Porosity evolution during the saturation process is evaluated in order to check if the final saturated density accomplishes the homogenization requirements. This report also describes the effect of the existence of an air-filled gap located between the canister and the bentonite block rings in thermo-hydro-mechanical behaviour of the future spent nuclear fuel

  6. Implementing the countercyclical capital buffer in South Africa: Practical considerations

    Directory of Open Access Journals (Sweden)

    Pravin Burra

    2015-03-01

    Full Text Available The Basel II regulatory framework significantly increased the resilience of the banking system, but proved ineffective in preventing the 2008/9 financial crisis. The subsequent introduction of Basel III aimed, inter alia, to supplement bank capital using buffers. The countercyclical buffer boosts existing minimum capital requirements when systemic risk surges are detected. Bolstering capital in favourable economic conditions cushions losses in unfavourable conditions, thereby addressing capital requirement procyclicality. This paper contains an overview of the countercyclical capital buffer and a critical discussion of its implementation as proposed in Basel III. Consequences of the buffer's introduction for South African banks are explored, and in particular, potential systemic risk indicator variables are identified that may be used by the South African Reserve Bank (SARB as early warning indicators of imminent systemic financial distress. These indicators may be of value to the SARB, which could use them in taking decisions on the build-up and release of the countercyclical buffer for South African banks.

  7. Carbon dioxide dissociation and buffering in chicken blood during development.

    Science.gov (United States)

    Tazawa, H; Piiper, J

    1984-07-01

    Carbon dioxide dissociation curves of oxygenated and deoxygenated bloods, the Haldane effect, the buffer value and other blood and true plasma buffering indices, O2 capacity and hematocrit were determined in bloods withdrawn from chicks before, during and after hatching and 8-month-old hens. Blood CO2 dissociation curves shifted upwards in the developing embryo till pipping, and moved downwards after pipping and hatching. In accordance with the position of the CO2 dissociation curves, the true plasma bicarbonate and red cell CO2 standardized to PCO2 = 40 torr changed. The Haldane factor at standard PCO2 increased from 0.12-0.13 on days 10-14 of incubation to 0.34 in young hens. The buffering power changed in parallel with O2 capacity and hematocrit, increasing steadily during incubation, dropping at hatching and then increasing again to the adult value. The observed changes in the CO2 dissociation curves and buffering variables during the development enable the chick to minimize the changes in the acid-base status and are favorable for coping with the increasing demand for CO2 transport and buffering of the developing bird.

  8. A readout buffer prototype for ATLAS high-level triggers

    CERN Document Server

    Calvet, D; Huet, M; Le Dû, P; Mandjavidze, I D; Mur, M

    2001-01-01

    Readout buffers are critical components in the dataflow chain of the ATLAS trigger/data-acquisition system. At up to 75 kHz, after each Level-1 trigger accept signal, these devices receive and store digitized data from groups of front-end electronic channels. Several readout buffers are grouped to form a readout buffer complex that acts as a data server for the high-level trigger selection algorithms and for the final data-collection system. This paper describes a functional prototype of a readout buffer based on a custom-made PCI mezzanine card that is designed to accept input data at up to 160 MB /s, to store up to 8 MB of data, and to distribute data chunks at the desired request rate. We describe the hardware of the card that is based on an Intel 1960 processor and complex programmable logic devices. We present the integration of several of these cards in a readout buffer complex. We measure various performance figures and discuss to which extent these can fulfil ATLAS needs. (5 refs).

  9. Evaluation of amine inhibitors for suitability as crevice buffering agents

    International Nuclear Information System (INIS)

    Jayaweera, P.; Hettiarachchi, S.

    1994-03-01

    This report describes the results of a research effort to evaluate the suitability of some selected amines and amino acids as a crevice-buffering agents in pressurized water reactor (PWR) steam generators. The amines may be useful for buffering acid crevices, and the amino acids, because they contain both acidic and basic groups, may be useful for acidic and caustic crevices. Five commercially available amines and two amino acids were studied during this research. The study involved (1) the hydrolysis of these commercially available amines and amino acids, including measurement of their kinetics of decomposition, in simulated steam generator bulk water at 290 C, and (2) determination of their thermal stability in a simulated crevice environment. The study showed that, although the high-molecular-weight amines undergo hydrothermal decomposition, they have a better buffering capacity than their low-molecular-weight counterparts at 290 C. The amines provide effective crevice buffering by increasing the pH of the crevice solution by as much as 2.84 and to 4.24 units in the experimental setup used in this program. It was concluded that polyamines provide excellent buffering of the simulated crevice environment at 290 C and morpholine remains the best low-molecular-weight amine investigated. However, detailed volatility studies of the amines were not considered in this work. Such data would be needed before in-plant testing to ensure that the amines can concentrate in steam generator crevices to the levels assumed in this study

  10. Novel nuclei isolation buffer for flow cytometric genome size estimation of Zingiberaceae: a comparison with common isolation buffers.

    Science.gov (United States)

    Sadhu, Abhishek; Bhadra, Sreetama; Bandyopadhyay, Maumita

    2016-11-01

    Cytological parameters such as chromosome numbers and genome sizes of plants are used routinely for studying evolutionary aspects of polyploid plants. Members of Zingiberaceae show a wide range of inter- and intrageneric variation in their reproductive habits and ploidy levels. Conventional cytological study in this group of plants is severely hampered by the presence of diverse secondary metabolites, which also affect their genome size estimation using flow cytometry. None of the several nuclei isolation buffers used in flow cytometry could be used very successfully for members of Zingiberaceae to isolate good quality nuclei from both shoot and root tissues. The competency of eight nuclei isolation buffers was compared with a newly formulated buffer, MB01, in six different genera of Zingiberaceae based on the fluorescence intensity of propidium iodide-stained nuclei using flow cytometric parameters, namely coefficient of variation of the G 0 /G 1 peak, debris factor and nuclei yield factor. Isolated nuclei were studied using fluorescence microscopy and bio-scanning electron microscopy to analyse stain-nuclei interaction and nuclei topology, respectively. Genome contents of 21 species belonging to these six genera were determined using MB01. Flow cytometric parameters showed significant differences among the analysed buffers. MB01 exhibited the best combination of analysed parameters; photomicrographs obtained from fluorescence and electron microscopy supported the superiority of MB01 buffer over other buffers. Among the 21 species studied, nuclear DNA contents of 14 species are reported for the first time. Results of the present study substantiate the enhanced efficacy of MB01, compared to other buffers tested, in the generation of acceptable cytograms from all species of Zingiberaceae studied. Our study facilitates new ways of sample preparation for further flow cytometric analysis of genome size of other members belonging to this highly complex polyploid family

  11. Piping and erosion in buffer and backfill materials. Current knowledge

    International Nuclear Information System (INIS)

    Boergesson, Lennart; Sanden, Torbjoern

    2006-09-01

    The water inflow into the deposition holes and tunnels in a repository will mainly take place through fractures in the rock and will lead to that the buffer and backfill will be wetted and homogenised. But in general the buffer and backfill cannot absorb all water that runs through a fracture, which leads to that a water pressure will be generated in the fracture when the inflow is hindered. If the counter pressure and strength of the buffer or backfill is insufficiently high, piping and subsequent erosion may take place. The processes and consequences of piping and erosion have been studied in some projects and several laboratory test series in different scales have been carried through. This brief report describes these tests and the results and conclusions that have emerged. The knowledge of piping and erosion is insufficient today and additional studies are needed and running

  12. On the risk of liquefaction of buffer and backfill

    Energy Technology Data Exchange (ETDEWEB)

    Pusch, R. [Geodevelopment AB, Lund (Sweden)

    2000-10-01

    The necessary prerequisites for liquefaction of buffers and backfills in a KBS-3 repository exist but the stress conditions and intended densities practically eliminate the risk of liquefaction for single earthquakes with magnitudes up to M=8 and normal duration. For buffers rich in expandable minerals it would be possible to reduce the density at water saturation to 1,700 - 1,800 kg/m{sup 3} or even less without any significant risk of liquefaction, while the density at saturation of backfills with 10 - 15% expandable clay should not be reduced to less than about 1,900 kg/m{sup 3}. Since the proposed densities of both buffers and backfills will significantly exceed these minimum values it is concluded that there is no risk of liquefaction of the engineered soil barriers in a KBS-3 repository even for very significant earthquakes.

  13. On the risk of liquefaction of buffer and backfill

    International Nuclear Information System (INIS)

    Pusch, R.

    2000-10-01

    The necessary prerequisites for liquefaction of buffers and backfills in a KBS-3 repository exist but the stress conditions and intended densities practically eliminate the risk of liquefaction for single earthquakes with magnitudes up to M=8 and normal duration. For buffers rich in expandable minerals it would be possible to reduce the density at water saturation to 1,700 - 1,800 kg/m 3 or even less without any significant risk of liquefaction, while the density at saturation of backfills with 10 - 15% expandable clay should not be reduced to less than about 1,900 kg/m 3 . Since the proposed densities of both buffers and backfills will significantly exceed these minimum values it is concluded that there is no risk of liquefaction of the engineered soil barriers in a KBS-3 repository even for very significant earthquakes

  14. Hydrothermal stability of bentonite-based buffer materials

    International Nuclear Information System (INIS)

    Johnston, R.M.; Miller, H.G.

    1985-02-01

    The reactions expected in bentonite-based buffer materials under conditions typical of a nuclear fuel waste disposal include mineral transformations (e.g. smectite to illite; smectite to zeolite) and a range of low-temperature cementation reactions. The probable extent and significance of these reactions are reviewed, and other reactions involving proposed filler sands are also examined briefly. The effects of mineral transformations on buffer performance will be insignificant if disposal vault temperatures do not exceed 100-120 degrees C and pH remains in the range 4 to 8. At pH > 9, zeolitization and silica dissolution may occur and buffer stability cannot be assured. The effects of cementation reactions may be significant, but are difficult to predict and require further investigation

  15. Capacity gains of buffer-aided moving relays

    KAUST Repository

    Zafar, Ammar

    2017-03-14

    This work investigates the gain due to reduction in path loss by deploying buffer-aided moving relaying. In particular, the increase in gain due to moving relays is studied for dual-hop broadcast channels and the bidirectional relay channel. It is shown that the exploited gains in these channels due to buffer-aided relaying can be enhanced by utilizing the fact that a moving relay can communicate with the terminal closest to it and store the data in the buffer and then forward the data to the intended destination when it comes in close proximity with the destination. Numerical results show that for both the considered channels the achievable rates are increased as compared to the case of stationary relays. Numerical results also show that more significant increase in performance is seen when the relay moves to-and-fro between the source and the relay.

  16. Buffer thermal energy storage for an air Brayton solar engine

    Science.gov (United States)

    Strumpf, H. J.; Barr, K. P.

    1981-01-01

    The application of latent-heat buffer thermal energy storage to a point-focusing solar receiver equipped with an air Brayton engine was studied. To demonstrate the effect of buffer thermal energy storage on engine operation, a computer program was written which models the recuperator, receiver, and thermal storage device as finite-element thermal masses. Actual operating or predicted performance data are used for all components, including the rotating equipment. Based on insolation input and a specified control scheme, the program predicts the Brayton engine operation, including flows, temperatures, and pressures for the various components, along with the engine output power. An economic parametric study indicates that the economic viability of buffer thermal energy storage is largely a function of the achievable engine life.

  17. Database on gas migration tests through bentonite buffer material

    International Nuclear Information System (INIS)

    Tanai, Kenji

    2009-02-01

    Carbon steel is a candidate material for an overpack for geological disposal of high-level radioactive waste in Japan. The corrosion of the carbon steel overpack in aqueous solution under anoxic conditions will cause the generation of hydrogen gas, which may affect hydrological and mechanical properties of the bentonite buffer. To evaluate such an effect of gas generation, it is necessary to develop a model of gas migration through bentonite buffer material taking account of data obtained from experiments. The gas migration experiments under both unsaturated and saturated conditions have been carried out to clarify the fundamental characteristics of bentonite for gas migration. This report compiles the experimental data obtained from gas migration tests for buffer material which has been conducted by JAEA until December, 2007. A CD-ROM is attached as an appendix. (author)

  18. Qualitative and quantitative assessment of interior moisture buffering by enclosures

    DEFF Research Database (Denmark)

    Janssen, Hans; Roels, Staf

    2009-01-01

    The significance of interior humidity in attaining sustainable, durable, healthy and comfortable buildings is increasingly recognised. Given their significant interaction, interior humidity appraisals need a qualitative and/or quantitative assessment of interior moisture buffering. While the effe......The significance of interior humidity in attaining sustainable, durable, healthy and comfortable buildings is increasingly recognised. Given their significant interaction, interior humidity appraisals need a qualitative and/or quantitative assessment of interior moisture buffering. While...... the effective moisture penetration depth and effective capacitance models allow quantified assessment, their reliance on the ‘moisture penetration depth’ necessitates comprehensive material properties and hampers their application to multi-dimensional interior objects. On the other hand, while various recently...... an alternative basis for quantitative evaluation of interior moisture buffering by the effective moisture penetration depth and effective capacitance models. The presented methodology uses simple and fast measurements only and can also be applied to multimaterial and/or multidimensional interior elements....

  19. Clay colloid formation and release from MX-80 buffer

    International Nuclear Information System (INIS)

    Pusch, R.

    1999-12-01

    Flowing groundwater can tear off clay colloids from buffer clay that has penetrated into fractures and transport them and bring sorbed radionuclides up to the biosphere. The colloids are 2-50 μm particle aggregates that are liberated from expanded, softened buffer if the water flow rate in the fractures exceeds a few centimeters per second. Except for the first few months or years after application of the buffer in the deposition holes the flow rate will not be as high as that. The aperture of the fractures will not hinder transport of colloids but most of the fractures contain clastic fillings, usually chlorite, that attract and immobilize them. This condition and the flow rate criterion combine to reduce the chance of radionuclide-bearing clay colloids to reach the biosphere to practically zero except for certain cases that need to be considered

  20. Capacity gains of buffer-aided moving relays

    KAUST Repository

    Zafar, Ammar; Shaqfeh, Mohammad; Alnuweiri, Hussein; Alouini, Mohamed-Slim

    2017-01-01

    This work investigates the gain due to reduction in path loss by deploying buffer-aided moving relaying. In particular, the increase in gain due to moving relays is studied for dual-hop broadcast channels and the bidirectional relay channel. It is shown that the exploited gains in these channels due to buffer-aided relaying can be enhanced by utilizing the fact that a moving relay can communicate with the terminal closest to it and store the data in the buffer and then forward the data to the intended destination when it comes in close proximity with the destination. Numerical results show that for both the considered channels the achievable rates are increased as compared to the case of stationary relays. Numerical results also show that more significant increase in performance is seen when the relay moves to-and-fro between the source and the relay.

  1. Agroforestry buffers for nonpoint source pollution reductions from agricultural watersheds.

    Science.gov (United States)

    Udawatta, Ranjith P; Garrett, Harold E; Kallenbach, Robert

    2011-01-01

    Despite increased attention and demand for the adoption of agroforestry practices throughout the world, rigorous long-term scientific studies confirming environmental benefits from the use of agroforestry practices are limited. The objective was to examine nonpoint-source pollution (NPSP) reduction as influenced by agroforestry buffers in watersheds under grazing and row crop management. The grazing study consists of six watersheds in the Central Mississippi Valley wooded slopes and the row crop study site consists of three watersheds in a paired watershed design in Central Claypan areas. Runoff water samples were analyzed for sediment, total nitrogen (TN), and total phosphorus (TP) for the 2004 to 2008 period. Results indicate that agroforestry and grass buffers on grazed and row crop management sites significantly reduce runoff, sediment, TN, and TP losses to streams. Buffers in association with grazing and row crop management reduced runoff by 49 and 19%, respectively, during the study period as compared with respective control treatments. Average sediment loss for grazing and row crop management systems was 13.8 and 17.9 kg ha yr, respectively. On average, grass and agroforestry buffers reduced sediment, TN, and TP losses by 32, 42, and 46% compared with the control treatments. Buffers were more effective in the grazing management practice than row crop management practice. These differences could in part be attributed to the differences in soils, management, and landscape features. Results from this study strongly indicate that agroforestry and grass buffers can be designed to improve water quality while minimizing the amount of land taken out of production. American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America.

  2. Exposure to buffer solution alters tendon hydration and mechanics.

    Science.gov (United States)

    Safa, Babak N; Meadows, Kyle D; Szczesny, Spencer E; Elliott, Dawn M

    2017-08-16

    A buffer solution is often used to maintain tissue hydration during mechanical testing. The most commonly used buffer solution is a physiological concentration of phosphate buffered saline (PBS); however, PBS increases the tissue's water content and decreases its tensile stiffness. In addition, solutes from the buffer can diffuse into the tissue and interact with its structure and mechanics. These bathing solution effects can confound the outcome and interpretation of mechanical tests. Potential bathing solution artifacts, including solute diffusion, and their effect on mechanical properties, are not well understood. The objective of this study was to measure the effects of long-term exposure of rat tail tendon fascicles to several concentrations (0.9-25%) of NaCl, sucrose, polyethylene glycol (PEG), and SPEG (NaCl+PEG) solutions on water content, solute diffusion, and mechanical properties. We found that with an increase in solute concentration the apparent water content decreased for all solution types. Solutes diffused into the tissue for NaCl and sucrose, however, no solute diffusion was observed for PEG or SPEG. The mechanical properties changed for both NaCl solutions, in particular after long-term (8h) incubation the modulus and equilibrium stress decreased compared to short-term (15min) for 25% NaCl, and the cross sectional area increased for 0.9% NaCl. However, the mechanical properties were unchanged for both PEG and SPEG except for minor alterations in stress relaxation parameters. This study shows that NaCl and sucrose buffer solutions are not suitable for long-term mechanical tests. We therefore propose using PEG or SPEG as alternative buffer solutions that after long-term incubation can maintain tissue hydration without solute diffusion and produce a consistent mechanical response. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. MATHEMATICAL MODELLING OF QUAZISTEADY MODE OF BEARING AIR BUFFER FILLING

    Directory of Open Access Journals (Sweden)

    E. D. Chertov

    2014-01-01

    Full Text Available Summary. Today the only way to eliminate contact with the product during the manufacturing process is to provide a support surface under its support surface air buffer layer formed due to the expiration of the working environment through holes perforated gas distribution grids forms. There proposed the method of contactless formation of products consisting of composite materials by the means of air buffer in the article. The results of theoretical and experimental investigations of hydro-gas-dynamic processes occurring when casting of organic- mineral composite material onto the bearing air buffer expressed in the form of mathematical description realizing original hypotheses reflected in the choice of transformation algorithm and limiting conditions are presented. On the base of obtained mathematical model the algorithm of calculation of optimum parameters of transporting systems with discretely powered gas buffer is developed. The method of deduction of a semi-finished product on the gas buffer, which allows to level the pressure field under the bearing surface of the deduction object due to the usage of devices of pseudo fluidized granular material in pneumatic chambers is offered. The application of this method allows to eliminate the possibility of contact between the composite material and the working surface of the equipment and also to reduce the cost of production of pneumatic devices, to improve operational characteristics of this equipment. Submitted depending allowed to develop the methodology and implementation of engineering calculation device for non-contact casting composite materials on air buffer, semi-industrial and industrial variants were created and put into production.

  4. Silencing of cytosolic NADP+-dependent isocitrate dehydrogenase gene enhances ethanol-induced toxicity in HepG2 cells.

    Science.gov (United States)

    Yang, Eun Sun; Lee, Su-Min; Park, Jeen-Woo

    2010-07-01

    It has been shown that acute and chronic alcohol administrations increase the production of reactive oxygen species, lower cellular antioxidant levels and enhance oxidative stress in many tissues. We recently reported that cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) functions as an antioxidant enzyme by supplying NADPH to the cytosol. Upon exposure to ethanol, IDPc was susceptible to the loss of its enzyme activity in HepG2 cells. Transfection of HepG2 cells with an IDPc small interfering RNA noticeably downregulated IDPc and enhanced the cells' vulnerability to ethanol-induced cytotoxicity. Our results suggest that suppressing the expression of IDPc enhances ethanol-induced toxicity in HepG2 cells by further disruption of the cellular redox status.

  5. Cyclic GMP-AMP Synthase is a Cytosolic DNA Sensor that Activates the Type-I Interferon Pathway

    Science.gov (United States)

    Sun, Lijun; Wu, Jiaxi; Du, Fenghe; Chen, Xiang; Chen, Zhijian J.

    2013-01-01

    The presence of DNA in the cytoplasm of mammalian cells is a danger signal that triggers the host immune responses such as the production of type-I interferons (IFN). Cytosolic DNA induces IFN through the production of cyclic-GMP-AMP (cGAMP), which binds to and activates the adaptor protein STING. Through biochemical fractionation and quantitative mass spectrometry, we identified a cGAMP synthase (cGAS), which belongs to the nucleotidyltransferase family. Overexpression of cGAS activated the transcription factor IRF3 and induced IFNβ in a STING-dependent manner. Knockdown of cGAS inhibited IRF3 activation and IFNβ induction by DNA transfection or DNA virus infection. cGAS bound to DNA in the cytoplasm and catalyzed cGAMP synthesis. These results indicate that cGAS is a cytosolic DNA sensor that induces interferons by producing the second messenger cGAMP. PMID:23258413

  6. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway.

    Science.gov (United States)

    Sun, Lijun; Wu, Jiaxi; Du, Fenghe; Chen, Xiang; Chen, Zhijian J

    2013-02-15

    The presence of DNA in the cytoplasm of mammalian cells is a danger signal that triggers host immune responses such as the production of type I interferons. Cytosolic DNA induces interferons through the production of cyclic guanosine monophosphate-adenosine monophosphate (cyclic GMP-AMP, or cGAMP), which binds to and activates the adaptor protein STING. Through biochemical fractionation and quantitative mass spectrometry, we identified a cGAMP synthase (cGAS), which belongs to the nucleotidyltransferase family. Overexpression of cGAS activated the transcription factor IRF3 and induced interferon-β in a STING-dependent manner. Knockdown of cGAS inhibited IRF3 activation and interferon-β induction by DNA transfection or DNA virus infection. cGAS bound to DNA in the cytoplasm and catalyzed cGAMP synthesis. These results indicate that cGAS is a cytosolic DNA sensor that induces interferons by producing the second messenger cGAMP.

  7. Polysaccharide fraction from higher plants which strongly interacts with the cytosolic phosphorylase isozyme. I. Isolation and characterization

    International Nuclear Information System (INIS)

    Yang, Yi; Steup, M.

    1990-01-01

    From leaves of Spinacia oleracea L. or from Pisum sativum L. and from cotyledons of germinating pea seeds a high molecular weight polysaccharide fraction was isolated. The apparent size of the fraction, as determined by gel filtration, was similar to that of dextran blue. Following acid hydrolysis the monomer content of the polysaccharide preparation was studied using high pressure liquid and thin layer chromatography. Glucose, galactose, arabinose, and ribose were the main monosaccharide compounds. The native polysaccharide preparation interacted strongly with the cytosolic isozyme of phosphorylase (EC 2.4.1.1). Interaction with the plastidic phosphorylase isozyme(s) was by far weaker. Interaction with the cytosolic isozyme was demonstrated by affinity electrophoresis, kinetic measurements, and by 14 C-labeling experiments in which the glucosyl transfer from [ 14 C]glucose 1-phosphate to the polysaccharide preparation was monitored

  8. Differential compartmentation of sucrose and gentianose in the cytosol and vacuoles of storage root protoplasts from Gentiana Lutea L.

    Science.gov (United States)

    Keller, F; Wiemken, A

    1982-12-01

    The storage roots of perennial Gentiana lutea L.plants contain several sugars. The predominant carbohydrate reserve is gentianose (β-D-glucopyranosyl-(1 → 6)-α-D-glucopyranosyl-(1 ↔ 2)-β-D-fructofuranoside). Vacuoles were isolated from root protoplasts and purified through a betaine density gradient. The yield was about 75%. Gentianose and gentiobiose were localized to 100% in the vacuoles, fructose and glucose to about 80%, and sucrose to only about 50%. Taking the volumes of the vacuolar and extravacuolar (cytosolic) compartments into account it is inferred that gentianose is located exclusively in the vacuoles, whilst sucrose is much more concentrated in the cytosol where it may play a role as a cryoprotectant. The concentration of fructose and glucose appeared to be similar on both sides of the tonoplast.

  9. Thermo-Hydraulic Modelling of Buffer and Backfill

    International Nuclear Information System (INIS)

    Pintado, X.; Rautioaho, E.

    2013-09-01

    The temporal evolution of saturation, liquid pressure and temperature in the components of the engineered barrier system was studied using numerical methods. A set of laboratory tests was conducted to calibrate the parameters employed in the models. The modelling consisted of thermal, hydraulic and thermo-hydraulic analysis in which the significant thermo-hydraulic processes, parameters and features were identified. CODE B RIGHT was used for the finite element modelling and supplementary calculations were conducted with analytical methods. The main objective in this report is to improve understanding of the thermo-hydraulic processes and material properties that affect buffer behaviour in the Olkiluoto repository and to determine the parametric requirements of models for the accurate prediction of this behaviour. The analyses consisted of evaluating the influence of initial canister temperature and gaps in the buffer, and the role played by fractures and the rock mass located between fractures in supplying water for buffer and backfill saturation. In the thermo-hydraulic analysis, the primary processes examined were the effects of buffer drying near the canister on temperature evolution and the manner in which heat flow affects the buffer saturation process. Uncertainties in parameters and variations in the boundary conditions, modelling geometry and thermo-hydraulic phenomena were assessed with a sensitivity analysis. The material parameters, constitutive models, and assumptions made were carefully selected for all the modelling cases. The reference parameters selected for the simulations were compared and evaluated against laboratory measurements. The modelling results highlight the importance of understanding groundwater flow through the rock mass and from fractures in the rock in order to achieve reliable predictions regarding buffer saturation, since saturation times could range from a few years to tens of thousands of years depending on the hydrogeological

  10. Impact of Burst Buffer Architectures on Application Portability

    Energy Technology Data Exchange (ETDEWEB)

    Harms, Kevin [Argonne National Lab. (ANL), Argonne, IL (United States); Oral, H. Sarp [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). National Center for Computational Science; Atchley, Scott [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). National Center for Computational Science; Vazhkudai, Sudharshan S. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). National Center for Computational Science

    2016-09-30

    The Oak Ridge and Argonne Leadership Computing Facilities are both receiving new systems under the Collaboration of Oak Ridge, Argonne, and Livermore (CORAL) program. Because they are both part of the INCITE program, applications need to be portable between these two facilities. However, the Summit and Aurora systems will be vastly different architectures, including their I/O subsystems. While both systems will have POSIX-compliant parallel file systems, their Burst Buffer technologies will be different. This difference may pose challenges to application portability between facilities. Application developers need to pay attention to specific burst buffer implementations to maximize code portability.

  11. The Countercyclical Capital Buffer of Basel III: A Critical Assessment

    OpenAIRE

    Repullo, Rafael; Saurina, Jesús

    2011-01-01

    We provide a critical assessment of the countercyclical capital buffer in the new regulatory framework known as Basel III, which is based on the deviation of the creditto- GDP ratio with respect to its trend. We argue that a mechanical application of the buffer would tend to reduce capital requirements when GDP growth is high and increase them when GDP growth is low, so it may end up exacerbating the inherent pro-cyclicality of risk-sensitive bank capital regulation. We also note that Basel I...

  12. Survey of Swedish buffer material candidates and methods for characterization

    International Nuclear Information System (INIS)

    Erlstroem, M.; Pusch, R.

    1987-12-01

    The study has given a good overview of potential clay buffer candidates in the part of Sweden that offers the best possibilities to find large accessible quantities of smectitic materials. The most promising Scanian materials are those in the Kaageroed and Vallaakra (Margreteberg) areas since they represent the most smectitic ones, which may serve as raw material for the production of canister embedment. The moraine clays in the Lund-Landskrona region seem to be useful for backfilling purposes. A refined version of Reynolds technique is suggested as an SKB standard for prospecting and characterization of buffer materials. (orig./DG)

  13. Buffer capacity of the coelomic fluid in echinoderms.

    Science.gov (United States)

    Collard, Marie; Laitat, Kim; Moulin, Laure; Catarino, Ana I; Grosjean, Philippe; Dubois, Philippe

    2013-09-01

    The increase in atmospheric CO2 due to anthropogenic activity results in an acidification of the surface waters of the oceans. The impact of these chemical changes depends on the considered organisms. In particular, it depends on the ability of the organism to control the pH of its inner fluids. Among echinoderms, this ability seems to differ significantly according to species or taxa. In the present paper, we investigated the buffer capacity of the coelomic fluid in different echinoderm taxa as well as factors modifying this capacity. Euechinoidea (sea urchins except Cidaroidea) present a very high buffer capacity of the coelomic fluid (from 0.8 to 1.8mmolkg(-1) SW above that of seawater), while Cidaroidea (other sea urchins), starfish and holothurians have a significantly lower one (from -0.1 to 0.4mmolkg(-1) SW compared to seawater). We hypothesize that this is linked to the more efficient gas exchange structures present in the three last taxa, whereas Euechinoidea evolved specific buffer systems to compensate lower gas exchange abilities. The constituents of the buffer capacity and the factors influencing it were investigated in the sea urchin Paracentrotus lividus and the starfish Asterias rubens. Buffer capacity is primarily due to the bicarbonate buffer system of seawater (representing about 63% for sea urchins and 92% for starfish). It is also partly due to coelomocytes present in the coelomic fluid (around 8% for both) and, in P. lividus only, a compound of an apparent size larger than 3kDa is involved (about 15%). Feeding increased the buffer capacity in P. lividus (to a difference with seawater of about 2.3mmolkg(-1) SW compared to unfed ones who showed a difference of about 0.5mmolkg(-1) SW) but not in A. rubens (difference with seawater of about 0.2 for both conditions). In P. lividus, decreased seawater pH induced an increase of the buffer capacity of individuals maintained at pH7.7 to about twice that of the control individuals and, for those at pH7

  14. Evidence for the buffer zone in a plasma accelerator

    International Nuclear Information System (INIS)

    Thomas, K.A.; Clothiaux, E.J.

    1991-01-01

    The existence of an absorbing layer, or buffer zone, of weakly ionized gas between the rear of the projectile and the front of the plasma arc armature in a plasma accelerator has been postulated. In the studies reported here a technique for finding the position of the projectile as a function of time is given and compared to the plasma armature position as determined by inductive probes. Analyses of these signals provide the basis for a description of the in-bore motion of the projectile with respect to the plasma arc armature. The experimental evidence appears to support the existence of a buffer zone

  15. Moisture buffering and its consequence in whole building hygrothermal modeling

    DEFF Research Database (Denmark)

    Rode, Carsten; Grau, Karl

    2008-01-01

    Moisture absorption and desorption of materials in contact with indoor air of buildings can be used as a passive, i.e., nonmechanical, way to moderate the variation of indoor humidity. This phenomenon, which is recognized as,moisture buffering', could potentially be used as an attractive feature...... for ventilation if indoor humidity is a parameter for controlling ventilation rate, 2. it is possible to improve the perceived acceptability of indoor air, as judged by the temperature and humidity of the air, by using moisture buffering to control the indoor humidity. The results of the whole building...

  16. Biomechanical Response and Behavior of Users under Emergency Buffer Crash

    Directory of Open Access Journals (Sweden)

    R. Miralbes

    2013-01-01

    Full Text Available This paper aims to study the biomechanical effects on elevator users and the injuries sustained should an elevator crash happen. The analysis will focus on buffer impact, signaling that the earlier mentioned buffer is usually located at the bottom of the pit. In order to carry out this analysis, a numerical technique based on finite element method will be used, while elevator users will be simulated by means of automotive dummies. Two crash factors will be studied, namely, location of dummy and fall velocity. The analysis criteria will be damages sustained by the dummy, based on biomechanical index such as HIC, CSI, forces, and accelerations.

  17. Effect of buffer at nanoscale molecular recognition interfaces - electrostatic binding of biological polyanions.

    Science.gov (United States)

    Rodrigo, Ana C; Laurini, Erik; Vieira, Vânia M P; Pricl, Sabrina; Smith, David K

    2017-10-19

    We investigate the impact of an over-looked component on molecular recognition in water-buffer. The binding of a cationic dye to biological polyanion heparin is shown by isothermal calorimetry to depend on buffer (Tris-HCl > HEPES > PBS). The heparin binding of self-assembled multivalent (SAMul) cationic micelles is even more buffer dependent. Multivalent electrostatic molecular recognition is buffer dependent as a result of competitive interactions between the cationic binding interface and anions present in the buffer.

  18. Fully-coupled mathematical modeling of actomyosin-cytosolic two-phase flow in a highly deformable moving Keratocyte cell.

    Science.gov (United States)

    Nikmaneshi, M R; Firoozabadi, B; Saidi, M S

    2018-01-23

    Interaction between intracellular dynamics and extracellular matrix (ECM) generally occurred into very thin fragment of moving cell, namely lamellipodia, enables all movable cells to crawl on ECM. In fast-moving cells such as fish Keratocytes, Lamellipodia including most cell area finds a fan-like shape during migration, with a variety of aspect ratio function of fish type. In this work, our purpose is to present a novel and more complete two-dimensional continuum mathematical model of actomyosin-cytosolic two-phase flow of a self-deforming Keratocyte with circular spreaded to steady fan-like shape. In the new approach, in addition to the two-phase flow of the F-actin and cytosol, the G-actin transport was spatiotemporally modeled. We also for the first time modeled the effect of variable volume fraction of the moving F-actin porous network on solute transport in the cytosolic fluid. Our novel fully-coupled mathematical model provides a better understanding of intracellular dynamics of fast-migrating Keratocytes; such as the F-actin centripetal and cytosolic fountain-like flows, free-active myosin distribution, distribution sequence of the G-actin, F-actin, and myosin, and myosin-induced pressure flied of cytoplasm as well as the map of intracellular forces like myosin contraction and adhesion traction. All these results are qualitatively and quantitatively in good agreement with experimental observations. According to a range of value of parameters used in this model, our steady state of moving Keratocyte finds fan-like shape with the same aspect ratio as wide category of fish Keratocytes. This new model can predict shape of Keratocytes in other range of parameter values. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Negative Effect of Ellagic Acid on Cytosolic pH Regulation and Glycolytic Flux in Human Endometrial Cancer Cells

    OpenAIRE

    Khalid N. M. Abdelazeem; Yogesh Singh; Florian Lang; Madhuri S. Salker

    2017-01-01

    Background/Aims: Key properties of tumor cells include enhanced glycolytic flux with excessive consumption of glucose and formation of lactate. As glycolysis is highly sensitive to cytosolic pH, maintenance of glycolysis requires export of H+ ions, which is in part accomplished by Na+/H+ exchangers, such as NHE1. The carrier is sensitive to oxidative stress. Growth of tumor cells could be suppressed by the polyphenol Ellagic acid, which is found in various fruits and vegetables. An effect of ...

  20. Exchange of cytosolic content between T cells and tumor cells activates CD4 T cells and impedes cancer growth.

    Directory of Open Access Journals (Sweden)

    Matthias Hardtke-Wolenski

    Full Text Available BACKGROUND: T cells are known to participate in the response to tumor cells and react with cytotoxicity and cytokine release. At the same time tumors established versatile mechanisms for silencing the immune responses. The interplay is far from being completely understood. In this study we show contacts between tumor cells and lymphocytes revealing novel characteristics in the interaction of T cells and cancer cells in a way not previously described. METHODS/ FINDINGS: Experiments are based on the usage of a hydrophilic fluorescent dye that occurs free in the cytosol and thus transfer of fluorescent cytosol from one cell to the other can be observed using flow cytometry. Tumor cells from cell lines of different origin or primary hepatocellular carcinoma (HCC cells were incubated with lymphocytes from human and mice. This exposure provoked a contact dependent uptake of tumor derived cytosol by lymphocytes--even in CD4⁺ T cells and murine B cells--which could not be detected after incubation of lymphocytes with healthy cells. The interaction was a direct one, not requiring the presence of accessory cells, but independent of cytotoxicity and TCR engagement. Electron microscopy disclosed 100-200 nm large gaps in the cell membranes of connected cells which separated viable and revealed astonishing outcome. While the lymphocytes were induced to proliferate in a long term fashion, the tumor cells underwent a temporary break in cell division. The in vitro results were confirmed in vivo using a murine acute lymphoblastic leukemia (ALL model. The arrest of tumor proliferation resulted in a significant prolonged survival of challenged mice. CONCLUSIONS: The reported cell-cell contacts reveal new characteristics i.e. the enabling of cytosol flow between the cells including biological active proteins that influence the cell cycle and biological behaviour of the recipient cells. This adds a completely new aspect in tumor induced immunology.

  1. Inhibitor-induced oxidation of the nucleus and cytosol in Arabidopsis thaliana: implications for organelle to nucleus retrograde signalling.

    Science.gov (United States)

    Karpinska, Barbara; Alomrani, Sarah Owdah; Foyer, Christine H

    2017-09-26

    Concepts of organelle-to-nucleus signalling pathways are largely based on genetic screens involving inhibitors of chloroplast and mitochondrial functions such as norflurazon, lincomycin (LINC), antimycin A (ANT) and salicylhydroxamic acid. These inhibitors favour enhanced cellular oxidation, but their precise effects on the cellular redox state are unknown. Using the in vivo reduction-oxidation (redox) reporter, roGFP2, inhibitor-induced changes in the glutathione redox potentials of the nuclei and cytosol were measured in Arabidopsis thaliana root, epidermal and stomatal guard cells, together with the expression of nuclear-encoded chloroplast and mitochondrial marker genes. All the chloroplast and mitochondrial inhibitors increased the degree of oxidation in the nuclei and cytosol. However, inhibitor-induced oxidation was less marked in stomatal guard cells than in epidermal or root cells. Moreover, LINC and ANT caused a greater oxidation of guard cell nuclei than the cytosol. Chloroplast and mitochondrial inhibitors significantly decreased the abundance of LHCA1 and LHCB1 transcripts. The levels of WHY1 , WHY3 and LEA5 transcripts were increased in the presence of inhibitors. Chloroplast inhibitors decreased AOXA1 mRNA levels, while mitochondrial inhibitors had the opposite effect. Inhibitors that are used to characterize retrograde signalling pathways therefore have similar general effects on cellular redox state and gene expression.This article is part of the themed issue 'Enhancing photosynthesis in crop plants: targets for improvement'. © 2017 The Authors.

  2. Low glutathione regulates gene expression and the redox potentials of the nucleus and cytosol in Arabidopsis thaliana.

    Science.gov (United States)

    Schnaubelt, Daniel; Queval, Guillaume; Dong, Yingping; Diaz-Vivancos, Pedro; Makgopa, Matome Eugene; Howell, Gareth; De Simone, Ambra; Bai, Juan; Hannah, Matthew A; Foyer, Christine H

    2015-02-01

    Reduced glutathione (GSH) is considered to exert a strong influence on cellular redox homeostasis and to regulate gene expression, but these processes remain poorly characterized. Severe GSH depletion specifically inhibited root meristem development, while low root GSH levels decreased lateral root densities. The redox potential of the nucleus and cytosol of Arabidopsis thaliana roots determined using roGFP probes was between -300 and -320 mV. Growth in the presence of the GSH-synthesis inhibitor buthionine sulfoximine (BSO) increased the nuclear and cytosolic redox potentials to approximately -260 mV. GSH-responsive genes including transcription factors (SPATULA, MYB15, MYB75), proteins involved in cell division, redox regulation (glutaredoxinS17, thioredoxins, ACHT5 and TH8) and auxin signalling (HECATE), were identified in the GSH-deficient root meristemless 1-1 (rml1-1) mutant, and in other GSH-synthesis mutants (rax1-1, cad2-1, pad2-1) as well as in the wild type following the addition of BSO. Inhibition of auxin transport had no effect on organ GSH levels, but exogenous auxin decreased the root GSH pool. We conclude that GSH depletion significantly increases the redox potentials of the nucleus and cytosol, and causes arrest of the cell cycle in roots but not shoots, with accompanying transcript changes linked to altered hormone responses, but not oxidative stress. © 2013 John Wiley & Sons Ltd.

  3. [Effects of cytosolic bacteria on cyclic GMP-AMP synthase expression in human gingival tissues and periodontal ligament cells].

    Science.gov (United States)

    Xiaojun, Yang; Yongmei, Tan; Zhihui, Tian; Ting, Zhou; Wanghong, Zhao; Jin, Hou

    2017-04-01

    This work aims to determine the effect of cytosolic bacteria on the expression of cyclic GMP-AMP synthase (cGAS) in human periodontal ligament cells (hPDLCs) and gingival tissues. The ability of Porphyromonas gingivalis (P. gingivalis) to invade hPDLCs was detected using laser scanning confocal microscope assay at a multiplicity of infection of 10. P. gingivalis-infected cells were sorted by fluorescence-activated cell sorting (FACS). Then, quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect cGAS expression in infected cells. Finally, the location and expression of cGAS in inflammatory and normal gingival tissues were investigated by immunohistochemistry. P. gingivalis actively invaded hPDLCs. Moreover, cGAS expression significantly increased in P. gingivalis-infected cells. Although cGAS was expressed in the epithelial and subepithelial cells of both inflamed and normal gingival tissues, cGAS expression significantly increased in inflamed gingival tissues. Cytosolic bacteria can upregulate cGAS expression in infected cells. These data suggest that cGAS may act as pattern-recognition receptors and participate in recognizing cytosolic nucleic acid pathogen-associated molecular patterns.
.

  4. Cytosolic calcium transients are a determinant of contraction-induced HSP72 transcription in single skeletal muscle fibers.

    Science.gov (United States)

    Stary, Creed M; Hogan, Michael C

    2016-05-15

    The intrinsic activating factors that induce transcription of heat shock protein 72 (HSP72) in skeletal muscle following exercise remain unclear. We hypothesized that the cytosolic Ca(2+) transient that occurs with depolarization is a determinant. We utilized intact, single skeletal muscle fibers from Xenopus laevis to test the role of the cytosolic Ca(2+) transient and several other exercise-related factors (fatigue, hypoxia, AMP kinase, and cross-bridge cycling) on the activation of HSP72 transcription. HSP72 and HSP60 mRNA levels were assessed with real-time quantitative PCR; cytosolic Ca(2+) concentration ([Ca(2+)]cyt) was assessed with fura-2. Both fatiguing and nonfatiguing contractions resulted in a significant increase in HSP72 mRNA. As expected, peak [Ca(2+)]cyt remained tightly coupled with peak developed tension in contracting fibers. Pretreatment with N-benzyl-p-toluene sulfonamide (BTS) resulted in depressed peak developed tension with stimulation, while peak [Ca(2+)]cyt remained largely unchanged from control values. Despite excitation-contraction uncoupling, BTS-treated fibers displayed a significant increase in HSP72 mRNA. Treatment of fibers with hypoxia (Po2: skeletal muscle depolarization provides a sufficient activating stimulus for HSP72 transcription. Metabolic or mechanical factors associated with fatigue development and cross-bridge cycling likely play a more limited role. Copyright © 2016 the American Physiological Society.

  5. Cytosolic Pellino-1-Mediated K63-Linked Ubiquitination of IRF5 in M1 Macrophages Regulates Glucose Intolerance in Obesity

    Directory of Open Access Journals (Sweden)

    Donghyun Kim

    2017-07-01

    Full Text Available IRF5 is a signature transcription factor that induces M1 macrophage polarization. However, little is known regarding cytosolic proteins that induce IRF5 activation for M1 polarization. Here, we report the interaction between ubiquitin E3 ligase Pellino-1 and IRF5 in the cytoplasm, which increased nuclear translocation of IRF5 by K63-linked ubiquitination in human and mouse M1 macrophages. LPS and/or IFN-γ increased Pellino-1 expression, and M1 polarization was attenuated in Pellino-1-deficient macrophages in vitro and in vivo. Defective M1 polarization in Pellino-1-deficient macrophages improved glucose intolerance in mice fed a high-fat diet. Furthermore, macrophages in adipose tissues from obese humans exhibited increased Pellino-1 expression and IRF5 nuclear translocation compared with nonobese subjects, and these changes are associated with insulin resistance index. This study demonstrates that cytosolic Pellino-1-mediated K63-linked ubiquitination of IRF5 in M1 macrophages regulates glucose intolerance in obesity, suggesting a cytosolic mediator function of Pellino-1 in TLR4/IFN-γ receptor-IRF5 axis during M1 polarization.

  6. Nicotine-evoked cytosolic Ca2+ increase and cell depolarization in capillary endothelial cells of the bovine adrenal medulla

    Directory of Open Access Journals (Sweden)

    RAÚL VINET

    2009-01-01

    Full Text Available Endothelial cells are directly involved in many functions of the cardiovascular system by regulating blood flow and blood pressure through Ca2+ dependent exocitosis of vasoactive compounds. Using the Ca2+ indicator Fluo-3 and the patch-clamp technique, we show that bovine adrenal medulla capillary endothelial cells (B AMCECs respond to acetylcholine (ACh with a cytosolic Ca2+ increase and depolarization of the membrane potential (20.3±0.9 mV; n=23. The increase in cytosolic Ca2+ induced by 10µM ACh was mimicked by the same concentration of nicotine but not by muscarine and was blocked by 100 µM of hexamethonium. On the other hand, the increase in cytosolic Ca2+ could be depressed by nifedipine (0.01 -100 µM or withdrawal of extracellular Ca2+. Taken together, these results give evidence for functional nicotinic receptors (nAChRs in capillary endothelial cells of the adrenal medulla. It suggests that nAChRs in B AMCECs may be involved in the regulation of the adrenal gland's microcirculation by depolarizing the membrane potential, leading to the opening of voltage-activated Ca2+ channels, influx of external Ca2+ and liberation of vasoactive compounds.

  7. Liver cytosolic 1 antigen-antibody system in type 2 autoimmune hepatitis and hepatitis C virus infection.

    Science.gov (United States)

    Lenzi, M; Manotti, P; Muratori, L; Cataleta, M; Ballardini, G; Cassani, F; Bianchi, F B

    1995-01-01

    Within the multiform liver/kidney microsomal (LKM) family, a subgroup of sera that reacts with a liver cytosolic (LC) protein has been isolated and the new antigen-antibody system is called LC1. Unlike LKM antibody type 1 (anti-LKM1), anti-LC1 is said to be unrelated to hepatitis C virus (HCV) infection and has therefore been proposed as a marker of 'true' autoimmune hepatitis type 2. Altogether 100 LKM1 positive sera were tested by immunodiffusion (ID). Twenty five gave a precipitation line with human liver cytosol; 17 of the 25 also reacted with rat liver cytosol. Thirteen of the 25 sera were anti-HCV positive by second generation ELISA: anti-HCV positive patients were significantly older (p LKM1, and that it is an additional marker of juvenile autoimmune hepatitis type 2. It does not, however, discriminate between patients with and without HCV infection. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:7797126

  8. Hsp70 facilitates trans-membrane transport of bacterial ADP-ribosylating toxins into the cytosol of mammalian cells.

    Science.gov (United States)

    Ernst, Katharina; Schmid, Johannes; Beck, Matthias; Hägele, Marlen; Hohwieler, Meike; Hauff, Patricia; Ückert, Anna Katharina; Anastasia, Anna; Fauler, Michael; Jank, Thomas; Aktories, Klaus; Popoff, Michel R; Schiene-Fischer, Cordelia; Kleger, Alexander; Müller, Martin; Frick, Manfred; Barth, Holger

    2017-06-02

    Binary enterotoxins Clostridium (C.) botulinum C2 toxin, C. perfringens iota toxin and C. difficile toxin CDT are composed of a transport (B) and a separate non-linked enzyme (A) component. Their B-components mediate endocytic uptake into mammalian cells and subsequently transport of the A-components from acidic endosomes into the cytosol, where the latter ADP-ribosylate G-actin resulting in cell rounding and cell death causing clinical symptoms. Protein folding enzymes, including Hsp90 and peptidyl-prolyl cis/trans isomerases facilitate transport of the A-components across endosomal membranes. Here, we identified Hsp70 as a novel host cell factor specifically interacting with A-components of C2, iota and CDT toxins to facilitate their transport into the cell cytosol. Pharmacological Hsp70-inhibition specifically prevented pH-dependent trans-membrane transport of A-components into the cytosol thereby protecting living cells and stem cell-derived human miniguts from intoxication. Thus, Hsp70-inhibition might lead to development of novel therapeutic strategies to treat diseases associated with bacterial ADP-ribosylating toxins.

  9. Investigation on performance of all optical buffer with large dynamical delay time based on cascaded double loop optical buffers

    International Nuclear Information System (INIS)

    Yong-Jun, Wang; Xiang-Jun, Xin; Xiao-Lei, Zhang; Chong-Qing, Wu; Kuang-Lu, Yu

    2010-01-01

    Optical buffers are critical for optical signal processing in future optical packet-switched networks. In this paper, a theoretical study as well as an experimental demonstration on a new optical buffer with large dynamical delay time is carried out based on cascaded double loop optical buffers (DLOBs). It is found that pulse distortion can be restrained by a negative optical control mode when the optical packet is in the loop. Noise analysis indicates that it is feasible to realise a large variable delay range by cascaded DLOBs. These conclusions are validated by the experiment system with 4-stage cascaded DLOBs. Both the theoretical simulations and the experimental results indicate that a large delay range of 1–9999 times the basic delay unit and a fine granularity of 25 ns can be achieved by the cascaded DLOBs. The performance of the cascaded DLOBs is suitable for the all optical networks. (classical areas of phenomenology)

  10. Molding method of buffer material for underground disposal of radiation-contaminated material, and molded buffer material

    International Nuclear Information System (INIS)

    Akasaka, Hidenari; Shimura, Satoshi; Kawakami, Susumu; Ninomiya, Nobuo; Yamagata, Junji; Asano, Eiichi

    1995-01-01

    Upon molding of a buffer material to be used upon burying a vessel containing radiation-contaminated materials in a sealed state, a powdery buffer material to be molded such as bentonite is disposed at the periphery of a mandrel having a cylindrical portion somewhat larger than contaminate container to be subjected to underground disposal. In addition, it is subjected to integration-molding such as cold isotropic press with a plastic film being disposed therearound, to form a molding product at high density. The molding product is released and taken out with the plastic film being disposed thereon. Releasability from an elastic mold is improved by the presence of the plastic film. In addition, if it is stored or transported while having the plastic film being disposed thereon, swelling of the buffer material due to water absorption or moisture absorption can be suppressed. (T.M.)

  11. The Buffer and Backfill Handbook. Part 2: Materials and techniques

    International Nuclear Information System (INIS)

    Pusch, Roland

    2001-12-01

    Improved technology and prospection yielding more pure and homogeneous raw materials for preparing buffers and backfills will ultimately outdate the clays and ballast materials described in the present part of the Handbook. It describes experimentally investigated materials of potential use in repositories but other, more suitable materials will replace them in the future. The Handbook will hence have to be reviewed regularly, making room for superior materials in future, upgraded Handbook versions. Buffer is the term for dense clay used for embedment of canisters with highly radioactive waste, while backfill is soil used for filling tunnels and shafts in repositories. Examples of soil materials of potential use as buffers and backfills in repositories of KBS-3 type are described in this part of the Handbook. They are: smectitic clay materials intended for preparation of buffers (canister-embedding clay) and used as clay component in artificially prepared tunnel and shaft backfills consisting of mixtures of clay and ballast. Ballast materials intended for backfilling of tunnels and shafts and used as components of artificially prepared backfills. Smectitic natural clay soils intended for use as buffers and backfills. Very fine-grained smectite clay used as grout for sealing rock fractures. In this part of the Handbook for Buffers and Backfills, description of various candidate materials will be made with respect to their mineral composition and physical properties, with respect to the groundwater chemistry that can be expected in a deep repository in Swedish bedrock. Chapter 3 deals with smectitic clay materials intended for embedment of heat-producing canisters with highly radioactive waste. Focus is on the nature of the buffer constituents, i. e. the smectite content, the non-expanding clay minerals colloidal and the accessory non-clay minerals as well as amorphous matter and organic substances. The dominant part of the chapter describes the occurrence and origin

  12. The Buffer and Backfill Handbook. Part 2: Materials and techniques

    Energy Technology Data Exchange (ETDEWEB)

    Pusch, Roland [Geodevelopment AB, Lund (Sweden)

    2001-12-01

    Improved technology and prospection yielding more pure and homogeneous raw materials for preparing buffers and backfills will ultimately outdate the clays and ballast materials described in the present part of the Handbook. It describes experimentally investigated materials of potential use in repositories but other, more suitable materials will replace them in the future. The Handbook will hence have to be reviewed regularly, making room for superior materials in future, upgraded Handbook versions. Buffer is the term for dense clay used for embedment of canisters with highly radioactive waste, while backfill is soil used for filling tunnels and shafts in repositories. Examples of soil materials of potential use as buffers and backfills in repositories of KBS-3 type are described in this part of the Handbook. They are: smectitic clay materials intended for preparation of buffers (canister-embedding clay) and used as clay component in artificially prepared tunnel and shaft backfills consisting of mixtures of clay and ballast. Ballast materials intended for backfilling of tunnels and shafts and used as components of artificially prepared backfills. Smectitic natural clay soils intended for use as buffers and backfills. Very fine-grained smectite clay used as grout for sealing rock fractures. In this part of the Handbook for Buffers and Backfills, description of various candidate materials will be made with respect to their mineral composition and physical properties, with respect to the groundwater chemistry that can be expected in a deep repository in Swedish bedrock. Chapter 3 deals with smectitic clay materials intended for embedment of heat-producing canisters with highly radioactive waste. Focus is on the nature of the buffer constituents, i. e. the smectite content, the non-expanding clay minerals colloidal and the accessory non-clay minerals as well as amorphous matter and organic substances. The dominant part of the chapter describes the occurrence and origin

  13. High-speed digital imaging of cytosolic Ca2+ and contraction in single cardiomyocytes.

    Science.gov (United States)

    O'Rourke, B; Reibel, D K; Thomas, A P

    1990-07-01

    A charge-coupled device (CCD) camera, with the capacity for simultaneous spatially resolved photon counting and rapid frame transfer, was utilized for high-speed digital image collection from an inverted epifluorescence microscope. The unique properties of the CCD detector were applied to an analysis of cell shortening and the Ca2+ transient from fluorescence images of fura-2-loaded [corrected] cardiomyocytes. On electrical stimulation of the cell, a series of sequential subimages was collected and used to create images of Ca2+ within the cell during contraction. The high photosensitivity of the camera, combined with a detector-based frame storage technique, permitted collection of fluorescence images 10 ms apart. This rate of image collection was sufficient to resolve the rapid events of contraction, e.g., the upstroke of the Ca2+ transient (less than 40 ms) and the time to peak shortening (less than 80 ms). The technique was used to examine the effects of beta-adrenoceptor activation, fura-2 load, and stimulus frequency on cytosolic Ca2+ transients and contractions of single cardiomyocytes. beta-Adrenoceptor stimulation resulted in pronounced increases in peak Ca2+, maximal rates of rise and decay of Ca2+, extent of shortening, and maximal velocities of shortening and relaxation. Raising the intracellular load of fura-2 had little effect on the rising phase of Ca2+ or the extent of shortening but extended the duration of the Ca2+ transient and contraction. In related experiments utilizing differential-interference contrast microscopy, the same technique was applied to visualize sarcomere dynamics in contracting cells. This newly developed technique is a versatile tool for analyzing the Ca2+ transient and mechanical events in studies of excitation-contraction coupling in cardiomyocytes.

  14. Botanical Polyphenols Mitigate Microglial Activation and Microglia-Induced Neurotoxicity: Role of Cytosolic Phospholipase A2.

    Science.gov (United States)

    Chuang, Dennis Y; Simonyi, Agnes; Cui, Jiankun; Lubahn, Dennis B; Gu, Zezong; Sun, Grace Y

    2016-09-01

    Microglia play a significant role in the generation and propagation of oxidative/nitrosative stress, and are the basis of neuroinflammatory responses in the central nervous system. Upon stimulation by endotoxins such as lipopolysaccharides (LPS), these cells release pro-inflammatory factors which can exert harmful effects on surrounding neurons, leading to secondary neuronal damage and cell death. Our previous studies demonstrated the effects of botanical polyphenols to mitigate inflammatory responses induced by LPS, and highlighted an important role for cytosolic phospholipase A2 (cPLA2) upstream of the pro-inflammatory pathways (Chuang et al. in J Neuroinflammation 12(1):199, 2015. doi: 10.1186/s12974-015-0419-0 ). In this study, we investigate the action of botanical compounds and assess whether suppression of cPLA2 in microglia is involved in the neurotoxic effects on neurons. Differentiated SH-SY5Y neuroblastoma cells were used to test the neurotoxicity of conditioned medium from stimulated microglial cells, and WST-1 assay was used to assess for the cell viability of SH-SY5Y cells. Botanicals such as quercetin and honokiol (but not cyanidin-3-O-glucoside, 3CG) were effective in inhibiting LPS-induced nitric oxide (NO) production and phosphorylation of cPLA2. Conditioned medium from BV-2 cells stimulated with LPS or IFNγ caused neurotoxicity to SH-SY5Y cells. Decrease in cell viability could be ameliorated by pharmacological inhibitors for cPLA2 as well as by down-regulating cPLA2 with siRNA. Botanicals effective in inhibition of LPS-induced NO and cPLA2 phosphorylation were also effective in ameliorating microglial-induced neurotoxicity. Results demonstrated cytotoxic factors from activated microglial cells to cause damaging effects to neurons and potential use of botanical polyphenols to ameliorate the neurotoxic effects.

  15. PGC-1α accelerates cytosolic Ca2+ clearance without disturbing Ca2+ homeostasis in cardiac myocytes

    International Nuclear Information System (INIS)

    Chen, Min; Wang, Yanru; Qu, Aijuan

    2010-01-01

    Energy metabolism and Ca 2+ handling serve critical roles in cardiac physiology and pathophysiology. Peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) is a multi-functional coactivator that is involved in the regulation of cardiac mitochondrial functional capacity and cellular energy metabolism. However, the regulation of PGC-1α in cardiac Ca 2+ signaling has not been fully elucidated. To address this issue, we combined confocal line-scan imaging with off-line imaging processing to characterize calcium signaling in cultured adult rat ventricular myocytes expressing PGC-1α via adenoviral transduction. Our data shows that overexpressing PGC-1α improved myocyte contractility without increasing the amplitude of Ca 2+ transients, suggesting that myofilament sensitivity to Ca 2+ increased. Interestingly, the decay kinetics of global Ca 2+ transients and Ca 2+ waves accelerated in PGC-1α-expressing cells, but the decay rate of caffeine-elicited Ca 2+ transients showed no significant change. This suggests that sarcoplasmic reticulum (SR) Ca 2+ -ATPase (SERCA2a), but not Na + /Ca 2+ exchange (NCX) contribute to PGC-1α-induced cytosolic Ca 2+ clearance. Furthermore, PGC-1α induced the expression of SERCA2a in cultured cardiac myocytes. Importantly, overexpressing PGC-1α did not disturb cardiac Ca 2+ homeostasis, because SR Ca 2+ load and the propensity for Ca 2+ waves remained unchanged. These data suggest that PGC-1α can ameliorate cardiac Ca 2+ cycling and improve cardiac work output in response to physiological stress. Unraveling the PGC-1α-calcium handing pathway sheds new light on the role of PGC-1α in the therapy of cardiac diseases.

  16. Regulation of plant cytosolic glyceraldehyde 3-phosphate dehydrogenase isoforms by thiol modifications.

    Science.gov (United States)

    Holtgrefe, Simone; Gohlke, Jochen; Starmann, Julia; Druce, Samantha; Klocke, Susanne; Altmann, Bianca; Wojtera, Joanna; Lindermayr, Christian; Scheibe, Renate

    2008-06-01

    Cytosolic NAD-dependent glyceraldehyde 3-P dehydrogenase (GAPDH; GapC; EC 1.2.1.12) catalyzes the oxidation of triose phosphates during glycolysis in all organisms, but additional functions of the protein has been put forward. Because of its reactive cysteine residue in the active site, it is susceptible to protein modification and oxidation. The addition of GSSG, and much more efficiently of S-nitrosoglutathione, was shown to inactivate the enzymes from Arabidopsis thaliana (isoforms GapC1 and 2), spinach, yeast and rabbit muscle. Inactivation was fully or at least partially reversible upon addition of DTT. The incorporation of glutathione upon formation of a mixed disulfide could be shown using biotinylated glutathione ethyl ester. Furthermore, using the biotin-switch assay, nitrosylated thiol groups could be shown to occur after treatment with nitric oxide donors. Using mass spectrometry and mutant proteins with one cysteine lacking, both cysteines (Cys-155 and Cys-159) were found to occur as glutathionylated and as nitrosylated forms. In preliminary experiments, it was shown that both GapC1 and GapC2 can bind to a partial gene sequence of the NADP-dependent malate dehydrogenase (EC 1.2.1.37; At5g58330). Transiently expressed GapC-green fluorescent protein fusion proteins were localized to the nucleus in A. thaliana protoplasts. As nuclear localization and DNA binding of GAPDH had been shown in numerous systems to occur upon stress, we assume that such mechanism might be part of the signaling pathway to induce increased malate-valve capacity and possibly other protective systems upon overreduction and initial formation of reactive oxygen and nitrogen species as well as to decrease and protect metabolism at the same time by modification of essential cysteine residues.

  17. Cytosolic chaperones mediate quality control of higher-order septin assembly in budding yeast.

    Science.gov (United States)

    Johnson, Courtney R; Weems, Andrew D; Brewer, Jennifer M; Thorner, Jeremy; McMurray, Michael A

    2015-04-01

    Septin hetero-oligomers polymerize into cytoskeletal filaments with essential functions in many eukaryotic cell types. Mutations within the oligomerization interface that encompasses the GTP-binding pocket of a septin (its "G interface") cause thermoinstability of yeast septin hetero-oligomer assembly, and human disease. When coexpressed with its wild-type counterpart, a G interface mutant is excluded from septin filaments, even at moderate temperatures. We show that this quality control mechanism is specific to G interface mutants, operates during de novo septin hetero-oligomer assembly, and requires specific cytosolic chaperones. Chaperone overexpression lowers the temperature permissive for proliferation of cells expressing a G interface mutant as the sole source of a given septin. Mutations that perturb the septin G interface retard release from these chaperones, imposing a kinetic delay on the availability of nascent septin molecules for higher-order assembly. Un-expectedly, the disaggregase Hsp104 contributes to this delay in a manner that does not require its "unfoldase" activity, indicating a latent "holdase" activity toward mutant septins. These findings provide new roles for chaperone-mediated kinetic partitioning of non-native proteins and may help explain the etiology of septin-linked human diseases. © 2015 Johnson et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  18. A Comparison of Two Yeast MnSODs: Mitochondrial Saccharomyces cerevisiae versus Cytosolic Candida albicans

    International Nuclear Information System (INIS)

    Sheng, Y.; Cabelli, D.; Stich, T.A.; Barnese, K.; Gralla, E.B.; Cascio, D.; Britt, R.D.; Valentine, J.S.

    2011-01-01

    Human MnSOD is significantly more product-inhibited than bacterial MnSODs at high concentrations of superoxide (O 2 - ). This behavior limits the amount of H 2 O 2 produced at high [O 2 - ]; its desirability can be explained by the multiple roles of H 2 O 2 in mammalian cells, particularly its role in signaling. To investigate the mechanism of product inhibition in MnSOD, two yeast MnSODs, one from Saccharomyces cerevisiae mitochondria (ScMnSOD) and the other from Candida albicans cytosol (CaMnSODc), were isolated and characterized. ScMnSOD and CaMnSODc are similar in catalytic kinetics, spectroscopy, and redox chemistry, and they both rest predominantly in the reduced state (unlike most other MnSODs). At high [O 2 - ], the dismutation efficiencies of the yeast MnSODs surpass those of human and bacterial MnSODs, due to very low level of product inhibition. Optical and parallel-mode electron paramagnetic resonance (EPR) spectra suggest the presence of two Mn 3+ species in yeast Mn 3+ SODs, including the well-characterized 5-coordinate Mn 3+ species and a 6-coordinate L-Mn 3+ species with hydroxide as the putative sixth ligand (L). The first and second coordination spheres of ScMnSOD are more similar to bacterial than to human MnSOD. Gln154, an H-bond donor to the Mn-coordinated solvent molecule, is slightly further away from Mn in yeast MnSODs, which may result in their unusual resting state. Mechanistically, the high efficiency of yeast MnSODs could be ascribed to putative translocation of an outer-sphere solvent molecule, which could destabilize the inhibited complex and enhance proton transfer from protein to peroxide. Our studies on yeast MnSODs indicate the unique nature of human MnSOD in that it predominantly undergoes the inhibited pathway at high [O 2 - ].

  19. Cytosolic proteome profiling of aminoglycosides resistant Mycobacterium tuberculosis clinical isolates using MALDI-TOF/MS

    Directory of Open Access Journals (Sweden)

    Divakar Sharma

    2016-11-01

    Full Text Available Emergence of extremely drug resistant tuberculosis (XDR-TB is the consequence of the failure of second line TB treatment. Aminoglycosides are the important second line anti-TB drugs used to treat the multi drug resistant tuberculosis (MDR-TB. Main known mechanism of action of aminoglycosides is to inhibit the protein synthesis by inhibiting the normal functioning of ribosome. Primary target of aminoglycosides are the ribosomal RNA and its associated proteins. Various mechanisms have been proposed for aminoglycosides resistance but still some are unsolved. As proteins are involved in most of the biological processes, these act as a potential diagnostic markers and drug targets. In the present study we analyzed the purely cytosolic proteome of amikacin (AK and kanamycin (KM resistant Mycobacterium tuberculosis isolates by proteomic and bioinformatic approaches. Twenty protein spots were found to have over expressed in resistant isolates and were identified. Among these Rv3208A, Rv2623, Rv1360, Rv2140c, Rv1636 and Rv2185c are six proteins with unknown functions or undefined role. Docking results showed that AK and KM binds to the conserved domain (DUF, USP-A, Luciferase, PEBP and Polyketidecyclase/dehydrase domain of these hypothetical proteins and over expression of these proteins might neutralize/modulate the effect of drug molecules. TBPred and GPS-PUP predicted cytoplasmic nature and potential pupylation sites within these identified proteins respectively. String analysis also suggested that over expressed proteins along with their interactive partners might be involved in aminoglycosides resistance. Cumulative effect of these over expressed proteins could be involved in AK and KM resistance by mitigating the toxicity, repression of drug target and neutralizing affect. These findings need further exploitation for the expansion of newer therapeutics or diagnostic markers against AK and KM resistance so that an extreme condition like XDR-TB can

  20. Cytosolic Proteome Profiling of Aminoglycosides Resistant Mycobacterium tuberculosis Clinical Isolates Using MALDI-TOF/MS.

    Science.gov (United States)

    Sharma, Divakar; Lata, Manju; Singh, Rananjay; Deo, Nirmala; Venkatesan, Krishnamurthy; Bisht, Deepa

    2016-01-01

    Emergence of extensively drug resistant tuberculosis (XDR-TB) is the consequence of the failure of second line TB treatment. Aminoglycosides are the important second line anti-TB drugs used to treat the multi drug resistant tuberculosis (MDR-TB). Main known mechanism of action of aminoglycosides is to inhibit the protein synthesis by inhibiting the normal functioning of ribosome. Primary target of aminoglycosides are the ribosomal RNA and its associated proteins. Various mechanisms have been proposed for aminoglycosides resistance but still some are unsolved. As proteins are involved in most of the biological processes, these act as a potential diagnostic markers and drug targets. In the present study we analyzed the purely cytosolic proteome of amikacin (AK) and kanamycin (KM) resistant Mycobacterium tuberculosis isolates by proteomic and bioinformatic approaches. Twenty protein spots were found to have over expressed in resistant isolates and were identified. Among these Rv3208A, Rv2623, Rv1360, Rv2140c, Rv1636, and Rv2185c are six proteins with unknown functions or undefined role. Docking results showed that AK and KM binds to the conserved domain (DUF, USP-A, Luciferase, PEBP and Polyketidecyclase/dehydrase domain) of these hypothetical proteins and over expression of these proteins might neutralize/modulate the effect of drug molecules. TBPred and GPS-PUP predicted cytoplasmic nature and potential pupylation sites within these identified proteins, respectively. String analysis also suggested that over expressed proteins along with their interactive partners might be involved in aminoglycosides resistance. Cumulative effect of these over expressed proteins could be involved in AK and KM resistance by mitigating the toxicity, repression of drug target and neutralizing affect. These findings need further exploitation for the expansion of newer therapeutics or diagnostic markers against AK and KM resistance so that an extreme condition like XDR-TB can be prevented.

  1. Cytosolic NADP+-dependent isocitrate dehydrogenase plays a key role in lipid metabolism.

    Science.gov (United States)

    Koh, Ho-Jin; Lee, Su-Min; Son, Byung-Gap; Lee, Soh-Hyun; Ryoo, Zae Young; Chang, Kyu-Tae; Park, Jeen-Woo; Park, Dong-Chan; Song, Byoung J; Veech, Richard L; Song, Hebok; Huh, Tae-Lin

    2004-09-17

    NADPH is an essential cofactor for many enzymatic reactions including glutathione metabolism and fat and cholesterol biosynthesis. We have reported recently an important role for mitochondrial NADP(+)-dependent isocitrate dehydrogenase in cellular defense against oxidative damage by providing NADPH needed for the regeneration of reduced glutathione. However, the role of cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDPc) is still unclear. We report here for the first time that IDPc plays a critical role in fat and cholesterol biosynthesis. During differentiation of 3T3-L1 adipocytes, both IDPc enzyme activity and its protein content were increased in parallel in a time-dependent manner. Increased expression of IDPc by stable transfection of IDPc cDNA positively correlated with adipogenesis of 3T3-L1 cells, whereas decreased IDPc expression by an antisense IDPc vector retarded adipogenesis. Furthermore, transgenic mice with overexpressed IDPc exhibited fatty liver, hyperlipidemia, and obesity. In the epididymal fat pads of the transgenic mice, the expressions of adipocyte-specific genes including peroxisome proliferator-activated receptor gamma were markedly elevated. The hepatic and epididymal fat pad contents of acetyl-CoA and malonyl-CoA in the transgenic mice were significantly lower, whereas the total triglyceride and cholesterol contents were markedly higher in the liver and serum of transgenic mice compared with those measured in wild type mice, suggesting that the consumption rate of those lipogenic precursors needed for fat biosynthesis must be increased by elevated IDPc activity. Taken together, our findings strongly indicate that IDPc would be a major NADPH producer required for fat and cholesterol synthesis.

  2. Cytosolic NADP(+)-dependent isocitrate dehydrogenase status modulates oxidative damage to cells.

    Science.gov (United States)

    Lee, Su Min; Koh, Ho-Jin; Park, Dong-Chan; Song, Byoung J; Huh, Tae-Lin; Park, Jeen-Woo

    2002-06-01

    NADPH is an important cofactor in many biosynthesis pathways and the regeneration of reduced glutathione, critically important in cellular defense against oxidative damage. It is mainly produced by glucose 6-phosphate dehydrogenase (G6PD), malic enzyme, and the cytosolic form of NADP(+)-dependent isocitrate dehydrogenase (IDPc). Little information is available about the role of IDPc in antioxidant defense. In this study we investigated the role of IDPc against cytotoxicity induced by oxidative stress by comparing the relative degree of cellular responses in three different NIH3T3 cells with stable transfection with the cDNA for mouse IDPc in sense and antisense orientations, where IDPc activities were 3-4-fold higher and 35% lower, respectively, than that in the parental cells carrying the vector alone. Although the activities of other antioxidant enzymes, such as superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, and G6PD, were comparable in all transformed cells, the ratio of GSSG to total glutathione was significantly higher in the cells expressing the lower level of IDPc. This finding indicates that IDPc is essential for the efficient glutathione recycling. Upon transient exposure to increasing concentrations of H(2)O(2) or menadione, an intracellular source of free radicals and reactive oxygen species, the cells with low levels of IDPc became more sensitive to oxidative damage by H(2)O(2) or menadione. Lipid peroxidation, oxidative DNA damage, and intracellular peroxide generation were higher in the cell-line expressing the lower level of IDPc. However, the cells with the highly over-expressed IDPc exhibited enhanced resistance against oxidative stress, compared to the control cells. This study provides direct evidence correlating the activities of IDPc and the maintenance of the cellular redox state, suggesting that IDPc plays an important role in cellular defense against oxidative stress.

  3. A bacterial cyclic dinucleotide activates the cytosolic surveillance pathway and mediates innate resistance to tuberculosis.

    Science.gov (United States)

    Dey, Bappaditya; Dey, Ruchi Jain; Cheung, Laurene S; Pokkali, Supriya; Guo, Haidan; Lee, Jong-Hee; Bishai, William R

    2015-04-01

    Detection of cyclic-di-adenosine monophosphate (c-di-AMP), a bacterial second messenger, by the host cytoplasmic surveillance pathway (CSP) is known to elicit type I interferon (IFN) responses, which are crucial to antimicrobial defense. However, the mechanisms and role of c-di-AMP signaling in Mycobacterium tuberculosis virulence remain unclear. Here we show that resistance to tuberculosis requires CSP-mediated detection of c-di-AMP produced by M. tuberculosis and that levels of c-di-AMP modulate the fate of infection. We found that a di-adenylate cyclase (disA or dacA)-overexpressing M. tuberculosis strain that secretes excess c-di-AMP activates the interferon regulatory factor (IRF) pathway with enhanced levels of IFN-β, elicits increased macrophage autophagy, and exhibits substantial virulence attenuation in mice. We show that c-di-AMP-mediated IFN-β induction during M. tuberculosis infection requires stimulator of interferon genes (STING)-signaling. We observed that c-di-AMP induction of IFN-β is independent of the cytosolic nucleic acid receptor cyclic GMP-AMP (cGAMP) synthase (cGAS), but cGAS nevertheless contributes substantially to the overall IFN-β response to M. tuberculosis infection. In sum, our results reveal c-di-AMP to be a key mycobacterial pathogen-associated molecular pattern (PAMP) driving host type I IFN responses and autophagy. These findings suggest that modulating the levels of this small molecule may lead to novel immunotherapeutic strategies against tuberculosis.

  4. An alpha-glucose-1-phosphate phosphodiesterase is present in rat liver cytosol

    International Nuclear Information System (INIS)

    Srisomsap, C.; Richardson, K.L.; Jay, J.C.; Marchase, R.B.

    1989-01-01

    UDP-glucose:glycoprotein glucose-1-phosphotransferase (Glc-phosphotransferase) catalyzes the transfer of alpha-Glc-1-P from UDP-Glc to mannose residues on acceptor glycoproteins. The predominant acceptor for this transfer in both mammalian cells and Paramecium is a cytoplasmic glycoprotein of 62-63 kDa. When cytoplasmic proteins from rat liver were fractionated by preparative isoelectric focusing following incubation of a liver homogenate with the 35S-labeled phosphorothioate analogue of UDP-Glc ([beta-35S]UDP-Glc), the acceptor was found to have a pI of about 6.0. This fraction, when not labeled prior to the focusing, became very heavily labeled when mixed with [beta-35S]. UDP-Glc and intact liver microsomes, a rich source of the Glc-phosphotransferase. In addition, it was observed that the isoelectric fractions of the cytosol having pI values of 2-3.2 contained a degradative activity, alpha-Glc-1-P phosphodiesterase, that was capable of removing alpha-Glc-1-P, monitored through radioactive labeling both in the sugar and the phosphate, as an intact unit from the 62-kDa acceptor. Identification of the product of this cleavage was substantiated by its partial transformation to UDP-Glc in the presence of UTP and UDP-Glc pyrophosphorylase. The alpha-Glc-1-P phosphodiesterase had a pH optimum of 7.5 and was not effectively inhibited by any of the potential biochemical inhibitors that were tested. Specificity for the Glc-alpha-1-P-6-Man diester was suggested by the diesterase's inability to degrade UDP-Glc or glucosylphosphoryldolichol. This enzyme may be important in the regulation of secretion since the alpha-Glc-1-P present on the 62-kDa phosphoglycoprotein appears to be removed and then rapidly replaced in response to secretagogue

  5. Cytosolic 5′-nucleotidase 1A autoantibody profile and clinical characteristics in inclusion body myositis

    Science.gov (United States)

    Rietveld, A; Pye, S R; Mariampillai, K; Benveniste, O; Peeters, M T J; Miller, J A L; Hanna, M G; Machado, P M; Parton, M J; Gheorghe, K R; Badrising, U A; Lundberg, I E; Sacconi, S; Herbert, M K; McHugh, N J; Lecky, B R F; Brierley, C; Hilton-Jones, D; Lamb, J A; Roberts, M E; Cooper, R G; Saris, C G J; Pruijn, G J M; Chinoy, H; van Engelen, B G M

    2017-01-01

    Objectives Autoantibodies directed against cytosolic 5′-nucleotidase 1A have been identified in many patients with inclusion body myositis. This retrospective study investigated the association between anticytosolic 5′-nucleotidase 1A antibody status and clinical, serological and histopathological features to explore the utility of this antibody to identify inclusion body myositis subgroups and to predict prognosis. Materials and methods Data from various European inclusion body myositis registries were pooled. Anticytosolic 5′-nucleotidase 1A status was determined by an established ELISA technique. Cases were stratified according to antibody status and comparisons made. Survival and mobility aid requirement analyses were performed using Kaplan-Meier curves and Cox proportional hazards regression. Results Data from 311 patients were available for analysis; 102 (33%) had anticytosolic 5′-nucleotidase 1A antibodies. Antibody-positive patients had a higher adjusted mortality risk (HR 1.89, 95% CI 1.11 to 3.21, p=0.019), lower frequency of proximal upper limb weakness at disease onset (8% vs 23%, adjusted OR 0.29, 95% CI 0.12 to 0.68, p=0.005) and an increased prevalence of excess of cytochrome oxidase deficient fibres on muscle biopsy analysis (87% vs 72%, adjusted OR 2.80, 95% CI 1.17 to 6.66, p=0.020), compared with antibody-negative patients. Interpretation Differences were observed in clinical and histopathological features between anticytosolic 5′-nucleotidase 1A antibody positive and negative patients with inclusion body myositis, and antibody-positive patients had a higher adjusted mortality risk. Stratification of inclusion body myositis by anticytosolic 5′-nucleotidase 1A antibody status may be useful, potentially highlighting a distinct inclusion body myositis subtype with a more severe phenotype. PMID:28122761

  6. Cytosolic 5'-nucleotidase 1A autoantibody profile and clinical characteristics in inclusion body myositis.

    Science.gov (United States)

    Lilleker, J B; Rietveld, A; Pye, S R; Mariampillai, K; Benveniste, O; Peeters, M T J; Miller, J A L; Hanna, M G; Machado, P M; Parton, M J; Gheorghe, K R; Badrising, U A; Lundberg, I E; Sacconi, S; Herbert, M K; McHugh, N J; Lecky, B R F; Brierley, C; Hilton-Jones, D; Lamb, J A; Roberts, M E; Cooper, R G; Saris, C G J; Pruijn, G J M; Chinoy, H; van Engelen, B G M

    2017-05-01

    Autoantibodies directed against cytosolic 5'-nucleotidase 1A have been identified in many patients with inclusion body myositis. This retrospective study investigated the association between anticytosolic 5'-nucleotidase 1A antibody status and clinical, serological and histopathological features to explore the utility of this antibody to identify inclusion body myositis subgroups and to predict prognosis. Data from various European inclusion body myositis registries were pooled. Anticytosolic 5'-nucleotidase 1A status was determined by an established ELISA technique. Cases were stratified according to antibody status and comparisons made. Survival and mobility aid requirement analyses were performed using Kaplan-Meier curves and Cox proportional hazards regression. Data from 311 patients were available for analysis; 102 (33%) had anticytosolic 5'-nucleotidase 1A antibodies. Antibody-positive patients had a higher adjusted mortality risk (HR 1.89, 95% CI 1.11 to 3.21, p=0.019), lower frequency of proximal upper limb weakness at disease onset (8% vs 23%, adjusted OR 0.29, 95% CI 0.12 to 0.68, p=0.005) and an increased prevalence of excess of cytochrome oxidase deficient fibres on muscle biopsy analysis (87% vs 72%, adjusted OR 2.80, 95% CI 1.17 to 6.66, p=0.020), compared with antibody-negative patients. Differences were observed in clinical and histopathological features between anticytosolic 5'-nucleotidase 1A antibody positive and negative patients with inclusion body myositis, and antibody-positive patients had a higher adjusted mortality risk. Stratification of inclusion body myositis by anticytosolic 5'-nucleotidase 1A antibody status may be useful, potentially highlighting a distinct inclusion body myositis subtype with a more severe phenotype. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  7. The cytosolic glyoxalases of Plasmodium falciparum are dispensable during asexual blood-stage development

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    Cletus A. Wezena

    2017-11-01

    Full Text Available The enzymes glyoxalase 1 and 2 (Glo1 and Glo2 are found in most eukaryotes and catalyze the glutathione-dependent conversion of 2-oxoaldehydes to 2-hydroxycarboxylic acids. Four glyoxalases are encoded in the genome of the malaria parasite Plasmodium falciparum, the cytosolic enzymes PfGlo1 and PfcGlo2, the apicoplast enzyme PftGlo2, and an inactive Glo1-like protein that also carries an apicoplast-targeting sequence. Inhibition or knockout of the Plasmodium glyoxalases was hypothesized to lead to an accumulation of 2-oxoaldehydes and advanced glycation end-products (AGE in the host-parasite unit and to result in parasite death. Here, we generated clonal P. falciparum strain 3D7 knockout lines for PFGLO1 and PFcGLO2 using the CRISPR-Cas9 system. Although 3D7Δglo1 knockout clones had an increased susceptibility to external glyoxal, all 3D7Δglo1 and 3D7Δcglo2 knockout lines were viable and showed no significant growth phenotype under standard growth conditions. Furthermore, the lack of PfcGlo2, but not PfGlo1, increased gametocyte commitment in the knockout lines. In summary, PfGlo1 and PfcGlo2 are dispensable during asexual blood-stage development while the loss of PfcGlo2 may induce the formation of transmissible gametocytes. These combined data show that PfGlo1 and PfcGlo2 are most likely not suited as targets for selective drug development.

  8. The Yeast Nbp35-Cfd1 Cytosolic Iron-Sulfur Cluster Scaffold Is an ATPase.

    Science.gov (United States)

    Camire, Eric J; Grossman, John D; Thole, Grace J; Fleischman, Nicholas M; Perlstein, Deborah L

    2015-09-25

    Nbp35 and Cfd1 are prototypical members of the MRP/Nbp35 class of iron-sulfur (FeS) cluster scaffolds that function to assemble nascent FeS clusters for transfer to FeS-requiring enzymes. Both proteins contain a conserved NTPase domain that genetic studies have demonstrated is essential for their cluster assembly activity inside the cell. It was recently reported that these proteins possess no or very low nucleotide hydrolysis activity in vitro, and thus the role of the NTPase domain in cluster biogenesis has remained uncertain. We have reexamined the NTPase activity of Nbp35, Cfd1, and their complex. Using in vitro assays and site-directed mutagenesis, we demonstrate that the Nbp35 homodimer and the Nbp35-Cfd1 heterodimer are ATPases, whereas the Cfd1 homodimer exhibited no or very low ATPase activity. We ruled out the possibility that the observed ATP hydrolysis activity might result from a contaminating ATPase by showing that mutation of key active site residues reduced activity to background levels. Finally, we demonstrate that the fluorescent ATP analog 2'/3'-O-(N'-methylanthraniloyl)-ATP (mantATP) binds stoichiometrically to Nbp35 with a KD = 15.6 μM and that an Nbp35 mutant deficient in ATP hydrolysis activity also displays an increased KD for mantATP. Together, our results demonstrate that the cytosolic iron-sulfur cluster assembly scaffold is an ATPase and pave the way for interrogating the role of nucleotide hydrolysis in cluster biogenesis by this large family of cluster scaffolding proteins found across all domains of life. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. O-GlcNAcylation mediates the control of cytosolic phosphoenolpyruvate carboxykinase activity via Pgc1α.

    Directory of Open Access Journals (Sweden)

    Pedro Latorre

    Full Text Available PGC1α is a coactivator of many transcription factors and cytosolic phosphoenolpyruvate carboxykinase (PCK1 is a key enzyme for gluconeogenesis. PGC1α interacts with the transcription factor PPARγ to stimulate PCK1 expression and thus de novo glucose synthesis. These proteins are not only important for central energy metabolism but also for supplying intermediates for other metabolic pathways, including lipidogenesis and protein synthesis and might therefore be important factors in the ethiopathogenesis of metabolic disorders like diabetes but also in other pathologies like cancer. Since polymorphisms in these proteins have been related to some phenotypic traits in animals like pigs and PGC1α G482S polymorphism increases fat deposition in humans, we have investigated the molecular basis of such effects focusing on a commonly studied polymorphism in pig Pgc1α, which changes a cysteine at position 430 (WT of the protein to a serine (C430S. Biochemical analyses show that Pgc1α WT stimulates higher expression of human PCK1 in HEK293T and HepG2 cells. Paradoxically, Pgc1α WT is less stable than Pgc1α p.C430S in HEK293T cells. However, the study of different post-translational modifications shows a higher O-GlcNAcylation level of Pgc1α p.C430S. This higher O-GlcNAcylation level significantly decreases the interaction between Pgc1α and PPARγ demonstrating the importance of post-translational glycosylation of PGC1α in the regulation of PCK1 activity. This, furthermore, could explain at least in part the observed epistatic effects between PGC1α and PCK1 in pigs.

  10. Cytosolic streaming in vegetative mycelium and aerial structures of Aspergillus niger.

    Science.gov (United States)

    Bleichrodt, R; Vinck, A; Krijgsheld, P; van Leeuwen, M R; Dijksterhuis, J; Wösten, H A B

    2013-03-15

    Aspergillus niger forms aerial hyphae and conidiophores after a period of vegetative growth. The hyphae within the mycelium of A. niger are divided by septa. The central pore in these septa allows for cytoplasmic streaming. Here, we studied inter- and intra-compartmental streaming of the reporter protein GFP in A. niger. Expression of the gene encoding nuclear targeted GFP from the gpdA or glaA promoter resulted in strong fluorescence of nuclei within the vegetative hyphae and weak fluorescence in nuclei within the aerial structures. These data and nuclear run on experiments showed that gpdA and glaA are higher expressed in the vegetative mycelium when compared to aerial hyphae, conidiophores and conidia. Notably, gpdA or glaA driven expression of the gene encoding cytosolic GFP resulted in strongly fluorescent vegetative hyphae and aerial structures. Apparently, GFP streams from vegetative hyphae into aerial structures. This was confirmed by monitoring fluorescence of photo-activatable GFP (PA-GFP). In contrast, PA-GFP did not stream from aerial structures to vegetative hyphae. Streaming of PA-GFP within vegetative hyphae or within aerial structures of A. niger occurred at a rate of 10-15 μm s(-1). Taken together, these results not only show that GFP streams from the vegetative mycelium to aerial structures but it also indicates that its encoding RNA is not streaming. Absence of RNA streaming would explain why distinct RNA profiles were found in aerial structures and the vegetative mycelium by nuclear run on analysis and micro-array analysis.

  11. PHYSICAL CONTACT BETWEEN HUMAN VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS MODULATES CYTOSOLIC AND NUCLEAR CALCIUM HOMEOSTASIS.

    Science.gov (United States)

    Hassan, Ghada S; Jacques, Danielle; D'Orleans-Juste, Pedro; Magder, Sheldon; Bkaily, Ghassan

    2018-05-14

    The interaction between vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs) plays an important role in the modulation of vascular tone. There is however no information on whether direct physical communication regulates the intracellular calcium levels of human VECs (hVECs) and/or hVSMCs . Thus, the objective of the study is to verify whether co-culture of hVECs and hVSMCs modulates cytosolic ([Ca2+]c) and nuclear calcium ([Ca2+]n) levels via physical contact and/or factors released by both cell types. Quantitative 3D confocal microscopy for [Ca2+]c and [Ca2+]n measurement was performed in cultured hVECs or hVSMCs or in co-culture of hVECs-hVSMCs. Our results show that: 1) physical contact between hVECs-hVECs or hVSMCs-hVSMCs does not affect [Ca2+]c and [Ca2+]n in these two cell types; 2) physical contact between hVECs and hVSMCs induces a significant increase only of [Ca2+]n of hVECs without affecting the level of [Ca2+]c and [Ca2+]n of hVSMCs; and 3) preconditioned culture medium of hVECs or hVSMCs does not affect [Ca2+]c and [Ca2+]n of both types of cells. We concluded that physical contact between hVECs and hVSMCs only modulates [Ca2+]n in hVECs. The increase of [Ca2+]n in hVECs may modulate nuclear functions that are calcium dependent.

  12. Cytosolic Phospholipase A2-α: A Potential Therapeutic Target for Prostate Cancer

    Science.gov (United States)

    Patel, Manish I.; Singh, Jaskirat; Niknami, Marzieh; Kurek, Caroline; Yao, Mu; Lu, Sasa; Maclean, Fiona; King, Nicholas J.C.; Gelb, Michael H.; Scott, Kieran F.; Russell, Pamela J.; Boulas, John; Dong., Qihan

    2008-01-01

    Purpose Cytosolic Phospholipase A2-α (cPLA2-α) provides intracellular arachidonic acid to supply both cyclooxygenase and lipoxygenase pathways. We aim to determine the expression and activation of cPLA2-α in prostate cancer (PC) cell line and tissue and the effect of targeting cPLA2-α in-vitro and in-vivo. Experimental Design The expression of cPLA2-α was determined in PC cells by RT-PCR, Western blot and immunocytochemistry. Growth inhibition, apoptosis and cPLA2-α activity were determined after inhibition with cPLA2-α siRNA or inhibitor (Wyeth-1). cPLA2-α inhibitor or vehicle was also administered to PC xenograft mouse models. Finally the expression of phospho-cPLA2-α was determined by immunohistochemistry in human normal, androgen sensitive and insensitive PC specimens. Results cPLA2-α is present in all PC cells lines, but increased in androgen insensitive cells. Inhibition with siRNA or Wyeth-1 results in significant reductions in PC cell numbers, as a result of reduced proliferation as well as increased apoptosis and this was also associated with a reduction in cPLA2-α activity. Expression of cyclin D1 and phosphorylation of Akt were also observed to decrease. Wyeth-1 inhibited PC3 xenograft growth by approximately 33% and again, also reduced cyclin D1. Immunohistochemistry of human prostate tissue revealed that phospho-cPLA2-α is increased when hormone refractory is reached. Conclusions cPLA2-α expression and activation is increased in the androgen insensitive cancer cell line and tissue. Inhibition of cPLA2-α results in cells and xenograft tumor growth inhibition and serves as a potentially effective therapy for hormone refractory PC. PMID:19088022

  13. Real-time Recording of Cytosolic Calcium Levels in Arabidopsis thaliana Cell Cultures during Parabolic Flights

    Science.gov (United States)

    Neef, Maren; Ecke, Margret; Hampp, Rüdiger

    2015-07-01

    In plants, like in other organisms, calcium (Ca2+) is an important second messenger which participates in the conversion of environmental signals into molecular responses. There is increasing evidence, that sensing of changes in gravitation or reorientation of tissues is an example for such signaling cascades in which Ca2+ is involved. In order to determine g-dependent changes in the cytosolic calcium (Ca^{2+}_{ {cyt}}) concentration of plant cells, semisolid transgenic callus cell cultures of Arabidopsis thaliana (A.t.), expressing the calcium sensor YC3.6 (cameleon), were exposed to g-forces between 1.8 g and μ g during parabolic flights. Using such cells, intracellular calcium transients can be monitored by FRET in vivo and in real-time. Interestingly we observed a slight decrease of the Ca^{2+}_{ {cyt}} level during the hypergravity phases of a parabola but a significant increase of the Ca^{2+}_{ {cyt}} concentration during microgravity. Application of known Ca2+ inhibitors and antagonists yielded the following effects: nifedipine (Ca2+ channel blocker) showed no effect, whereas LaCl3, GdCl3 (both inhibitors of uptake at the plasma membrane), DPI (inhibitor of NADP oxidase), and DMSO (solvent) diminished the gravity-alteration-related Ca^{2+}_{ {cyt}} response. EGTA (binding of Ca2+) and eosin yellow (inhibitor of a plasma membrane-located Ca2+ pump) suppressed the respective Ca^{2+}_{ {cyt}} changes entirely. We thus conclude that the significant increase in Ca^{2+}_{ {cyt}} under microgravity is largely due to extracellular Ca2+ sources.

  14. Imidacloprid sorption and transport in cropland, grass buffer and riparian buffer soils

    Science.gov (United States)

    Satkowski, Laura E.; Goyne, Keith W.; Anderson, Stephen H.; Lerch, Robert N.; Allen, Craig R.; Snow, Daniel D.

    2018-01-01

    An understanding of neonicotinoid sorption and transport in soil is critical for determining and mitigating environmental risk associated with the most widely used class of insecticides. The objective of this study was to evaluate mobility and transport of the neonicotinoid imidacloprid (ICD) in soils collected from cropland, grass vegetative buffer strip (VBS), and riparian VBS soils. Soils were collected at six randomly chosen sites within grids that encompassed all three land uses. Single-point equilibrium batch sorption experiments were conducted using radio-labeled (14C) ICD to determine solid–solution partition coefficients (Kd). Column experiments were conducted using soils collected from the three vegetation treatments at one site by packing soil into glass columns. Water flow was characterized by applying Br− as a nonreactive tracer. A single pulse of 14C-ICD was then applied, and ICD leaching was monitored for up to 45 d. Bromide and ICD breakthrough curves for each column were simulated using CXTFIT and HYDRUS-1D models. Sorption results indicated that ICD sorbs more strongly to riparian VBS (Kd = 22.6 L kg−1) than crop (Kd = 11.3 L kg−1) soils. Soil organic C was the strongest predictor of ICD sorption (p < 0.0001). The column transport study found mean peak concentrations of ICD at 5.83, 10.84, and 23.8 pore volumes for crop, grass VBS, and riparian VBS soils, respectively. HYDRUS-1D results indicated that the two-site, one-rate linear reversible model best described results of the breakthrough curves, indicating the complexity of ICD sorption and demonstrating its mobility in soil. Greater sorption and longer retention by the grass and riparian VBS soils than the cropland soil suggests that VBS may be a viable means to mitigate ICD loss from agroecosystems, thereby preventing ICD transport into surface water, groundwater, or drinking water resources.

  15. LeftyA sensitive cytosolic pH regulation and glycolytic flux in Ishikawa human endometrial cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Salker, Madhuri S.; Zhou, Yuetao; Singh, Yogesh [Department of Physiology, University of Tuebingen, 72076 Tuebingen (Germany); Brosens, Jan [Division of Reproductive Health, Warwick Medical School, Clinical Sciences Research Laboratories, University Hospital, Coventry CV2 2DX (United Kingdom); Lang, Florian, E-mail: florian.lang@uni-tuebingen.de [Department of Physiology, University of Tuebingen, 72076 Tuebingen (Germany)

    2015-05-08

    Objective: LeftyA, a powerful regulator of stemness, embryonic differentiation, and reprogramming of cancer cells, counteracts cell proliferation and tumor growth. Key properties of tumor cells include enhanced glycolytic flux, which is highly sensitive to cytosolic pH and thus requires export of H{sup +} and lactate. H{sup +} extrusion is in part accomplished by Na{sup +}/H{sup +} exchangers, such as NHE1. An effect of LeftyA on transport processes has, however, never been reported. The present study thus explored whether LeftyA modifies regulation of cytosolic pH (pHi) in Ishikawa cells, a well differentiated endometrial carcinoma cell model. Methods: NHE1 transcript levels were determined by qRT-PCR, NHE1 protein abundance quantified by Western blotting, pH{sub i} estimated utilizing (2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein [BCECF] fluorescence, Na{sup +}/H{sup +} exchanger activity from Na{sup +} dependent realkalinization after an ammonium pulse, and lactate concentration in the supernatant utilizing an enzymatic assay and subsequent colorimetry. Results: A 2 h treatment with LeftyA (8 ng/ml) significantly decreased NHE1 transcript levels (by 99.6%), NHE1 protein abundance (by 71%), Na{sup +}/H{sup +} exchanger activity (by 55%), pHi (from 7.22 ± 0.02 to 7.05 ± 0.02), and lactate release (by 41%). Conclusions: LeftyA markedly down-regulates NHE1 expression, Na{sup +}/H{sup +} exchanger activity, pHi, and lactate release in Ishikawa cells. Those effects presumably contribute to cellular reprogramming and growth inhibition. - Highlights: • LeftyA, an inhibitor of tumor growth, reduces Na{sup +}/H{sup +}-exchanger activity by 55%. • LeftyA decreases NHE1 transcripts by 99.6% and NHE1 protein by 71%. • LeftyA decreases cytosolic pH from 7.22 ± 0.02 to 7.05 ± 0.02. • Cytosolic acidification by Lefty A decreases glycolysis by 41%. • Cytosolic acidification by Lefty A compromises energy production of tumor cells.

  16. Runtime buffer management to improve the performance in irregular ...

    Indian Academy of Sciences (India)

    S¯adhan¯a Vol. 40, Part 4, June 2015, pp. 1117–1137. c Indian Academy of Sciences. Runtime buffer management to improve the performance in irregular Network-on-Chip architecture. UMAMAHESWARI S1,∗, MEGANATHAN D2 and. RAJA PAUL PERINBAM J3. 1Department of Information Technology, Anna University, ...

  17. Optimizing the HLT Buffer Strategy with Monte Carlo Simulations

    CERN Document Server

    AUTHOR|(CDS)2266763

    2017-01-01

    This project aims to optimize the strategy of utilizing the disk buffer for the High Level Trigger (HLT) of the LHCb experiment with the help of Monte-Carlo simulations. A method is developed, which simulates the Event Filter Farm (EFF) -- a computing cluster for the High Level Trigger -- as a compound of nodes with different performance properties. In this way, the behavior of the computing farm can be analyzed at a deeper level than before. It is demonstrated that the current operating strategy might be improved when data taking is reaching a mid-year scheduled stop or the year-end technical stop. The processing time of the buffered data can be lowered by distributing the detector data according to the processing power of the nodes instead of the relative disk size as long as the occupancy level of the buffer is low enough. Moreover, this ensures that data taken and stored on the buffer at the same time is processed by different nodes nearly simultaneously, which reduces load on the infrastructure.

  18. The function of buffer layer in resistive switching device.

    Czech Academy of Sciences Publication Activity Database

    Zhang, B.; Prokop, V.; Střižík, L.; Zima, Vítězslav; Kutálek, P.; Vlček, Milan; Wágner, T.

    2017-01-01

    Roč. 14, č. 8 (2017), s. 291-295 ISSN 1584-8663 Institutional support: RVO:61389013 Keywords : resistive switching * chalcogenide glasses * buffer layer Subject RIV: CA - Inorganic Chemistry Impact factor: 0.732, year: 2016 http://www.chalcogen.ro/291_ZhangB.pdf

  19. The effect of buffer zone width on biodiversity

    DEFF Research Database (Denmark)

    Navntoft, Søren; Sigsgaard, Lene; Kristensen, Kristian Morten

    2012-01-01

    Field margin management for conservation purposes is a way to protect both functional biodiversity and biodiversity per se without considerable economical loss as field margins are less productive. However, the effect of width of the buffer zone on achievable biodiversity gains has received littl...

  20. Moisture buffering phenomenon and its impact on building energy consumption

    DEFF Research Database (Denmark)

    Zhang, Mingjie; Qin, Menghao; Rode, Carsten

    2017-01-01

    buffering on building energy consumption in different climate conditions is assessed by using numerical simulations. The results show that the potential energy saving rate could be up to 25–30% when using proper hygroscopic materials in the test building in temperate climates and semi-arid climates. Finally......, the relationship between MBV and potential energy saving rate is also discussed....

  1. Optimal Maintenance of a Production System with Intermediate Buffers

    Directory of Open Access Journals (Sweden)

    Constantinos C. Karamatsoukis

    2012-01-01

    Full Text Available We consider a production-inventory system that consists of an input-generating installation, a production unit and L intermediate buffers. It is assumed that the installation transfers the raw material to buffer and the production unit pulls the raw material from buffer We consider the problem of the optimal preventive maintenance of the installation if the installation deteriorates stochastically with usage and the production unit is always in operative condition. We also consider the problem of the optimal preventive maintenance of the production unit if the production unit deteriorates stochastically with usage and the installation is always in operative condition. Under a suitable cost structure and for given contents of the buffers, it is proved that the average-cost optimal policy for the first (second problem initiates a preventive maintenance of the installation (production unit if and only if the degree of deterioration of the installation (production unit exceeds some critical level. Numerical results are presented for both problems.

  2. Study of buffer layer thickness on bulk heterojunction solar cell.

    Science.gov (United States)

    Noh, Seunguk; Suman, C K; Lee, Donggu; Kim, Seohee; Lee, Changhee

    2010-10-01

    We studied the effect of the buffer layer (molybdenum-oxide (MoO3)) thickness on the performance of organic solar cell based on blends of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C61 butyric acid methyl ester fullerene derivative (PCBM). The thickness of MoO3 was varied from 1 nm to 30 nm for optimization of device performance. The photocurrent-voltage and impedance spectroscopy were measured under dark and AM1.5G solar simulated illumination of 100 mW/cm2 for exploring the role of the buffer layer thickness on carrier collection at an anode. The MoO3 thickness of the optimized device (efficiency approximately 3.7%) was found to be in the range of 5 approximately 10 nm. The short-circuit current and the shunt resistance decrease gradually for thicker MoO3 layer over 5 nm. The device can be modeled as the combination of three RC parallel circuits (each one for the active layer, buffer layer and interface between the buffer layer and the active layer) in series with contact resistance (Rs approximately 60 ohm).

  3. Buffering Implications for the Design Space of Streaming MEMS Storage

    NARCIS (Netherlands)

    Khatib, M.G.; Abelmann, Leon; Preas, Kathy

    2011-01-01

    Emerging nanotechnology-based systems encounter new non-functional requirements. This work addresses MEMS storage, an emerging technology that promises ultrahigh density and energy-efficient storage devices. We study the buffering requirement of MEMS storage in streaming applications. We show that

  4. Nitrogen transformation and retention in riparian buffer zones

    NARCIS (Netherlands)

    Hefting, Maria Margaretha

    2003-01-01

    Diffuse pollution of nutrients and pesticides from agricultural areas is increasingly recognised as a major problem in water management. Ecotechnological measures as constructed wetlands and riparian buffer zones clearly have an important role in the reduction of diffuse pollution by removing and

  5. Challenges of Buffer Zone Management in Cross River National ...

    African Journals Online (AJOL)

    These developments might be adduced to the low level of awareness of Park laws and buffer zone management policies by these communities. It is therefore recommended that public enlightenment campaigns should be stepped up to educate the populace on the need to abide by the laws and policies governing the Park ...

  6. Effects of Terrestrial Buffer Zones on Amphibians on Golf Courses

    Science.gov (United States)

    Puglis, Holly J.; Boone, Michelle D.

    2012-01-01

    A major cause of amphibian declines worldwide is habitat destruction or alteration. Public green spaces, such as golf courses and parks, could serve as safe havens to curb the effects of habitat loss if managed in ways to bolster local amphibian communities. We reared larval Blanchard's cricket frogs (Acris blanchardi) and green frogs (Rana clamitans) in golf course ponds with and without 1 m terrestrial buffer zones, and released marked cricket frog metamorphs at the golf course ponds they were reared in. Larval survival of both species was affected by the presence of a buffer zone, with increased survival for cricket frogs and decreased survival for green frogs when reared in ponds with buffer zones. No marked cricket frog juveniles were recovered at any golf course pond in the following year, suggesting that most animals died or migrated. In a separate study, we released cricket frogs in a terrestrial pen and allowed them to choose between mown and unmown grass. Cricket frogs had a greater probability of using unmown versus mown grass. Our results suggest that incorporating buffer zones around ponds can offer suitable habitat for some amphibian species and can improve the quality of the aquatic environment for some sensitive local amphibians. PMID:22761833

  7. SDRAM-based packet buffer model for high speed switches

    DEFF Research Database (Denmark)

    Rasmussen, Anders; Ruepp, Sarah Renée; Berger, Michael Stübert

    2011-01-01

    based on the specifications of a real-life DDR3-SDRAM chip. Based on this model the performance of different schemes for optimizing the performance of such a packet buffer can be evaluated. The purpose of this study is to find efficient schemes for memory mapping of the packet queues and I/O traffic...

  8. The Edinburgh approach to urban heritage : why a buffer zone?

    NARCIS (Netherlands)

    Bennink, R.H.J.; van Niel, W.; Veldpaus, L.; Pereira Roders, A.R.

    2013-01-01

    The architectural ensemble of Old and New Towns of Edinburgh, UNESCO World Heritage since 1995, is among the sites affected by urban development. The absence of a buffer zone is assumed to be one of the causes of the impact and the recently adopted Historic Urban Landscape approach aims to assist on

  9. VLSI scaling methods and low power CMOS buffer circuit

    International Nuclear Information System (INIS)

    Sharma Vijay Kumar; Pattanaik Manisha

    2013-01-01

    Device scaling is an important part of the very large scale integration (VLSI) design to boost up the success path of VLSI industry, which results in denser and faster integration of the devices. As technology node moves towards the very deep submicron region, leakage current and circuit reliability become the key issues. Both are increasing with the new technology generation and affecting the performance of the overall logic circuit. The VLSI designers must keep the balance in power dissipation and the circuit's performance with scaling of the devices. In this paper, different scaling methods are studied first. These scaling methods are used to identify the effects of those scaling methods on the power dissipation and propagation delay of the CMOS buffer circuit. For mitigating the power dissipation in scaled devices, we have proposed a reliable leakage reduction low power transmission gate (LPTG) approach and tested it on complementary metal oxide semiconductor (CMOS) buffer circuit. All simulation results are taken on HSPICE tool with Berkeley predictive technology model (BPTM) BSIM4 bulk CMOS files. The LPTG CMOS buffer reduces 95.16% power dissipation with 84.20% improvement in figure of merit at 32 nm technology node. Various process, voltage and temperature variations are analyzed for proving the robustness of the proposed approach. Leakage current uncertainty decreases from 0.91 to 0.43 in the CMOS buffer circuit that causes large circuit reliability. (semiconductor integrated circuits)

  10. Buffered Simulation Games for Büchi Automata

    Directory of Open Access Journals (Sweden)

    Milka Hutagalung

    2014-05-01

    Full Text Available Simulation relations are an important tool in automata theory because they provide efficiently computable approximations to language inclusion. In recent years, extensions of ordinary simulations have been studied, for instance multi-pebble and multi-letter simulations which yield better approximations and are still polynomial-time computable. In this paper we study the limitations of approximating language inclusion in this way: we introduce a natural extension of multi-letter simulations called buffered simulations. They are based on a simulation game in which the two players share a FIFO buffer of unbounded size. We consider two variants of these buffered games called continuous and look-ahead simulation which differ in how elements can be removed from the FIFO buffer. We show that look-ahead simulation, the simpler one, is already PSPACE-hard, i.e. computationally as hard as language inclusion itself. Continuous simulation is even EXPTIME-hard. We also provide matching upper bounds for solving these games with infinite state spaces.

  11. Comparison of Five Nuclear Isolation Buffers for Flow Cytometric ...

    African Journals Online (AJOL)

    Jocky

    2012-02-23

    Feb 23, 2012 ... species. A systematic comparison of nuclear lysis buffers has been ... All experiments were carried out with 3 replicates (n = 3) per treatment. ... Na2EDTA, 0.5 mM spermine.4HCl, 80 mM KCl, 20 mM NaCl, 0.1%. (v/v) Triton ...

  12. Rare earth zirconium oxide buffer layers on metal substrates

    Science.gov (United States)

    Williams, Robert K.; Paranthaman, Mariappan; Chirayil, Thomas G.; Lee, Dominic F.; Goyal, Amit; Feenstra, Roeland

    2001-01-01

    A laminate article comprises a substrate and a biaxially textured (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer over the substrate, wherein 0layer can be deposited using sol-gel or metal-organic decomposition. The laminate article can include a layer of YBCO over the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer. A layer of CeO.sub.2 between the YBCO layer and the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer can also be include. Further included can be a layer of YSZ between the CeO.sub.2 layer and the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer. The substrate can be a biaxially textured metal, such as nickel. A method of forming the laminate article is also disclosed.

  13. Buffer provisioning for large-scale data-acquisition systems

    CERN Document Server

    AUTHOR|(SzGeCERN)756497; The ATLAS collaboration; Garcia Garcia, Pedro Javier; Froening, Holger; Vandelli, Wainer

    2018-01-01

    The data acquisition system of the ATLAS experiment, a major experiment of the Large Hadron Collider (LHC) at CERN, will go through a major upgrade in the next decade. The upgrade is driven by experimental physics requirements, calling for increased data rates on the order of 6~TB/s. By contrast, the data rate of the existing system is 160~GB/s. Among the changes in the upgraded system will be a very large buffer with a projected size on the order of 70 PB. The buffer role will be decoupling of data production from on-line data processing, storing data for periods of up to 24~hours until it can be analyzed by the event processing system. The larger buffer will allow a new data recording strategy, providing additional margins to handle variable data rates. At the same time it will provide sensible trade-offs between buffering space and on-line processing capabilities. This compromise between two resources will be possible since the data production cycle includes time periods where the experiment will not produ...

  14. Moisture Buffer Performance of a Fully Furnished Room

    DEFF Research Database (Denmark)

    Svennberg, Kaisa; Hedegaard, Lone Grønbæk; Rode, Carsten

    2004-01-01

    of the indoor air, such as growth of house dust mites, emissions from materials and mould growth it is anticipated that the moisture buffer effect can help to ensure healthier indoor environments. The building materials as well as furniture and other furnishing materials exposed to the indoor air...

  15. Unusual buffer action of free-standing nanoscopically confined water.

    Science.gov (United States)

    Liao, Kylin; Xu, Xiaozhou; Du, Xuezhong

    2010-01-15

    The acid-base properties of nanoscopic water confined in the black soap films (BSFs), which were prepared from aqueous solutions of sodium dodecylsulphate (SDS) with the dye neutral red (NR) as a pH probe, were investigated using a combination of UV-vis and FTIR spectroscopy. For the SDS micellar solutions at pH 1.0-9.5 adjusted with HCl/NaOH solutions and at pH 9.4 with ammonium buffered solution, the aqueous core thicknesses in the corresponding BSFs ranged from 2.7 to 6.2 nm, and the nanoscopically confined water exhibits unusual buffer action resistant not only to acidic/alkaline solutions but also to standard buffer solution. In the heavily water-depleted confined zones, it is most likely that charge pairs in proton-transfer reactions could not be formed effectively and proton transfer was prohibited in the absence of sufficient solvating ability. Theoretical analyzes indicated that the buffer action of the nanoscopic water originated from the confinement effect of two charged surfaces of the BSFs. These results might inspire deeper understanding and further studies of biobuffering, enzyme superactivity, acid-catalyzed reactions, and Nafion fuel cell membranes.

  16. Spirituality and optimism as buffer of sense of coherence among ...

    African Journals Online (AJOL)

    This study examined the buffering influence of spirituality and optimism on sense of coherence among imminent retirees in Lagos State, Nigeria. The study adopted a descriptive survey utilizing an expost-facto research design. Six hundred and five imminent retirees constituted the participants for the study. Through the use ...

  17. Evaluation of buffering potency of oyster ( Crassostrea gasar ) shells ...

    African Journals Online (AJOL)

    ) shells on acidic borehole water for fish farming. Eighteen (18), 100-litre shaded plastic tanks (3/4- filled, pH 4.0) of three replicates per treatment of 270 g buffer agent were used. There were four trials of six treatments (T) namely control (C) ...

  18. Multiple function benefit - cost comparison of conservation buffer placement strategies

    Science.gov (United States)

    Z. Qiu; M.G. Dosskey

    2012-01-01

    Conservation buffers are considered to be effective practices for repairing impaired streams and restoring multiple ecosystem functions in degraded agricultural watersheds. Six different planning strategies for targeting their placement within watersheds were compared in terms of cost-effectiveness for environmental improvement in the 144 km² Neshanic River...

  19. TRIO: Burst Buffer Based I/O Orchestration

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Teng [Auburn University; Oral, H Sarp [ORNL; Pritchard, Michael [Auburn University; Wang, Bin [Auburn University; Yu, Weikuan [Auburn University

    2015-01-01

    The growing computing power on leadership HPC systems is often accompanied by ever-escalating failure rates. Checkpointing is a common defensive mechanism used by scientific applications for failure recovery. However, directly writing the large and bursty checkpointing dataset to parallel filesystem can incur significant I/O contention on storage servers. Such contention in turn degrades the raw bandwidth utilization of storage servers and prolongs the average job I/O time of concurrent applications. Recently burst buffer has been proposed as an intermediate layer to absorb the bursty I/O traffic from compute nodes to storage backend. But an I/O orchestration mechanism is still desired to efficiently move checkpointing data from bursty buffers to storage backend. In this paper, we propose a burst buffer based I/O orchestration framework, named TRIO, to intercept and reshape the bursty writes for better sequential write traffic to storage severs. Meanwhile, TRIO coordinates the flushing orders among concurrent burst buffers to alleviate the contention on storage server bandwidth. Our experimental results reveal that TRIO can deliver 30.5% higher bandwidth and reduce the average job I/O time by 37% on average for data-intensive applications in various checkpointing scenarios.

  20. Substrate-induced magnetism in epitaxial graphene buffer layers.

    Science.gov (United States)

    Ramasubramaniam, A; Medhekar, N V; Shenoy, V B

    2009-07-08

    Magnetism in graphene is of fundamental as well as technological interest, with potential applications in molecular magnets and spintronic devices. While defects and/or adsorbates in freestanding graphene nanoribbons and graphene sheets have been shown to cause itinerant magnetism, controlling the density and distribution of defects and adsorbates is in general difficult. We show from first principles calculations that graphene buffer layers on SiC(0001) can also show intrinsic magnetism. The formation of graphene-substrate chemical bonds disrupts the graphene pi-bonds and causes localization of graphene states near the Fermi level. Exchange interactions between these states lead to itinerant magnetism in the graphene buffer layer. We demonstrate the occurrence of magnetism in graphene buffer layers on both bulk-terminated as well as more realistic adatom-terminated SiC(0001) surfaces. Our calculations show that adatom density has a profound effect on the spin distribution in the graphene buffer layer, thereby providing a means of engineering magnetism in epitaxial graphene.