Sample records for high salt buffer

  1. Controlled Assembly of Ag Nanoparticles and Carbon Nanotube Hybrid Structures for Biosensing

    DTIC Science & Technology

    2010-01-01

    to∼190 kΩ. The same device was again washed with DI water and treated with the thiolated ssDNA in high salt buffer. After a 2 h treatment, the device...after the cleaning only thiolated DNA should be present on the device, whereas the nonspecifically bound DNA as well as the buffer salts should be...ssDNA molecules for 2 h. Specific immobiliza- tion of thiolated ssDNA (sequence: 50thiol_TCATAC AGCTAGATA ACC AAAGA) was carried out in high salt

  2. Effect of glycine on pH changes and protein stability during freeze-thawing in phosphate buffer systems.

    PubMed

    Pikal-Cleland, Katherine A; Cleland, Jeffrey L; Anchordoquy, Thomas J; Carpenter, John F

    2002-09-01

    Previous studies have established that the selective precipitation of a less soluble buffer component during freezing can induce a significant pH shift in the freeze concentrate. During freezing of sodium phosphate solutions, crystallization of the disodium salt can produce a pH decrease as great as 3 pH units which can dramatically affect protein stability. The objective of our study was to determine how the presence of glycine (0-500 mM), a commonly used bulking agent in pharmaceutical protein formulations, affects the pH changes normally observed during freezing in sodium phosphate buffer solutions and to determine whether these pH changes contribute to instability of model proteins in glycine/phosphate formulations. During freezing in sodium phosphate buffers, the presence of glycine significantly influenced the pH. Glycine at the lower concentrations (< or = 50 mM) suppressed the pH decrease normally observed during freezing in 10 and 100 mM sodium phosphate buffer, possibly by reducing the nucleation rate of salt and thereby decreasing the extent of buffer salt crystallization. The presence of glycine at higher concentration (> 100 mM) in the sodium phosphate buffer resulted in a more complete crystallization of the disodium salt as indicated by the frozen pH values closer to the equilibrium value (pH 3.6). Although high concentrations of glycine can facilitate more buffer salt crystallization and these pH shifts may prove to be potentially damaging to the protein, glycine, in its amorphous state, can also act to stabilize a protein via the preferential exclusion mechanism. Copyright 2002 Wiley-Liss Inc.

  3. Evidence for Intramolecular Antiparallel Beta-Sheet Structure in Alpha-Synuclein Fibrils from a Combination of Two-Dimensional Infrared Spectroscopy and Atomic Force Microscopy

    NASA Astrophysics Data System (ADS)

    Roeters, Steven J.; Iyer, Aditya; Pletikapić, Galja; Kogan, Vladimir; Subramaniam, Vinod; Woutersen, Sander

    2017-01-01

    The aggregation of the intrinsically disordered protein alpha-synuclein (αS) into amyloid fibrils is thought to play a central role in the pathology of Parkinson’s disease. Using a combination of techniques (AFM, UV-CD, XRD, and amide-I 1D- and 2D-IR spectroscopy) we show that the structure of αS fibrils varies as a function of ionic strength: fibrils aggregated in low ionic-strength buffers ([NaCl] ≤ 25 mM) have a significantly different structure than fibrils grown in higher ionic-strength buffers. The observations for fibrils aggregated in low-salt buffers are consistent with an extended conformation of αS molecules, forming hydrogen-bonded intermolecular β-sheets that are loosely packed in a parallel fashion. For fibrils aggregated in high-salt buffers (including those prepared in buffers with a physiological salt concentration) the measurements are consistent with αS molecules in a more tightly-packed, antiparallel intramolecular conformation, and suggest a structure characterized by two twisting stacks of approximately five hydrogen-bonded intermolecular β-sheets each. We find evidence that the high-frequency peak in the amide-I spectrum of αS fibrils involves a normal mode that differs fundamentally from the canonical high-frequency antiparallel β-sheet mode. The high sensitivity of the fibril structure to the ionic strength might form the basis of differences in αS-related pathologies.

  4. Evidence for Intramolecular Antiparallel Beta-Sheet Structure in Alpha-Synuclein Fibrils from a Combination of Two-Dimensional Infrared Spectroscopy and Atomic Force Microscopy

    PubMed Central

    Roeters, Steven J.; Iyer, Aditya; Pletikapić, Galja; Kogan, Vladimir; Subramaniam, Vinod; Woutersen, Sander

    2017-01-01

    The aggregation of the intrinsically disordered protein alpha-synuclein (αS) into amyloid fibrils is thought to play a central role in the pathology of Parkinson’s disease. Using a combination of techniques (AFM, UV-CD, XRD, and amide-I 1D- and 2D-IR spectroscopy) we show that the structure of αS fibrils varies as a function of ionic strength: fibrils aggregated in low ionic-strength buffers ([NaCl] ≤ 25 mM) have a significantly different structure than fibrils grown in higher ionic-strength buffers. The observations for fibrils aggregated in low-salt buffers are consistent with an extended conformation of αS molecules, forming hydrogen-bonded intermolecular β-sheets that are loosely packed in a parallel fashion. For fibrils aggregated in high-salt buffers (including those prepared in buffers with a physiological salt concentration) the measurements are consistent with αS molecules in a more tightly-packed, antiparallel intramolecular conformation, and suggest a structure characterized by two twisting stacks of approximately five hydrogen-bonded intermolecular β-sheets each. We find evidence that the high-frequency peak in the amide-I spectrum of αS fibrils involves a normal mode that differs fundamentally from the canonical high-frequency antiparallel β-sheet mode. The high sensitivity of the fibril structure to the ionic strength might form the basis of differences in αS-related pathologies. PMID:28112214

  5. Hybridization parameters revisited: solutions containing SDS.

    PubMed

    Rose, Ken; Mason, John O; Lathe, Richard

    2002-07-01

    Salt concentration governs nucleic acid hybridization according to the Schildkraut-Lifson equation. High concentrations of SDS are used in some common protocols, but the effects of SDS on hybridization stringency have not been reported. We investigated hybridization parameters in solutions containing SDS. With targets immobilized on nylon membranes and PCR- or transcription-generated probes, we report that the 50% dissociation temperature (Tm*) in the absence of SDS was 15 degrees C-17degrees C lower than the calculated Tm. SDS had only modest effects on Tm* [1% (w/v) equating to 8 mM NaCl]. RNA/DNA hybrids were approximately 11 degrees C more stable than DNA/DNA hybrids. Incomplete homology (69%) significantly reduced the Tm* for DNA/DNA hybrids (approximately /4degrees C; 0.45 degrees C/% nonhomology) but far less so for RNA/DNA hybrids (approximately 2.3 degrees C; approximately 0.07 degrees C/% non-homology); incomplete homology also markedly reduced the extent of hybridization. On these nylonfilters, SDS had a major effect on nonspecific binding. Buffers lacking SDS, or with low salt concentration, gave high hybridization backgrounds; buffers containing SDS, or high-salt buffers, gave reproducibly low backgrounds.

  6. Effect of protein solution components in the adsorption of Herbaspirillum seropedicae GlnB protein on mica.

    PubMed

    Ferreira, Cecília F G; Benelli, Elaine M; Klein, Jorge J; Schreiner, Wido; Camargo, Paulo C

    2009-10-15

    The adsorption of proteins and its buffer solution on mica surfaces was investigated by atomic force microscopy (AFM). Different salt concentration of the Herbaspirillum seropedicae GlnB protein (GlnB-Hs) solution deposited on mica was investigated. This protein is a globular, soluble homotrimer (36kDa), member of PII-like proteins family involved in signal transducing in prokaryote. Supramolecular structures were formed when this protein was deposited onto bare mica surface. The topographic AFM images of the GlnB-Hs films showed that at high salt concentration the supramolecular structures are spherical-like, instead of the typical doughnut-like shape for low salt concentration. AFM images of NaCl and Tris from the buffer solution showed structures with the same pattern as those observed for high salt protein solution, misleading the image interpretation. XPS experiments showed that GlnB protein film covers the mica surface without chemical reaction.

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

    PubMed

    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.

  8. Cryo-electron microscopy of vitrified SV40 minichromosomes: the liquid drop model.

    PubMed

    Dubochet, J; Adrian, M; Schultz, P; Oudet, P

    1986-03-01

    The structure of SV40 minichromosomes has been studied by cryo-electron microscopy of vitrified thin layers of solution. In high-salt buffer (130 mM NaCl), freshly prepared minichromosomes are condensed into globules 30 nm or more in diameter. On the micrograph, they appear to be formed by the close packing of 10 nm granules which give rise to a 10 nm reflection in the optical diffractogram. The globules can adopt many different conformations. At high concentration, they fuse into a homogeneous 'sea' of closely packed 10 nm granules. In low-salt buffer (less than 10 mM NaCl), the globules open, first into 10 nm filaments, and then into nucleosome-strings. The 'liquid drop' model is proposed to explain the condensed structure of the minichromosome in high-salt buffer: nucleosomes stack specifically on top of one another, thus forming the 10 nm filaments. 10 nm filaments in turn, tend to aggregate laterally. Optimizing both these interactions results in the condensation of 10 nm filaments or portions thereof into a structure similar to that of a liquid. Some implications of this model for the structure of cellular chromatin are discussed.

  9. Fundamental and Applied Studies of Polymer Membranes

    NASA Astrophysics Data System (ADS)

    Imbrogno, Joseph

    Four major areas have been studied in this research: 1) synthesizing novel monomers, e.g. chiral monomers, to produce new types of functionalized membranes for the biotechnology and pharmaceutical industries, 2) hydrophobic brush membranes for desalinating brackish water, sea water, and separating organics, 3) fundamental studies of water interactions at surfaces using sum frequency generation (SFG), and 4) discovering new surface chemistries that will control the growth and differentiation of stem cells. We have developed a novel synthesis method in order to increase the breadth of our high throughput screening library. This library was generated using maleimide chemistry to react a common methacrylate linker with a variety of different functions groups (R groups) in order to form new monomers that were grafted from the surface of PES ultrafiltration membranes. From this work, we discovered that the chirality of a membrane can affect performance when separating chiral feed streams. This effect was observed when filtering bovine serum albumin (BSA) and ovalbumin in a high salt phosphate buffered saline (PBS, 150 mM salt). The Phe grafted membranes showed a large difference in performance when filtering BSA with selectivity of 1.13 and 1.00 for (S) and (R) Phe, respectively. However, when filtering ovalbumin, the (S) and (R) modified surfaces showed selectivity of 2.06 and 2.31, respectively. The higher selectivity enantiomer switched for the two different proteins. Permeability when filtering BSA was 3.06 LMH kPa-1 and 4.31 LMH kPa -1 for (S)- and (R)- Phe, respectively, and 2.65 LMH kPa -1 and 2.10 LMH kPa-1 when filtering ovalbumin for (S)- and (R)- Phe, respectively. Additionally, these effects were no longer present when using a low salt phosphate buffer (PB, 10 mM salt). Since, to our knowledge, membrane chirality is not considered in current industrial systems, this discovery could have a large impact on the pharmaceutical and biotechnology industries. We have developed hydrophobic brush membranes that were able to selectively separate valuable organics (isobutanol) from water, while rejecting other undesirable species, such as enzymes, using pervaporation (PV). These membranes (grafted from nanofiltration (NF) support membranes) had a selectivity ˜1.5x higher than the current industrial standard, polydimethylsiloxane (PDMS), with alpha = 10.1 +/- 0.9 for our brush membranes and alpha = 6.7 +/- 0.1 for PDMS membranes. Since the mechanism of pervaporation is based on the solution diffusion (SD) model, these membranes may be used to desalinate water or fractionate gases since they are also based on the SD mechanism. We have discovered that hydrophobic brush membranes are able to reject monovalent salt ions. This type of membrane is analogous to carbon nanotubes (CNTs), which are believed to have extremely high water fluxes through them due to near frictionless flow caused by a lack of hydrogen bonding. Using these brush membranes we were able to achieve 42% monovalent (NaCl) salt rejection of simulated seawater (32,000 ppm salt). These membranes are easier to scale-up than current composite membranes produced using interfacial polymerization. We have been using SFG to study interfacial water on membrane surfaces. We believe that water interactions with the membrane surface and with the feed species, e.g. proteins, play a critical role during the fouling process. Relevant buffers, such as phosphate buffered saline (PBS) and phosphate buffer, contain ions that are known to restructure water at interfaces. Sum frequency generation spectroscopy (SFG) was used to characterize interfacial water structure at poly(ether sulfone) (PES) thin films in the presence of 0.01 M phosphate buffer (low salt) and 0.01 M phosphate buffered saline (high salt). Three model surfaces were studied: unmodified PES, hydrophobic alkane (C18) modified PES, and poly(ethylene glycol) (PEG) modified PES. In the presence of the low salt phosphate buffer (10 mM salt), phosphate anions were excluded from the PEG-modified PES film. This led to a charge separation between the phosphate anions and sodium cations, creating a surface potential which strongly ordered water molecules into the bulk. When using high salt PBS (138 mM salt) the sodium chloride ions screened this charge and reduced water ordering. Interestingly, this effect was the greatest for the PEG modified surface, with minor or no effects observed for the C18 modified PES and unmodified PES, respectively. Using our high throughput screening platform, we were able to determine that (N-[3-(dimethylamino)propyl] methacrylamide), DMAPMA, supported strong attachment and long-term self-renewal of mouse embryonic stem (ES) cells while preventing differentiation (maintaining pluripotency). After developing this platform, it was used to screen for a surface that could instead induce differentiation of bovine and human retinal pigment epithelium (RPE) cells while promoting cell growth. Several PEG based surfaces were able to induce cobblestone morphology of the RPE cells, which is indicative of differentiation. (Abstract shortened by UMI.).

  10. Method for improving performance of high temperature superconductors within a magnetic field

    DOEpatents

    Wang, Haiyan; Foltyn, Stephen R.; Maiorov, Boris A.; Civale, Leonardo

    2010-01-05

    The present invention provides articles including a base substrate including a layer of an oriented cubic oxide material having a rock-salt-like structure layer thereon; and, a buffer layer upon the oriented cubic oxide material having a rock-salt-like structure layer, the buffer layer having an outwardly facing surface with a surface morphology including particulate outgrowths of from 10 nm to 500 run in size at the surface, such particulate outgrowths serving as flux pinning centers whereby the article maintains higher performance within magnetic fields than similar articles without the necessary density of such outgrowths.

  11. 7 CFR 1410.30 - Signup.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... highly under § 1410.31(b) and may include high priority practices such as filter strips, riparian buffers, shelterbelts, field windbreaks, and living snow fences, grass waterways, shallow water areas for wildlife, salt...

  12. 7 CFR 1410.30 - Signup.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... highly under § 1410.31(b) and may include high priority practices such as filter strips, riparian buffers, shelterbelts, field windbreaks, and living snow fences, grass waterways, shallow water areas for wildlife, salt...

  13. 7 CFR 1410.30 - Signup.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... highly under § 1410.31(b) and may include high priority practices such as filter strips, riparian buffers, shelterbelts, field windbreaks, and living snow fences, grass waterways, shallow water areas for wildlife, salt...

  14. 7 CFR 1410.30 - Signup.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... highly under § 1410.31(b) and may include high priority practices such as filter strips, riparian buffers, shelterbelts, field windbreaks, and living snow fences, grass waterways, shallow water areas for wildlife, salt...

  15. 7 CFR 1410.30 - Signup.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... highly under § 1410.31(b) and may include high priority practices such as filter strips, riparian buffers, shelterbelts, field windbreaks, and living snow fences, grass waterways, shallow water areas for wildlife, salt...

  16. Effect of Pressure-Induced Changes in the Ionization Equilibria of Buffers on Inactivation of Escherichia coli and Staphylococcus aureus by High Hydrostatic Pressure

    PubMed Central

    Gayán, Elisa; Condón, Santiago; Álvarez, Ignacio; Nabakabaya, Maria

    2013-01-01

    Survival rates of Escherichia coli and Staphylococcus aureus after high-pressure treatment in buffers that had large or small reaction volumes (ΔV°), and which therefore underwent large or small changes in pH under pressure, were compared. At a low buffer concentration of 0.005 M, survival was, as expected, better in MOPS (morpholinepropanesulfonic acid), HEPES, and Tris, whose ΔV° values are approximately 5.0 to 7.0 cm3 mol−1, than in phosphate or dimethyl glutarate (DMG), whose ΔV° values are about −25 cm3 mol−1. However, at a concentration of 0.1 M, survival was unexpectedly better in phosphate and DMG than in MOPS, HEPES, or Tris. This was because the baroprotective effect of phosphate and DMG increased much more rapidly with increasing concentration than it did with MOPS, HEPES, or Tris. Further comparisons of survival in solutions of salts expected to cause large electrostriction effects (Na2SO4 and CaCl2) and those causing lower electrostriction (NaCl and KCl) were made. The salts with divalent ions were protective at much lower concentrations than salts with monovalent ions. Buffers and salts both protected against transient membrane disruption in E. coli, but the molar concentrations necessary for membrane protection were much lower for phosphate and Na2SO4 than for HEPES and NaCl. Possible protective mechanisms discussed include effects of electrolytes on water compressibility and kosmotropic and specific ion effects. The results of this systematic study will be of considerable practical significance in studies of pressure inactivation of microbes under defined conditions but also raise important fundamental questions regarding the mechanisms of baroprotection by ionic solutes. PMID:23624471

  17. Inactivation of viruses using novel protein A wash buffers.

    PubMed

    Bolton, Glen R; Selvitelli, Keith R; Iliescu, Ionela; Cecchini, Douglas J

    2015-01-01

    Low pH viral inactivation is typically performed in the eluate pool following the protein A capture step during the manufacturing of monoclonal antibodies and Fc-fusion proteins. However, exposure to low pH has the potential to alter protein quality. To avoid these difficulties, novel wash buffers capable of inactivating viruses while antibodies or Fc-fusion proteins were bound to protein A or mixed mode resins were developed. By equilibrating the column in high salt buffer (2 M ammonium sulfate or 3 M sodium chloride) after loading, the hydrophobic interactions between antibodies and protein A ligands were increased enough to prevent elution at pH 3. The ammonium sulfate was also found to cause binding of an antibody to a mixed mode cation exchange and a mixed mode anion exchange resin at pH values that caused elution in conventional cation and anion exchange resins (pH 3.5 for Capto Adhere and pH 8.0 for Capto MMC), indicating that retention was due to enhanced hydrophobic interactions. The potential of the 2 M ammonium sulfate pH 3 buffer, a 1 M arginine buffer, and a buffer containing the detergent LDAO to inactivate XMuLV virus when used as protein A wash buffers with a 1 hour contact time were studied. The high salt and detergent containing wash buffers provided about five logs of removal, determined using PCR, and complete combined removal and inactivation (> 6 logs), determined by measuring infectivity. The novel protein A washes could provide more rapid, automated viral inactivation steps with lower pool conductivities. © 2014 American Institute of Chemical Engineers.

  18. Electrolytic method to make alkali alcoholates using ion conducting alkali electrolyte/separator

    DOEpatents

    Joshi, Ashok V [Salt Lake City, UT; Balagopal, Shekar [Sandy, UT; Pendelton, Justin [Salt Lake City, UT

    2011-12-13

    Alkali alcoholates, also called alkali alkoxides, are produced from alkali metal salt solutions and alcohol using a three-compartment electrolytic cell. The electrolytic cell includes an anolyte compartment configured with an anode, a buffer compartment, and a catholyte compartment configured with a cathode. An alkali ion conducting solid electrolyte configured to selectively transport alkali ions is positioned between the anolyte compartment and the buffer compartment. An alkali ion permeable separator is positioned between the buffer compartment and the catholyte compartment. The catholyte solution may include an alkali alcoholate and alcohol. The anolyte solution may include at least one alkali salt. The buffer compartment solution may include a soluble alkali salt and an alkali alcoholate in alcohol.

  19. Desalting by crystallization: detection of attomole biomolecules in picoliter buffers by mass spectrometry.

    PubMed

    Gong, Xiaoyun; Xiong, Xingchuang; Wang, Song; Li, Yanyan; Zhang, Sichun; Fang, Xiang; Zhang, Xinrong

    2015-10-06

    Sensitive detection of biomolecules in small-volume samples by mass spectrometry is, in many cases, challenging because of the use of buffers to maintain the biological activities of proteins and cells. Here, we report a highly effective desalting method for picoliter samples. It was based on the spontaneous separation of biomolecules from salts during crystallization of the salts. After desalting, the biomolecules were deposited in the tip of the quartz pipet because of the evaporation of the solvent. Subsequent detection of the separated biomolecules was achieved using solvent assisted electric field induced desorption/ionization (SAEFIDI) coupled with mass spectrometry. It allowed for direct desorption/ionization of the biomolecules in situ from the tip of the pipet. The organic component in the assistant solvent inhibited the desorption/ionization of salts, thus assured successful detection of biomolecules. Proteins and peptides down to 50 amol were successfully detected using our method even if there were 3 × 10(5) folds more amount of salts in the sample. The concentration and ion species of the salts had little influence on the detection results.

  20. The influence of bile salts on the distribution of simvastatin in the octanol/buffer system.

    PubMed

    Đanić, Maja; Pavlović, Nebojša; Stanimirov, Bojan; Vukmirović, Saša; Nikolić, Katarina; Agbaba, Danica; Mikov, Momir

    2016-01-01

    Distribution coefficient (D) is useful parameter for evaluating drugs permeability properties across biological membranes, which are of importance for drugs bioavailability. Given that bile acids are intensively studied as drug permeation-modifying and -solubilizing agents, the aim of this study was to estimate the influence of sodium salts of cholic (CA), deoxycholic (DCA) and 12-monoketocholic acids (MKC) on distribution coefficient of simvastatin (SV) (lactone [SVL] and acid form [SVA]) which is a highly lipophilic compound with extremely low water solubility and bioavailability. LogD values of SVA and SVL with or without bile salts were measured by liquid-liquid extraction in n-octanol/buffer systems at pH 5 and 7.4. SV concentrations in aqueous phase were determined by HPLC-DAD. Chem3D Ultra program was applied for computation of physico-chemical properties of analyzed compounds and their complexes. Statistically significant decrease in both SVA and SVL logD was observed for all three studied bile salts at both selected pH. MKC exerted the most pronounced effect in the case of SVA while there were no statistically significant differences between observed bile salts for SVL. The calculated physico-chemical properties of analyzed compounds and their complexes supported experimental results. Our data indicate that the addition of bile salts into the n-octanol/buffer system decreases the values of SV distribution coefficient at both studied pH values. This may be the result of the formation of hydrophilic complexes increasing the solubility of SV that could consequently impact the pharmacokinetic parameters of SV and the final drug response in patients.

  1. Near Wall Velocity and Vorticity Measurements, In A Very High R(theta) Turbulent Boundary Layer

    DTIC Science & Technology

    2006-06-15

    Fluctuation in temperature 7 Angle of the oncoming flow with respect to the axis of the probe body Superscripts + Variables scaled on inner parameters Denotes...at high Reynolds numbers (Re = 5* 106). They examined flow in the viscous sublayer and buffer layer to explore the influence of very high Reynolds...Great Salt Lake and surrounding "salt flats" sit in the drying basin of what is believed to have once been a much larger body of water covering the

  2. Intertidal salt marshes as an important source of inorganic carbon to the coastal ocean

    USGS Publications Warehouse

    Wang, Zhaohui Aleck; Kroeger, Kevin D.; Ganju, Neil K.; Gonneea, Meagan; Chu, Sophie N.

    2016-01-01

    Dynamic tidal export of dissolved inorganic carbon (DIC) to the coastal ocean from highly productive intertidal marshes and its effects on seawater carbonate chemistry are thoroughly evaluated. The study uses a comprehensive approach by combining tidal water sampling of CO2parameters across seasons, continuous in situ measurements of biogeochemically-relevant parameters and water fluxes, with high-resolution modeling in an intertidal salt marsh of the U.S. northeast region. Salt marshes can acidify and alkalize tidal water by injecting CO2 (DIC) and total alkalinity (TA). DIC and TA generation may also be decoupled due to differential effects of marsh aerobic and anaerobic respiration on DIC and TA. As marsh DIC is added to tidal water, the buffering capacity first decreases to a minimum and then increases quickly. Large additions of marsh DIC can result in higher buffering capacity in ebbing tide than incoming tide. Alkalization of tidal water, which mostly occurs in the summer due to anaerobic respiration, can further modify buffering capacity. Marsh exports of DIC and alkalinity may have complex implications for the future, more acidified ocean. Marsh DIC export exhibits high variability over tidal and seasonal cycles, which is modulated by both marsh DIC generation and by water fluxes. The marsh DIC export of 414 g C m−2 yr−1, based on high-resolution measurements and modeling, is more than twice the previous estimates. It is a major term in the marsh carbon budget and translates to one of the largest carbon fluxes along the U.S. East Coast.

  3. Buffer salt effects in off-line coupling of capillary electrophoresis and mass spectrometry.

    PubMed

    Marák, Jozef; Stanová, Andrea

    2014-05-01

    In this work, the impact of buffer salts/matrix effects on the signal in direct injection MS with an electrospray interface (DI-ESI-MS) following pITP fractionation of the sample was studied. A range of buffers frequently used in CE analyses (pH 3-10) was prepared containing 10, 50, and 90% v/v of ACN, respectively. The sets of calibration solutions of cetirizine (an antihistaminic drug with an amphiprotic character) within a 0.05-2.0 mg/L concentration range were prepared in different buffers. The greatest enhancements in the MS signal (in terms of change in the slope of the calibration line) were obtained for the beta-alanine buffer (pH 3.5) in positive ionization and for the borate buffer (pH 9.2) in negative ionization, respectively. The procedure was successfully applied to the analysis of buserelin (a peptidic drug). The slope of the calibration line for solutions containing the beta-alanine buffer with 50% of ACN was 4 times higher than for water or urine, respectively. This study clearly demonstrates that the buffer salt/matrix effects in an offline combination of pITP and DI-ESI-MS can also play a positive role, as they can enhance the signal in MS. A similar influence of the above effects can also be presumed in the CE techniques combined on-line with ESI-MS.

  4. Effect of modulator sorption on gradient shape in ion-exchange chromatography

    NASA Technical Reports Server (NTRS)

    Velayudhan, A.; Ladisch, M. R.; Mitchell, C. A. (Principal Investigator)

    1995-01-01

    Mobile phase additives, or modulators, are used in gradient elution chromatography to facilitate separation and reduce separation time. The modulators are usually assumed to be linearly adsorbed or unadsorbed. Here, the consequences of nonlinear modulator adsorption are examined for ion-exchange gradient elution through a series of simulations. Even when the buffer salt is identical to the modulator salt, gradient deformation is observed; the extent of deformation increases as the volume of the feed is increased. When the modulator salt is different from the buffer salt, unusual effects are observed, and the chromatograms are quite different from those predicted by classical gradient elution theory. In particular, local increases in the buffer concentration are found between feed bands, and serve to improve the separation. These effects become more pronounced as the feed volume increases, and could therefore prove valuable in preparative applications.

  5. Dahl SS rats demonstrate enhanced aortic perivascular adipose tissue-mediated buffering of vasoconstriction through activation of NOS in the endothelium

    PubMed Central

    Spradley, Frank T.; Ho, Dao H.

    2015-01-01

    Perivascular adipose tissue (PVAT) mediates buffering of vasoconstriction through activation of endothelium-derived factors. We hypothesized that the PVAT of Dahl salt-sensitive (Dahl SS) rats has reduced ability to buffer vasoconstriction. Vascular reactivity experiments were performed on aortic rings with PVAT intact (+PVAT) or removed (−PVAT), and endothelium intact (+ENDO) or removed (−ENDO) from Dahl SS rats and control SS.13BN rats (Dahl SS rats that have had chromosome 13 completely replaced with that of the Brown Norway rat, rendering this strain insensitive to high-salt or high-fat diet-induced hypertension). Endothelial dysfunction, assessed by ACh-mediated vasorelaxation, was confirmed in aortic rings of Dahl SS rats. The +PVAT+ENDO aortic rings had indistinguishable phenylephrine-induced vasoconstriction between genotypes. In both strains, removal of PVAT significantly enhanced vasoconstriction. Dahl SS rat −PVAT+ENDO aortic rings displayed exaggerated vasoconstriction to phenylephrine vs. SS.13BN rats, indicating that PVAT-mediated buffering of vasoconstriction was greater in Dahl SS rats. Removal of both the ENDO and PVAT restored vasoconstriction in both strains. The nitric oxide synthase (NOS) inhibitor, Nω-nitro-l-arginine methyl ester (l-NAME), produced a similar effect as that seen with −ENDO. These data indicate that the function of the PVAT to activate endothelium-derived NOS is enhanced in Dahl SS compared with SS.13BN rats and, most likely, occurs through a pathway that is distinct from ACh-mediated activation of NOS. PVAT weight and total PVAT leptin levels were greater in Dahl SS rats. Leptin induced a significantly decreased vasoconstriction in −PVAT+ENDO aortic rings from Dahl SS rats, but not SS.13BN rats. In contrast to our initial hypothesis, PVAT in Dahl SS rats buffers vasoconstriction by activating endothelial NOS via mechanisms that may include the involvement of leptin. Thus, the PVAT serves a vasoprotective role in Dahl SS rats on normal-salt diet. PMID:26608658

  6. Intrinsic electric fields and proton diffusion in immobilized protein membranes. Effects of electrolytes and buffers.

    PubMed Central

    Zabusky, N J; Deem, G S

    1979-01-01

    We present a theory for proton diffusion through an immobilized protein membrane perfused with an electrolyte and a buffer. Using a Nernst-Planck equation for each species and assuming local charge neutrality, we obtain two coupled nonlinear diffusion equations with new diffusion coefficients dependent on the concentration of all species, the diffusion constants or mobilities of the buffers and salts, the pH-derivative of the titration curves of the mobile buffer and the immobilized protein, and the derivative with respect to ionic strength of the protein titration curve. Transient time scales are locally pH-dependent because of protonation-deprotonation reactions with the fixed protein and are ionic strength-dependent because salts provide charge carriers to shield internal electric fields. Intrinsic electric fields arise proportional to the gradient of an "effective" charge concentration. The field may reverse locally if buffer concentrations are large (greater to or equal to 0.1 M) and if the diffusivity of the electrolyte species is sufficiently small. The "ideal" electrolyte case (where each species has the same diffusivity) reduces to a simple form. We apply these theoretical considerations to membranes composed of papain and bovine serum albumin (BSA) and show that intrinsic electric fields greatly enhance the mobility of protons when the ionic strength of the salts is smaller than 0.1 M. These results are consistent with experiments where pH changes are observed to depend strongly on buffer, salt, and proton concentrations in baths adjacent to the membranes. PMID:233570

  7. Salt Stability - The Effect of pHmax on Salt to Free Base Conversion.

    PubMed

    Hsieh, Yi-Ling; Merritt, Jeremy M; Yu, Weili; Taylor, Lynne S

    2015-09-01

    The aim of this study was to investigate how the disproportionation process can be impacted by the properties of the salt, specifically pHmax. Five miconazole salts and four sertraline salts were selected for this study. The extent of conversion was quantified using Raman spectroscopy. A mathematical model was utilized to estimate the theoretical amount of conversion. A trend was observed that for a given series of salts of a particular basic compound (both sertraline and miconazole are bases), the extent of disproportionation increases as pHmax decreases. Miconazole phosphate monohydrate and sertraline mesylate, although exhibiting significantly different pHmax values (more than 2 units apart), underwent a similar extent of disproportionation, which may be attributed to the lower buffering capacity of sertraline salts. This work shows that the disproportionation tendency can be influenced by pHmax and buffering capacity and thus highlights the importance of selecting the appropriate salt form during the screening process in order to avoid salt-to-free form conversion.

  8. Substrate Structures For Growth Of Highly Oriented And/Or Epitaxial Layers Thereon

    DOEpatents

    Arendt, Paul N.; Foltyn, Stephen R.; Groves, James R.; Jia, Quanxi

    2005-07-26

    A composite substrate structure including a substrate, a layer of a crystalline metal oxide or crystalline metal oxynitride material upon the substrate, a layer of an oriented cubic oxide material having a rock-salt-like structure upon the crystalline metal oxide or crystalline metal oxynitride material layer is provided together with additional layers such as one or more layers of a buffer material upon the oriented cubic oxide material layer. Jc's of 2.3×106 A/cm2 have been demonstrated with projected Ic's of 320 Amperes across a sample 1 cm wide for a superconducting article including a flexible polycrystalline metallic substrate, an inert oxide material layer upon the surface of the flexible polycrystalline metallic substrate, a layer of a crystalline metal oxide or crystalline metal oxynitride material upon the layer of the inert oxide material, a layer of an oriented cubic oxide material having a rock-salt-like structure upon the crystalline metal oxide or crystalline metal oxynitride material layer, a layer of a buffer material upon the oriented cubic oxide material layer, and, a top-layer of a high temperature superconducting material upon the layer of a buffer material.

  9. Effects of buffers on milk fatty acids and mammary arteriovenous differences in dairy cows fed Ca salts of fatty acids.

    PubMed

    Thivierge, M C; Chouinard, P Y; Lévesque, J; Girard, V; Seoane, J R; Brisson, G J

    1998-07-01

    Ten Holstein cows in early lactation were used in a replicated 5 x 5 Latin square design to study the effects of MgO and three buffers added to diets containing Ca salts of canola oil fatty acids. Treatments were 1) control (basal diet; no buffer). 2) 1.1% NaHCO3 plus 1.1% KHCO3, 3) 1.9% NaHCO3, 4) 0.5% MgO, and 5) 2.0% Na sesquicarbonate (percentage of dry matter). The control diet contained 53% grass silage, 43% concentrate, and 4% Ca salts. Body weight, intake, milk yield, and percentages of milk fat, protein, and lactose were unaffected by treatments. Buffers and MgO tended to increase triacylglycerol extraction by the mammary gland and changed the proportions of some fatty acids in milk. Arterial concentrations of acetate and triacylglycerol were correlated with their respective arteriovenous differences. Extraction by the mammary gland was high for acetate (approximately equal to 58.2%), triacylglycerol (approximately equal to 47.3%) propionate (approximately equal to 34.6%), and glucose (approximately equal to 24.3%). Extraction of free fatty acids, phospholipids, or cholesterol was negligible. Mammary triacylglycerol arteriovenous difference tended to be higher than when MgO was fed than when NaHCO3 was fed. Sodium sesquicarbonate, NaHCO3, and the blend of bicarbonate buffers increased C18:2 in milk fat when compared with the control treatment. The concentration of C18:2 in milk fat decreased when MgO was fed, but the ratio of cis-C18:1 to trans-C18:1 increased compared with effects of dietary NaHCO3. Medium-chain fatty acids in milk fat tended to be higher than Na sesquicarbonate than with NaHCO3. Buffers and MgO modified the profiles of fatty acids in milk.

  10. Extraction and LC determination of lysine clonixinate salt in water/oil microemulsions.

    PubMed

    Pineros, I; Ballesteros, P; Lastres, J L

    2002-02-01

    A new reversed-phase high performance liquid chromatography method has been developed and validated for the quantitative determination of lysine clonixinate salt in water/oil microemulsions. The mobile phase was acetonitrile-buffer phosphate pH 3.3. Detection was UV absorbance at 252 nm. The precision and accurately of the method were excellent. The established linearity range was 5-60 microg ml(-1) (r(2)=0.999). Microemulsions samples were dispersed with chloroform and extracted lysine clonixinate salt with water. This easy method employing chloroformic extraction has been done three times. The recovery of lysine clonixinate salt from spiked placebo and microemulsion were >90% over the linear range.

  11. Architecture for coated conductors

    DOEpatents

    Foltyn, Stephen R.; Arendt, Paul N.; Wang, Haiyan; Stan, Liliana

    2010-06-01

    Articles are provided including a base substrate having a layer of an oriented cubic oxide material with a rock-salt-like structure layer thereon, and, a layer of epitaxial titanium nitride upon the layer of an oriented cubic oxide material having a rock-salt-like structure. Such articles can further include thin films of high temperature superconductive oxides such as YBCO upon the layer of epitaxial titanium nitride or upon a intermediate buffer layer upon the layer of epitaxial titanium nitride.

  12. Quantitation of yeast total proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for uniform loading.

    PubMed

    Sheen, Hyukho

    2016-04-01

    Proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer are difficult to quantitate due to SDS and reducing agents being in the buffer. Although acetone precipitation has long been used to clean up proteins from detergents and salts, previous studies showed that protein recovery from acetone precipitation varies from 50 to 100% depending on the samples tested. Here, this article shows that acetone precipitates proteins highly efficiently from SDS-PAGE sample buffer and that quantitative recovery is achieved in 5 min at room temperature. Moreover, precipitated proteins are resolubilized with urea/guanidine, rather than with SDS. Thus, the resolubilized samples are readily quantifiable with Bradford reagent without using SDS-compatible assays. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Hydrology in a peaty high marsh: hysteretic flow and biogeochemical implications

    EPA Science Inventory

    Terrestrial nutrient input to coastal waters is a critical water quality problem worldwide, and salt marshes may provide a valuable nutrient buffer (either by removal or by smoothing out pulse inputs) between terrestrial sources and sensitive estuarine habitats. One of the major...

  14. Capillary Electrophoresis Analysis of Organic Amines and Amino Acids in Saline and Acidic Samples Using the Mars Organic Analyzer

    NASA Astrophysics Data System (ADS)

    Stockton, Amanda M.; Chiesl, Thomas N.; Lowenstein, Tim K.; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A.

    2009-11-01

    The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pKa values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

  15. Capillary electrophoresis analysis of organic amines and amino acids in saline and acidic samples using the Mars organic analyzer.

    PubMed

    Stockton, Amanda M; Chiesl, Thomas N; Lowenstein, Tim K; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A

    2009-11-01

    The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pK(a) values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

  16. Iron clad wetlands: Soil iron-sulfur buffering determines coastal wetland response to salt water incursion

    NASA Astrophysics Data System (ADS)

    Schoepfer, Valerie A.; Bernhardt, Emily S.; Burgin, Amy J.

    2014-12-01

    Coastal freshwater wetland chemistry is rapidly changing due to increased frequency of salt water incursion, a consequence of global change. Seasonal salt water incursion introduces sulfate, which microbially reduces to sulfide. Sulfide binds with reduced iron, producing iron sulfide (FeS), recognizable in wetland soils by its characteristic black color. The objective of this study is to document iron and sulfate reduction rates, as well as product formation (acid volatile sulfide (AVS) and chromium reducible sulfide (CRS)) in a coastal freshwater wetland undergoing seasonal salt water incursion. Understanding iron and sulfur cycling, as well as their reduction products, allows us to calculate the degree of sulfidization (DOS), from which we can estimate how long soil iron will buffer against chemical effects of sea level rise. We show that soil chloride, a direct indicator of the degree of incursion, best predicted iron and sulfate reduction rates. Correlations between soil chloride and iron or sulfur reduction rates were strongest in the surface layer (0-3 cm), indicative of surface water incursion, rather than groundwater intrusion at our site. The interaction between soil moisture and extractable chloride was significantly related to increased AVS, whereas increased soil chloride was a stronger predictor of CRS. The current DOS in this coastal plains wetland is very low, resulting from high soil iron content and relatively small degree of salt water incursion. However, with time and continuous salt water exposure, iron will bind with incoming sulfur, creating FeS complexes, and DOS will increase.

  17. Interaction of two diclofenac acid salts with copolymers of ammoniomethacrylate: effect of additives and release profiles.

    PubMed

    Khalil, E; Sallam, A

    1999-04-01

    The copolymer of ammoniomethacrylate Eudragit RL (ERL) interacted with diclofenac acid salts (sodium and diethylamine salts) in aqueous solutions, forming a complex. Sorption experiments were done in aqueous solutions of either sodium lauryl sulfate (SLS), Tween 20, or Tween 80. The SLS competed strongly with the drug, even at low concentrations, and reduced significantly the amount of drug sorbed by ERL. Tweens at high concentrations exhibited two phase profiles: the sorption phase, which was short and during which drug concentration dropped sharply, and the release phase, during which the drug was released slowly over 24 hr and which was accompanied by dispersion of ERL particles into the colloidal dispersion. The interaction was dependent on temperature, ionic strength, and nature of the additives. The extent of interaction in water and phosphate buffer solutions was in the following order: water > pH 6 > pH 7-8. In-vitro dissolution studies of the dried complex were done over 24 hr. In water, the drug remained bound to the polymer. In aqueous surfactant solutions (SLS, Tween 20, and Tween 80) and phosphate buffer at pH 6.8, a linear relationship between drug concentration and the square root of time was obtained, indicating a matrix diffusion-controlled mechanism. However, 100% release was not reached, and resorption was observed in the phosphate buffer solution.

  18. Sampling and RNA quality for successful diagnostics using quantitative PCR

    USDA-ARS?s Scientific Manuscript database

    Diagnostic analyses of RNA targets are widely used in honey bee pathology. These diagnostics can be compromised by the actions of endogenous RNA-degrading enzymes activated upon bee death. RNA degradation can be minimized by storage at ultra-cold temperatures or by immersion in high-salt buffers. H...

  19. DENITRIFICATION ENZYME ACTIVITY OF FRINGE SALT MARSHES IN NEW ENGLAND (USA)

    EPA Science Inventory

    Coastal salt marshes are a buffer between the uplands and adjacent coastal waters in New England (USA). With increasing N loads from developed watersheds, salt marshes could play an important role in the water quality maintenance of coastal waters. In this study we examined seaso...

  20. Electrophoretic mobilities of erythrocytes in various buffers

    NASA Technical Reports Server (NTRS)

    Plank, L. D.; Kunze, M. E.; Todd, P. W.

    1985-01-01

    The calibration of space flight equipment depends on a source of standard test particles, this test particle of choice is the fixed erythrocyte. Erythrocytes from different species have different electrophoretic mobilities. Electrophoretic mobility depends upon zeta potential, which, in turn depends upon ionic strength. Zeta potential decreases with increasing ionic strength, so cells have high electrophoretic mobility in space electrophoresis buffers than in typical physiological buffers. The electrophoretic mobilities of fixed human, rat, and rabbit erythrocytes in 0.145 M salt and buffers of varying ionic strength, temperature, and composition, to assess the effects of some of the unique combinations used in space buffers were characterized. Several effects were assessed: glycerol or DMSO (dimethylsulfoxide) were considered for use as cryoprotectants. The effect of these substances on erythrocyte electrophoretic mobility was examined. The choice of buffer depended upon cell mobility. Primary experiments with kidney cells established the choice of buffer and cryoprotectant. A nonstandard temperature of EPM in the suitable buffer was determined. A loss of ionic strength control occurs in the course of preparing columns for flight, the effects of small increases in ionic strength over the expected low values need to be evaluated.

  1. Effect of various halide salts on the incompatibility of cyanocobalamin and ascorbic acid in aqueous solution.

    PubMed

    Ichikawa, Makoto; Ide, Nagatoshi; Shiraishi, Sumihiro; Ono, Kazuhisa

    2005-06-01

    Combination of cyanocobalamin (VB12) and ascorbic acid (VC) has been widely seen in pharmaceutical products and dietary supplements. However, VB12 has been reported that its behavior in stability in aqueous solution is quite different when VC is mixed. In the present study, we examined the stabilities of these vitamins in acetate buffer (pH 4.8) using high performance liquid chromatography. Degradation of VB12 was not observed in the absence of VC in the buffer. However, when VC was mixed in the VB12 solution, VB12 concentrations decreased in accordance with VC degradation. VB12 and VC degradations were inhibited by adding sodium halides to acetate buffer at pH 4.8. These stabilization effects were also observed in the range from pH 3.5 to 5.3 and by adding potassium, magnesium, and calcium halides. Furthermore, our data demonstrated that increases in the halide anion concentrations and atomic number (Cl-

  2. Effect of pH buffer molecules on the light-induced currents from oriented purple membrane.

    PubMed Central

    Liu, S Y; Kono, M; Ebrey, T G

    1991-01-01

    The effect of pH buffers on the microsecond photocurrent component, B2, of oriented purple membranes has been studied. We found that under low salt conditions (less than 10 mM monovalent cationic salt) pH buffers can dramatically alter the waveform of the B2 component. The effect is induced by the protonation process of the buffer molecules by protons expelled from the membrane. These effects can be classified according to the charge transition upon protonation of the buffer. Buffers that carry two positive charges in their protonated form add a negative current component (N component) to B2. Almost all of the other buffers add a positive current component (P component) to B2, which is essentially a mirror image of the N component. Buffers with a pK less than 5.5 have only a small positive buffer component. The pH dependence of the buffer effect is closely related to the pK of the buffer; it requires that the buffer be in its unprotonated form. The rise time of the buffer component increases with the concentration of the buffer molecules. All the buffer effects can be inhibited by the addition of 5 mM of a divalent cation such as Ca2+. Reducing the surface potential slows down the N component but accelerates the P component without affecting the amplitude of the buffer effect significantly. Many of the buffer effects can be explained if we assume that upon protonation of the buffer by a proton expelled from the membrane by light, the buffer molecules move toward the membrane. This backward movement of buffer molecules forms a counter current very similar to that due to cations discussed in Liu, S. Y., R. Govindjee, and T. G. Ebrey. (1990. Biophys. J. 57:951-963). PMID:1883939

  3. Anions in Electrothermal Supercharging of Proteins with Electrospray Ionization Follow a Reverse Hofmeister Series

    PubMed Central

    2015-01-01

    The effects of different anions on the extent of electrothermal supercharging of proteins from aqueous ammonium and sodium salt solutions were investigated. Sulfate and hydrogen phosphate are the most effective anions at producing high charge state protein ions from buffered aqueous solution, whereas iodide and perchlorate are ineffective with electrothermal supercharging. The propensity for these anions to produce high charge state protein ions follows the following trend: sulfate > hydrogen phosphate > thiocyanate > bicarbonate > chloride > formate ≈ bromide > acetate > iodide > perchlorate. This trend correlates with the reverse Hofmeister series over a wide range of salt concentrations (1 mM to 2 M) and with several physical properties, including solvent surface tension, anion viscosity B-coefficient, and anion surface/bulk partitioning coefficient, all of which are related to the Hofmeister series. The effectiveness of electrothermal supercharging does not depend on bubble formation, either from thermal degradation of the buffer or from coalescence of dissolved gas. These results provide evidence that the effect of different ions in the formation of high charge state ions by electrothermal supercharging is largely a result of Hofmeister effects on protein stability leading to protein unfolding in the heated ESI droplet. PMID:24410546

  4. Inhibition of clostridium perfringens spore germination and outgrowth by buffered vinegar and lemon juice concentrate during chilling.....of ground turkey road containing minimal ingredients

    USDA-ARS?s Scientific Manuscript database

    Inhibition of Clostridium perfringens spore germination and outgrowth in ground turkey roast containing minimal ingredients (salt and sugar), by buffered vinegar (MoStatin V) and a blend (buffered) of lemon juice concentrate and vinegar (MoStatin LV) was evaluated. Ground turkey roast was formulat...

  5. Buffers more than buffering agent: introducing a new class of stabilizers for the protein BSA.

    PubMed

    Gupta, Bhupender S; Taha, Mohamed; Lee, Ming-Jer

    2015-01-14

    In this study, we have analyzed the influence of four biological buffers on the thermal stability of bovine serum albumin (BSA) using dynamic light scattering (DLS). The investigated buffers include 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), 4-(2-hydroxyethyl)-1-piperazine-propanesulfonic acid (EPPS), 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid sodium salt (HEPES-Na), and 4-morpholinepropanesulfonic acid sodium salt (MOPS-Na). These buffers behave as a potential stabilizer for the native structure of BSA against thermal denaturation. The stabilization tendency follows the order of MOPS-Na > HEPES-Na > HEPES ≫ EPPS. To obtain an insight into the role of hydration layers and peptide backbone in the stabilization of BSA by these buffers, we have also explored the phase transition of a thermoresponsive polymer, poly(N-isopropylacrylamide (PNIPAM)), a model compound for protein, in aqueous solutions of HEPES, EPPS, HEPES-Na, and MOPS-Na buffers at different concentrations. It was found that the lower critical solution temperatures (LCST) of PNIPAM in the aqueous buffer solutions substantially decrease with increase in buffer concentration. The mechanism of interactions between these buffers and protein BSA was probed by various techniques, including UV-visible, fluorescence, and FTIR. The results of this series of studies reveal that the interactions are mainly governed by the influence of the buffers on the hydration layers surrounding the protein. We have also explored the possible binding sites of BSA with these buffers using a molecular docking technique. Moreover, the activities of an industrially important enzyme α-chymotrypsin (α-CT) in 0.05 M, 0.5 M, and 1.0 M of HEPES, EPPS, HEPES-Na, and MOPS-Na buffer solutions were analyzed at pH = 8.0 and T = 25 °C. Interestingly, the activities of α-CT were found to be enhanced in the aqueous solutions of these investigated buffers. Based upon the Jones-Dole viscosity parameters, the kosmotropic or chaotropic behaviors of the investigated buffers at 25 °C have been examined.

  6. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

    Studies of a zwitterionic buffer formulated for cell electrophoresis were done using the McDonnell-Douglas Continuous Flow Electrophoresis System. Standard buffers were analyzed for their stability in the electrical field and the results showed that both buffers tested were inherently unstable. Further, titration studies showed that the standards buffers buffered poorly at the pH employed for electrophoresis. The zwitterionic buffer buffered well at its nominal pH and was shown to be stable in the electrical field. Comparative studies of the buffer with standard cell separation buffers using formalin fixed rabbit and goose red blood cells showed that the zwitterionic buffer gave better resolution of the fixed cells. Studies with viable hybridoma cells showed that buffer Q supported cell viability equal to Hank's Balanced Salt Solution and that hybridoma cells in different stages of the growth cycle demonstrated reproducible differences in electrophoretic mobility.

  7. The impact of reduced gastric acid secretion on dissolution of salts of weak bases in the fasted upper gastrointestinal lumen: Data in biorelevant media and in human aspirates.

    PubMed

    Litou, Chara; Vertzoni, Maria; Xu, Wei; Kesisoglou, Filippos; Reppas, Christos

    2017-06-01

    To propose media for simulating the intragastric environment under reduced gastric acid secretion in the fasted state at three levels of simulation of the gastric environment and evaluate their usefulness in evaluating the intragastric dissolution of salts of weak bases. To evaluate the importance of bicarbonate buffer in biorelevant in vitro dissolution testing when using Level II biorelevant media simulating the environment in the fasted upper small intestine, regardless of gastric acid secretions. Media for simulating the hypochlorhydric and achlorhydric conditions in stomach were proposed using phosphates, maleates and bicarbonates buffers. The impact of bicarbonates in Level II biorelevant media simulating the environment in upper small intestine was evaluated so that pH and bulk buffer capacity were maintained. Dissolution data were collected using two model compounds, pioglitazone hydrochloride and semifumarate cocrystal of Compound B, and the mini-paddle dissolution apparatus in biorelevant media and in human aspirates. Simulated gastric fluids proposed in this study were in line with pH, buffer capacity, pepsin content, total bile salt/lecithin content and osmolality of the fasted stomach under partial and under complete inhibition of gastric acid secretion. Fluids simulating the conditions under partial inhibition of acid secretion were useful in simulating concentrations of both model compounds in gastric aspirates. Bicarbonates in Level III biorelevant gastric media and in Level II biorelevant media simulating the composition in the upper intestinal lumen did not improve simulation of concentrations in human aspirates. Level III biorelevant media for simulating the intragastric environment under hypochlorhydric conditions were proposed and their usefulness in the evaluation of concentrations of two model salts of weak bases in gastric aspirates was shown. Level II biorelevant media for simulating the environment in upper intestinal lumen led to underestimation of concentrations in aspirates, even when bicarbonate buffer was used. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. The effects of buffers and pH on the thermal stability, unfolding and substrate binding of RecA.

    PubMed

    Metrick, Michael A; Temple, Joshua E; MacDonald, Gina

    2013-12-31

    The Escherichia coli protein RecA is responsible for catalysis of the strand transfer reaction used in DNA repair and recombination. Previous studies in our lab have shown that high concentrations of salts stabilize RecA in a reverse-anionic Hofmeister series. Here we investigate how changes in pH and buffer alter the thermal unfolding and cofactor binding. RecA in 20mM HEPES, MES, Tris and phosphate buffers was studied in the pH range from 6.5 to 8.5 using circular dichroism (CD), infrared (IR) and fluorescence spectroscopies. The results show all of the buffers studied stabilize RecA up to 50°C above the Tris melting temperature and influence RecA's ability to nucleate on double-stranded DNA. Infrared and CD spectra of RecA in the different buffers do not show that secondary structural changes are associated with increased stability or decreased ability to nucleate on dsDNA. These results suggest the differences in stability arise from decreasing positive charge and/or buffer interactions. © 2013. Published by Elsevier B.V. All rights reserved.

  9. One-step large-scale deposition of salt-free DNA origami nanostructures

    PubMed Central

    Linko, Veikko; Shen, Boxuan; Tapio, Kosti; Toppari, J. Jussi; Kostiainen, Mauri A.; Tuukkanen, Sampo

    2015-01-01

    DNA origami nanostructures have tremendous potential to serve as versatile platforms in self-assembly -based nanofabrication and in highly parallel nanoscale patterning. However, uniform deposition and reliable anchoring of DNA nanostructures often requires specific conditions, such as pre-treatment of the chosen substrate or a fine-tuned salt concentration for the deposition buffer. In addition, currently available deposition techniques are suitable merely for small scales. In this article, we exploit a spray-coating technique in order to resolve the aforementioned issues in the deposition of different 2D and 3D DNA origami nanostructures. We show that purified DNA origamis can be controllably deposited on silicon and glass substrates by the proposed method. The results are verified using either atomic force microscopy or fluorescence microscopy depending on the shape of the DNA origami. DNA origamis are successfully deposited onto untreated substrates with surface coverage of about 4 objects/mm2. Further, the DNA nanostructures maintain their shape even if the salt residues are removed from the DNA origami fabrication buffer after the folding procedure. We believe that the presented one-step spray-coating method will find use in various fields of material sciences, especially in the development of DNA biochips and in the fabrication of metamaterials and plasmonic devices through DNA metallisation. PMID:26492833

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

    Sawyer, Thomas W., E-mail: Thomas.Sawyer@drdc-rddc.gc.ca; Nelson, Peggy; Bjarnason, Stephen

    The effect of ionic environment on sulphur mustard (bis 2-chloroethyl sulphide; HD) toxicity was examined in CHO-K1 cells. Cultures were treated with HD in different ionic environments at constant osmolar conditions (320 mOsM, pH 7.4). The cultures were refed with fresh culture medium 1 h after HD exposure, and viability was assessed. Little toxicity was apparent when HD exposures were carried out in ion-free sucrose buffer compared to LC{sub 50} values of {approx} 100-150 {mu}M when the cultures were treated with HD in culture medium. Addition of NaCl to the buffer increased HD toxicity in a salt concentration-dependent manner tomore » values similar to those obtained in culture medium. HD toxicity was dependent on both cationic and anionic species with anionic environment playing a much larger role in determining toxicity. Substitution of NaI for NaCl in the treatment buffers increased HD toxicity by over 1000%. The activity of the sodium hydrogen exchanger (NHE) in recovering from cytosolic acidification in salt-free and in different chloride salts did not correlate with the HD-induced toxicity in these buffers. However, the inhibition by HD of intracellular pH regulation correlated with its toxicity in NaCl, NaI and sucrose buffers. Analytical chemical studies and the toxicity of the iodine mustard derivative ruled out the role of chemical reactions yielding differentially toxic species as being responsible for the differences in HD toxicity observed. This work demonstrates that the early events that HD sets into motion to cause toxicity are dependent on ionic environment, possibly due to intracellular pH deregulation.« less

  11. Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting.

    PubMed

    Heda, Ghanshyam D; Omotola, Oluwabukola B; Heda, Rajiv P; Avery, Jamie

    2016-09-01

    SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin's transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting.

  12. Stability of polymer encapsulated quantum dots in cell culture media

    NASA Astrophysics Data System (ADS)

    Ojea-Jiménez, I.; Piella, J.; Nguyen, T.-L.; Bestetti, A.; Ryan, A. D.; Puntes, V.

    2013-04-01

    The unique optical properties of Quantum Dots have attracted a great interest to use these nanomaterials in diverse biological applications. The synthesis of QDs by methods from the literature permits one to obtain nanocrystals coated by hydrophobic alkyl coordinating ligands and soluble in most of the cases in organic solvents. The ideal biocompatible QD must be homogeneously dispersed and colloidally stable in aqueous solvents, exhibit pH and salt stability, show low levels of nonspecific binding to biological components, maintain a high quantum yield, and have a small hydrodynamic diameter. Polymer encapsulation represents an excellent scaffold on which to build additional biological function, allowing for a wide range of grafting approaches for biological ligands. As these QD are functionalized with poly(ethylene)glycol (PEG) derivatives on their surface, they show long term stability without any significant change in the optical properties, and they are also highly stable in the most common buffer solutions such as Phosphate Buffer Saline (PBS) or borate. However, as biological studies are normally done in more complex biological media which contain a mixture of amino acids, salts, glucose and vitamins, it is essential to determine the stability of our synthesized QDs under these conditions before tackling biological studies.

  13. WATER LEVEL AND OXYGEN DELIVERY/UTILIZATION IN POROUS SALT MARSH SEDIMENTS

    EPA Science Inventory

    Increasing terrestrial nutrient inputs to coastal waters is a global water quality issue worldwide, and salt marshes may provide a valuable nutrient buffer, either by direct removal or by smoothing out pulse inputs between sources and sensitive estuarine habitats. A major challen...

  14. Electrodeposition of amorphous ternary nickel-chromium-phosphorus alloy

    DOEpatents

    Guilinger, Terry R.

    1990-01-01

    Amorphous ternary nickel-chromium-phosphorus alloys are electrodeposited from a bath comprising a nickel salt, a chromium salt, a phosphorus source such as sodium hypophosphite, a complexing agent for the nickel ions, supporting salts to increase conductivity, and a buffering agent. The process is carried out at about room temperature and requires a current density between about 20 to 40 A/dm.sup.2.

  15. Chaotropic salts: novel modifiers for the capillary electrophoretic analysis of benzodiazepines.

    PubMed

    Su, Hsiu-Li; Lan, Min-Tsu; Lin, Kuan-Wen; Hsieh, You-Zung

    2008-08-01

    This paper describes a CE method for analyzing benzodiazepines using the chaotropic salts lithium trifluoromethanesulfonate (LiOTf), lithium hexafluorophosphate (LiPF(6)), and lithium bis(trifluoromethanesulfonyl)imide (LiNTf(2)) as modifiers in the running buffer. Although adequate resolution of seven benzodiazepine analytes occurred under the influence of each of the chaotropic anions, the separation efficiency was highest when bis(trifluoromethanesulfonyl)imide (Tf(2)N(-)) was the modifier. We applied affinity CE in conjunction with linear analysis to determine the association constants for the formation of complexes between the Tf(2)N(-) anion and the benzodiazepines. According to the estimated Gibbs free energies, the interactions between this chaotropic anion and the benzodiazepines were either ion-dipole or ion-induced dipole interactions. Adding chaotropic salts as modifiers into CE buffers is a simple and reproducible technique for separating benzodiazepines.

  16. Novel Mechanism for Buffering Dietary Salt in Humans: Effects of Salt Loading on Skin Sodium, Vascular Endothelial Growth Factor C, and Blood Pressure.

    PubMed

    Selvarajah, Viknesh; Mäki-Petäjä, Kaisa M; Pedro, Liliana; Bruggraber, Sylvaine F A; Burling, Keith; Goodhart, Anna K; Brown, Morris J; McEniery, Carmel M; Wilkinson, Ian B

    2017-11-01

    High dietary sodium intake triggers increased blood pressure (BP). Animal studies show that dietary salt loading results in dermal Na + accumulation and lymphangiogenesis mediated by VEGF-C (vascular endothelial growth factor C), both attenuating the rise in BP. Our objective was to determine whether these mechanisms function in humans. We assessed skin electrolytes, BP, and plasma VEGF-C in 48 healthy participants randomized to placebo (70 mmol sodium/d) and slow sodium (200 mmol/d) for 7 days. Skin Na + and K + concentrations were measured in mg/g of wet tissue and expressed as the ratio Na + :K + to correct for variability in sample hydration. Skin Na + :K + increased between placebo and slow sodium phases (2.91±0.08 versus 3.12±0.09; P =0.01). In post hoc analysis, there was a suggestion of a sex-specific effect, with a significant increase in skin Na + :K + in men (2.59±0.09 versus 2.88±0.12; P =0.008) but not women (3.23±0.10 versus 3.36±0.12; P =0.31). Women showed a significant increase in 24-hour mean BP with salt loading (93±1 versus 91±1 mm Hg; P <0.001) while men did not (96±2 versus 96±2 mm Hg; P =0.91). Skin Na + :K + correlated with BP, stroke volume, and peripheral vascular resistance in men but not in women. No change was noted in plasma VEGF-C. These findings suggest that the skin may buffer dietary Na + , reducing the hemodynamic consequences of increased salt, and this may be influenced by sex. © 2017 The Authors.

  17. Redox buffered hydrofluoric acid etchant for the reduction of galvanic attack during release etching of MEMS devices having noble material films

    DOEpatents

    Hankins, Matthew G [Albuquerque, NM

    2009-10-06

    Etchant solutions comprising a redox buffer can be used during the release etch step to reduce damage to the structural layers of a MEMS device that has noble material films. A preferred redox buffer comprises a soluble thiophosphoric acid, ester, or salt that maintains the electrochemical potential of the etchant solution at a level that prevents oxidation of the structural material. Therefore, the redox buffer preferentially oxidizes in place of the structural material. The sacrificial redox buffer thereby protects the exposed structural layers while permitting the dissolution of sacrificial oxide layers during the release etch.

  18. Measurement of europium (III)/europium (II) couple in fluoride molten salt for redox control in a molten salt reactor concept

    NASA Astrophysics Data System (ADS)

    Guo, Shaoqiang; Shay, Nikolas; Wang, Yafei; Zhou, Wentao; Zhang, Jinsuo

    2017-12-01

    The fluoride molten salt such as FLiNaK and FLiBe is one of the coolant candidates for the next generation nuclear reactor concepts, for example, the fluoride salt cooled high temperature reactor (FHR). For mitigating corrosion of structural materials in molten fluoride salt, the redox condition of the salts needs to be monitored and controlled. This study investigates the feasibility of applying the Eu3+/Eu2+ couple for redox control. Cyclic voltammetry measurements of the Eu3+/Eu2+ couple were able to obtain the concentrations ratio of Eu3+/Eu2+ in the melt. Additionally, the formal standard potential of Eu3+/Eu2+ was characterized over the FHR's operating temperatures allowing for the application of the Nernst equation to establish a Eu3+/Eu2+ concentration ratio below 0.05 to prevent corrosion of candidate structural materials. A platinum quasi-reference electrode with potential calibrated by potassium reduction potential is shown as reliable for the redox potential measurement. These results show that the Eu3+/Eu2+ couple is a feasible redox buffering agent to control the redox condition in molten fluoride salts.

  19. Adsorption equilibrium and kinetics of monomer-dimer monoclonal antibody mixtures on a cation exchange resin.

    PubMed

    Reck, Jason M; Pabst, Timothy M; Hunter, Alan K; Wang, Xiangyang; Carta, Giorgio

    2015-07-10

    Adsorption equilibrium and kinetics are determined for a monoclonal antibody (mAb) monomer and dimer species, individually and in mixtures, on a macroporous cation exchange resin both under the dilute limit of salt gradient elution chromatography and at high protein loads and low salt based on batch adsorption equilibrium and confocal laser scanning microscopy (CLSM) experiments. In the dilute limit and weak binding conditions, the dimer/monomer selectivity in 10mM phosphate at pH 7 varies between 8.7 and 2.3 decreasing with salt concentration in the range of 170-230mM NaCl. At high protein loads and strong binding conditions (0-60mM NaCl), the selectivity in the same buffer is near unity with no NaCl added, but increases gradually with salt concentration reaching high values between 2 and 15 with 60mM added NaCl. For these conditions, the two-component adsorption kinetics is controlled by pore diffusion and is predicted approximately by a dual shrinking core model using parameters based on single component equilibrium and kinetics measurements. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Visible‐Light‐Mediated Selective Arylation of Cysteine in Batch and Flow

    PubMed Central

    Bottecchia, Cecilia; Rubens, Maarten; Gunnoo, Smita B.; Hessel, Volker; Madder, Annemieke

    2017-01-01

    Abstract A mild visible‐light‐mediated strategy for cysteine arylation is presented. The method relies on the use of eosin Y as a metal‐free photocatalyst and aryldiazonium salts as arylating agents. The reaction can be significantly accelerated in a microflow reactor, whilst allowing the in situ formation of the required diazonium salts. The batch and flow protocol described herein can be applied to obtain a broad series of arylated cysteine derivatives and arylated cysteine‐containing dipeptides. Moreover, the method was applied to the chemoselective arylation of a model peptide in biocompatible reaction conditions (room temperature, phosphate‐buffered saline (PBS) buffer) within a short reaction time. PMID:28805276

  1. DNA adsorption to and elution from silica surfaces: influence of amino acid buffers.

    PubMed

    Vandeventer, Peter E; Mejia, Jorge; Nadim, Ali; Johal, Malkiat S; Niemz, Angelika

    2013-09-19

    Solid phase extraction and purification of DNA from complex samples typically requires chaotropic salts that can inhibit downstream polymerase amplification if carried into the elution buffer. Amino acid buffers may serve as a more compatible alternative for modulating the interaction between DNA and silica surfaces. We characterized DNA binding to silica surfaces, facilitated by representative amino acid buffers, and the subsequent elution of DNA from the silica surfaces. Through bulk depletion experiments, we found that more DNA adsorbs to silica particles out of positively compared to negatively charged amino acid buffers. Additionally, the type of the silica surface greatly influences the amount of DNA adsorbed and the final elution yield. Quartz crystal microbalance experiments with dissipation monitoring (QCM-D) revealed multiphasic DNA adsorption out of stronger adsorbing conditions such as arginine, glycine, and glutamine, with DNA more rigidly bound during the early stages of the adsorption process. The DNA film adsorbed out of glutamate was more flexible and uniform throughout the adsorption process. QCM-D characterization of DNA elution from the silica surface indicates an uptake in water mass during the initial stage of DNA elution for the stronger adsorbing conditions, which suggests that for these conditions the DNA film is partly dehydrated during the prior adsorption process. Overall, several positively charged and polar neutral amino acid buffers show promise as an alternative to methods based on chaotropic salts for solid phase DNA extraction.

  2. Optimization of protein buffer cocktails using Thermofluor.

    PubMed

    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.

  3. Deconvoluting the effects of buffer salt concentration in hydrophilic interaction chromatography on a zwitterionic stationary phase.

    PubMed

    West, Caroline; Auroux, Emeline

    2016-08-26

    Quantitative structure-retention relationships (QSRRs) furnish a detailed and reliable description of the role and extent of different molecular interactions that can be established between the analytes and the chromatographic system. Among QSRRs, the solvation parameter model using Abraham descriptors has gained acceptance as a general tool to explore the factors affecting retention in chromatographic systems. We have previously shown how a modified version of the solvation parameter model, with two extra terms to take account of interactions occurring with ionic and ionizable species (with positive and/or negative charges), could be applied to the characterization of hydrophilic interaction chromatographic (HILIC) systems. In the present study, we will show how this methodology can be used to evaluate the effects of increasing buffer salt concentration on retention and separation in a HILIC system. A commercial stationary phase possessing a sulfobetaine zwitterionic bonded ligand (Nucleodur HILIC) was used with a mobile phase composed of 80% acetonitrile and 20% pwwH4 ammonium acetate buffer, with aqueous buffer concentrations varying from 10 to 100mM, resulting in overall concentrations ranging from 2 to 20mM in the mobile phase. Retention factors were measured for a selection of 76 probe analytes. The chosen compounds are small molecules presenting a wide diversity of molecular structures and are relevant to biomedical and pharmaceutical applications. The QSRR models obtained allow for a rationalization of the interactions contributing to retention and separation in the HILIC system considered and shed some light on the effect of varying buffer salt concentration, namely the progressive transition from ion-exchange and electrostatic-repulsion mechanisms to hydrophilic partitioning. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Canine intestinal contents vs. simulated media for the assessment of solubility of two weak bases in the human small intestinal contents.

    PubMed

    Kalantzi, Lida; Persson, Eva; Polentarutti, Britta; Abrahamsson, Bertil; Goumas, Konstantinos; Dressman, Jennifer B; Reppas, Christos

    2006-06-01

    This study was conducted to assess the relative usefulness of canine intestinal contents and simulated media in the prediction of solubility of two weak bases (dipyridamole and ketoconazole) in fasted and fed human intestinal aspirates that were collected under conditions simulating those in bioavailability/bioequivalence studies. After administration of 250 mL of water or 500 mL of Ensure plus [both containing 10 mg/mL polyethylene glycol (PEG) 4000 as nonabsorbable marker], intestinal aspirates were collected from the fourth part of the duodenum of 12 healthy adults and from the mid-jejunum of four Labradors. Pooled samples were analyzed for PEG, pH, buffer capacity, osmolality, surface tension, pepsin, total carbohydrates, total protein content, bile salts, phospholipids, and neutral lipids. The shake-flask method was used to measure the solubility of dipyridamole and ketoconazole in pooled human and canine intestinal contents and in fasted-state-simulating intestinal fluid (FaSSIF) and fed-state-simulating intestinal fluid (FeSSIF) containing various bile salts and pH-buffering agents. For both compounds, solubility in canine contents may be predictive of human intralumenal solubility in the fasting state but not in the fed state. The poor agreement of results in canine and human aspirates can be attributed to the higher bile salt content in canine bile. Solubility in FaSSIF containing a mixture of bile salts from crude bile predicted satisfactorily the intralumenal solubility of both drugs in the fasted state in humans. Solubility in FeSSIF, regardless of the identity of bile salts or of the buffering species, deviated from intralumenal values in the fed human aspirates by up to 40%. This was attributed to the lack of lipolytic products in FeSSIF, the higher bile salt content of FeSSIF, and the lower pH of FeSSIF. FaSSIF containing a mixture of bile salts from crude bile, and FeSSIF containing lipolytic products and, perhaps, having lower bile salt content but slightly higher pH, should be more useful than canine intestinal aspirates for predicting intralumenal solubilities in humans.

  5. Selective Sulfidation of Lead Smelter Slag with Sulfur

    NASA Astrophysics Data System (ADS)

    Han, Junwei; Liu, Wei; Wang, Dawei; Jiao, Fen; Qin, Wenqing

    2016-02-01

    The selective sulfidation of lead smelter slag with sulfur was studied. The effects of temperature, sulfur dosage, carbon, and Na salts additions were investigated based on thermodynamic calculation. The results indicated that more than 96 pct of zinc in the slag could be converted into sulfides. Increasing temperature, sulfur dosage, or Na salts dosage was conducive to the sulfidation of the zinc oxides in the slag. High temperature and excess Na salts would result in the more consumption of carbon and sulfur. Carbon addition not only promoted the selective sulfidation but reduced the sulfur dosage and eliminated the generation of SO2. Iron oxides had a buffering role on the sulfur efficient utilization. The transformation of sphalerite to wurtzite was feasible under reducing condition at high temperature, especially above 1273 K (1000 °C). The growth of ZnS particles largely depended upon the roasting temperature. They were significantly increased when the temperature was above 1273 K (1000 °C), which was attributed to the formation of a liquid phase.

  6. Physicochemical parameters influencing coaggregation between the freshwater bacteria Sphingomonas natatoria 2.1 and Micrococcus luteus 2.13.

    PubMed

    Min, K R; Zimmer, M N; Rickard, A H

    2010-11-01

    The aim of this study was to explore the physicochemical parameters that influence coaggregation between the freshwater bacteria Sphingomonas natatoria 2.1 and Micrococcus luteus 2.13. Using visual coaggregation assays, the effect of different buffers, solutions of differing ionic strength, pH, temperature, and viscosity on the degree of coaggregation was assessed. Coaggregation occurred maximally in distilled water but was inhibited when coaggregates were suspended in a commonly-used oral bacterial coaggregation buffer, saline solutions, and Tris-Cl buffers. Coaggregation was weakly expressed in standard laboratory buffers. The ionic strength of inorganic salt solutions required to inhibit coaggregation depended upon the inorganic salt being tested. Coaggregation occurred at a pH of 3-10, between 5 and 80°C and was inhibited in solutions with a viscosity of 22.5 centipoises at 20°C. Inhibition of coaggregation with NaCl impaired biofilm development. When developing buffers to test for coaggregation, the natural liquid environment should be considered. Coaggregation between S. natatoria 2.1 and M. luteus 2.13 is only affected by physicochemical conditions beyond those typically found in natural freshwater ecosystems. Such a robust ability to coaggregate may enhance the ability of S. natatoria 2.1 and M. luteus 2.13 to develop a niche in freshwater biofilms.

  7. Salt Sensitivity and Hypertension: A Paradigm Shift from Kidney Malfunction to Vascular Endothelial Dysfunction

    PubMed Central

    Choi, Hoon Young; Park, Hyeong Cheon

    2015-01-01

    Hypertension is a complex trait determined by both genetic and environmental factors and is a major public health problem due to its high prevalence and concomitant increase in the risk for cardiovascular disease. With the recent large increase of dietary salt intake in most developed countries, the prevalence of hypertension increases tremendously which is about 30% of the world population. There is substantial evidence that suggests some people can effectively excrete high dietary salt intake without an increase in arterial BP, and another people cannot excrete effectively without an increase in arterial BP. Salt sensitivity of BP refers to the BP responses for changes in dietary salt intake to produce meaningful BP increases or decreases. The underlying mechanisms that promote salt sensitivity are complex and range from genetic to environmental influences. The phenotype of salt sensitivity is therefore heterogeneous with multiple mechanisms that potentially link high salt intake to increases in blood pressure. Moreover, excess salt intake has functional and pathological effects on the vasculature that are independent of blood pressure. Epidemiologic data demonstrate the role of high dietary salt intake in mediating cardiovascular and renal morbidity and mortality. Almost five decades ago, Guyton and Coleman proposed that whenever arterial pressure is elevated, pressure natriuresis enhances the excretion of sodium and water until blood volume is reduced sufficiently to return arterial pressure to control values. According to this hypothesis, hypertension can develop only when something impairs the excretory ability of sodium in the kidney. However, recent studies suggest that nonosmotic salt accumulation in the skin interstitium and the endothelial dysfunction which might be caused by the deterioration of vascular endothelial glycocalyx layer (EGL) and the epithelial sodium channel on the endothelial luminal surface (EnNaC) also play an important role in nonosmotic storage of salt. These new concepts emphasize that sodium homeostasis and salt sensitivity seem to be related not only to the kidney malfunction but also to the endothelial dysfunction. Further investigations will be needed to assess the extent to which changes in the sodium buffering capacity of the skin interstitium and develop the treatment strategy for modulating the endothelial dysfunction. PMID:26240595

  8. Systematic interpolation method predicts protein chromatographic elution with salt gradients, pH gradients and combined salt/pH gradients.

    PubMed

    Creasy, Arch; Barker, Gregory; Carta, Giorgio

    2017-03-01

    A methodology is presented to predict protein elution behavior from an ion exchange column using both individual or combined pH and salt gradients based on high-throughput batch isotherm data. The buffer compositions are first optimized to generate linear pH gradients from pH 5.5 to 7 with defined concentrations of sodium chloride. Next, high-throughput batch isotherm data are collected for a monoclonal antibody on the cation exchange resin POROS XS over a range of protein concentrations, salt concentrations, and solution pH. Finally, a previously developed empirical interpolation (EI) method is extended to describe protein binding as a function of the protein and salt concentration and solution pH without using an explicit isotherm model. The interpolated isotherm data are then used with a lumped kinetic model to predict the protein elution behavior. Experimental results obtained for laboratory scale columns show excellent agreement with the predicted elution curves for both individual or combined pH and salt gradients at protein loads up to 45 mg/mL of column. Numerical studies show that the model predictions are robust as long as the isotherm data cover the range of mobile phase compositions where the protein actually elutes from the column. Copyright © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Understanding the Underground Hydrous Minerals on Mars: Stability Field, Phase Transitions, and Environmental Implications

    NASA Astrophysics Data System (ADS)

    Wang, A.; Chou, I. M.; Ling, Z.; Sobron, P.

    2017-12-01

    Three types of studies form the bases of our understandings: (1) systematic laboratory experiments on the thermodynamics and kinetic properties of hydrous (Mg, Fe2+, Fe3+, Ca, Al, Na) -sulfates, -chlorides, and -perchlorates, made by this and many other teams. (2) the thermal modeling of two-layer regolith with very different thermal inertia (TI) and its validating observation on Mars [Mellon et al., 2004, 2009]. (3) the mission observations on Mars and the field investigations at analog sites. Following are some examples of these understandings, with more to be presented at AGU. Hydrous salts (sulfates, chlorides, perchlorates) in an enclosure could keep a relatively stable RH%, i.e., they are environmental RH buffers. Underground layers of hydrous salty soils (high TI) on Mars could be considered as a quasi-closed system, equilibrated within their environments. The RH% range kept by them would help to stabilize many hydrous salts. For example, Mg- & Fe3+-sulfates with high hydration degrees (6-20 H2O) were observed in the subsurface layers in a terrestrial hyperarid region and at Gusev on Mars. A general trend was found that the RH% levels kept by hydrous sulfates in an enclosure are much higher than those by hydrous perchlorates and by hydrous chlorides. This implies that in an underground layer of mixed hydrous salts, one type of salts (e.g. sulfates) can provide the necessary RH-buffering for the phase transition of other types,, e.g., the deliquescence of perchlorates or chlorides, to trigger RSL or to provide liquid H2O at relatively warm T. The dehydration rates of hydrous sulfates have a high dependence on cation types. Among them, Mg and Fe2+-sulfate have higher dehydration rates, and ferric sulfates dehydrate much slow. This lab-observation was validated by MER mission observation, i.e., the finding of highly hydrated ferric sulfates, i.e. Fe4.67(SO4)6(OH)2.20H2O, in subsurface at Gusev crater. However, the dehydration rate of hydrous sulfates can also be affected by their structural details and by the catalysis of co-existing phases. Overall, underground hydrous salt layers not only provide a stable, long-lived habitable environments, their phase changes would contribute to the current hydrological (and S, Cl) circulation, and they would certainly represent a major portion of the water budget on Mars.

  10. Zero lattice mismatch and twin-free single crystalline ScN buffer layers for GaN growth on silicon

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

    Lupina, L.; Zoellner, M. H.; Dietrich, B.

    2015-11-16

    We report the growth of thin ScN layers deposited by plasma-assisted molecular beam epitaxy on Sc{sub 2}O{sub 3}/Y{sub 2}O{sub 3}/Si(111) substrates. Using x-ray diffraction, Raman spectroscopy, and transmission electron microscopy, we find that ScN films grown at 600 °C are single crystalline, twin-free with rock-salt crystal structure, and exhibit a direct optical band gap of 2.2 eV. A high degree of crystalline perfection and a very good lattice matching between ScN and GaN (misfit < 0.1%) makes the ScN/Sc{sub 2}O{sub 3}/Y{sub 2}O{sub 3} buffer system a very promising template for the growth of high quality GaN layers on silicon.

  11. Acid/Salt/pH Gradient Improved Resolution and Sensitivity in Proteomics Study Using 2D SCX-RP LC-MS.

    PubMed

    Zhu, Ming-Zhi; Li, Na; Wang, Yi-Tong; Liu, Ning; Guo, Ming-Quan; Sun, Bao-Qing; Zhou, Hua; Liu, Liang; Wu, Jian-Lin

    2017-09-01

    The usage of strong cation exchange (SCX) chromatography in proteomics is limited by its poor resolution and nonspecific hydrophobic interactions with peptides, which lead to peptide overlap across fractions and change of peptide retention, respectively. The application of high concentration of salt (up to 1000 mM) in SCX also restricted its use in online 2D SCX-RP LC. In the present research, we first exploited the chromatographic ability of online 2D SCX-RP LC by combination of acid, salt, and pH gradient, three relatively independent modes of eluting peptides from SCX column. 50% ACN was added to elution buffer for eliminating hydrophobic interactions between SCX matrix and peptides, and the concentration of volatile salt was reduced to 50 mM. Acid/salt/pH gradient showed superior resolution and sensitivity as well as uniform distribution across fractions, consequently leading to significant improvements in peptide and protein identification. 112 191 unique peptides and 7373 proteins were identified by acid/salt/pH fractionation, while 69 870 unique peptides and 4536 proteins were identified by salt elution, that is, 62.5 and 60.6% more proteins and unique peptides, respectively, identified by the former. Fraction overlap was also significantly minimized by acid/salt/pH approach. Furthermore, acid/salt/pH elution showed more identification for acidic peptides and hydrophilic peptides.

  12. Pyrroloquinoline quinone (PQQ) is reduced to pyrroloquinoline quinol (PQQH2) by vitamin C, and PQQH2 produced is recycled to PQQ by air oxidation in buffer solution at pH 7.4.

    PubMed

    Mukai, Kazuo; Ouchi, Aya; Nagaoka, Shin-ichi; Nakano, Masahiko; Ikemoto, Kazuto

    2016-01-01

    Measurements of the reaction of sodium salt of pyrroloquinoline quinone (PQQNa2) with vitamin C (Vit C) were performed in phosphate-buffered solution (pH 7.4) at 25 °C under nitrogen atmosphere, using UV-vis spectrophotometry. The absorption spectrum of PQQNa2 decreased in intensity due to the reaction with Vit C and was changed to that of pyrroloquinoline quinol (PQQH2, a reduced form of PQQ). One molecule of PQQ was reduced by two molecules of Vit C producing a molecule of PQQH2 in the buffer solution. PQQH2, thus produced, was recycled to PQQ due to air oxidation. PQQ and Vit C coexist in many biological systems, such as vegetables, fruits, as well as in human tissues. The results obtained suggest that PQQ is reduced by Vit C and functions as an antioxidant in biological systems, because it has been reported that PQQH2 shows very high free-radical scavenging and singlet-oxygen quenching activities in buffer solutions.

  13. Isolation of intact RNA from murine CD4+ T cells after intracellular cytokine staining and fluorescence-activated cell sorting.

    PubMed

    Kunnath-Velayudhan, Shajo; Porcelli, Steven A

    2018-05-01

    Intracellular cytokine staining (ICS) is a powerful method for identifying functionally distinct lymphocyte subsets, and for isolating these by fluorescence activated cell sorting (FACS). Although transcriptomic analysis of cells sorted on the basis of ICS has many potential applications, this is rarely performed because of the difficulty in isolating intact RNA from cells processed using standard fixation and permeabilization buffers for ICS. To address this issue, we compared three buffers shown previously to preserve RNA in nonhematopoietic cells subjected to intracellular staining for their effects on RNA isolated from T lymphocytes processed for ICS. Our results showed that buffers containing the recombinant ribonuclease inhibitor RNasin or high molar concentrations of salt yielded intact RNA from fixed and permeabilized T cells. As proof of principle, we successfully used the buffer containing RNasin to isolate intact RNA from CD4 + T cells that were sorted by FACS on the basis of specific cytokine production, thus demonstrating the potential of this approach for coupling ICS with transcriptomic analysis. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Calorimetric and Diffractometric Evidence for the Sequential Crystallization of Buffer Components and the Consequential pH Swing in Frozen Solutions

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

    Sundaramurthi, Prakash; Shalaev, Evgenyi; Suryanarayanan, Raj

    2010-06-22

    Sequential crystallization of succinate buffer components in the frozen solution has been studied by differential scanning calorimetry and X-ray diffractometry (both laboratory and synchrotron sources). The consequential pH shifts were monitored using a low-temperature electrode. When a solution buffered to pH < pK{sub a2} was cooled from room temperature (RT), the freeze-concentrate pH first increased and then decreased. This was attributed to the sequential crystallization of succinic acid, monosodium succinate, and finally disodium succinate. When buffered to pH > pK{sub a2}, the freeze-concentrate pH first decreased and then increased due to the sequential crystallization of the basic (disodium succinate) followedmore » by the acidic (monosodium succinate and succinic acid) buffer components. XRD provided direct evidence of the crystallization events in the frozen buffer solutions, including the formation of disodium succinate hexahydrate [Na{sub 2}(CH{sub 2}COO){sub 2} {center_dot} 6H{sub 2}O]. When the frozen solution was warmed in a differential scanning calorimeter, multiple endotherms attributable to the melting of buffer components and ice were observed. When the frozen solutions were dried under reduced pressure, ice sublimation was followed by dehydration of the crystalline hexahydrate to a poorly crystalline anhydrate. However, crystalline succinic acid and monosodium succinate were retained in the final lyophiles. The pH and the buffer salt concentration of the prelyo solution influenced the crystalline salt content in the final lyophile. The direction and magnitude of the pH shift in the frozen solution depended on both the initial pH and the buffer concentration. In light of the pH-sensitive nature of a significant fraction of pharmaceuticals (especially proteins), extreme care is needed in both the buffer selection and its concentration.« less

  15. DNA Adsorption to and Elution from Silica Surfaces: Influence of Amino Acid Buffers

    PubMed Central

    Vandeventer, Peter E.; Mejia, Jorge; Nadim, Ali; Johal, Malkiat S.; Niemz, Angelika

    2014-01-01

    Solid phase extraction and purification of DNA from complex samples typically requires chaotropic salts that can inhibit downstream polymerase amplification if carried into the elution buffer. Amino acid buffers may serve as a more compatible alternative for modulating the interaction between DNA and silica surfaces. We characterized DNA binding to silica surfaces, facilitated by representative amino acid buffers, and the subsequent elution of DNA from the silica surfaces. Through bulk depletion experiments, we found that more DNA adsorbs to silica particles out of positively compared to negatively charged amino acid buffers. Additionally, the type of the silica surface greatly influences the amount of DNA adsorbed, and the final elution yield. Quartz crystal microbalance experiments with dissipation monitoring (QCM-D) revealed multiphasic DNA adsorption out of stronger adsorbing conditions such as arginine, glycine, and glutamine, with DNA more rigidly bound during the early stages of the adsorption process. The DNA film adsorbed out of glutamate was more flexible and uniform throughout the adsorption process. QCM-D characterization of DNA elution from the silica surface indicates an uptake in water mass during the initial stage of DNA elution for the stronger adsorbing conditions, which suggests that for these conditions the DNA film is partly dehydrated during the prior adsorption process. Overall, several positively charged and polar neutral amino acid buffers show promise as an alternative to methods based on chaotropic salts for solid phase DNA extraction. PMID:23931415

  16. THE FLOCCULATION MAXIMUM (pH) OF FIBRINOGEN AND SOME OTHER BLOOD-CLOTTING REAGENTS. (RELATIVE TURBIDIMETRY WITH THE EVELYN PHOTOELECTRIC COLORIMETER).

    PubMed

    Ferguson, J H

    1942-03-20

    By means of a novel adaptation of the Evelyn photoelectric colorimeter to the measurement of relative turbidities, the question of the flocculation maximum (F.M.) in acetate buffer solutions of varying pH and salt content has been studied on (a) an exceptionally stable prothrombin-free fibrinogen and its solutions after incipient thermal denaturation and incomplete tryptic proteolysis, (b) plasma, similarly treated, (c) prothrombin, thrombin, and (brain) thromboplastin solutions. All the fibrinogens show a remarkable uniformity of the precipitation pattern, viz. F.M. =4.7 (+/-0.2) pH in salt-containing buffer solutions and pH = 5.3 (+/-0.2) in salt-poor buffer (N/100 acetate). The latter approximates the isoelectric point (5.4) obtained by cataphoresis (14). There is no evidence that denaturation or digestion can produce any "second maximum." The data support the view that fibrin formation (under the specific influence of thrombin) is intrinsically unrelated to denaturation and digestion phenomena, although all three can proceed simultaneously in crude materials. A criticism is offered, therefore, of Wöhlisch's blood clotting theory. Further applications of the photoelectric colorimeter to coagulation problems are suggested, including kinetic study of fibrin formation and the assay of fibrinogen, with a possible sensitivity of 7.5 mg. protein in 100 cc. solution.

  17. Nitrogen Over-enrichment Effects on Belowground Structure in Coastal Wetlands and Implications for Delivery of Ecosysytem Services

    EPA Science Inventory

    Salt marshes supply many ecosystem services, such as fish, crab and bird habitat, flood abatement and carbon sequestration. Since salt marshes function as a moderating buffer between ocean and land, they are one of the first lines of defense to absorb the effects of sea level ris...

  18. Preservation of corals in salt-saturated DMSO buffer is superior to ethanol for PCR experiments

    NASA Astrophysics Data System (ADS)

    Gaither, M. R.; Szabó, Z.; Crepeau, M. W.; Bird, C. E.; Toonen, R. J.

    2011-06-01

    Specimen collection is time consuming and expensive, yet few laboratories test preservation methods before setting out on field expeditions. The most common preservation buffer used for coral specimens is >70% EtOH. However, alternatives exist that are less flammable, easier to ship, and are widely used in other taxa. Here, we compare the effects of salt-saturated DMSO (SSD) and EtOH preservation buffers on post-extraction DNA quantity and quality. We found that soft tissue integrity was better maintained and higher quantities of DNA were extracted from EtOH-preserved specimens; however, by all other measures, SSD was a superior preservative to EtOH. Extractions of SSD-preserved specimens resulted in higher molecular weight DNA, higher PCR success, and more efficient amplification than specimens preserved in EtOH. Our results show that SSD is generally a superior preservative to EtOH for specimens destined for PCR studies, but species-specific differences indicate that preservation comparisons should be undertaken before collection and storage of samples.

  19. Alternative for Perchlorates in Incendiary and Pyrotechnic Formulations for Projectiles

    DTIC Science & Technology

    2009-08-01

    although other copper salts including chloride, acetate, sulfate and carbonate are highly bioavailable in animal nutrition. The toxicity of a single oral...6 hours to 820 mg m-3 after 2 hr. In experimental animals, single inhalation exposures to hydrogen sulfide result in death and respiratory...is in general regulated by the equilibria between CO2, HCO3 - and CO3 2-. The buffer capacity depends on the concentration of these substances

  20. Novel Mechanism for Buffering Dietary Salt in Humans

    PubMed Central

    Mäki-Petäjä, Kaisa M.; Pedro, Liliana; Bruggraber, Sylvaine F.A.; Burling, Keith; Goodhart, Anna K.; Brown, Morris J.; McEniery, Carmel M.; Wilkinson, Ian B.

    2017-01-01

    High dietary sodium intake triggers increased blood pressure (BP). Animal studies show that dietary salt loading results in dermal Na+ accumulation and lymphangiogenesis mediated by VEGF-C (vascular endothelial growth factor C), both attenuating the rise in BP. Our objective was to determine whether these mechanisms function in humans. We assessed skin electrolytes, BP, and plasma VEGF-C in 48 healthy participants randomized to placebo (70 mmol sodium/d) and slow sodium (200 mmol/d) for 7 days. Skin Na+ and K+ concentrations were measured in mg/g of wet tissue and expressed as the ratio Na+:K+ to correct for variability in sample hydration. Skin Na+:K+ increased between placebo and slow sodium phases (2.91±0.08 versus 3.12±0.09; P=0.01). In post hoc analysis, there was a suggestion of a sex-specific effect, with a significant increase in skin Na+:K+ in men (2.59±0.09 versus 2.88±0.12; P=0.008) but not women (3.23±0.10 versus 3.36±0.12; P=0.31). Women showed a significant increase in 24-hour mean BP with salt loading (93±1 versus 91±1 mm Hg; P<0.001) while men did not (96±2 versus 96±2 mm Hg; P=0.91). Skin Na+:K+ correlated with BP, stroke volume, and peripheral vascular resistance in men but not in women. No change was noted in plasma VEGF-C. These findings suggest that the skin may buffer dietary Na+, reducing the hemodynamic consequences of increased salt, and this may be influenced by sex. PMID:28974570

  1. Dialysis buffer with different ionic strength affects the antigenicity of cultured nervous necrosis virus (NNV) suspensions.

    PubMed

    Gye, Hyun Jung; Nishizawa, Toyohiko

    2016-09-02

    Nervous necrosis virus (NNV) belongs to the genus Betanodavirus (Nodaviridae). It is highly pathogenic to various marine fishes. Here, we investigated the antigenicity changes of cultured NNV suspensions during 14days of dialyses using a dialysis tube at 1.4×10(4) molecular weight cut off (MWCO) in three different buffers (Dulbecco's phosphate buffered saline (D-PBS), 15mM Tris-HCl (pH 8.0), and deionized water (DIW)). Total NNV antigen titers of cultured NNV suspension varied depending on different dialysis buffers. For example, total NNV antigen titer during D-PBS dialysis was increased once but then decreased. During Tris-HCl dialysis, it was relatively stable. During dialysis in DIW, total NNV antigen titer was increased gradually. These antigenicity changes in NNV suspension might be due to changes in the aggregation state of NNV particles and/or coat proteins (CPs). ELISA values of NNV suspension changed due to changing aggregates state of NNV antigens. NNV particles in suspension were aggregated at a certain level. These aggregates were progressive after D-PBS dialysis, but regressive after Tris-HCl dialysis. The purified NNV particles self-aggregated after dialysis in D-PBS or in Tris-HCl containing 600mM NaCl, but not after dialysis in Tris-HCl or DIW. Quantitative analysis is merited to determine NNV antigens in the highly purified NNV particles suspended in buffer at low salt condition. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Apparatus for molecular weight separation

    DOEpatents

    Smith, Richard D.; Liu, Chuanliang

    2001-01-01

    The present invention relates generally to an apparatus and method for separating high molecular weight molecules from low molecular weight molecules. More specifically, the invention relates to the use of microdialysis for removal of the salt (low molecular weight molecules) from a nucleotide sample (high molecular weight molecules) for ESI-MS analysis. The dialysis or separation performance of the present invention is improved by (1) increasing dialysis temperature thereby increasing desalting efficiency and improving spectrum quality; (2) adding piperidine and imidazole to the dialysis buffer solution and reducing charge states and further increasing detection sensitivity for DNA; (3) using low concentrations (0-2.5 mM NH4OAc) of dialysis buffer and shifting the DNA negative ions to higher charge states, producing a nearly 10-fold increase in detection sensitivity and a slightly decreased desalting efficiency, (4) conducting a two-stage separation or (5) any combination of (1), (2), (3) and (4).

  3. Microdialysis unit for molecular weight separation

    DOEpatents

    Smith, Richard D.; Liu, Chuanliang

    1999-01-01

    The present invention relates generally to an apparatus and method for separating high molecular weight molecules from low molecular weight molecules. More specifically, the invention relates to the use of microdialysis for removal of the salt (low molecular weight molecules) from a nucleotide sample (high molecular weight molecules) for ESI-MS analysis. The dialysis or separation performance of the present invention is improved by (1) increasing dialysis temperature thereby increasing desalting efficiency and improving spectrum quality; (2) adding piperidine and imidazole to the dialysis buffer solution and reducing charge states and further increasing detection sensitivity for DNA; (3) using low concentrations (0-2.5 mM NH4OAc) of dialysis buffer and shifting the DNA negative ions to higher charge states, producing a nearly 10-fold increase in detection sensitivity and a slightly decreased desalting efficiency, or (4) any combination of (1), (2), and (3).

  4. Magnesium degradation influenced by buffering salts in concentrations typical of in vitro and in vivo models.

    PubMed

    Agha, Nezha Ahmad; Feyerabend, Frank; Mihailova, Boriana; Heidrich, Stefanie; Bismayer, Ulrich; Willumeit-Römer, Regine

    2016-01-01

    Magnesium and its alloys have considerable potential for orthopedic applications. During the degradation process the interface between material and tissue is continuously changing. Moreover, too fast or uncontrolled degradation is detrimental for the outcome in vivo. Therefore in vitro setups utilizing physiological conditions are promising for the material/degradation analysis prior to animal experiments. The aim of this study is to elucidate the influence of inorganic salts contributing to the blood buffering capacity on degradation. Extruded pure magnesium samples were immersed under cell culture conditions for 3 and 10 days. Hank's balanced salt solution without calcium and magnesium (HBSS) plus 10% of fetal bovine serum (FBS) was used as the basic immersion medium. Additionally, different inorganic salts were added with respect to concentration in Dulbecco's modified Eagle's medium (DMEM, in vitro model) and human plasma (in vivo model) to form 12 different immersion media. Influences on the surrounding environment were observed by measuring pH and osmolality. The degradation interface was analyzed by electron-induced X-ray emission (EIXE) spectroscopy, including chemical-element mappings and electron microprobe analysis, as well as Fourier transform infrared reflection micro-spectroscopy (FTIR). Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Statistical investigation of simulated fed intestinal media composition on the equilibrium solubility of oral drugs.

    PubMed

    Zhou, Zhou; Dunn, Claire; Khadra, Ibrahim; Wilson, Clive G; Halbert, Gavin W

    2017-03-01

    Gastrointestinal fluid is a complex milieu and it is recognised that gut drug solubility is different to that observed in simple aqueous buffers. Simulated gastrointestinal media have been developed covering fasted and fed states to facilitate in vitro prediction of gut solubility and product dissolution. However, the combination of bile salts, phospholipids, fatty acids and proteins in an aqueous buffered system creates multiple phases and drug solubility is therefore a complex interaction between these components, which may create unique environments for each API. The impact on solubility can be assessed through a statistical design of experiment (DoE) approach, to determine the influence and relationships between factors. In this paper DoE has been applied to fed simulated gastrointestinal media consisting of eight components (pH, bile salt, lecithin, sodium oleate, monoglyceride, buffer, salt and pancreatin) using a two level D-optimal design with forty-four duplicate measurements and four centre points. The equilibrium solubility of a range of poorly soluble acidic (indomethacin, ibuprofen, phenytoin, valsartan, zafirlukast), basic (aprepitant, carvedilol, tadalafil, bromocriptine) and neutral (fenofibrate, felodipine, probucol, itraconazole) drugs was investigated. Results indicate that the DoE provides equilibrium solubility values that are comparable to literature results for other simulated fed gastrointestinal media systems or human intestinal fluid samples. For acidic drugs the influence of pH predominates but other significant factors related to oleate and bile salt or interactions between them are present. For basic drugs pH, oleate and bile salt have equal significance along with interactions between pH and oleate and lecithin and oleate. Neutral drugs show diverse effects of the media components particularly with regard to oleate, bile salt, pH and lecithin but the presence of monoglyceride, pancreatin and buffer have significant but smaller effects on solubility. There are fourteen significant interactions between factors mainly related to the surfactant components and pH, indicating that the solubility of neutral drugs in fed simulated media is complex. The results also indicate that the equilibrium solubility of each drug can exhibit individualistic behaviour associated with the drug's chemical structure, physicochemical properties and interaction with media components. The utility of DoE for fed simulated media has been demonstrated providing equilibrium solubility values comparable with similar in vitro systems whilst also providing greater information on the influence of media factors and their interactions. The determination of a drug's gastrointestinal solubility envelope provides useful limits that can potentially be applied to in silico modelling and in vivo experiments. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Induced binding of proteins by ammonium sulfate in affinity and ion-exchange column chromatography.

    PubMed

    Arakawa, Tsutomu; Tsumoto, Kouhei; Ejima, Daisuke; Kita, Yoshiko; Yonezawa, Yasushi; Tokunaga, Masao

    2007-04-10

    In general, proteins bind to affinity or ion-exchange columns at low salt concentrations, and the bound proteins are eluted by raising the salt concentration, changing the solvent pH, or adding competing ligands. Blue-Sepharose is often used to remove bovine serum albumin (BSA) from samples, but when we applied BSA to Blue-Sepharose in 20 mM phosphate, pH 7.0, 50%-60% of the protein flowed through the column; however, complete binding of BSA was achieved by the addition of 2 M ammonium sulfate (AS) to the column equilibration buffer and the sample. The bound protein was eluted by decreasing the AS concentration or by adding 1 M NaCl or arginine. AS at high concentrations resulted in binding of BSA even to an ion-exchange column, Q-Sepharose, at pH 7.0. Thus, although moderate salt concentrations elute proteins from Blue-Sepharose or ion-exchange columns, proteins can be bound to these columns under extreme salting-out conditions. Similar enhanced binding of proteins by AS was observed with an ATP-affinity column.

  7. THE FLOCCULATION MAXIMUM (pH) OF FIBRINOGEN AND SOME OTHER BLOOD-CLOTTING REAGENTS. (RELATIVE TURBIDIMETRY WITH THE EVELYN PHOTOELECTRIC COLORIMETER)

    PubMed Central

    Ferguson, John H.

    1942-01-01

    By means of a novel adaptation of the Evelyn photoelectric colorimeter to the measurement of relative turbidities, the question of the flocculation maximum (F.M.) in acetate buffer solutions of varying pH and salt content has been studied on (a) an exceptionally stable prothrombin-free fibrinogen and its solutions after incipient thermal denaturation and incomplete tryptic proteolysis, (b) plasma, similarly treated, (c) prothrombin, thrombin, and (brain) thromboplastin solutions. All the fibrinogens show a remarkable uniformity of the precipitation pattern, viz. F.M. =4.7 (±0.2) pH in salt-containing buffer solutions and pH = 5.3 (±0.2) in salt-poor buffer (N/100 acetate). The latter approximates the isoelectric point (5.4) obtained by cataphoresis (14). There is no evidence that denaturation or digestion can produce any "second maximum." The data support the view that fibrin formation (under the specific influence of thrombin) is intrinsically unrelated to denaturation and digestion phenomena, although all three can proceed simultaneously in crude materials. A criticism is offered, therefore, of Wöhlisch's blood clotting theory. Further applications of the photoelectric colorimeter to coagulation problems are suggested, including kinetic study of fibrin formation and the assay of fibrinogen, with a possible sensitivity of 7.5 mg. protein in 100 cc. solution. PMID:19873299

  8. Titration Calorimetry Standards and the Precision of Isothermal Titration Calorimetry Data

    PubMed Central

    Baranauskienė, Lina; Petrikaitė, Vilma; Matulienė, Jurgita; Matulis, Daumantas

    2009-01-01

    Current Isothermal Titration Calorimetry (ITC) data in the literature have relatively high errors in the measured enthalpies of protein-ligand binding reactions. There is a need for universal validation standards for titration calorimeters. Several inorganic salt co-precipitation and buffer protonation reactions have been suggested as possible enthalpy standards. The performances of several commercial calorimeters, including the VP-ITC, ITC200, and Nano ITC-III, were validated using these suggested standard reactions. PMID:19582227

  9. Tensile strain effect in ferroelectric perovskite oxide thin films on spinel magnesium aluminum oxide substrate

    NASA Astrophysics Data System (ADS)

    Zhou, Xiaolan

    Ferroelectrics are used in FeRAM (Ferroelectric random-access memory). Currently (Pb,Zr)TiO3 is the most common ferroelectric material. To get lead-free and high performance ferroelectric material, we investigated perovskite ferroelectric oxides (Ba,Sr)TiO3 and BiFeO3 films with strain. Compressive strain has been investigated intensively, but the effects of tensile strain on the perovskite films have yet to be explored. We have deposited (Ba,Sr)TiO3, BiFeO3 and related films by pulsed laser deposition (PLD) and analyzed the films by X-ray diffractometry (XRD), atomic force microscopy (AFM), etc. To obtain inherently fully strained films, the selection of the appropriate substrates is crucial. MgAl2O4 matches best with good quality and size, yet the spinel structure has an intrinsic incompatibility to that of perovskite. We introduced a rock-salt structure material (Ni 1-xAlxO1+delta) as a buffer layer to mediate the structural mismatch for (Ba,Sr)TiO3 films. With buffer layer Ni1-xAlxO1+delta, we show that the BST films have high quality crystallization and are coherently epitaxial. AFM images show that the films have smoother surfaces when including the buffer layer, indicating an inherent compatibility between BST-NAO and NAO-MAO. In-plane Ferroelectricity measurement shows double hysteresis loops, indicating an antiferroelectric-like behavior: pinned ferroelectric domains with antiparallel alignments of polarization. The Curie temperatures of the coherent fully strained BST films are also measured. It is higher than 900°C, at least 800°C higher than that of bulk. The improved Curie temperature makes the use of BST as FeRAM feasible. We found that the special behaviors of ferroelectricity including hysteresis loop and Curie temperature are due to inherent fully tensile strain. This might be a clue of physics inside ferroelectric stain engineering. An out-of-plane ferroelectricity measurement would provide a full whole story of the tensile strain. However, a well suited electrode material that is both conducting, and full strained on the MgAl2O4 substrate is quite rare. We will supply some answers to this unique problem. XRD results show that Ni1-xAlxO1+delta (x=0.3, 0.4 & 0.5) film, although highly mixed with Al2O3, still takes rock-salt structure and is grown very well on the spinel MgAl 2O4 substrate, with perfect crystallization and a smooth surface. Ni0.7Al0.3O1+ delta and Ni 0.6Al0.4O1+ delta are good buffer layers for perovskite film on spinel MgAl2O4 substrate. Ni 0.5Al0.5O1+ delta could also be a good buffer layer. The structural transition from rock-salt to spinel was found at x=0.67. Tensile strain effects from thermal expansion difference of BiFeO3 films were found. Thermal expansion difference caused strain does not change the ferroelectric property greatly, due to film relaxation. BiFeO3 film with NAO buffer exhibit much larger strain.

  10. Use of immobilized exopeptidases and volatile buffers for analysis of peptides by fast atom bombardment mass spectrometry.

    PubMed

    Wagner, R M; Fraser, B A

    1987-05-01

    beta-Lipotrophin (62-77) or Ac-gastrin releasing peptide was incubated with immobilized carboxypeptidase Y or aminopeptidase M. Subsequent aliquots of each incubation mixture were analysed by fast atom bombardment mass spectrometry using a dithiothreitol/dithioerythritol liquid matrix. The use of immobilized enzymes and volatile buffers for exopeptidase digestions enabled rapid and facile separation of enzyme from digestion products. This approach to mass spectral peptide analysis reduced spectral background arising from a glycerol matrix, buffer salts, or enzyme proteins and contaminants, enabling analysis of as little as 200 picomoles of a suitable peptide.

  11. Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

    PubMed Central

    Murphy, Patrick J. M.; Stone, Orrin J.; Anderson, Michelle E.

    2011-01-01

    In contrast to other chromatographic methods for purifying proteins (e.g. gel filtration, affinity, and ion exchange), hydrophobic interaction chromatography (HIC) commonly requires experimental determination (referred to as screening or "scouting") in order to select the most suitable chromatographic medium for purifying a given protein 1. The method presented here describes an automated approach to scouting for an optimal HIC media to be used in protein purification. HIC separates proteins and other biomolecules from a crude lysate based on differences in hydrophobicity. Similar to affinity chromatography (AC) and ion exchange chromatography (IEX), HIC is capable of concentrating the protein of interest as it progresses through the chromatographic process. Proteins best suited for purification by HIC include those with hydrophobic surface regions and able to withstand exposure to salt concentrations in excess of 2 M ammonium sulfate ((NH4)2SO4). HIC is often chosen as a purification method for proteins lacking an affinity tag, and thus unsuitable for AC, and when IEX fails to provide adequate purification. Hydrophobic moieties on the protein surface temporarily bind to a nonpolar ligand coupled to an inert, immobile matrix. The interaction between protein and ligand are highly dependent on the salt concentration of the buffer flowing through the chromatography column, with high ionic concentrations strengthening the protein-ligand interaction and making the protein immobile (i.e. bound inside the column) 2. As salt concentrations decrease, the protein-ligand interaction dissipates, the protein again becomes mobile and elutes from the column. Several HIC media are commercially available in pre-packed columns, each containing one of several hydrophobic ligands (e.g. S-butyl, butyl, octyl, and phenyl) cross-linked at varying densities to agarose beads of a specific diameter 3. Automated column scouting allows for an efficient approach for determining which HIC media should be employed for future, more exhaustive optimization experiments and protein purification runs 4. The specific protein being purified here is recombinant green fluorescent protein (GFP); however, the approach may be adapted for purifying other proteins with one or more hydrophobic surface regions. GFP serves as a useful model protein, due to its stability, unique light absorbance peak at 397 nm, and fluorescence when exposed to UV light 5. Bacterial lysate containing wild type GFP was prepared in a high-salt buffer, loaded into a Bio-Rad DuoFlow medium pressure liquid chromatography system, and adsorbed to HiTrap HIC columns containing different HIC media. The protein was eluted from the columns and analyzed by in-line and post-run detection methods. Buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection increased the functionality of the system and reproducibility of the experimental approach. PMID:21968976

  12. Increased degradation rate of nitrososureas in media containing carbonate.

    PubMed

    Seidegård, Janeric; Grönquist, Lena; Tuvesson, Helen; Gunnarsson, Per-Olov

    2009-01-01

    The stability of two nitrosoureas, tauromustine and lomustine, has been investigated in different media and buffers. All media tested, except Leibovitz's L-15 medium, significantly increased the degradation rate of the investigated nitrosoureas at pH 7.4. Sodium bicarbonate seems to be the cause of the observed increase of the degradation rate, since it provides the main buffering capacity of all the media except for Leibovitz's L-15 medium, which is based on phosphate buffer. Other ingredients in the media, such as amino acids, vitamins, and inorganic salts, or the ionic strength of a buffer, did not have any major effect on the degradation rate of the nitrosoureas. These results suggest that media containing carbonated buffer should be avoided when the anti-tumor effect of nitrosoureas is to be investigated in different cell cultures.

  13. Arrestin binds to different phosphorylated regions of the thyrotropin-releasing hormone receptor with distinct functional consequences.

    PubMed

    Jones, Brian W; Hinkle, Patricia M

    2008-07-01

    Arrestin binding to agonist-occupied phosphorylated G protein-coupled receptors typically increases the affinity of agonist binding, increases resistance of receptor-bound agonist to removal with high acid/salt buffer, and leads to receptor desensitization and internalization. We tested whether thyrotropin-releasing hormone (TRH) receptors lacking phosphosites in the C-terminal tail could form stable and functional complexes with arrestin. Fibroblasts from mice lacking arrestins 2 and 3 were used to distinguish between arrestin-dependent and -independent effects. Arrestin did not promote internalization or desensitization of a receptor that had key Ser/Thr phosphosites mutated to Ala (4Ala receptor). Nevertheless, arrestin greatly increased acid/salt resistance and the affinity of 4Ala receptor for TRH. Truncation of 4Ala receptor just distal to the key phosphosites (4AlaStop receptor) abolished arrestin-dependent acid/salt resistance but not the effect of arrestin on agonist affinity. Arrestin formed stable complexes with activated wild-type and 4Ala receptors but not with 4AlaStop receptor, as measured by translocation of arrestin-green fluorescent protein to the plasma membrane or chemical cross-linking. An arrestin mutant that does not interact with clathrin and AP2 did not internalize receptor but still promoted high affinity TRH binding, acid/salt resistance, and desensitization. A sterically restricted arrestin mutant did not cause receptor internalization or desensitization but did promote acid/salt resistance and high agonist affinity. The results demonstrate that arrestin binds to proximal or distal phosphosites in the receptor tail. Arrestin binding at either site causes increased agonist affinity and acid/salt resistance, but only the proximal phosphosites evoke the necessary conformational changes in arrestin for receptor desensitization and internalization.

  14. Mechanism by which ammonium bicarbonate and ammonium sulfate inhibit mycotoxigenic fungi.

    PubMed Central

    DePasquale, D A; Montville, T J

    1990-01-01

    In this study we examined the mechanism by which ammonium bicarbonate inhibits mycotoxigenic fungi. Elevated extracellular pH, alone, was not responsible for the antifungal activity. Although conidia of Penicillium griseofulvum and Fusarium graminearum had internal pH (pHi) values as high as 8.0 in buffer at an external pH (pHo) of 9.5, their viability was not markedly affected. The pHi values from conidia equilibrated in glycine-NaOH-buffered treatments without ammonium bicarbonate or ammonium sulfate were similar to values obtained from buffered treatments containing the ammonium salts. Thus, inhibition did not appear to be directly related to increased pHi. Ammonium sulfate in buffered media at pH greater than or equal to 8.7 was as inhibitory as ammonium bicarbonate, but was completely ineffective at pH less than or equal to 7.8. The hypothesis that free ammonia caused the fungal inhibition was tested by using ammonium sulfate as a model for ammonium bicarbonate. Viability, expressed as log CFU/ml, and percent germination of P. griseofulvum and F. graminearum decreased dramatically as the free ammonia concentration increased. Germination rate ratios (the germination rate in buffered ammonium sulfate divided by the germination rate in buffer alone) decreased linearly as the free ammonia concentration increased, further establishing NH3 as the toxic agent. Ammonium bicarbonate inhibits fungi because the bicarbonate anion supplies the alkalinity necessary to establish an antifungal concentration of free ammonia. PMID:2082821

  15. Sodium and chloride levels in rainfall, mist. streamwater and groundwater at the Plynlimon catchments, mid-Wales: inferences on hydrological and chemical controls

    NASA Astrophysics Data System (ADS)

    Neal, C.; Kirchner, J. W.

    Variations in sodium and chloride in atmospheric inputs (rainfall and mist), stream runoff and groundwater stores are documented for the upper Severn River (Afon Hafren and Afon Hore catchments), Plynlimon, mid-Wales. The results show five salient features.

    1. Sodium and chloride concentrations are highly variable and highly correlated in rainfall and mist. The sodium-chloride relationship in rainfall has a slope close to the sodium/chloride ratio in sea-water, and an intercept that is not significantly different from zero. This indicates that sea-salt is the dominant source of both sodium and chloride in rainfall, which would be expected given the maritime nature of the metrology. For mist, there is also a straight line with near-zero intercept, but with a slightly higher gradient than the sea-salt ratio, presumably due to small additional sodium inputs from other sources.
    2. There is an approximate input-output balance for both sodium and chloride, with the exception of one groundwater well, in which high chemical weathering results in an anomalous high Na/Cl ratio. Thus, atmospheric deposition is the dominant source of both sodium and chloride in groundwater and streamflow.
    3. The fluctuations in sodium and chloride concentrations in the streams and groundwaters are strongly damped compared to those in the rain and the mist, reflecting the storage and mixing of waters in the subsurface.
    4. On all timescales, from weeks to years, sodium fluctuations are more strongly damped than chloride fluctuations in streamflow. The additional damping of sodium is consistent with ion exchange buffering of sodium in the catchment soils.
    5. Sodium and chloride concentrations are linearly correlated in the streams and groundwaters, but the slope is almost universally less than the sea-salt ratio and there is a non-zero intercept. The Na/Cl ratio in streamflow and groundwater is higher than the sea-salt ratio when salinity is low and lower than the sea-salt ratio when salinity is high. This pattern of behaviour is again consistent with ion exchange buffering of sodium in the catchment soils.
    The core features of this study are two fold. Firstly, sodium and chloride concentrations are highly damped within the streams and groundwaters relative to the atmospheric input. Secondly, streamflow sodium and chloride respond in similar ways across the catchments, except for the added cation exchange damping of the sodium signal. These findings are remarkable given the heterogeneous nature of the catchments and the complexity of the chemical time series signals in the streams.

  16. Ca2+ transients in cardiac myocytes measured with high and low affinity Ca2+ indicators.

    PubMed Central

    Berlin, J R; Konishi, M

    1993-01-01

    Intracellular calcium ion ([Ca2+]i) transients were measured in voltage-clamped rat cardiac myocytes with fura-2 or furaptra to quantitate rapid changes in [Ca2+]i. Patch electrode solutions contained the K+ salt of fura-2 (50 microM) or furaptra (300 microM). With identical experimental conditions, peak amplitude of stimulated [Ca2+]i transients in furaptra-loaded myocytes was 4- to 6-fold greater than that in fura-2-loaded cells. To determine the reason for this discrepancy, intracellular fura-2 Ca2+ buffering, kinetics of Ca2+ binding, and optical properties were examined. Decreasing cellular fura-2 concentration by lowering electrode fura-2 concentration 5-fold, decreased the difference between the amplitudes of [Ca2+]i transients in fura-2 and furaptra-loaded myocytes by twofold. Thus, fura-2 buffers [Ca2+]i under these conditions; however, Ca2+ buffering is not the only factor that explains the different amplitudes of the [Ca2+]i transients measured with these indicators. From the temporal comparison of the [Ca2+]i transients measured with fura-2 and furaptra, the apparent reverse rate constant for Ca2+ binding of fura-2 was at least 65s-1, much faster than previously reported in skeletal muscle fibers. These binding kinetics do not explain the difference in the size of the [Ca2+]i transients reported by fura-2 and furaptra. Parameters for fura-2 calibration, Rmin, Rmax, and beta, were obtained in salt solutions (in vitro) and in myocytes exposed to the Ca2+ ionophore, 4-Br A23187, in EGTA-buffered solutions (in situ). Calibration of fura-2 fluorescence signals with these in situ parameters yielded [Ca2+]i transients whose peak amplitude was 50-100% larger than those calculated with in vitro parameters. Thus, in vitro calibration of fura-2 fluorescence significantly underestimates the amplitude of the [Ca2+]i transient. These data suggest that the difference in amplitude of [Ca2+]i transients in fura-2 and furaptra-loaded myocytes is due, in part, to Ca2+ buffering by fura-2 and use of in vitro calibration parameters. PMID:8274651

  17. Pyrolytic sugars from cellulosic biomass.

    PubMed

    Kuzhiyil, Najeeb; Dalluge, Dustin; Bai, Xianglan; Kim, Kwang Ho; Brown, Robert C

    2012-11-01

    Depolymerization of cellulose offers the prospect of inexpensive sugars from biomass. Breaking the glycosidic bonds of cellulose to liberate glucose has usually been pursued by acid or enzymatic hydrolysis although a purely thermal depolymerization route to sugars is also possible. Fast pyrolysis of pure cellulose yields primarily the anhydrosugar levoglucosan (LG) whereas the presence of naturally occurring alkali and alkaline earth metals (AAEMs) in biomass strongly catalyzes ring-breaking reactions that favor formation of light oxygenates. Here, we show a method of significantly increasing the yield of sugars from biomass by purely thermal means through infusion of certain mineral acids (phosphoric and sulfuric acid) into the biomass to convert the AAEMs into thermally stable salts (particularly potassium sulfates and phosphates). These salts not only passivate AAEMs that normally catalyze fragmentation of pyranose rings, but also buffer the system at pH levels that favor glycosidic bond breakage. It appears that AAEM passivation contributes to 80 % of the enhancement in LG yield while the buffering effect of the acid salts contributes to the balance of the enhancement. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Good’s buffers as a basis for developing self-buffering and biocompatible ionic liquids for biological research†

    PubMed Central

    Taha, Mohamed; e Silva, Francisca A.; Quental, Maria V.; Ventura, Sónia P. M.; Freire, Mara G.; Coutinho, João A. P.

    2014-01-01

    This work reports a promising approach to the development of novel self-buffering and biocompatible ionic liquids for biological research in which the anions are derived from biological buffers (Good’s buffers, GB). Five Good’s buffers (Tricine, TES, CHES, HEPES, and MES) were neutralized with four suitable hydroxide bases (1-ethyl-3-methylimidazolium, tetramethylammonium, tetraethylammonium, and tetrabutylammonium) producing 20 Good’s buffer ionic liquids (GB-ILs). The presence of the buffering action of the synthesized GB-ILs was ascertained by measuring their pH-profiles in water. Moreover, a series of mixed GB-ILs with wide buffering ranges were formulated as universal buffers. The impact of GB-ILs on bovine serum albumin (BSA), here used as a model protein, is discussed and compared with more conventional ILs using spectroscopic techniques, such as infrared and dynamic light scattering. They appear to display, in general, a greater stabilizing effect on the protein secondary structure than conventional ILs. A molecular docking study was also carried out to investigate on the binding sites of GB-IL ions to BSA. We further used the QSAR-human serum albumin binding model, log K(HSA), to calculate the binding affinity of some conventional ILs/GB-ILs to HSA. The toxicity of the GB and GB-ILs was additionally evaluated revealing that they are non-toxic against Vitro fischeri. Finally, the GB-ILs were also shown to be able to form aqueous biphasic systems when combined with aqueous solutions of inorganic or organic salts, and we tested their extraction capability for BSA. These systems were able to extract BSA with an outstanding extraction efficiency of 100% in a single step for the GB-IL-rich phase, and, as a result, the use of GB-IL-based ABS for the separation and extraction of other added-value biomolecules is highly encouraging and worthy of further investigation. PMID:25729325

  19. Identification of a haloalkaliphilic and thermostable cellulase with improved ionic liquid tolerance

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

    Zhang, Tao; Datta, Supratim; Eichler, Jerry

    2011-02-17

    Some ionic liquids (ILs) have been shown to be very effective solvents for biomass pretreatment. It is known that some ILs can have a strong inhibitory effect on fungal cellulases, making the digestion of cellulose inefficient in the presence of ILs. The identification of IL-tolerant enzymes that could be produced as a cellulase cocktail would reduce the costs and water use requirements of the IL pretreatment process. Due to their adaptation to high salinity environments, halophilic enzymes are hypothesized to be good candidates for screening and identifying IL-resistant cellulases. Using a genome-based approach, we have identified and characterized a halophilicmore » cellulase (Hu-CBH1) from the halophilic archaeon, Halorhabdus utahensis. Hu-CBH1 is present in a gene cluster containing multiple putative cellulolytic enzymes. Sequence and theoretical structure analysis indicate that Hu-CBH1 is highly enriched with negatively charged acidic amino acids on the surface, which may form a solvation shell that may stabilize the enzyme, through interaction with salt ions and/or water molecules. Hu-CBH1 is a heat tolerant haloalkaliphilic cellulase and is active in salt concentrations up to 5 M NaCl. In high salt buffer, Hu-CBH1 can tolerate alkali (pH 11.5) conditions and, more importantly, is tolerant to high levels (20percent w/w) of ILs, including 1-allyl-3-methylimidazolium chloride ([Amim]Cl). Interestingly, the tolerances to heat, alkali and ILs are found to be salt-dependent, suggesting that the enzyme is stabilized by the presence of salt. Our results indicate that halophilic enzymes are good candidates for the screening of IL-tolerant cellulolytic enzymes.« less

  20. Quantitative Correlation between Viscosity of Concentrated MAb Solutions and Particle Size Parameters Obtained from Small-Angle X-ray Scattering.

    PubMed

    Fukuda, Masakazu; Moriyama, Chifumi; Yamazaki, Tadao; Imaeda, Yoshimi; Koga, Akiko

    2015-12-01

    To investigate the relationship between viscosity of concentrated MAb solutions and particle size parameters obtained from small-angle X-ray scattering (SAXS). The viscosity of three MAb solutions (MAb1, MAb2, and MAb3; 40-200 mg/mL) was measured by electromagnetically spinning viscometer. The protein interactions of MAb solutions (at 60 mg/mL) was evaluated by SAXS. The phase behavior of 60 mg/mL MAb solutions in a low-salt buffer was observed after 1 week storage at 25°C. The MAb1 solutions exhibited the highest viscosity among the three MAbs in the buffer containing 50 mM NaCl. Viscosity of MAb1 solutions decreased with increasing temperature, increasing salt concentration, and addition of amino acids. Viscosity of MAb1 solutions was lowest in the buffer containing histidine, arginine, and aspartic acid. Particle size parameters obtained from SAXS measurements correlated very well with the viscosity of MAb solutions at 200 mg/mL. MAb1 exhibited liquid-liquid phase separation at a low salt concentration. Simultaneous addition of basic and acidic amino acids effectively suppressed intermolecular attractive interactions and decreased viscosity of MAb1 solutions. SAXS can be performed using a small volume of samples; therefore, the particle size parameters obtained from SAXS at intermediate protein concentration could be used to screen for low viscosity antibodies in the early development stage.

  1. Exclusive nuclear location of estrogen receptors in Squalus testis.

    PubMed Central

    Callard, G V; Mak, P

    1985-01-01

    An estrogen (E)-binding molecule having both occupied and unoccupied sites is restricted to nuclear subfractions in the testis of the spiny dogfish (Squalus acanthias). We investigated the hypothesis that a species characterized by high body-fluid osmolarity (1010 mosM) has an estrogen receptor (ER) that binds to chromatin with high affinity and consequently resists redistribution during tissue processing. Although the steroid binding and sedimentation properties of the Squalus nuclear ER conformed to those of classical ER, its elution maximum from DNA-cellulose was unusually high (0.55 M NaCl). A tendency to adhere tightly to cell nuclei was reflected in the high salt concentration (0.43 M KCl) required to extract 50% of the receptors from the nuclear compartment during homogenization and in the stability of the nuclear ER population in the presence of high concentrations of a nonionic solute (urea) or increased buffer volume. Mixing and redistribution experiments showed that nuclear ER could be quantitatively and qualitatively measured in cytosolic extracts, ruling out the possibility that soluble receptors were being masked. Although Squalus oviduct ER was similar to that of testis, ER in the testis and liver of a related elasmobranch (Potamotrygon) that maintains osmotic equilibrium at 300 mosM more closely resembled mammalian ER in its elution maximum from DNA-cellulose (0.22 M NaCl) and cytosolic/nuclear ratios in low-salt buffers. We conclude that Squalus testis has a single ER pool located exclusively in the nuclear compartment. These observations support a revised concept of steroid action and further indicate that the chromatin affinity of the hormone-ER complex is an important factor in determining subfractional distribution during tissue processing. PMID:3856265

  2. An evaluation of MES (2(N-Morpholino)ethanesulfonic acid) and Amberlite IRC-50 as pH buffers for nutrient solution studies

    NASA Technical Reports Server (NTRS)

    Bugbee, B. G.; Salisbury, F. B.

    1985-01-01

    All buffering agents used to stabilize pH in hydroponic research have disadvantages. Inorganic buffers are absorbed and may become phytotoxic. Solid carbonate salts temporarily mitigate decreasing pH but provide almost no protection against increasing pH, and they alter nutrient absorption. Exchange resins are more effective, but we find that they remove magnesium and manganese from solution. We have tested 2(N-Morpholino)ethanesulfonic acid (MES) as a buffering agent at concentrations of 1 and 10 mol m-3 (1 and 10 mM) with beans, corn, lettuce, tomatoes, and wheat. MES appears to be biologically inert and does not interact significantly with other solution ions. Relative growth rates among controls and MES treatments were nearly identical for each species during the trial period. The pH was stabilized by 1 mol m-3 MES. This buffer warrants further consideration in nutrient research.

  3. Characterization of lipid-protein interactions in acetylcholinesterase lipoprotein extracted from bovine erythrocytes.

    PubMed Central

    Beauregard, G; Roufogalis, B D

    1979-01-01

    Acetylcholinesterase was released from bovine erythrocytes in hypo-osmotic sodium phosphate buffer. Initially, about 30% of the enzyme was released in a soluble lipoprotein form, and further incubation resulted in the progressive release of the enzyme in a particulate form. Solubilization of the acetylcholinesterase in the particulate fraction with Lubrol WX (2 mg/ml) resulted in the loss of all lipids except a non-exchangeable fraction identified as cardiolipin. Addition of a mixture of erythrocyte phospholipids to the soluble forms and to the Lubrol WX-solubilized enzyme resulted in the formation of particulate forms of the enzyme with increased partial specific volume and Stokes radius, and a break in the Arrhenius plot of the enzyme activity around 20 degrees C. The break in the Arrhenius plot was abolished by treatment of a soluble enzyme preparation with 1.8 M salt (NaCl) in phosphate buffer, conditions that allowed the extraction of cardiolipin from the enzyme by chloroform/methanol. Failure of the high-salt treatment to decrease the Stokes radius made it unlikely that the bound cardiolipin formed a boundary layer or annulus around the protein. It is suggested that cardiolipin is bound to the core of the dimeric protein structure, thereby controlling the acetylcholinesterase activity. PMID:475749

  4. On the hydrophilicity of polyzwitterion poly (N,N-dimethyl-N-(3-(methacrylamido)propyl)ammoniopropane sulfonate) in water, deuterated water, and aqueous salt solutions.

    PubMed

    Hildebrand, Viet; Laschewsky, André; Zehm, Daniel

    2014-01-01

    A series of zwitterionic model polymers with defined molar masses up to 150,000 Da and defined end groups are prepared from sulfobetaine monomer N,N-dimethyl-N-(3-(methacrylamido)propyl)ammoniopropanesulfonate (SPP). Polymers are synthesized by reversible addition-fragmentation chain transfer polymerization (RAFT) using a functional chain transfer agent labeled with a fluorescent probe. Their upper critical solution temperature-type coil-to-globule phase transition in water, deuterated water, and various salt solutions is studied by turbidimetry. Cloud points increase with polyzwitterion concentration and molar mass, being considerably higher in D2O than in H2O. Moreover, cloud points are strongly affected by the amount and nature of added salts. Typically, they increase with increasing salt concentration up to a maximum value, whereas further addition of salt lowers the cloud points again, mostly down to below freezing point. The different salting-in and salting-out effects of the studied anions can be correlated with the Hofmeister series. In physiological sodium chloride solution and in phosphate buffered saline (PBS), the cloud point is suppressed even for high molar mass samples. Accordingly, SPP-polymers behave strongly hydrophilic under most conditions encountered in biomedical applications. However, the direct transfer of results from model studies in D2O, using, e.g. (1)H NMR or neutron scattering techniques, to 'normal' systems in H2O is not obvious.

  5. ZnS/Zn(O,OH)S-based buffer layer deposition for solar cells

    DOEpatents

    Bhattacharya, Raghu N [Littleton, CO

    2009-11-03

    The invention provides CBD ZnS/Zn(O,OH)S and spray deposited ZnS/Zn(O,OH)S buffer layers prepared from a solution of zinc salt, thiourea and ammonium hydroxide dissolved in a non-aqueous/aqueous solvent mixture or in 100% non-aqueous solvent. Non-aqueous solvents useful in the invention include methanol, isopropanol and triethyl-amine. One-step deposition procedures are described for CIS, CIGS and other solar cell devices.

  6. SALICYLATE PROCESS FOR THORIUM SEPARATION FROM RARE EARTHS

    DOEpatents

    Cowan, G.A.

    1959-08-25

    The separation of thorium from rare earths is accomplished by forming an aqueous solution of salts of thorium and rare earths and sufficient acetate buffer to provide a pH of between 2 and 5, adding an ammonium salicylate to the aqueous buffered solution, contacting the resultant solution with a substantially water-immiscible organic solvent mixture of an ether and an ester, and separating the solvent extract phase containing thorium salicylate from the aqueous phase containing the rare earths.

  7. Common buffers, media, and stock solutions.

    PubMed

    2001-05-01

    This appendix describes the preparation of selected bacterial media and of buffers and reagents used in the manipulation of nucleic acids and proteins. Recipes for cell culture media and reagents are located elsewhere in the manual. RECIPES: Acids, concentrated stock solutions; Ammonium acetate, 10 M; Ammonium hydroxide, concentrated stock solution; ATP, 100 mM; BCIP, 5% (w/v); BSA (bovine serum albumin), 10% (100 mg/ml); Denhardt solution, 100x; dNTPs: dATP, dTTP, dCTP, and dGTP; DTT, 1 M; EDTA, 0.5 M (pH 8.0); Ethidium bromide solution; Formamide loading buffer, 2x; Gel loading buffer, 6x; HBSS (Hanks balanced salt solution); HCl, 1 M; HEPES-buffered saline, 2x; KCl, 1 M; LB medium; LB plates; Loading buffer; 2-ME, (2-mercaptoethanol)50 mM; MgCl(2), 1 M; MgSO(4), 1 M; NaCl, 5 M; NaOH, 10 M; NBT (nitroblue tetrazolium chloride), 5% (w/v); PCR amplification buffer, 10x; Phosphate-buffered saline (PBS), pH approximately 7.3; Potassium acetate buffer, 0.1 M; Potassium phosphate buffer, 0.1 M; RNase a stock solution (DNase-free), 2 mg/ml; SDS, 20%; SOC medium; Sodium acetate, 3 M; Sodium acetate buffer, 0.1 M; Sodium phosphate buffer, 0.1 M; SSC (sodium chloride/sodium citrate), 20x; SSPE (sodium chloride/sodium phosphate/EDTA), 20x; T4 DNA ligase buffer, 10x; TAE buffer, 50x; TBE buffer, 10x; TBS (Tris-buffered saline); TCA (trichloroacetic acid), 100% (w/v); TE buffer; Terrific broth (TB); TrisCl, 1 M; TY medium, 2x; Urea loading buffer, 2x.

  8. Corneal protection with high-molecular-weight hyaluronan against in vitro and in vivo sodium lauryl sulfate-induced toxic effects.

    PubMed

    Pauloin, Thierry; Dutot, Mélody; Liang, Hong; Chavinier, Emilie; Warnet, Jean-Michel; Rat, Patrice

    2009-10-01

    The aim of this study was to investigate high-molecular-weight hyaluronan (HA-HMW) corneal protection against sodium lauryl sulfate (SLS)-induced toxic effects with in vitro and in vivo experimental approaches. In vitro experiments consisted of a human corneal epithelial cell line incubated with HA-HMW, rinsed, and incubated with SLS. Cell viability, oxidative stress, chromatin condensation, caspase-3, -8, -9, and P2X7 cell death receptor activation, interleukin-6, and interleukin-8 production were investigated. In vivo experiments consisted of 36 New Zealand white rabbits treated for 3 days, 3 times per day, with HA-HMW or phosphate-buffered salt solution. At day 4, eyes were treated with SLS. Clinical observation and in vivo confocal microscopy using the Rostock Cornea Module of the Heidelberg Retina Tomograph-II were performed to evaluate and to compare SLS-induced toxicity between eyes treated with HA-HMW and eyes treated with phosphate-buffered salt solution. In vitro data indicate that exposure of human corneal epithelial cells to HA-HMW significantly decreased SLS-induced oxidative stress, apoptosis, and inflammation cytokine production. In vivo data indicate that SLS cornea injuries, characterized by damaged corneal epithelium, damaged anterior stroma, and inflammatory infiltrations, were attenuated with HA-HMW treatment. A good correlation was seen between in vitro and in vivo findings showing that HA-HMW decreases SLS-induced toxic effects and protects cornea.

  9. Purification, crystallization and preliminary crystallographic study of low oxygen-affinity haemoglobin from cat (Felis silvestris catus) in two different crystal forms.

    PubMed

    Balasubramanian, M; Moorthy, Pon Sathya; Neelagandan, K; Ponnuswamy, M N

    2009-03-01

    Haemoglobin is a metalloprotein which plays a major role in the transportation of oxygen from the lungs to tissues and of carbon dioxide back to the lungs. The present work reports the preliminary crystallographic study of low oxygen-affinity haemoglobin from cat in different crystal forms. Cat blood was collected, purified by anion-exchange chromatography and crystallized in two different conditions by the hanging-drop vapour-diffusion method under unbuffered low-salt and buffered high-salt concentrations using PEG 3350 as a precipitant. Intensity data were collected using MAR345 and MAR345dtb image-plate detector systems. Cat haemoglobin crystallizes in monoclinic and orthorhombic crystal forms with one and two whole biological molecules (alpha(2)beta(2)), respectively, in the asymmetric unit.

  10. Influence of organic buffers on bacteriocin production by Streptococcus thermophilus ST110.

    PubMed

    Somkuti, George A; Gilbreth, Stefanie E

    2007-08-01

    The effect of the organic buffer salts MES, MOPS, and PIPES on the growth of S. thermophilus ST110, medium pH, and accumulation of the antipediococcal bacteriocin thermophilin 110 were evaluated in whey permeate media over a period of 24 h. In nonbuffered medium, thermophilin 110 production at 37 degrees C paralleled the growth of S. thermophilus ST110 and reached a maximum after 8-10 h. Addition of organic buffer salts decreased the drop in medium pH and resulted in increased biomass (dry cells; microg/mL) and higher yields of thermophilin 110 (units/microg cells). The best results were obtained by the addition of 1% (w/v) MES to the medium, which reduced the pH drop to 1.8 units after 10 h of growth (compared to 2.3 pH units in the control) and resulted in a 1.5-fold increase in cell mass (495 microg/mL) and a 7-fold increase in thermophilin 110 yield (77 units/microg dry cells) over the control. The results showed that whey permeate-based media may be suitable for producing large amounts of thermophilin 110 needed for controlling spoilage pediococci in industrial wine and beer fermentations.

  11. High-performance cation-exchange chromatofocusing of proteins.

    PubMed

    Kang, Xuezhen; Frey, Douglas D

    2003-03-28

    Chromatofocusing using high-performance cation-exchange column packings, as opposed to the more commonly used anion-exchange column packings, is investigated with regard to the performance achieved and the range of applications possible. Linear or convex gradients in the range from pH 2.6 to 9 were formed using a variety of commercially available column packings that provide a buffering capacity in different pH ranges, and either polyampholytes or simple mixtures having a small number (three or fewer) of buffering species as the elution buffer. The resolutions achieved using cation-exchange or anion-exchange chromatofocusing were in general comparable, although for certain pairs of proteins better resolution could be achieved using one type of packing as compared to the other, evidently due to the way electrostatic charges are distributed on the protein surface. Several chromatofocusing methods were investigated that take advantage of the acid-base properties of commercially available cation-exchange column packings. These include the use of gradients with a composite shape, the use of very low pH ranges, and the use of elution buffers containing a single buffering species. The advantages of chromatofocusing over ion-exchange chromatography using a salt gradient at constant pH were illustrated by employing the former method and a cation-exchange column packing to separate beta-lactoglobulins A and B, which is a separation reported to be impossible using the latter method and a cation-exchange column packing. Trends in the apparent isoelectric points determined using cation- and anion-exchange chromatofocusing were interpreted using applicable theories. Results of this study indicate that cation-exchange chromatofocusing is a useful technique which is complementary to anion-exchange chromatofocusing and isoelectric focusing for separating proteins at both the analytical and preparative scales.

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

    Streltsov, Victor A.; Titmuss, Stephen J.; Epa, V. Chandana

    Neurodegeneration observed in Alzheimer disease (AD) is believed to be related to the toxicity from reactive oxygen species (ROS) produced in the brain by the amyloid-{beta} (A{beta}) protein bound primarily to copper ions. The evidence for an oxidative stress role of A{beta}-Cu redox chemistry is still incomplete. Details of the copper binding site in A{beta} may be critical to the etiology of AD. Here we present the structure determined by combining x-ray absorption spectroscopy (XAS) and density functional theory analysis of A{beta} peptides complexed with Cu{sup 2+} in solution under a range of buffer conditions. Phosphate-buffered saline buffer salt (NaCl)more » concentration does not affect the high-affinity copper binding mode but alters the second coordination sphere. The XAS spectra for truncated and full-length A{beta}-Cu{sup 2+} peptides are similar. The novel distorted six-coordinated (3N3O) geometry around copper in the A{beta}-Cu{sup 2+} complexes include three histidines: glutamic, or/and aspartic acid, and axial water. The structure of the high-affinity Cu{sup 2+} binding site is consistent with the hypothesis that the redox activity of the metal ion bound to A{beta} can lead to the formation of dityrosine-linked dimers found in AD.« less

  13. MEDULLARY THICK ASCENDING LIMB BUFFER VASOCONSTRICTION OF RENAL OUTER-MEDULLARY VASA RECTA IN SALT-RESISTANT BUT NOT SALT-SENSITIVE RATS

    PubMed Central

    O’Connor, Paul M.; Cowley, Allen W.

    2013-01-01

    We have previously demonstrated that paracrine signaling occurs between medullary thick ascending limb (mTAL) and the contractile pericytes of outer-medullary vasa recta (VR) termed ‘tubular-vascular cross talk’. The aim of the current study was to determine whether tubular-vascular cross talk has a functional effect on vasoconstrictor responses to angiotensin II, and to determine whether this is altered in the Dahl salt-sensitive (SS) rat. Studies were performed on salt-resistant consomic SS.13BN and SS rats using a novel outer medullary tissue strip preparation in which freshly isolated VR within VR bundles were perfused either alone or in combination with nearby mTAL. In VR from SS.13BN rats, angiotensin II (1μM) increased VR bundle intracellular Ca2+ concentration ([Ca2+]VR) 19±9nM (n=8) and reduced focal diameter in perfused VR by (−20±7%;n=5). In the presence of nearby mTAL however, [Ca2+]VR (−9±8nM; n=8) and VR diameter (−1±4%, n=7) in SS.13BN rats was unchanged by angiotensin II. In contrast, in Dahl SS rats, angiotensin II resulted in rapid and sustained increase in [Ca2+]VR (89±48 n=7;50±24% n=8) and a reduction in VR diameter of (−17±7;n=7 and −11±4%;n=5) in both isolated VR and VR with nearby mTAL, respectively. In VR with mTAL from SS13BN rats, inhibiton of purinergic receptors resulted in an increase in [Ca2+]VR, indicating purinergic signaling buffers vasoconstriction. Importantly, our in vitro data were able to predict medullary blood flow responses to angiotensin II in SS and SS.13BN rats in vivo. We conclude that paracrine signaling from mTAL buffers angiotensin II vasoconstriction in Dahl salt-resistant SS.13BN rats but not SS rats. PMID:22926950

  14. A Core Facility for the Study of Neurotoxins of Biological Origin

    DTIC Science & Technology

    1990-06-15

    toxicity of 5 x 10-8 MLD/mg protein. Sodium 125 Iodine and the Bolton-Hunter Reagent - 1251odine were purchased from Amersham. Chloramine- T , glycine...tyrosine and all salts and buffers were from Sigma Chemical Co. and Fisher. Iodination procedures. The chloramine- T method was used essentially as...previously described. Tetanus toxin (100 ig) in sodium phosphate buffer (100 mM, pH 7.4) was mixed with chloramine- T (0.5 mM) and Na 1251 (1 mCi) for 30

  15. [Systematic evaluation of retention behavior of carbohydrates in hydrophilic interaction liquid chromatography].

    PubMed

    Fu, Qing; Wang, Jun; Liang, Tu; Xu, Xiaoyong; Jin, Yu

    2013-11-01

    A systematic evaluation of retention behavior of carbohydrates in hydrophilic interaction liquid chromatography (HILIC) was performed. The influences of mobile phase, stationary phase and buffer salt on the retention of carbohydrates were investigated. According to the results, the retention time of carbohydrates decreased as the proportion of acetonitrile in mobile phase decreased. Increased time of carbohydrates was observed as the concentration of buffer salt in mobile phase increased. The retention behavior of carbohydrates was also affected by organic solvent and HILIC stationary phase. Furthermore, an appropriate retention equation was used in HILIC mode. The retention equation lnk = a + blnC(B) + cC(B) could quantitatively describe the retention factors of carbohydrates of plant origin with good accuracy: the relative error of the predicted time to actual time was less than 0.3%. The evaluation results could provide guidance for carbohydrates to optimize the experimental conditions in HILIC method development especially for carbohydrate separation

  16. Light Scattering Characterization of Elastin-Like Polypeptide Trimer Micelles

    NASA Astrophysics Data System (ADS)

    Tsuper, Ilona; Terrano, Daniel; Maraschky, Adam; Holland, Nolan; Streletzky, Kiril

    The elastin-like polypeptides (ELP) nanoparticles are composed of three-armed star polypeptides connected by a negatively charged foldon. Each of the three arms extending from the foldon domain includes 20 repeats of the (GVGVP) amino acid sequence. The ELP polymer chains are soluble at room temperature and become insoluble at the transition temperature (close to 50 ° C), forming micelles. The size and shape of the micelle are dependent on the temperature and the pH of the solution, and on the concentration of the phosphate buffered saline (PBS). The depolarized dynamic light scattering (DDLS) was employed to study the structure and dynamics of micelles at 62 ° C. The solution was maintained at an approximate pH level of 7.3 - 7.5, while varying PBS concentration. At low salt concentrations (<15 mM), the micelle radius was about 10nm but not very reproducible on account of unstable pH levels arising from low buffer concentrations. At intermediate salt concentrations (15 - 60 mM), the system formed spherically-shaped micelles, exhibiting a steady growth in the hydrodynamic radius (Rh) from 10 to 21 nm, with increasing PBS concentration. Interestingly, higher salt concentrations (>60 mM) displayed an apparent elongation of the micelles evident by a significant VH signal, along with a surge in the apparent Rh. A model of micelle growth (and potential elongation) with increase in salt concentration is considered.

  17. THE KINETICS OF SAPONIFICATION OF IODOACETIC ACID BY SODIUM HYDROXIDE AND BY CERTAIN ALKALINE BUFFER SOLUTIONS.

    PubMed

    Brdicka, R

    1936-07-20

    1. The rate of the saponification of iodoacetic acid in sodium hydroxide and alkaline buffer solutions yielding glycollic acid was measured by means of Heyrovský's polarographic method. 2. From the bimolecular velocity constants, increasing with the ionic strength of the solution, the Brönsted factor, F, which characterizes the primary salt effect, was calculated. 3. In the borate buffer solutions the monomolecular constants of the saponification were determined which, at values above the pH of neutralization of boric acid, show a proportionality to the concentration of hydroxyl anions. Below the pH of neutralization of boric acid, they are proportional to the concentration of borate anions.

  18. High Temperature Superconducting Thick Films

    DOEpatents

    Arendt, Paul N.; Foltyn, Stephen R.; Groves, James R.; Holesinger, Terry G.; Jia, Quanxi

    2005-08-23

    An article including a substrate, a layer of an inert oxide material upon the surface of the substrate, (generally the inert oxide material layer has a smooth surface, i.e., a RMS roughness of less than about 2 nm), a layer of an amorphous oxide or oxynitride material upon the inert oxide material layer, a layer of an oriented cubic oxide material having a rock-salt-like structure upon the amorphous oxide material layer is provided together with additional layers such as at least one layer of a buffer material upon the oriented cubic oxide material layer or a HTS top-layer of YBCO directly upon the oriented cubic oxide material layer. With a HTS top-layer of YBCO upon at least one layer of a buffer material in such an article, Jc's of 1.4×106 A/cm2 have been demonstrated with projected Ic's of 210 Amperes across a sample 1 cm wide.

  19. Phenomenological Partial Specific Volumes for G-Quadruplex DNAs

    PubMed Central

    Hellman, Lance M.; Rodgers, David W.; Fried, Michael G.

    2009-01-01

    Accurate partial specific volume (ν̄) values are required for sedimentation velocity and sedimentation equilibrium analyses. For nucleic acids, the estimation of these values is complicated by the fact that ν̄ depends on base composition, secondary structure, solvation and the concentrations and identities of ions in the surrounding buffer. Here we describe sedimentation equilibrium measurements of the apparent isopotential partial specific volume φ′ for two G-quadruplex DNAs and a single-stranded DNA of similar molecular weight and base composition. The G-quadruplex DNAs are a 22 nucleotide fragment of the human telomere consensus sequence and a 27 nucleotide fragment from the human c-myc promoter. The single-stranded DNA is 26 nucleotides long and is designed to have low propensity to form secondary structures. Parallel measurements were made in buffers containing NaCl and in buffers containing KCl, spanning the range 0.09M ≤ [salt] ≤ 2.3M. Limiting values of φ′, extrapolated to [salt] = 0M, were: 22-mer (NaCl-form), 0.525 ± 0.004 mL/g; 22-mer (KCl-form), 0.531 ± 0.006 mL/g; 27-mer (NaCl-form), 0.548 ± 0.005 mL/g; 27-mer (KCl-form), 0.557 ± 0.006 mL/g; 26-mer (NaCl-form), 0.555 ± 0.004 mL/g; 26-mer (KCl-form), 0.564 ± 0.006 mL/g. Small changes in φ′ with [salt] suggest that large changes in counterion association or hydration are unlikely to take place over these concentration ranges. PMID:19238377

  20. Quantifying the sensitivity of G. oxydans ATCC 621H and DSM 3504 to osmotic stress triggered by soluble buffers.

    PubMed

    Luchterhand, B; Fischöder, T; Grimm, A R; Wewetzer, S; Wunderlich, M; Schlepütz, T; Büchs, J

    2015-04-01

    In Gluconobacter oxydans cultivations on glucose, CaCO3 is typically used as pH-buffer. This buffer, however, has disadvantages: suspended CaCO3 particles make the medium turbid, thereby, obstructing analysis of microbial growth via optical density and scattered light. Upon searching for alternative soluble pH-buffers, bacterial growth and productivity was inhibited most probably due to osmotic stress. Thus, this study investigates in detail the osmotic sensitivity of G. oxydans ATCC 621H and DSM 3504 using the Respiratory Activity MOnitoring System. The tested soluble pH-buffers and other salts attained osmolalities of 0.32-1.19 osmol kg(-1). This study shows that G. oxydans ATCC 621H and DSM 3504 respond quite sensitively to increased osmolality in comparison to other microbial strains of industrial interest. Osmolality values of >0.5 osmol kg(-1) should not be exceeded to avoid inhibition of growth and product formation. This osmolality threshold needs to be considered when working with soluble pH-buffers.

  1. Spectroscopic investigation of the effect of salt on binding of tartrazine with two homologous serum albumins: quantification by use of the Debye-Hückel limiting law and observation of enthalpy-entropy compensation.

    PubMed

    Bolel, Priyanka; Datta, Shubhashis; Mahapatra, Niharendu; Halder, Mintu

    2012-08-30

    Formation of ion pair between charged molecule and protein can lead to interesting biochemical phenomena. We report the evolution of thermodynamics of the binding of tartrazine, a negatively charged azo colorant, and serum albumins with salt. The dye binds predominantly electrostatically in low buffer strengths; however, on increasing salt concentration, affinity decreases considerably. The calculated thermodynamic parameters in high salt indicate manifestation of nonelectrostatic interactions, namely, van der Waals force and hydrogen bonding. Site-marker competitive binding studies and docking simulations indicate that the dye binds with HSA in the warfarin site and with BSA at the interface of warfarin and ibuprofen binding sites. The docked poses indicate nearby amino acid positive side chains, which are possibly responsible for electrostatic interaction. Using the Debye-Hückel interionic attraction theory for binding equilibria, it is shown that, for electrostatic binding the calculated free energy change increases linearly with square root of ionic strength. Also UV-vis, fluorescence, CD data indicate a decrease of interaction with salt concentration. This study quantitatively relates how ionic strength modulates the strength of the protein-ligand electrostatic interaction. The binding enthalpy and entropy have been found to compensate one another. The enthalpy-entropy compensation (EEC), general property of weak intermolecular interactions, has been discussed.

  2. pH-Dependent Solubility and Dissolution Behavior of Carvedilol--Case Example of a Weakly Basic BCS Class II Drug.

    PubMed

    Hamed, Rania; Awadallah, Areeg; Sunoqrot, Suhair; Tarawneh, Ola; Nazzal, Sami; AlBaraghthi, Tamadur; Al Sayyad, Jihan; Abbas, Aiman

    2016-04-01

    The objective of this study was to investigate the pH-dependent solubility and dissolution of weakly basic Biopharmaceutical Classification Systems (BCS) class II drugs, characterized by low solubility and high permeability, using carvedilol, a weak base with a pK a value of 7.8, as a model drug. A series of solubility and in vitro dissolution studies was carried out using media that simulate the gastric and intestinal fluids and cover the physiological pH range of the GI from 1.2 to 7.8. The effect of ionic strength, buffer capacity, and buffer species of the dissolution media on the solubility and dissolution behavior of carvedilol was also investigated. The study revealed that carvedilol exhibited a typical weak base pH-dependent solubility profile with a high solubility at low pH (545.1-2591.4 μg/mL within the pH range 1.2-5.0) and low solubility at high pH (5.8-51.9 μg/mL within the pH range 6.5-7.8). The dissolution behavior of carvedilol was consistent with the solubility results, where carvedilol release was complete (95.8-98.2% released within 60 min) in media simulating the gastric fluid (pH 1.2-5.0) and relatively low (15.9-86.2% released within 240 min) in media simulating the intestinal fluid (pH 6.5-7.8). It was found that the buffer species of the dissolution media may influence the solubility and consequently the percentage of carvedilol released by forming carvedilol salts of varying solubilities. Carvedilol solubility and dissolution decreased with increasing ionic strength, while lowering the buffer capacity resulted in a decrease in carvedilol solubility and dissolution rate.

  3. Southern blotting.

    PubMed

    Brown, T

    2001-05-01

    Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support (the properties and advantages of the different types of membrane, transfer buffer, and transfer method are discussed in detail), resulting in immobilization of the DNA fragments, so the membrane carries a semipermanent reproduction of the banding pattern of the gel. After immobilization, the DNA can be subjected to hybridization analysis, enabling bands with sequence similarity to a labeled probe to be identified. This appendix describes Southern blotting via upward capillary transfer of DNA from an agarose gel onto a nylon or nitrocellulose membrane, using a high-salt transfer buffer to promote binding of DNA to the membrane. With the high-salt buffer, the DNA becomes bound to the membrane during transfer but not permanently immobilized. Immobilization is achieved by UV irradiation (for nylon) or baking (for nitrocellulose). A Support Protocol describes how to calibrate a UV transilluminator for optimal UV irradiation of a nylon membrane. An alternate protocol details transfer using nylon membranes and an alkaline buffer, and is primarily used with positively charged nylon membranes. The advantage of this combination is that no post-transfer immobilization step is required, as the positively charged membrane binds DNA irreversibly under alkaline transfer conditions. The method can also be used with neutral nylon membranes but less DNA will be retained. A second alternate protocol describes a transfer method based on a different transfer-stack setup. The traditional method of upward capillary transfer of DNA from gel to membrane described in the first basic and alternate protocols has certain disadvantages, notably the fact that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it. This slows down the blotting process and may reduce the amount of DNA that can be transferred. The downward capillary method described in the second alternate protocol is therefore more rapid than the basic protocol and can result in more complete transfer. Although the ease and reliability of capillary transfer methods makes this far and away the most popular system for Southern blotting with agarose gels, it unfortunately does not work with polyacrylamide gels, whose smaller pore size impedes the transverse movement of the DNA molecules. The third alternate protocol describes an electroblotting procedure that is currently the most reliable method for transfer of DNA from a polyacrylamide gel. Dot and slot blotting are also described.

  4. Vanadate inhibition of stomatal opening in epidermal peels of Commelina communis

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

    Schwartz, A.; Illan, N.; Assmann, S.M.

    There are conflicting reports on the effectiveness of the plasmamembrane H{sup +} ATPase inhibitor, vanadate, in inhibiting stomatal opening. We have observed that vanadate inhibited light-stimulated stomatal opening in epidermal peels of Commelina communis only at KCl concentrations lower than 50 mM. When KCl was replaced with n-methylglucamine chloride, vanadate was still ineffective at high salt concentrations. However, in the absence of Cl{sup {minus}}, when KOH was buffered with V{sub 2}O{sub 5}, vanadate inhibition of stomatal opening occurred even at high salt concentrations (K{sup +} = 70 mM). An inhibitor of anion uptake, anthracene-9-carboxylic acid (200 {mu}M), partially prevented vanadatemore » inhibition of stomatal opening; other inhibitors (DIDS, SITS, Zn{sup 2+}) were ineffective. These results suggest that inhibition of stomatal opening by vanadate requires its entry into guard cells through an anion uptake system. Decreasing vanadate inhibition at high Cl{sup {minus}}/vanadate ratios may result from competition between vanadate and Cl{sup {minus}} for a common uptake mechanism.« less

  5. Study of surface-bonded dicationic ionic liquids as stationary phases for hydrophilic interaction chromatography.

    PubMed

    Qiao, Lizhen; Li, Hua; Shan, Yuanhong; Wang, Shuangyuan; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2014-02-21

    In the present study, several geminal dicationic ionic liquids based on 1,4-bis(3-allylimidazolium)butane and 1,8-bis(3-allylimidazolium)octane in combination with different anions bromide and bis(trifluoromethanesulphonyl)imide were prepared and then bonded to the surface of 3-mercaptopropyl modified silica materials through the "thiol-ene" click chemistry as stationary phases for hydrophilic interaction chromatography (HILIC). Compared with their monocationic analogues, the dicationic ionic liquids stationary phases presented effective retention and good selectivity for typical hydrophilic compounds under HILIC mode with the column efficiency as high as 130,000 plates/m. Moreover, the influence of different alkyl chain spacer between dications and combined anions on the retention behavior and selectivity of the dicationic ionic liquids stationary phases under HILIC mode was displayed. The results indicated that the longer linkage chain would decrease the hydrophilicity and retention on the dicationic ionic liquid stationary phase, and while differently combined anions had no difference due to the exchangeability under the common HILIC mobile phase with buffer salt. Finally, the retention mechanism was investigated by evaluating the effect of chromatographic factors on retention, including the water content in the mobile phase, the mobile phase pH and buffer salt concentration. The results showed that the dicationic ionic liquids stationary phases presented a mixed-mode retention behavior with HILIC mechanism and anion exchange. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. 76 FR 57951 - Glycine From the People's Republic of China: Continuation of Antidumping Duty Order

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-19

    ... the order is glycine, which is a free- flowing crystalline material, like salt or sugar. Glycine is produced at varying levels of purity and is used as a sweetener/taste enhancer, a buffering agent...

  7. Buffer layers on metal alloy substrates for superconducting tapes

    DOEpatents

    Jia, Quanxi; Foltyn, Stephen R.; Arendt, Paul N.; Groves, James R.

    2004-06-29

    An article including a substrate, a layer of an inert oxide material upon the surface of the substrate, a layer of an amorphous oxide or oxynitride material upon the inert oxide material layer, a layer of an oriented cubic oxide material having a rock-salt-like structure upon the amorphous oxide material layer, and a layer of a SrRuO.sub.3 buffer material upon the oriented cubic oxide material layer is provided together with additional layers such as a HTS top-layer of YBCO directly upon the layer of a SrRuO.sub.3 buffer material layer. With a HTS top-layer of YBCO upon at least one layer of the SrRuO.sub.3 buffer material in such an article, J.sub.c 's of up to 1.3.times.10.sup.6 A/cm.sup.2 have been demonstrated with projected IC's of over 200 Amperes across a sample 1 cm wide.

  8. Buffer layers on metal alloy substrates for superconducting tapes

    DOEpatents

    Jia, Quanxi; Foltyn, Stephen R.; Arendt, Paul N.; Groves, James R.

    2004-10-05

    An article including a substrate, at least one intermediate layer upon the surface of the substrate, a layer of an oriented cubic oxide material having a rock-salt-like structure upon the at least one intermediate layer, and a layer of a SrRuO.sub.3 buffer material upon the oriented cubic oxide material layer is provided together with additional layers such as a HTS top-layer of YBCO directly upon the layer of a SrRuO.sub.3 buffer material layer. With a HTS top-layer of YBCO upon at least one layer of the SrRuO.sub.3 buffer material in such an article, J.sub.c 's of up to 1.3.times.10.sup.6 A/cm.sup.2 have been demonstrated with projected I.sub.c 's of over 200 Amperes across a sample 1 cm wide.

  9. THE KINETICS OF SAPONIFICATION OF IODOACETIC ACID BY SODIUM HYDROXIDE AND BY CERTAIN ALKALINE BUFFER SOLUTIONS

    PubMed Central

    Brdička, R.

    1936-01-01

    1. The rate of the saponification of iodoacetic acid in sodium hydroxide and alkaline buffer solutions yielding glycollic acid was measured by means of Heyrovský's polarographic method. 2. From the bimolecular velocity constants, increasing with the ionic strength of the solution, the Brönsted factor, F, which characterizes the primary salt effect, was calculated. 3. In the borate buffer solutions the monomolecular constants of the saponification were determined which, at values above the pH of neutralization of boric acid, show a proportionality to the concentration of hydroxyl anions. Below the pH of neutralization of boric acid, they are proportional to the concentration of borate anions. PMID:19872968

  10. Determination of melanterite-rozenite and chalcanthite-bonattite equilibria by humidity measurements at 0.1 MPa

    USGS Publications Warehouse

    Chou, I.-Ming; Seal, R.R.; Hemingway, B.S.

    2002-01-01

    Melanterite (FeSO4??7H2O)-rozenite (FeSO4??4H2O) and chalcanthite (CuSO4??5H2O)-bonattite (CuSO4??3H2O) equilibria were determined by humidity measurements at 0.1 MPa. Two methods were used; one is the gas-flow-cell method (between 21 and 98 ??C), and the other is the humidity-buffer method (between 21 and 70 ??C). The first method has a larger temperature uncertainty even though it is more efficient. With the aid of humidity buffers, which correspond to a series of saturated binary salt solutions, the second method yields reliable results as demonstrated by very tight reversals along each humidity buffer. These results are consistent with those obtained by the first method, and also with the solubility data reported in the literature. Thermodynamic analysis of these data yields values of 29.231 ?? 0.025 and 22.593 ?? 0.040 kJ/mol for standard Gibbs free energy of reaction at 298.15 K and 0.1 MPa for melanterite-rozenite and chalcanthite-bonattite equilibria, respectively. The methods used in this study hold great potential for unraveling the thermodynamic properties of sulfate salts involved in dehydration reactions at near ambient conditions.

  11. Extraction and purification of capsaicin from capsicum oleoresin using an aqueous two-phase system combined with chromatography.

    PubMed

    Fan, Yong; Lu, Yan-Min; Yu, Bin; Tan, Cong-Ping; Cui, Bo

    2017-09-15

    Capsaicin was extracted from capsicum oleoresin using an aqueous two-phase system (ATPS) composed of an ethylene oxide-propylene oxide (EOPO) copolymer, salt and ethanol. Capsaicin was concentrated in the top polymer-rich phase. To determine the optimal conditions, the partitioning of capsaicin in the ATPS was investigated, considering a single-factor experiment including the salt concentration, polymer concentration, buffer pH, ethanol concentration, sample loading and extraction duration. Response surface methodology was applied to investigate the effects of the polymer concentration, buffer pH and sample loading on capsaicin partitioning. A capsaicin yield of 95.5% was obtained using the optimal extraction system, which consisted of 16.3% UCON 50-HB-5100/10% K 2 HPO 4 /1% ethanol, a buffer pH of 4.35 and 0.24g of capsicum oleoresin. Capsaicin was purified from the capsaicinoid extract using a two-step macroporous adsorption resin (MAR) method. After purification using non-polar MAR ADS-17, the recovery and purity of capsaicin were 83.7% and 50.3%, respectively. After purification using weakly polar MAR AB-8, the recovery and purity of capsaicin were 88.0% and 85.1%, respectively. Copyright © 2017. Published by Elsevier B.V.

  12. The effect of EDTA, cations, and various buffers on the morphology of erythrocyte membranes: an electron-microscopic study.

    PubMed

    Pinteric, L; Manery, J F; Chaudry, I H; Madapallimattam, G

    1975-05-01

    Membranes of human erythrocytes were prepared by stepwise osmotic hemolysis in Ca2+-free solutions. Examination with the electron microscope after negative staining showed some short, conelike protuberances on the surface of about 20 percent of the ghosts, while 80 percent were round, intact spheres. After Ca2+ treatment, all membranes were round and intact. After exposure to ethylenediaminetetraacetic acid (EDTA) (1.0 mM, pH 7.4), the entire ghost surface was covered with long, thin extrusions called stromalytic forms (about 460 per cell). Their sizes, shapes, and fine structure are described. Exposure to ionic calcium (1.4 times 10-minus 4M) abolished the EDTA-induced stromalytic forms. A second exposure to EDTA reversed this Ca2+ effect. ATP, like EDTA, produced stromalytic forms. EDTA-induced stromalytic forms were also abolished by Zn2+, La3+, and Nd3+ at concentrations of 1-5 times 10-minus 4 M. Mg2+ at 10-minus 2 M was ineffective. Ghosts were prepared by graded lysis in various buffers. Those prepared in phosphate were the most stable and provided consistent EDTA effects and Ca2+ reversal. Ghosts in Tris-HCl showed breakdown unless salt was added. Moderately satisfactory ghosts were also obtained in Hepes-NaOH buffer and salt.

  13. Salt marsh persistence is threatened by predicted sea-level rise

    NASA Astrophysics Data System (ADS)

    Crosby, Sarah C.; Sax, Dov F.; Palmer, Megan E.; Booth, Harriet S.; Deegan, Linda A.; Bertness, Mark D.; Leslie, Heather M.

    2016-11-01

    Salt marshes buffer coastlines and provide critical ecosystem services from storm protection to food provision. Worldwide, these ecosystems are in danger of disappearing if they cannot increase elevation at rates that match sea-level rise. However, the magnitude of loss to be expected is not known. A synthesis of existing records of salt marsh elevation change was conducted in order to consider the likelihood of their future persistence. This analysis indicates that many salt marshes did not keep pace with sea-level rise in the past century and kept pace even less well over the past two decades. Salt marshes experiencing higher local sea-level rise rates were less likely to be keeping pace. These results suggest that sea-level rise will overwhelm most salt marshes' capacity to maintain elevation. Under the most optimistic IPCC emissions pathway, 60% of the salt marshes studied will be gaining elevation at a rate insufficient to keep pace with sea-level rise by 2100. Without mitigation of greenhouse gas emissions this potential loss could exceed 90%, which will have substantial ecological, economic, and human health consequences.

  14. The vesicle-to-micelle transition of phosphatidylcholine vesicles induced by nonionic detergents: effects of sodium chloride, sucrose and urea.

    PubMed

    Walter, A; Kuehl, G; Barnes, K; VanderWaerdt, G

    2000-11-23

    The vesicle-to-micelle transition of egg phosphatidylcholine LUVs induced by octylglucoside was studied in buffers with 0-4 M sodium chloride, sucrose or urea. We used both light scattering and fluorescent probes to follow the lipid-detergent complexes in these buffers. The vesicle-to-micelle transition process was fundamentally the same in each solute. However, the detergent-to-lipid ratio required for micelle formation shifted in ways that depended on the aqueous solute. The partitioning of octylglucoside between the vesicles and the aqueous phase was primarily determined by the change in its critical micelle concentration (cmc) induced by each solute. Specifically, the cmc decreased in high salt and sucrose buffers but increased in high concentrations of urea. Cmc for two additional nonionic detergents, decyl- and dodecyl-maltoside, and three zwittergents (3-12, 3-14 and 3-16) were determined as a function of concentration for each of the solutes. In all cases NaCl and sucrose decreased the solubility of the detergents, whereas urea increased their solubilities. The effects clearly depended on acyl chain length in urea-containing solutions, but this dependence was less clear with increasing NaCl and sucrose concentrations. The contributions of these solutes to solubility and to interfacial interactions in the bilayers, pure and mixed micelles are considered.

  15. Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: pH buffering properties of cheese.

    PubMed

    Upreti, P; Bühlmann, P; Metzger, L E

    2006-03-01

    The pH buffering capacity of cheese is an important determinant of cheese pH. However, the effects of different constituents of cheese on its pH buffering capacity have not been fully clarified. The objective of this study was to characterize the chemical species and chemical equilibria that are responsible for the pH buffering properties of cheese. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), residual lactose (2.4 vs. 0.78%), and salt-to-moisture ratio (6.4 vs. 4.8%) were manufactured. The pH-titration curves for these cheeses were obtained by titrating cheese:water (1:39 wt/wt) dispersions with 1 N HCl, and backtitrating with 1 N NaOH. To understand the role of different chemical equilibria and the respective chemical species in controlling the pH of cheese, pH buffering was modeled mathematically. The 36 chemical species that were found to be relevant for modeling can be classified as cations (Na+, Ca2+, Mg2+), anions (phosphate, citrate, lactate), protein-bound amino acids with a side-chain pKa in the range of 3 to 9 (glutamate, histidine, serine phosphate, aspartate), metal ion complexes (phosphate, citrate, and lactate complexes of Na+, Ca2+, and Mg2+), and calcium phosphate precipitates. A set of 36 corresponding equations was solved to give the concentrations of all chemical species as a function of pH, allowing the prediction of buffering curves. Changes in the calculated species concentrations allowed the identification of the chemical species and chemical equilibria that dominate the pH buffering properties of cheese in different pH ranges. The model indicates that pH buffering in the pH range from 4.5 to 5.5 is predominantly due to a precipitate of Ca and phosphate, and the protonation equilibrium involving the side chains of protein-bound glutamate. In the literature, the precipitate is often referred to as amorphous colloidal calcium phosphate. A comparison of experimental data and model predictions shows that the buffering properties of the precipitate can be explained, assuming that it consists of hydroxyapatite [Ca5(OH)(PO4)3] or Ca3(PO4)2. The pH buffering in the region from pH 3.5 to 4.5 is due to protonation of side-chain carboxylates of protein-bound glutamate, aspartate, and lactate, in order of decreasing significance. In addition, pH buffering between pH 5 to 8 in the backtitration results from the reprecipitation of calcium and phosphate either as CaHPO4 or Ca4H(PO4)3.

  16. A comparison of constant-load and constant-deflection stress-corrosion tests on precracked DCB specimens. [Double Cantilever Beam

    NASA Technical Reports Server (NTRS)

    Dorward, R. C.; Hasse, K. R.

    1978-01-01

    A comparison is made between measurements of stress-corrosion crack propagation made by a constant-load procedure and by a constant-deflection procedure. Precracked double cantilever beam specimens from 7075 aluminum alloy plate were used. The specimens were oriented in such a way that cracking would begin in the short-transverse plane and would propagate in the rolling direction. The specimens were subjected to a buffered salt-chromate solution and a 3.6% synthetic sea salt solution. The measurements were made optically with a binocular microscope. Stress intensities and crack lengths were calculated and crack velocities were obtained. Velocity was plotted against the average calculated stress intensity. Good agreement between the two methods was found for the salt-chromate solution, although some descrepancies were noted for the artificial sea salt solution.

  17. Spatial variability in denitrification rates in an Oregon tidal salt marsh

    EPA Science Inventory

    Modeling denitrification (DeN) is particularly challenging in tidal systems, which play a vital role in buffering adjacent coastal waters from nitrogen inputs. These systems are hydrologically and biogeochemically complex, varying on fine temporal and spatial scales. As part of a...

  18. Effect of the ionic strength of a mobile phase on the chromatographic retention and thermodynamic characteristics of the adsorption of enantiomers of α-phenylcarboxylic acids on a chiral adsorbent with grafted antibiotic eremomycin

    NASA Astrophysics Data System (ADS)

    Reshetova, E. N.

    2017-01-01

    The effect the ionic strength of an aqueous ethanol mobile phase containing buffer salt has the on retention and thermodynamics of adsorption of optical isomers of some α-phenylcarboxylic acids on chiral adsorbent Nautilus-E with grafted antibiotic eremomycin is investigated. It is shown that ion exchange processes participate in the adsorption of enantiomers of α-phenylcarboxylic acids. It is established that electrostatic interactions contribute to the retention of enantiomers of α-phenylcarboxylic acids and affect selectivity only slightly. The dependences of retention characteristics, selectivity, and thermodynamic parameters on the concentration of the buffer salt in the eluent are determined. A statistical analysis of enthalpy-entropy compensation is performed, and the compensation effect is shown to be true. It is found that the points corresponding to the investigated adsorbates are distributed over the compensation dependence according to the spatial structural characteristics of molecules.

  19. Fe-SAPONITE and Chlorite Growth on Stainless Steel in Hydrothermal Engineered Barrier Experiments

    NASA Astrophysics Data System (ADS)

    Cheshire, M. C.; Caporuscio, F. A.; McCarney, M.

    2012-12-01

    The United States recently has initiated the Used Fuel Disposition campaign to evaluate various generic geological repositories for the disposal of high-level, spent nuclear fuel within environments ranging from hard-rock, salt/clay, to deep borehole settings. Previous work describing Engineered Barrier Systems (EBS) for repositories focused on low temperature and pressure conditions. The focus of this experimental work is to characterize the stability and alteration of a bentonite-based EBS with different waste container materials in brine at higher heat loads and pressures. All experiments were run at ~150 bar and 125 to 300 C for ~1 month. Unprocessed bentonite from Colony, Wyoming was used in the experiments as the clay buffer material. The redox conditions for each system were buffered along the magnetite-iron oxygen fugacity univariant curve using Fe3O4 and Feo filings. A K-Na-Ca-Cl-based salt solution was chosen to replicate deep groundwater compositions. The experimental mixtures were 1) salt solution-clay; 2) salt solution -clay-304 stainless steel; and 3) salt solution -clay-316 stainless steel with a water/bentonite ratio of ~9. Mineralogy and aqueous geochemistry of each experiment was evaluated to monitor the reactions that took place. No smectite illitization was observed in these reactions. However, it appears that K-smectite was produced, possibly providing a precursor to illitization. It is unclear whether reaction times were sufficient for bentonite illitization at 212 and 300 C or whether conditions conducive to illite formation were obtained. The more notable clay mineral reactions occurred at the stainless steel surfaces. Authigenic chlorite and Fe-saponite grew with their basal planes near perpendicular to the steel plate, forming a 10 - 40 μm thick 'corrosion' layer. Partial dissolution of the steel plates was the likely iron source for chlorite/saponite formation; however, dissolution of the Feo/Fe3O4 may also have acted as an iron source, with the steel plates acting as a substrate for chlorite/saponite growth. Trace amounts of pyrite in the bentonite appeared to have reacted to form H2S gas and pentlandite ((Ni,Fe)8S9). Mineral growth on the waste containers was influenced by the container, buffer, and fluid compositions, in addition to pressure and temperature conditions. No significant mineralogical changes were apparent away from the steel-smectite interface. Results of this research show that the waste container may act as a substrate for mineral growth in response to corrosion. However, it is presently unknown whether chlorite and Fe-saponite will act as passivating agents or whether their presence will facilitate further corrosion of the waste containers. The role of these Fe-rich minerals on the stability of steel canisters at elevated heat loads is currently under investigation. LA-UR-12-23845

  20. Atom-scale covalent electrochemical modification of single-layer graphene on SiC substrates by diaryliodonium salts

    DOE PAGES

    Gearba, Raluca I.; Mueller, Kory M.; Veneman, Peter A.; ...

    2015-05-09

    Owing to its high conductivity, graphene holds promise as an electrode for energy devices such as batteries and photovoltaics. However, to this end, the work function and doping levels in graphene need to be precisely tuned. One promising route for modifying graphene’s electronic properties is via controlled covalent electrochemical grafting of molecules. We show that by employing diaryliodonium salts instead of the commonly used diazonium salts, spontaneous functionalization is avoided. This then allows for precise tuning of the grafting density. Moreover, by employing bis(4-nitrophenyl)iodonium(III) tetrafluoroborate (DNP) salt calibration curves, the surface functionalization density (coverage) of glassy carbon was controlled usingmore » cyclic voltammetry in varying salt concentrations. These electro-grafting conditions and calibration curves translated directly over to modifying single layer epitaxial graphene substrates (grown on insulating 6H-SiC (0 0 0 1)). In addition to quantifying the functionalization densities using electrochemical methods, samples with low grafting densities were characterized by low-temperature scanning tunneling microscopy (LT-STM). We show that the use of buffer-layer free graphene substrates is required for clear observation of the nitrophenyl modifications. Furthermore, atomically-resolved STM images of single site modifications were obtained, showing no preferential grafting at defect sites or SiC step edges as supposed previously in the literature. Most of the grafts exhibit threefold symmetry, but occasional extended modifications (larger than 4 nm) were observed as well.« less

  1. EFFECTS OF NUTRIENT LOADING ON BIOGEOCHEMICAL AND MICROBIAL PROCESSES IN A NEW ENGLAND SALT MARSH

    EPA Science Inventory

    Coastal marshes represent an important transitional zone between uplands and estuaries. One important function of marshes is to assimilate nutrient inputs from uplands, thus providing a buffer for anthropogenic nutrient loads. We examined the effects of nitrogen (N) and phosphoru...

  2. Effect of High Hydrostatic Pressure Combined with Moderate Heat to Inactivate Pressure-Resistant Bacteria in Water-Boiled Salted Duck.

    PubMed

    Ye, Keping; Feng, Yulin; Wang, Kai; Bai, Yun; Xu, Xinglian; Zhou, Guanghong

    2015-06-01

    The objective of this work was to study the effect of high hydrostatic pressure combined with moderate heat to inactivate pressure-resistant bacteria in water-boiled salted duck meat (WBSDM), and to establish suitable procedures to improve the quality of WBSDM. The conditions (300 MPa/60 °C, 400 MPa/60 °C, and 500 MPa/50 °C) effectively inactivated the pressure-resistant bacteria (Bacillus cereus and Staphylococcus warneri) in WBSDM. Although more pressure-resistant than S. warneri, the above treatment conditions inactivated B. cereus more than 10(7) CFU/mL in buffer, and more than 10(6) CFU/g in WBSDM, and did not cause any changes in color, texture, or moisture content of products. The interaction between pressure and temperature is a more significant factor than only pressure in inactivating both B. cereus and S. warneri, the treatment of WBSDM at 400 MPa/ 60 °C/ 10 min is the most practical condition for postprocess of WBSDM after cooking. © 2015 Institute of Food Technologists®

  3. Separation of proteins by hydrophobic interaction chromatography at low salt concentration.

    PubMed

    Kato, Yoshio; Nakamura, Koji; Kitamura, Takashi; Moriyama, Hiroyuki; Hasegawa, Masazumi; Sasaki, Hiroo

    2002-09-20

    We investigated protein separation by hydrophobic interaction chromatography (HIC) at low salt concentration on the supports of various hydrophobicities. Hydrophobic proteins could be successfully separated with more than 90% recovery by gradient elution of ammonium sulfate from 0.3-0.5 M to 0 in 50 mM phosphate buffer (pH 6.8) by using supports whose hydrophobicities were properly adjusted individually for each protein. Satisfactory results were also obtained by isocratic elution without ammonium sulfate and gradient elution of ethanol from 0 to 10%. HIC at low salt concentration was compatible with other modes of liquid chromatography like ion-exchange chromatography. On the other hand, it was not successful to separate hydrophilic proteins at low salt concentration. Recoveries of hydrophilic proteins decreased before they were retained enough as support hydrophobicity increased. Therefore, it is inevitable to use a higher concentration of salt, e.g., 1-2 M ammonium sulfate, on hydrophilic or moderately hydrophobic support in order to retain hydrophilic proteins without decrease in recovery.

  4. Stabilization of photosystem II reaction centers: influence of bile salt detergents and low pH.

    PubMed

    Gall, B; Scheer, H

    1998-07-17

    Rapid deterioration of samples is a major obstacle in research on the isolated reaction center of photosystem II. Its stability was tested systematically using a wide range of detergents, varying pH and temperature. Stability and activity did not depend on ionic properties of detergents or on critical micellar concentration. However, both were significantly increased by bile salt detergents in the dark as well as in the light. Low pH (5.5) and low temperature further improved stability. The results suggest that in particular the zwitterionic bile salt detergent, CHAPS, in pH 5.5 buffers is a very useful detergent and even superior to dodecylmaltoside for work with photosystem II reaction centers.

  5. Halophilic Nuclease from a Moderately Halophilic Micrococcus varians

    PubMed Central

    Kamekura, Masahiro; Onishi, Hiroshi

    1974-01-01

    The moderately halophilic bacterium Micrococcus varians, isolated from soy sauce mash, produced extracellular nuclease when cultivated aerobically in media containing 1 to 4 M NaCl or KCl. The enzyme, purified to an electrophoretically homogeneous state, had both ribonuclease and deoxyribonuclease activities. The nuclease had maximal activity in the presence of 2.9 M NaCl or 2.1 M KCl at 40 C. The enzymatic activity was lost by dialysis against low-salt buffer, whereas when the inactivated enzyme was dialyzed against 3.4 M NaCl buffer as much as 77% of the initial activity could be restored. Images PMID:4852218

  6. Release of salicylic acid, diclofenac acid and diclofenac acid salts from isotropic and anisotropic nonionic surfactant systems across rat skin.

    PubMed

    Gabboun, N H; Najib, N M; Ibrahim, H G; Assaf, S

    2001-01-05

    Release of salicylic acid, diclofenac acid, diclofenac diethylamine and diclofenac sodium, from lyotropic structured systems, namely; neat and middle liquid crystalline phases, across mid-dorsal hairless rat skin into aqueous buffer were studied. Release results were compared with those from the isotropic systems. The donor systems composed of the surfactant polyoxyethylene (20) isohexadecyl ether, HCl buffer of pH 1 or distilled water and the specific drug. High performance liquid chromatography (HPLC) methods were used to monitor the transfer of the drugs across the skin barrier. Results indicated that the rate-determining step in the transport process was the release of the drug from the specified donor system. Further, apparent zero order release was demonstrated with all systems. Except for diclofenac sodium, drug fluxes decreased as the donor medium changed from isotropic to anisotropic. The decrease in fluxes was probably due to the added constrains on the movement of drug molecules. By changing the anisotropic donor medium from neat to middle phase, drug flux decreased in case of salicylic acid and diclofenac sodium. In the mean time, flux increased in case of the diethylamine salt and appeared nearly similar in case of diclofenac acid. Rates of drug transfer across the skin from the anisotropic donors seemed to be largely controlled by the entropy contribution to the transport process. The type and extent of drug-liquid crystal interactions probably influenced the latter.

  7. Direct Observation of Protein Microcrystals in Crystallization Buffer by Atmospheric Scanning Electron Microscopy

    PubMed Central

    Maruyama, Yuusuke; Ebihara, Tatsuhiko; Nishiyama, Hidetoshi; Konyuba, Yuji; Senda, Miki; Numaga-Tomita, Takuro; Senda, Toshiya; Suga, Mitsuo; Sato, Chikara

    2012-01-01

    X-ray crystallography requires high quality crystals above a given size. This requirement not only limits the proteins to be analyzed, but also reduces the speed of the structure determination. Indeed, the tertiary structures of many physiologically important proteins remain elusive because of the so-called “crystallization bottleneck”. Once microcrystals have been obtained, crystallization conditions can be optimized to produce bigger and better crystals. However, the identification of microcrystals can be difficult due to the resolution limit of optical microscopy. Electron microscopy has sometimes been utilized instead, with the disadvantage that the microcrystals usually must be observed in vacuum, which precludes the usage for crystal screening. The atmospheric scanning electron microscope (ASEM) allows samples to be observed in solution. Here, we report the use of this instrument in combination with a special thin-membrane dish with a crystallization well. It was possible to observe protein crystals of lysozyme, lipase B and a histone chaperone TAF-Iβ in crystallization buffers, without the use of staining procedures. The smallest crystals observed with ASEM were a few μm in width, and ASEM can be used with non-transparent solutions. Furthermore, the growth of salt crystals could be monitored in the ASEM, and the difference in contrast between salt and protein crystals made it easy to distinguish between these two types of microcrystals. These results indicate that the ASEM could be an important new tool for the screening of protein microcrystals. PMID:22949879

  8. Direct observation of protein microcrystals in crystallization buffer by atmospheric scanning electron microscopy.

    PubMed

    Maruyama, Yuusuke; Ebihara, Tatsuhiko; Nishiyama, Hidetoshi; Konyuba, Yuji; Senda, Miki; Numaga-Tomita, Takuro; Senda, Toshiya; Suga, Mitsuo; Sato, Chikara

    2012-01-01

    X-ray crystallography requires high quality crystals above a given size. This requirement not only limits the proteins to be analyzed, but also reduces the speed of the structure determination. Indeed, the tertiary structures of many physiologically important proteins remain elusive because of the so-called "crystallization bottleneck". Once microcrystals have been obtained, crystallization conditions can be optimized to produce bigger and better crystals. However, the identification of microcrystals can be difficult due to the resolution limit of optical microscopy. Electron microscopy has sometimes been utilized instead, with the disadvantage that the microcrystals usually must be observed in vacuum, which precludes the usage for crystal screening. The atmospheric scanning electron microscope (ASEM) allows samples to be observed in solution. Here, we report the use of this instrument in combination with a special thin-membrane dish with a crystallization well. It was possible to observe protein crystals of lysozyme, lipase B and a histone chaperone TAF-Iβ in crystallization buffers, without the use of staining procedures. The smallest crystals observed with ASEM were a few μm in width, and ASEM can be used with non-transparent solutions. Furthermore, the growth of salt crystals could be monitored in the ASEM, and the difference in contrast between salt and protein crystals made it easy to distinguish between these two types of microcrystals. These results indicate that the ASEM could be an important new tool for the screening of protein microcrystals.

  9. Salt-Dependent DNA-DNA Spacings in Intact Bacteriophage lambda Reflect Relative Importance of DNA Self-Repulsion and Bending Energies

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

    X Qiu; D Rau; V Parsegian

    2011-12-31

    Using solution synchrotron x-ray scattering, we measure the variation of DNA-DNA d spacings in bacteriophage {lambda} with mono-, di-, and polyvalent salt concentrations, for wild-type [48.5 x 10{sup 3} base pairs (bp)] and short-genome-mutant (37.8 kbp) strains. From the decrease in d spacings with increasing salt, we deduce the relative contributions of DNA self-repulsion and bending to the energetics of packaged phage genomes. We quantify the DNA-DNA interaction energies within the intact phage by combining the measured d spacings in the capsid with measurements of osmotic pressure in DNA assemblies under the same salt conditions in bulk solution. In themore » commonly used Tris-Mg buffer, the DNA-DNA interaction energies inside the phage capsids are shown to be about 1 kT/bp, an order of magnitude larger than the bending energies.« less

  10. Zebrafish as a Model for Systems Medicine R&D: Rethinking the Metabolic Effects of Carrier Solvents and Culture Buffers Determined by (1)H NMR Metabolomics.

    PubMed

    Akhtar, Muhammad T; Mushtaq, Mian Y; Verpoorte, Robert; Richardson, Michael K; Choi, Young H

    2016-01-01

    Zebrafish is a frequently employed model organism in systems medicine and biomarker discovery. A crosscutting fundamental question, and one that has been overlooked in the field, is the "system-wide" (omics) effects induced in zebrafish by metabolic solvents and culture buffers. Indeed, any bioactivity or toxicity test requires that the target compounds are dissolved in an appropriate nonpolar solvent or aqueous media. It is important to know whether the solvent or the buffer itself has an effect on the zebrafish model organism. We evaluated the effects of two organic carrier solvents used in research with zebrafish, as well as in drug screening: dimethyl sulfoxide (DMSO) and ethanol, and two commonly used aqueous buffers (egg water and Hank's balanced salt solution). The effects of three concentrations (0.01, 0.1, and 1%) of DMSO and ethanol were tested in the 5-day-old zebrafish embryo using proton nuclear magnetic resonance ((1)H NMR) based metabolomics. DMSO (1% and 0.1%, but not 0.01%) exposure significantly decreased the levels of adenosine triphosphate (ATP), betaine, alanine, histidine, lactate, acetate, and creatine (p < 0.05). By contrast, ethanol exposure did not alter the embryos' metabolome at any concentration tested. The two different aqueous media noted above impacted the zebrafish embryo metabolome as evidenced by changes in valine, alanine, lactate, acetate, betaine, glycine, glutamate, adenosine triphosphate, and histidine. These results show that DMSO has greater effects on the embryo metabolome than ethanol, and thus is used with caution as a carrier solvent in zebrafish biomarker research and oral medicine. Moreover, the DMSO concentration should not be higher than 0.01%. Careful attention is also warranted for the use of the buffers egg water and Hank's balanced salt solution in zebrafish. In conclusion, as zebrafish is widely used as a model organism in life sciences, metabolome changes induced by solvents and culture buffers warrant further attention for robust systems science, and precision biomarkers that will stand the test of time.

  11. On the Preparation of Buffer Solutions.

    ERIC Educational Resources Information Center

    Thomson, Bruce M.; Kessick, Michael A.

    1981-01-01

    Presents a method, suitable for use on programmable calculators, which allows calculation of the pH and ionic strength (I) of a mixed solution of salts of an acid or amounts necessary to produce a solution of a particular pH and I. Includes limitations when using the calculations described. (SK)

  12. EFFECTS OF NITROGEN SOURCE ON CRUDE OIL BIODEGRADATION

    EPA Science Inventory

    The effects of NH4Cl and KNO3 on biodegradation of light Arabian crude oil by an oil-degrading enrichment culture were studied in respirometers. In poorly buffered sea salts medium, the pH decreased dramatically in cultures that contained NH4Cl, b...

  13. Fermentation cover brine reformulation for cucumber processing with low salt to reduce bloater defect

    USDA-ARS?s Scientific Manuscript database

    Reformulation of calcium chloride cover brine for cucumber fermentation was explored as a mean to minimize the incidence of bloater defect. This study particularly focused on cover brine supplementation with calcium hydroxide, sodium chloride (NaCl), and acids to enhance buffer capacity, inhibit the...

  14. 40 CFR 180.920 - Inert ingredients used pre-harvest; exemptions from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... di- and monohydrogen phosphate esters and the corresponding ammonium, calcium, magnesium, monoethanolamine, potassium, sodium, and zinc salts of the phosphate esters; minimum oxyethylene content is 2 moles.... 14433-76-2) Emulsifier, solvent, cosolvent Diammonium phosphate (CAS Reg. No. 7783-28-0) Buffer...

  15. 40 CFR 180.920 - Inert ingredients used pre-harvest; exemptions from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... di- and monohydrogen phosphate esters and the corresponding ammonium, calcium, magnesium, monoethanolamine, potassium, sodium, and zinc salts of the phosphate esters; minimum oxyethylene content is 2 moles... phosphate (CAS Reg. No. 7783-28-0) Buffer, surfactant dibenzylidene sorbitol (32647-67-9) Thinning agent...

  16. 40 CFR 180.920 - Inert ingredients used pre-harvest; exemptions from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... di- and monohydrogen phosphate esters and the corresponding ammonium, calcium, magnesium, monoethanolamine, potassium, sodium, and zinc salts of the phosphate esters; minimum oxyethylene content is 2 moles... phosphate (CAS Reg. No. 7783-28-0) Buffer, surfactant dibenzylidene sorbitol (32647-67-9) Thinning agent...

  17. Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

    PubMed Central

    Sung, Nackmoon; Collins, Michael T.

    2000-01-01

    Low pH and salt are two factors contributing to the inactivation of bacterial pathogens during a 60-day curing period for cheese. The kinetics of inactivation for Mycobacterium avium subsp. paratuberculosis strains ATCC 19698 and Dominic were measured at 20°C under different pH and NaCl conditions commonly used in processing cheese. The corresponding D values (decimal reduction times; the time required to kill 1 log10 concentration of bacteria) were measured. Also measured were the D values for heat-treated and nonheated M. avium subsp. paratuberculosis in 50 mM acetate buffer (pH 5.0, 2% [wt/vol] NaCl) and a soft white Hispanic-style cheese (pH 6.0, 2% [wt/vol] NaCl). Samples were removed at various intervals until no viable cells were detected using the radiometric culture method (BACTEC) for enumeration of M. avium subsp. paratuberculosis. NaCl had little or no effect on the inactivation of M. avium subsp. paratuberculosis, and increasing NaCl concentrations were not associated with decreasing D values (faster killing) in the acetate buffer. Lower pHs, however, were significantly correlated with decreasing D values of M. avium subsp. paratuberculosis in the acetate buffer. The D values for heat-treated M. avium subsp. paratuberculosis ATCC 19698 in the cheese were higher than those predicted by studies done in acetate buffer. The heat-treated M. avium subsp. paratuberculosis strains had lower D values than the nonheated cells (faster killing) both in the acetate buffer (pH 5, 2% [wt/vol] NaCl) and in the soft white cheese. The D value for heat-treated M. avium subsp. paratuberculosis ATCC 19698 in the cheese (36.5 days) suggests that heat treatment of raw milk coupled with a 60-day curing period will inactivate about 103 cells of M. avium subsp. paratuberculosis per ml. PMID:10742208

  18. Salting-Out Assisted Liquid-Liquid Extraction for Quantification of Febuxostat in Plasma Using RP-HPLC and Its Pharmacokinetic Application.

    PubMed

    Tandel, Devang; Shah, Purvi; Patel, Kalpana; Thakkar, Vaishali; Patel, Kirti; Gandhi, Tejal

    2016-11-01

    A rapid and sensitive reversed-phase high-performance liquid chromatography (HPLC) method using novel salting-out assisted liquid-liquid extraction technique has been developed for the quantitative determination of febuxostat (FEB), used for the treatment of gout, in rat plasma. The method was validated according to US FDA guideline. Separation was achieved using a Phenomenex Luna-C 18 (250 × 4.60 mm, 5 µm) column and mobile phase composed of potassium dihydrogen orthophosphate buffer 25 mM, adjusted to pH 6.8 with triethylamine:methanol in a ratio of 35:65 (v/v) showing retention time 5.56 and 8.86 min for FEB and internal standard, respectively. The optimal salting-out parameters; 1 mL of acetonitrile and 200 µL of 2 M ammonium acetate salt showed extraction recovery >90% for FEB from plasma. This extraction procedure afforded clear samples resulting in convenient and cost-saving procedure and showed good linear relationship (r > 0.9997) between peak area ratio and concentration from 0.3 to 20 µg/mL. The results of pharmacokinetic study showed that absorption profile of spherical agglomerate of FEB compared to marketed formulation was higher indicating greater systemic absorption. In conclusion, the developed SALLE-HPLC method with simple ultraviolet detection offered a number of advantages including good quantitative ability, wide linear range, high recovery, short analysis time as well as low cost. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Investigation of salt formation between memantine and pamoic acid: Its exploitation in nanocrystalline form as long acting injection.

    PubMed

    Mittapelly, Naresh; Rachumallu, Ramakrishna; Pandey, Gitu; Sharma, Shweta; Arya, Abhishek; Bhatta, Rabi Shankar; Mishra, Prabhat Ranjan

    2016-04-01

    In the present work, we prepared memantine-pamoic acid (MEM-PAM) salt by counter ion exchange in the aqueous phase to reduce the water solubility of MEM hydrochloride (native form) to make it suitable for long acting injection. The ratio of MEM to PAM in salt formation was optimized to maximize the loading efficiency and complexation efficiency. The 2:1 molar ratio of MEM to PAM salt form displayed nearly 95% complexation efficiency and 50% drug loading. The solubility was decreased by a ∼1250 folds. Thermo Gravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC), and Powder X-ray Diffraction Analysis (PXRD) studies revealed the formation of new solid phase. Additionally, Nuclear Magnetic Resonance (NMR) spectroscopy confirmed the anhydrous nature of the salt form. Through Fourier transformation infrared spectroscopy (FT-IR) we identified the molecular interactions. Further, the microcrystals of the salt were transformed into nanocrystals (NCs) using high pressure homogenization. The particle size distribution and atomic force microscopy confirmed the monodispersed and spherical shape of the NCs. The in vitro dissolution studies were performed under sink condition in phosphate buffer saline pH 6.8. The results of MTT assay in murine fibroblast 3T3 cell line show that the NCs were less cytotoxic and more tolerable than plain MEM HCl. The in vivo performance of NCs administered as i.m. injection at three different doses in female Sprague-Dawley rats showed that the plasma levels lasted till the 24th day of the study. The pharmacokinetic parameters AUC0-∞ and Cmax increased linearly with increasing dose. Therefore, the results suggest that injectable NCs could represent a therapeutic alternative for the treatment of AD. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Conductive layer for biaxially oriented semiconductor film growth

    DOEpatents

    Findikoglu, Alp T.; Matias, Vladimir

    2007-10-30

    A conductive layer for biaxially oriented semiconductor film growth and a thin film semiconductor structure such as, for example, a photodetector, a photovoltaic cell, or a light emitting diode (LED) that includes a crystallographically oriented semiconducting film disposed on the conductive layer. The thin film semiconductor structure includes: a substrate; a first electrode deposited on the substrate; and a semiconducting layer epitaxially deposited on the first electrode. The first electrode includes a template layer deposited on the substrate and a buffer layer epitaxially deposited on the template layer. The template layer includes a first metal nitride that is electrically conductive and has a rock salt crystal structure, and the buffer layer includes a second metal nitride that is electrically conductive. The semiconducting layer is epitaxially deposited on the buffer layer. A method of making such a thin film semiconductor structure is also described.

  1. Fish Viruses: Buffers and Methods for Plaquing Eight Agents Under Normal Atmosphere

    PubMed Central

    Wolf, Ken; Quimby, M. C.

    1973-01-01

    A universal procedure was sought for plaque assay of eight fish viruses (bluegill myxovirus, channel catfish virus, eel virus, Egtved virus, infectious hematopoietic necrosis virus, infectious pancreatic necrosis virus, lymphocystis virus, and the agent of spring viremia of carp (Rhabdovirus carpio), in dish cultures of various fish cells. Eagle minimal essential medium with sodium bicarbonate-CO2 buffer (Earle’s salt solution) was compared with minimal essential medium buffered principally with tris (hydroxymethyl)aminomethane or N-2-hydroxyethylpiperazine-N′-2′-ethanesulfonic acid at a pH or in the range of 7.6 to 8.0 depending upon temperature. Five fish cell lines collectively capable of replicating all fish viruses thus far isolated were tested and quantitatively found to grow comparably well in the three media. Two-phase (gel-liquid) media incorporating the various buffer systems allowed plaquing at 15 to 33 C either in partial pressures of CO2 or in normal atmosphere, but greater efficiency and sensitivity were obtained with the organic buffers, and, overall, the best results were obtained with tris(hydroxymethyl)aminomethane. Epizootiological data, specific fish cell line response, and plaque morphology permit presumptive identification of most of the agents. At proper pH, use of organic buffers obviates the need for CO2 incubators. Images PMID:4349252

  2. The muscle protein dysferlin accumulates in the Alzheimer brain

    PubMed Central

    Palamand, Divya; Strider, Jeff; Milone, Margherita; Pestronk, Alan

    2006-01-01

    Dysferlin is a transmembrane protein that is highly expressed in muscle. Dysferlin mutations cause limb-girdle dystrophy type 2B, Miyoshi myopathy and distal anterior compartment myopathy. Dysferlin has also been described in neural tissue. We studied dysferlin distribution in the brains of patients with Alzheimer disease (AD) and controls. Twelve brains, staged using the Clinical Dementia Rating were examined: 9 AD cases (mean age: 85.9 years and mean disease duration: 8.9 years), and 3 age-matched controls (mean age: 87.5 years). Dysferlin is a cytoplasmic protein in the pyramidal neurons of normal and AD brains. In addition, there were dysferlin-positive dystrophic neurites within Aβ plaques in the AD brain, distinct from tau-positive neurites. Western blots of total brain protein (RIPA) and sequential extraction buffers (high salt, high salt/Triton X-100, SDS and formic acid) of increasing protein extraction strength were performed to examine solubility state. In RIPA fractions, dysferlin was seen as 230–272 kDa bands in normal and AD brains. In serial extractions, there was a shift of dysferlin from soluble phase in high salt/Triton X-100 to the more insoluble SDS fraction in AD. Dysferlin is a new protein described in the AD brain that accumulates in association with neuritic plaques. In muscle, dysferlin plays a role in the repair of muscle membrane damage. The accumulation of dysferlin in the AD brain may be related to the inability of neurons to repair damage due to Aβ deposits accumulating in the AD brain. PMID:17024495

  3. Impact of salt form and molecular weight of chitosan on swelling and drug release from chitosan matrix tablets.

    PubMed

    Huanbutta, Kampanart; Cheewatanakornkool, Kamonrak; Terada, Katsuhide; Nunthanid, Jurairat; Sriamornsak, Pornsak

    2013-08-14

    Magnetic resonance imaging (MRI) and gravimetric techniques were used to assess swelling and erosion behaviors of hydrophilic matrix tablets made of chitosan. The impact of salt form, molecular weight (MW) and dissolution medium on swelling behavior and drug (theophylline) release was studied. The matrix tablets made of chitosan glycolate (CGY) showed the greatest swelling in both acid and neutral media, compared to chitosan aspartate, chitosan glutamate and chitosan lactate. MRI illustrated that swelling region of CGY in both media was not different in the first 100 min but glassy region (dry core) in 0.1N HCl was less than in pH 6.8 buffer. The tablets prepared from chitosan with high MW swelled greater than those of low MW. Moreover, CGY can delay drug release in the acid condition due to thick swollen gel and low erosion rate. Therefore, CGY may be suitably applied as sustained drug release polymer or enteric coating material. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. High-performance liquid chromatography analysis methods developed for quantifying enzymatic esterification of flavonoids in ionic liquids.

    PubMed

    Lue, Bena-Marie; Guo, Zheng; Xu, Xuebing

    2008-07-11

    Methods using reversed-phase high-performance liquid chromatography (RP-HPLC) with ELSD were investigated to quantify enzymatic reactions of flavonoids with fatty acids in the presence of diverse room temperature ionic liquids (RTILs). A buffered salt (preferably triethylamine-acetate) was found essential for separation of flavonoids from strongly polar RTILs, whereby RTILs were generally visible as two major peaks identified based on an ion-pairing/exchanging hypothesis. C8 and C12 stationary phases were optimal while mobile phase pH (3-7) had only a minor influence on separation. The method developed was successfully applied for primary screening of RTILs (>20), with in depth evaluation of substrates in 10 RTILs, for their evaluation as reaction media.

  5. Design of Phosphonium-Type Zwitterion as an Additive to Improve Saturated Water Content of Phase-Separated Ionic Liquid from Aqueous Phase toward Reversible Extraction of Proteins

    PubMed Central

    Ito, Yoritsugu; Kohno, Yuki; Nakamura, Nobuhumi; Ohno, Hiroyuki

    2013-01-01

    We designed phosphonium-type zwitterion (ZI) to control the saturated water content of separated ionic liquid (IL) phase in the hydrophobic IL/water biphasic systems. The saturated water content of separated IL phase, 1-butyl-3-methyimidazolium bis(trifluoromethanesulfonyl)imide, was considerably improved from 0.4 wt% to 62.8 wt% by adding N,N,N-tripentyl-4-sulfonyl-1-butanephosphonium-type ZI (P555C4S). In addition, the maximum water content decreased from 62.8 wt% to 34.1 wt% by increasing KH2PO4/K2HPO4 salt content in upper aqueous phosphate buffer phase. Horse heart cytochrome c (cyt.c) was dissolved selectively in IL phase by improving the water content of IL phase, and spectroscopic analysis revealed that the dissolved cyt.c retained its higher ordered structure. Furthermore, cyt. c dissolved in IL phase was re-extracted again from IL phase to aqueous phase by increasing the concentration of inorganic salts of the buffer solution. PMID:24013379

  6. Isolation of isoelectrically pure cholera toxin for crystallization

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

    Spangler, B.D.; Westbrook, E.M.

    1989-01-01

    We have determined that the failure of cholera toxin to crystallize well results from its isoelectric heterogeneity, which is probably due to a post-translational process such as deamidation of its B subunit. Every sample of cholera toxin we have examined from commercial or academic suppliers has been heterogeneous; heterogeneous cholera toxin does not crystallize satisfactorily. We have overcome this problem by using ion-exchange fast protein liquid chromatography (FPLC) to obtain an isoelectrically homogeneous species of cholera toxin. Homogeneous cholera toxin crystallizes readily, forming single, nonmosaic crystals suitable for x-ray diffraction studies. For this process, protein was applied to a MonoQmore » ion-exchange column, then eluted with an isocratic low salt buffer followed by a linear salt gradient (0-100 mM NaCl). Column fractions were analyzed on isoelectric focusing gels, and those fractions containing the desired homogeneous species were pooled and concentrated. Crystals formed within 24 to 48 hours in a MOPS/PEG buffer, which made use of slow isoelectric precipitation to induce crystallization. 23 refs., 6 figs.« less

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

    Reilly, Sean Douglas; Smith, Paul Herrick; Jarvinen, Gordon D.

    Understanding the water solubility of plutonium and uranium compounds and residues at TA-55 is necessary to provide a technical basis for appropriate criticality safety, safety basis and accountability controls. Individual compound solubility was determined using published solubility data and solution thermodynamic modeling. Residue solubility was estimated using a combination of published technical reports and process knowledge of constituent compounds. The scope of materials considered includes all compounds and residues at TA-55 as of March 2016 that contain Pu-239 or U-235 where any single item in the facility has more than 500 g of nuclear material. This analysis indicates that themore » following materials are not appreciably soluble in water: plutonium dioxide (IDC=C21), plutonium phosphate (IDC=C66), plutonium tetrafluoride (IDC=C80), plutonium filter residue (IDC=R26), plutonium hydroxide precipitate (IDC=R41), plutonium DOR salt (IDC=R42), plutonium incinerator ash (IDC=R47), uranium carbide (IDC=C13), uranium dioxide (IDC=C21), U 3O 8 (IDC=C88), and uranium filter residue (IDC=R26). This analysis also indicates that the following materials are soluble in water: plutonium chloride (IDC=C19) and uranium nitrate (IDC=C52). Equilibrium calculations suggest that PuOCl is water soluble under certain conditions, but some plutonium processing reports indicate that it is insoluble when present in electrorefining residues (R65). Plutonium molten salt extraction residues (IDC=R83) contain significant quantities of PuCl 3, and are expected to be soluble in water. The solubility of the following plutonium residues is indeterminate due to conflicting reports, insufficient process knowledge or process-dependent composition: calcium salt (IDC=R09), electrorefining salt (IDC=R65), salt (IDC=R71), silica (IDC=R73) and sweepings/screenings (IDC=R78). Solution thermodynamic modeling also indicates that fire suppression water buffered with a commercially-available phosphate buffer would significantly reduce the solubility of PuCl 3 by the precipitation of PuPO 4.« less

  8. In vitro degradation of ZM21 magnesium alloy in simulated body fluids.

    PubMed

    Witecka, Agnieszka; Bogucka, Aleksandra; Yamamoto, Akiko; Máthis, Kristián; Krajňák, Tomáš; Jaroszewicz, Jakub; Święszkowski, Wojciech

    2016-08-01

    In vitro degradation behavior of squeeze cast (CAST) and equal channel angular pressed (ECAP) ZM21 magnesium alloy (2.0wt% Zn-0.98wt% Mn) was studied using immersion tests up to 4w in three different biological environments. Hanks' Balanced Salt Solution (Hanks), Earle's Balanced Salt Solution (Earle) and Eagle minimum essential medium supplemented with 10% (v/v) fetal bovine serum (E-MEM+10% FBS) were used to investigate the effect of carbonate buffer system, organic compounds and material processing on the degradation behavior of the ZM21 alloy samples. Corrosion rate of the samples was evaluated by their Mg(2+) ion release, weight loss and volume loss. In the first 24h, the corrosion rate sequence of the CAST samples was as following: Hanks>E-MEM+10% FBS>Earle. However, in longer immersion periods, the corrosion rate sequence was Earle>E-MEM+10% FBS≥Hanks. Strong buffering effect provided by carbonate buffer system helped to maintain the pH avoiding drastic increase of the corrosion rate of ZM21 in the initial stage of immersion. Organic compounds also contributed to maintain the pH of the fluid. Moreover, they adsorbed on the sample surface and formed an additional barrier on the insoluble salt layer, which was effective to retard the corrosion of CAST samples. In case of ECAP, however, this effect was overcome by the occurrence of strong localized corrosion due to the lower pH of the medium. Corrosion of ECAP samples was much greater than that of CAST, especially in Hanks, due to higher sensitivity of ECAP to localized corrosion and the presence of Cl(-). The present work demonstrates the importance of using an appropriate solution for a reliable estimation of the degradation rate of Mg-base degradable implants in biological environments, and concludes that the most appropriate solution for this purpose is E-MEM+10% FBS, which has the closest chemical composition to human blood plasma. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Ex vivo study of transdermal permeation of four diclofenac salts from different vehicles.

    PubMed

    Minghetti, Paola; Cilurzo, Francesco; Casiraghi, Antonella; Montanari, Luisa; Fini, Adamo

    2007-04-01

    The ex vivo permeation of diclofenac was studied using four different salts (sodium, potassium, diethylamine, and epolamine) dissolved in four different solvents (water, propylene glycol (PG), Transcutol, and oleic acid (OA)) as donor phases through a human skin membrane. The four salts show different solubility values and different behavior in the four solvents, which are also permeation enhancers and this fact further is connected to the permeation results. The same order of magnitude of fluxes through the membrane as those previously reported for acidic diclofenac released from buffer solutions of pH >7 were found, taking into account differences originated by different membranes and other parameters tested in the experiments. Saturation concentration for the four salts in different solvents, necessary to calculate permeation coefficients, was critically evaluated; a short discussion made it possible to explain that corrections in the solubility values must be considered, related to the complex behavior in solution of these salts. Statistical processing of the experimental data suggests that differences between the four salts in promoting absorption of the drug is unproven; while differences are evident between the solvents, water is the most effective enhancing vehicle. Aqueous formulations containing diclofenac salt with an organic base appear to be the best combination to promote permeation in topical applications. (c) 2007 Wiley-Liss, Inc.

  10. Insight into the Development of Dissolution Media for BCS Class II Drugs: A Review from Quality Control and Prediction of In Vivo Performance Perspectives.

    PubMed

    Wu, Chunnuan; Liu, Yan; He, Zhonggui; Sun, Jin

    2016-01-01

    To assess in vivo behavior through in vitro method, the dissolution test is mostly used, both for quality control (QC) and for development purpose. In view of the fact that a dissolution test can hardly achieve two goals at the same time, the design of dissolution testing generally varies along with the development stage of drug products and therefore the selection of dissolution media may change with the goals of the dissolution test. To serve the QC purpose, a dissolution medium is designed to provide a sink condition; for development purpose, the dissolution medium is required to simulate the physiological conditions in the gastrointestinal tract as far as possible. In this review, we intended to provide an initial introduction to the various dissolution media applied for QC and formulation development purposes for poorly water soluble drugs. We focused on these methods like addition of cosolvents, surfactants and utilization of biphasic media, applied to provide sink conditions which are difficult to be achieved by simple aqueous buffers for lipophilic drugs, and introduced the development of physiologically relevant media for human and animals like dog and rat with respect to the choice of buffers, bile salts, lipids and so on. In addition, we further discussed the influence of biorelevant dissolution media on the modification of drug Biopharmaceutical Classification System (BCS) classification, especially for BCS class II drugs with low solubility and high permeability, the solubility of which is relatively sensitive to the presence of bile salts and lipids.

  11. Electro-hydrodynamic generation of monodisperse nanoparticles in the sub-10 nm size range from strongly electrolytic salt solutions: governing parameters of scaling laws

    NASA Astrophysics Data System (ADS)

    Maißer, Anne; Attoui, Michel B.; Gañán-Calvo, Alfonso M.; Szymanski, Wladyslaw W.

    2013-01-01

    A charge reduced electro-hydrodynamic atomization (EHDA) device has been used to generate airborne salt clusters in the sub 10 nm size range. The focus of this study on that specific sub-micron range of electrospray droplets with relatively high electrical conductivities and permittivities aims to address the still existing controversy on the scaling laws of electrosprayed droplet diameters. In this study different concentrations of sodium chloride and potassium chloride—both show strong electrolytic behavior—have been electrosprayed from solutions in pure water, or from aqueous ammonium acetate buffer liquids of varying concentrations. The dry residue salt cluster diameter generated by the EHDA process have been measured using a differential mobility analyzer. The initial droplet diameter has been determined indirectly from the measured particle size following the steps of Chen et al. (J Aerosol Sci 26:963-977, 1995). Results have been compared to existing scaling laws valid for direct droplet measurements. They can be interpreted concisely on the basis of a realistic hypothesis on possible electrochemical effects taking place and affecting the droplet and thus nanoparticle formation in EHDA. The hypothesis developed in this work and the comparison with the experimental results are shown and discussed in the manuscript.

  12. The Effects of pH on the Growth and Aspect Ratio of Chicken Egg White Lysozyme Crystals Prepared in Different Buffers

    NASA Technical Reports Server (NTRS)

    Gibson, U. J.; Horrell, E. E.; Kou, Y.; Pusey, Marc

    2000-01-01

    We have measured the nucleation and aspect ratio of CEWL crystals grown by vapor diffusion in acetate, butyrate, carbonate, succinate, and phosphate buffers in a range of pH spanning the pK(sub a) of these buffers. The nucleation numbers drop off significantly in the vicinity of pK(sub a) for each of the buffers except the phosphate system, in which we used only the pH range around the second titration point(pK2). There is a concomitant increase in the sizes of the crystals. Some typical nucleation number results are shown. These data support and extend other observations. In addition, we have examined changes in aspect ratio which accompany the suppression of nucleation within each buffer system. The length of the face in the [001] direction was measured, and compared to the width of the (110) face in the [110] type directions. We find that while the aspect ratio of the crystals is affected by pH, it is dominated by a correlation with the size of the crystals. Small crystals are longer in the [0011 direction than crystals that are larger (higher pH within a buffer system). This relationship is found to hold independent of the choice of buffer. These results are consistent with those of Judge et al, who used a batch process which resulted in uniform sizing of crystals at each pH. In these experiments, we specifically avoid agitating the protein/salt buffer mixture when combining the two. This permits the formation of a range of sizes at a given pH. The results for a .05 M acetate 5% NaCl buffer are also shown. We will discuss these results in light of a growth model.

  13. Nogo goes in the pure water: solution structure of Nogo-60 and design of the structured and buffer-soluble Nogo-54 for enhancing CNS regeneration.

    PubMed

    Li, Minfen; Liu, Jingxian; Song, Jianxing

    2006-08-01

    The inability to determine the structure of the buffer-insoluble Nogo extracellular domain retarded further design of Nogo receptor (NgR) antagonists to treat CNS axonal injuries. Very surprisingly, we recently discovered that Nogo-60 was soluble and structured in salt-free water, thus allowing the determination of the first Nogo structure by heteronuclear NMR spectroscopy. Nogo-60 adopts an unusual helical structure with the N- and C-terminal helices connected by a long middle helix. While the N-helix has no contact with the rest of the molecule, the C-helix flips back to pack against the 20-residue middle helix. This packing appears to trigger the formation of the stable Nogo-60 structure because Nogo-40 with the last helix truncated is unstructured. The Nogo-60 structure offered us rationales for further design of the structured and buffer-soluble Nogo-54, which may be used as a novel NgR antagonist. Furthermore, our discovery may imply a general solution to solubilizing a category of buffer-insoluble proteins for urgent structural investigations.

  14. Tension in Skinned Frog Muscle Fibers in Solutions of Varying Ionic Strength and Neutral Salt Composition

    PubMed Central

    Gordon, A. M.; Godt, R. E.; Donaldson, S. K. B.; Harris, C. E.

    1973-01-01

    The maximal calcium-activated isometric tension produced by a skinned frog single muscle fiber falls off as the ionic strength of the solution bathing this fiber is elevated declining to zero near 0.5 M as the ionic strength is varied using KCl. When other neutral salts are used, the tension always declines at high ionic strength, but there is some difference between the various neutral salts used. The anions and cations can be ordered in terms of their ability to inhibit the maximal calcium-activated tension. The order of increasing inhibition of tension (decreasing tension) at high ionic strength for anions is propionate- ≃ SO4 -- < Cl- < Br-. The order of increasing inhibition of calcium-activated tension for cations is K+ ≃ Na+ ≃ TMA+ < TEA+ < TPrA+ < TBuA+. The decline of maximal calcium-activated isometric tension with elevated salt concentration (ionic strength) can quantitatively explain the decline of isometric tetanic tension of a frog muscle fiber bathed in a hypertonic solution if one assumes that the internal ionic strength of a muscle fiber in normal Ringer's solution is 0.14–0.17 M. There is an increase in the base-line tension of a skinned muscle fiber bathed in a relaxing solution (no added calcium and 3 mM EGTA) of low ionic strength. This tension, which has no correlate in the intact fiber in hypotonic solutions, appears to be a noncalcium-activated tension and correlates more with a declining ionic strength than with small changes in [MgATP], [Mg], pH buffer, or [EGTA]. It is dependent upon the specific neutral salts used with cations being ordered in increasing inhibition of this noncalcium-activated tension (decreasing tension) as TPrA+ < TMA+ < K+ ≃ Na+. Measurements of potentials inside these skinned muscle fibers bathed in relaxing solutions produced occasional small positive values (<6 mV) which were not significantly different from zero. PMID:4543066

  15. Comparative Evaluation of Small Molecular Additives and Their Effects on Peptide/Protein Identification.

    PubMed

    Gao, Jing; Zhong, Shaoyun; Zhou, Yanting; He, Han; Peng, Shuying; Zhu, Zhenyun; Liu, Xing; Zheng, Jing; Xu, Bin; Zhou, Hu

    2017-06-06

    Detergents and salts are widely used in lysis buffers to enhance protein extraction from biological samples, facilitating in-depth proteomic analysis. However, these detergents and salt additives must be efficiently removed from the digested samples prior to LC-MS/MS analysis to obtain high-quality mass spectra. Although filter-aided sample preparation (FASP), acetone precipitation (AP), followed by in-solution digestion, and strong cation exchange-based centrifugal proteomic reactors (CPRs) are commonly used for proteomic sample processing, little is known about their efficiencies at removing detergents and salt additives. In this study, we (i) developed an integrative workflow for the quantification of small molecular additives in proteomic samples, developing a multiple reaction monitoring (MRM)-based LC-MS approach for the quantification of six additives (i.e., Tris, urea, CHAPS, SDS, SDC, and Triton X-100) and (ii) systematically evaluated the relationships between the level of additive remaining in samples following sample processing and the number of peptides/proteins identified by mass spectrometry. Although FASP outperformed the other two methods, the results were complementary in terms of peptide/protein identification, as well as the GRAVY index and amino acid distributions. This is the first systematic and quantitative study of the effect of detergents and salt additives on protein identification. This MRM-based approach can be used for an unbiased evaluation of the performance of new sample preparation methods. Data are available via ProteomeXchange under identifier PXD005405.

  16. Monitoring the degrafting of polyelectrolyte brushes by using surface gradients

    NASA Astrophysics Data System (ADS)

    Ko, Yeongun; Genzer, Jan

    Polymer brushes comprise densely grafted polymer chains on surfaces, which possess high stability and high concentration of reactive centers per unit area compared to physisorbed polymer film. Polymer brushes are employed in many applications, including anti-fouling surfaces, cell adhesive surfaces, responsive surfaces, low-friction surfaces, etc. Recently, researchers reported that charged (or chargeable) polymer brushes can be degrafted from substrate while incubated in buffer solutions. Based on previous experiments conducted in our group and by others, we assume that chain degrafting results from the hydrolysis of Si-O groups in head-group of the initiator and/or the ester groups in main body of the initiator. The kinetic of hydrolysis is affected by mechanical forces acting on the initiator. Those forces depend on the molecular weight and the grafting density of the brush, and the concentration and distribution of charges along the macromolecule (tuned by pH - for weak electrolytes - and concentration of external salt). In this work, we study the stability of poly(2-dimethylaminoethyl methacrylate) (PDMAEMA) brushes in two solvents (ethanol and water) at various pH values in water and under different levels of external salt concentration. National Science Foundation.

  17. Cofactor-Dependent Aldose Dehydrogenase of Rhodopseudomonas spheroides

    PubMed Central

    Niederpruem, Donald J.; Doudoroff, Michael

    1965-01-01

    Niederpruem, Donald J. (University of California, Berkeley), and Michael Doudoroff. Cofactor-dependent aldose dehydrogenase of Rhodopseudomonas spheroides. J. Bacteriol. 89:697–705. 1965.—Particulate enzyme preparations of cell extracts of Rhodopseudomonas spheroides possess constitutive dehydrogenase and oxidase activities for aldose sugars, reduced nicotinamide adenine dinucleotide (NADH2), and succinate. The dehydrogenation of aldoses requires an unidentified cofactor which is not required for the oxidation of succinate nor of NADH2. The cofactor is present in the particulate fraction of aerobic cells, but is unavailable to the enzyme system. It can be liberated by boiling or by treatment with salts at high concentration. The cofactor also appears in the soluble fraction of aerobic cells, but only after exponential growth has ceased. Extracts of cells grown anaerobically in the light possess the apoenzyme, but not the cofactor, for aldose oxidation. Cofactor activity was found in extracts of Bacterium anitratum (= Moraxella sp.) but not in Escherichia coli, Pseudomonas fluorescens, yeast, or mouse liver. In 0.075 m tris(hydroxymethyl)aminomethane-phosphoric acid buffer (pH 7.3), the oxidation of NADH2 was stimulated and succinoxidase was inhibited by high salt concentrations. PMID:14273648

  18. Dominant Negative Mutants of the Estrogen Receptor as Probes of Estrogen Action and Inhibitors of Breast Cancer Growth

    DTIC Science & Technology

    1996-07-01

    tetrazolium, inner salt; MTS; Promega], 1.9 mg/ml, and an electron coupling reagent ( phenazine methosulfate; PMS; Sigma), 0.044 mg/ml, in Dulbecco’s...acids PBS, phosphate buffered saline PCR, polymerase chain reaction PMS, phenazine methosulfate poly A, polyadenylation s.e., standard error TAE, tris

  19. Excited Atomic Bromine Energy Transfer and Quenching Mechanisms

    DTIC Science & Technology

    1993-08-01

    slope, m, and intercept, b, given as MBr 2 ] + k (3.16) b= 1 By collecting the Br* emission intensity as a function of added buffer gas concentration... submerged in an H20 ice and NaCl salt slurrie. The temperature is monitored with a temperature probe to insure that it is below 00 C. Then with LN 2

  20. Protein solubility modeling

    NASA Technical Reports Server (NTRS)

    Agena, S. M.; Pusey, M. L.; Bogle, I. D.

    1999-01-01

    A thermodynamic framework (UNIQUAC model with temperature dependent parameters) is applied to model the salt-induced protein crystallization equilibrium, i.e., protein solubility. The framework introduces a term for the solubility product describing protein transfer between the liquid and solid phase and a term for the solution behavior describing deviation from ideal solution. Protein solubility is modeled as a function of salt concentration and temperature for a four-component system consisting of a protein, pseudo solvent (water and buffer), cation, and anion (salt). Two different systems, lysozyme with sodium chloride and concanavalin A with ammonium sulfate, are investigated. Comparison of the modeled and experimental protein solubility data results in an average root mean square deviation of 5.8%, demonstrating that the model closely follows the experimental behavior. Model calculations and model parameters are reviewed to examine the model and protein crystallization process. Copyright 1999 John Wiley & Sons, Inc.

  1. Adsorption equilibrium and kinetics of Immunoglobulin G on a mixed-mode adsorbent in batch and packed bed configuration.

    PubMed

    Gomes, Pedro Ferreira; Loureiro, José Miguel; Rodrigues, Alírio E

    2017-11-17

    It is commonly accepted that efficient protein separation and purification to the desired level of purity is one bottleneck in pharmaceutical industries. MabDirect MM is a new type of mixed mode adsorbent, especially designed to operate in expanded bed adsorption (EBA) mode. In this study, equilibrium and kinetics experiments were carried out for the adsorption of Human Immunoglobulin G (hIgG) protein on this new adsorbent. The effects of ionic strength and pH are assessed. Langmuir isotherms parameters are obtained along with the estimation of the effective pore diffusion coefficient (D pe ) by fitting the batch adsorption kinetics experiments with the pore diffusion model. The maximum adsorption of the IgG protein on the MabDirect MM adsorbent, 149.7±7.1mg·g dry -1 , was observed from a pH 5.0 buffer solution without salt addition. Adding salt to the buffer solution, and/or increasing pH, decreases the adsorption capacity which is 4.7±0.4mg·g dry -1 for pH 7.0 with 0.4M NaCl in solution. Regarding the D pe estimation, a value of 15.4×10 -6 cm 2 ·min -1 was obtained for a pH 5.0 solution without salt. Increasing the salt concentration and/or the pH value will decrease the effective pore diffusion, the lowest D pe (0.16×10 -6 cm 2 ·min -1 ) value being observed for an IgG solution at pH 7.0 with 0.4M NaCl. Fixed bed experiments were conducted with the purpose to validate the equilibrium and kinetic parameters obtained in batch. For a feed concentration of 0.5 g·L -1 of IgG in pH 5.0 buffer solution with 0.4M NaCl, a dynamic binding capacity at 10% of breakthrough of 5.3mg·g wet -1 (15.4mg IgG ·mL resin -1 ) was obtained, representing 62% of the saturation capacity. As far as the authors know, this study is the first one concerning the adsorption of hIgG on this type of mixed mode chromatography adsorbent. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Effect of salt addition on acid resistance response of Escherichia coli O157:H7 against acetic acid.

    PubMed

    Bae, Young-Min; Lee, Sun-Young

    2017-08-01

    A combination of salt and acid is commonly used in the production of many foods, such as pickles and fermented foods. However, in our previous studies, addition of salt significantly reduced the inhibitory effect of acetic acid against E. coli O157:H7 in laboratory media and pickled cucumbers. Therefore, this study was conducted to determine the effect of salt addition on the acid resistance (AR) response of E. coli O157:H7 after treatment with acetic acid. The combined effect of acetic acid and salt showed different results depending on media tested. Organic compounds such as yeast extract and tryptone were required to observe the antagonistic effect of salt and acetic acid in combination. However, use of an rpoS mutant or addition of chloramphenicol resulted in no changes in the antagonistic effect of acetic acid and salt. The addition of glutamate to phosphate buffer significantly increased the survival levels of E. coli O157:H7 after the acetic acid treatment; however, the survival levels were lower than those after the treatment with acetic acid alone. Thus, the addition of salt may increase the AR response of E. coli O157:H7; however, these survival mechanisms were not proven clearly. Therefore, further studies need to be performed to better understand the antagonism of acetic acid salt against E. coli O157:H7. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Ozone and photocatalytic processes to remove the antibiotic sulfamethoxazole from water.

    PubMed

    Beltrán, Fernando J; Aguinaco, Almudena; García-Araya, Juan F; Oropesa, Ana

    2008-08-01

    In this study, water containing the pharmaceutical compound sulfamethoxazole (SMT) was subjected to the various treatments of different oxidation processes involving ozonation, and photolysis and catalysis under different experimental conditions. Removal rates of SMT and total organic carbon (TOC), from experiments of simple UVA radiation, ozonation (O(3)), catalytic ozonation (O(3)/TiO(2)), ozone photolysis (O(3)/UVA), photocatalytic oxidation (O(2)/TiO(2)/UVA) and photocatalytic ozonation (O(3)/UVA/TiO(2)), have been compared. Photocatalytic ozonation leads to the highest SMT removal rate (pH 7 in buffered systems, complete removal is achieved in less than 5min) and total organic carbon (in unbuffered systems, with initial pH=4, 93% TOC removal is reached). Also, lowest ozone consumption per TOC removed and toxicity was achieved with the O(3)/UVA/TiO(2) process. Direct ozone and free radical reactions were found to be the principal mechanisms for SMT and TOC removal, respectively. In photocatalytic ozonation, with buffered (pH 7) aqueous solutions phosphates (buffering salts) and accumulation of bicarbonate scavengers inhibit the reactions completely on the TiO(2) surface. As a consequence, TOC removal diminishes. In all cases, hydrogen peroxide plays a key role in TOC mineralization. According to the results obtained in this work the use of photocatalytic ozonation is recommended to achieve a high mineralization degree of water containing SMT type compounds.

  4. Multiple, simultaneous, independent gradients for a versatile multidimensional liquid chromatography. Part II: Application 2: Computer controlled pH gradients in the presence of urea provide improved separation of proteins: Stability influenced anion and cation exchange chromatography.

    PubMed

    Hirsh, Allen G; Tsonev, Latchezar I

    2017-04-28

    This paper details the use of a method of creating controlled pH gradients (pISep) to improve the separation of protein isoforms on ion exchange (IEX) stationary phases in the presence of various isocratic levels of urea. The pISep technology enables the development of computer controlled pH gradients on both cationic (CEX) and anionic (AEX) IEX stationary phases over the very wide pH range from 2 to 12. In pISep, titration curves generated by proportional mixing of the acidic and basic pISep working buffers alone, or in the presence of non-buffering solutes such as the neutral salt NaCl (0-1M), polar organics such as urea (0-8M) or acetonitrile (0-80 Vol%), can be fitted with high fidelity using high order polynomials which, in turn allows construction of a mathematical manifold %A (% acidic pISep buffer) vs. pH vs. [non-buffering solute], permitting precise computer control of pH and the non-buffering solute concentration allowing formation of dual uncoupled liquid chromatographic (LC) gradients of arbitrary shape (Hirsh and Tsonev, 2012 [1]). The separation of protein isoforms examined in this paper by use of such pH gradients in the presence of urea demonstrates the fractionation power of a true single step two dimensional liquid chromatography which we denote as Stability-Influenced Ion Exchange Chromatography (SIIEX). We present evidence that SIIEX is capable of increasing the resolution of protein isoforms difficult to separate by ordinary pH gradient IEX, and potentially simplifying the development of laboratory and production purification strategies involving on-column simultaneous pH and urea unfolding or refolding of targeted proteins. We model some of the physics implied by the dynamics of the observed protein fractionations as a function of both urea concentration and pH assuming that urea-induced native state unfolding competes with native state electrostatic interaction binding to an IEX stationary phase. Implications for in vivo protein-membrane interactions are discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Tetragonal Chicken Egg White Lysozyme Solubility in Sodium Chloride Solutions

    NASA Technical Reports Server (NTRS)

    Forsythe, Elizabeth L.; Judge, Russell A.; Pusey, Marc L.

    1998-01-01

    The solubility of chicken egg white lysozyme, crystallized in the tetragonal form was measured in sodium chloride solutions from 1.6 to 30.7 C, using a miniature column solubility apparatus. Sodium chloride solution concentrations ranged from 1 to 7% (w/v). The solutions were buffered with 0.1 M sodium acetate buffer with the solubility being measured at pH values in 0.2 pH unit increments in the range pH 4.0 to 5.4, with data also included at pH 4.5. Lysozyme solubility was found to increase with increases in temperature and decreasing salt concentration. Solution pH has a varied and unpredictable effect on solubility.

  6. Enhancement of DNaseI Salt Tolerance by Mimicking the Domain Structure of DNase from an Extremely Halotolerant Bacterium Thioalkalivibrio sp. K90mix

    PubMed Central

    Alzbutas, Gediminas; Kaniusaite, Milda; Lagunavicius, Arunas

    2016-01-01

    In our previous work we showed that DNaseI-like protein from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix retained its activity at salt concentrations as high as 4 M NaCl and the key factor allowing this was the C-terminal DNA-binding domain, which comprised two HhH (helix-hairpin-helix) motifs. The further investigations revealed that this domain originated from proteins related to bacterial competence ComEA/ComE proteins. It is likely that in the course of evolution the DNA-binding domain from these proteins was fused to a metallo-β-lactamase superfamily domain. Very likely such domain organization having proteins subsequently “donated” the DNA-binding domain to bacterial DNases. In this study we have mimicked this evolutionary step by fusing bovine DNaseI and DNA-binding domains. We have created two fusions: one harboring the DNA-binding domain of DNaseI-like protein from Thioalkalivibrio sp. K90mix and the second one harboring the DNA-binding domain of bacterial competence protein ComEA from Bacillus subtilis. Both domains enhanced salt tolerance of DNaseI, albeit to different extent. Molecular modeling revealed the essential differences between their interaction with DNA shedding some light on the differences in salt tolerance. In this study we have enhanced salt tolerance of bovine DNaseI; thus, we successfully mimicked the Nature’s evolutionary engineering that created the extremely halotolerant bacterial DNase. We have demonstrated that the newly engineered DNaseI variants can be successfully used in applications where activity of the wild type bovine DNaseI is impeded by buffers used. PMID:26939122

  7. Salt marsh as a coastal filter for the oceans: changes in function with experimental increases in nitrogen loading and sea-level rise.

    PubMed

    Nelson, Joanna L; Zavaleta, Erika S

    2012-01-01

    Coastal salt marshes are among Earth's most productive ecosystems and provide a number of ecosystem services, including interception of watershed-derived nitrogen (N) before it reaches nearshore oceans. Nitrogen pollution and climate change are two dominant drivers of global-change impacts on ecosystems, yet their interacting effects at the land-sea interface are poorly understood. We addressed how sea-level rise and anthropogenic N additions affect the salt marsh ecosystem process of nitrogen uptake using a field-based, manipulative experiment. We crossed simulated sea-level change and ammonium-nitrate (NH(4)NO(3))-addition treatments in a fully factorial design to examine their potentially interacting effects on emergent marsh plants in a central California estuary. We measured above- and belowground biomass and tissue nutrient concentrations seasonally and found that N-addition had a significant, positive effect on a) aboveground biomass, b) plant tissue N concentrations, c) N stock sequestered in plants, and d) shoot:root ratios in summer. Relative sea-level rise did not significantly affect biomass, with the exception of the most extreme sea-level-rise simulation, in which all plants died by the summer of the second year. Although there was a strong response to N-addition treatments, salt marsh responses varied by season. Our results suggest that in our site at Coyote Marsh, Elkhorn Slough, coastal salt marsh plants serve as a robust N trap and coastal filter; this function is not saturated by high background annual N inputs from upstream agriculture. However, if the marsh is drowned by rising seas, as in our most extreme sea-level rise treatment, marsh plants will no longer provide the ecosystem service of buffering the coastal ocean from eutrophication.

  8. Salt Marsh as a Coastal Filter for the Oceans: Changes in Function with Experimental Increases in Nitrogen Loading and Sea-Level Rise

    PubMed Central

    Nelson, Joanna L.; Zavaleta, Erika S.

    2012-01-01

    Coastal salt marshes are among Earth's most productive ecosystems and provide a number of ecosystem services, including interception of watershed-derived nitrogen (N) before it reaches nearshore oceans. Nitrogen pollution and climate change are two dominant drivers of global-change impacts on ecosystems, yet their interacting effects at the land-sea interface are poorly understood. We addressed how sea-level rise and anthropogenic N additions affect the salt marsh ecosystem process of nitrogen uptake using a field-based, manipulative experiment. We crossed simulated sea-level change and ammonium-nitrate (NH4NO3)-addition treatments in a fully factorial design to examine their potentially interacting effects on emergent marsh plants in a central California estuary. We measured above- and belowground biomass and tissue nutrient concentrations seasonally and found that N-addition had a significant, positive effect on a) aboveground biomass, b) plant tissue N concentrations, c) N stock sequestered in plants, and d) shoot:root ratios in summer. Relative sea-level rise did not significantly affect biomass, with the exception of the most extreme sea-level-rise simulation, in which all plants died by the summer of the second year. Although there was a strong response to N-addition treatments, salt marsh responses varied by season. Our results suggest that in our site at Coyote Marsh, Elkhorn Slough, coastal salt marsh plants serve as a robust N trap and coastal filter; this function is not saturated by high background annual N inputs from upstream agriculture. However, if the marsh is drowned by rising seas, as in our most extreme sea-level rise treatment, marsh plants will no longer provide the ecosystem service of buffering the coastal ocean from eutrophication. PMID:22879873

  9. Basement inheritance and salt tectonics in the SE Barents Sea: Insights from new potential field data

    NASA Astrophysics Data System (ADS)

    Gernigon, L.; Broenner, M.; Dumais, M. A.; Gradmann, S.; Grønlie, A.; Nasuti, A.; Roberts, D.

    2017-12-01

    The tectonic evolution of the former `grey zone' between Russia and Norway has so far remained poorly constrained due to a lack of geophysical data. In 2014, we carried out a new aeromagnetic survey (BASAR-14) in the southern part of the new Norwegian offshore territory. Caledonian and Timanian structures, highlighted by the new potential field data, dominate the basement patterns and have exerted a strong influence on the structure and development of the overlying basins and basement highs. Clearly associated with NW-SE-oriented Timanian trends, the Tiddlybanken Basin represents an atypical sag basin that developed at the southern edge of the Fedynsky High. Regional extension and rapid sedimentation initiated the salt tectonics in the Barents Sea in the Early Triassic. Some of the pillows became diapiric during the Early Triassic and rejuvenated during subsequent Jurassic-Tertiary episodes of regional extension and/or compression. At present, quite a few large diapiric salt domes along the Nordkapp and Tiddlybanken basins are relatively shallow, locally reaching the seabed and thus show a clear bathymetric and magnetic signature. Quantitative modelling along 2D seismic transects was also carried out to constrain the structural and basement composition of the study area. The predominant NE-SW Mesozoic trend of the Nordkapp Basin represents a major crustal hinge zone between the Finnmark Platform, poorly affected by major crustal deformation, and the Bjarmeland Platform where Late Palaeozoic rifting controlled the widespread accumulation of salt deposits in Late Carboniferous-Early Permian time. The entire structure and segmentation of the Nordkapp Basin have been influenced by the inherited basement configuration highlighted by the new aeromagnetic data. Both the Nordkapp and the Tiddlybanken basins appear to lie at the edge of a peculiar thick and rigid crustal feature that coincides with a highly magnetic region. The abrupt termination of the eastern Nordkapp Basin at the edge of this magnetic domain suggests the presence of an old and thick Precambrian continental block. This magnetic and tectonic buffer controlled the Late Palaeozoic-Mesozoic rifting and the salt tectonic development of the southeastern Barents Sea.

  10. Escherichia coli growth changes by the mediated effects after low-intensity electromagnetic irradiation of extremely high frequencies.

    PubMed

    Torgomyan, Heghine; Hovnanyan, Karlen; Trchounian, Armen

    2013-04-01

    Water is the major constituent of environmental medium and biological systems. The effects occurring in water as a result of low-intensity electromagnetic irradiation (EMI) in extremely high frequencies are supposed to be the primary mechanism to create conditions for biological responses. The EMI effects on Escherichia coli, after irradiation of their suspension, are most probably water-mediated. Indirect effects of EMI at 51.8, 53, 70.6, and 73 GHz frequencies on bacteria, through water, assay buffer (Tris-phosphate buffer with inorganic salts at low or moderate concentrations), or peptone growth medium were studied. The mediated effects of 70.6 and 73 GHz irradiated water, assay buffer, and growth medium on E. coli growth characteristics were insignificant. But the results were different for 51.8 and 53 GHz. EMI mediated effects on bacterial growth were clearly demonstrated. The effects were more strongly expressed with 53 GHz. Moreover, it was shown that 70.6 and 73 GHz similarly suppressed the cell growth after direct irradiation of E. coli in water or on solid medium. Interestingly, for 51.8 and 53 GHz the bacterial growth decreases after suspension irradiation was less, compared to the direct irradiation of bacteria on solid medium. Especially, it was also more expressed in case of 53 GHz. Also with electron microscopy, EMI-induced bacterial cell sizes and structure different changes were detected. In addition, the distinguished changes in surface tension, oxidation-reduction potential and pH of water, assay buffer, growth medium, and bacterial suspension were determined. They depended on EMI frequency used. The differences could be associated with the partial absorbance of EMI energy by the surrounding medium, which depends on a specific frequency. The results are crucial to understand biophysical mechanisms of EMI effects on bacteria.

  11. Acemetacin cocrystals and salts: structure solution from powder X-ray data and form selection of the piperazine salt

    PubMed Central

    Sanphui, Palash; Bolla, Geetha; Nangia, Ashwini; Chernyshev, Vladimir

    2014-01-01

    Acemetacin (ACM) is a non-steroidal anti-inflammatory drug (NSAID), which causes reduced gastric damage compared with indomethacin. However, acemetacin has a tendency to form a less soluble hydrate in the aqueous medium. We noted difficulties in the preparation of cocrystals and salts of acemetacin by mechanochemical methods, because this drug tends to form a hydrate during any kind of solution-based processing. With the objective to discover a solid form of acemetacin that is stable in the aqueous medium, binary adducts were prepared by the melt method to avoid hydration. The coformers/salt formers reported are pyridine carboxamides [nicotinamide (NAM), isonicotinamide (INA), and picolinamide (PAM)], caprolactam (CPR), p-aminobenzoic acid (PABA), and piperazine (PPZ). The structures of an ACM–INA cocrystal and a binary adduct ACM–PABA were solved using single-crystal X-ray diffraction. Other ACM cocrystals, ACM–PAM and ACM–CPR, and the piperazine salt ACM–PPZ were solved from high-resolution powder X-ray diffraction data. The ACM–INA cocrystal is sustained by the acid⋯pyridine heterosynthon and N—H⋯O catemer hydrogen bonds involving the amide group. The acid⋯amide heterosynthon is present in the ACM–PAM cocrystal, while ACM–CPR contains carboxamide dimers of caprolactam along with acid–carbonyl (ACM) hydrogen bonds. The cocrystals ACM–INA, ACM–PAM and ACM–CPR are three-dimensional isostructural. The carboxyl⋯carboxyl synthon in ACM–PABA posed difficulty in assigning the position of the H atom, which may indicate proton disorder. In terms of stability, the salts were found to be relatively stable in pH 7 buffer medium over 24 h, but the cocrystals dissociated to give ACM hydrate during the same time period. The ACM–PPZ salt and ACM–nicotinamide cocrystal dissolve five times faster than the stable hydrate form, whereas the ACM–PABA adduct has 2.5 times faster dissolution rate. The pharmaceutically acceptable piperazine salt of acemetacin exhibits superior stability, faster dissolution rate and is able to overcome the hydration tendency of the reference drug. PMID:25075330

  12. Acemetacin cocrystals and salts: structure solution from powder X-ray data and form selection of the piperazine salt.

    PubMed

    Sanphui, Palash; Bolla, Geetha; Nangia, Ashwini; Chernyshev, Vladimir

    2014-03-01

    Acemetacin (ACM) is a non-steroidal anti-inflammatory drug (NSAID), which causes reduced gastric damage compared with indomethacin. However, acemetacin has a tendency to form a less soluble hydrate in the aqueous medium. We noted difficulties in the preparation of cocrystals and salts of acemetacin by mechanochemical methods, because this drug tends to form a hydrate during any kind of solution-based processing. With the objective to discover a solid form of acemetacin that is stable in the aqueous medium, binary adducts were prepared by the melt method to avoid hydration. The coformers/salt formers reported are pyridine carboxamides [nicotinamide (NAM), isonicotinamide (INA), and picolinamide (PAM)], caprolactam (CPR), p-aminobenzoic acid (PABA), and piperazine (PPZ). The structures of an ACM-INA cocrystal and a binary adduct ACM-PABA were solved using single-crystal X-ray diffraction. Other ACM cocrystals, ACM-PAM and ACM-CPR, and the piperazine salt ACM-PPZ were solved from high-resolution powder X-ray diffraction data. The ACM-INA cocrystal is sustained by the acid⋯pyridine heterosynthon and N-H⋯O catemer hydrogen bonds involving the amide group. The acid⋯amide heterosynthon is present in the ACM-PAM cocrystal, while ACM-CPR contains carboxamide dimers of caprolactam along with acid-carbonyl (ACM) hydrogen bonds. The cocrystals ACM-INA, ACM-PAM and ACM-CPR are three-dimensional isostructural. The carboxyl⋯carboxyl synthon in ACM-PABA posed difficulty in assigning the position of the H atom, which may indicate proton disorder. In terms of stability, the salts were found to be relatively stable in pH 7 buffer medium over 24 h, but the cocrystals dissociated to give ACM hydrate during the same time period. The ACM-PPZ salt and ACM-nicotinamide cocrystal dissolve five times faster than the stable hydrate form, whereas the ACM-PABA adduct has 2.5 times faster dissolution rate. The pharmaceutically acceptable piperazine salt of acemetacin exhibits superior stability, faster dissolution rate and is able to overcome the hydration tendency of the reference drug.

  13. [Experiments for physiologic improvement of disinfectants].

    PubMed

    Beyer, C; Wozniak, K D; Höhne, K; Laub, R

    1990-11-01

    The use of customary trade disinfectants lead to occupational disease no. 80 in many employers of public health. For that reason our aim is to improve the skin compatibility of this disinfectants. Its possible to achieve a pH-shift to a more physiological level eighter by buffer salt addition or by neutralization. Following this, disinfectants were investigated in a microbiological test.

  14. Complete Genome Sequence of Dehalobacterium formicoaceticum Strain DMC, a Strictly Anaerobic Dichloromethane-Degrading Bacterium

    DOE PAGES

    Chen, Gao; Murdoch, Robert W.; Mack, E. Erin; ...

    2017-09-14

    Dehalobacterium formicoaceticum utilizes dichloromethane as the sole energy source in defined anoxic bicarbonate-buffered mineral salt medium. The products are formate, acetate, inorganic chloride, and biomass. The bacterium’s genome was sequenced using PacBio, assembled, and annotated. The complete genome consists of one 3.77-Mb circular chromosome harboring 3,935 predicted protein-encoding genes.

  15. Inhibition of Clostridium perfringens spore germination and outgrowth by lemon juice and vinegar product in reduced NaCl roast beef

    USDA-ARS?s Scientific Manuscript database

    Inhibition of Clostridium perfringens spore germination and outgrowth in reduced sodium roast beef by a blend of buffered lemon juice concentrate and vinegar (MoStatin LV) during abusive exponential cooling was evaluated. Roast beef containing salt (NaCl; 1, 1.5, or 2%, wt/wt), blend of sodium pyro-...

  16. Complete Genome Sequence of Dehalobacterium formicoaceticum Strain DMC, a Strictly Anaerobic Dichloromethane-Degrading Bacterium

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

    Chen, Gao; Murdoch, Robert W.; Mack, E. Erin

    Dehalobacterium formicoaceticum utilizes dichloromethane as the sole energy source in defined anoxic bicarbonate-buffered mineral salt medium. The products are formate, acetate, inorganic chloride, and biomass. The bacterium’s genome was sequenced using PacBio, assembled, and annotated. The complete genome consists of one 3.77-Mb circular chromosome harboring 3,935 predicted protein-encoding genes.

  17. Kinetics of Pseudomonas aeruginosa adhesion to 304 and 316-L stainless steel: role of cell surface hydrophobicity.

    PubMed Central

    Vanhaecke, E; Remon, J P; Moors, M; Raes, F; De Rudder, D; Van Peteghem, A

    1990-01-01

    Fifteen different isolates of Pseudomonas aeruginosa were used to study the kinetics of adhesion to 304 and 316-L stainless steel. Stainless steel plates were incubated with approximately 1.5 X 10(7) CFU/ml in 0.01 M phosphate-buffered saline (pH 7.4). After the plates were rinsed with the buffer, the number of adhering bacteria was determined by a bioluminescence assay. Measurable adhesion, even to the electropolished surfaces, occurred within 30 s. Bacterial cell surface hydrophobicity, as determined by the bacterial adherence to hydrocarbons test and the contact angle measurement test, was the major parameter influencing the adhesion rate constant for the first 30 min of adhesion. A parabolic relationship between the CAM values and the logarithm of the adhesion rate constants (In k) was established. No correlation between either the salt aggregation or the improved salt aggregation values and the bacterial adhesion rate constants could be found. Since there was no significant correlation between the bacterial electrophoretic mobilities and the In k values, the bacterial cell surface charge seemed of minor importance in the process of adhesion of P. aeruginosa to 304 and 316-L stainless steel. PMID:2107796

  18. Chiral recognition ability of an (S)-naproxen- imprinted monolith by capillary electrochromatography.

    PubMed

    Xu, Yan-Li; Liu, Zhao-Sheng; Wang, He-Fang; Yan, Chao; Gao, Ru-Yu

    2005-02-01

    The racemic naproxen was selectively recognized by capillary electrochromatography (CEC) on an (S)-naproxen-imprinted monolith, which was prepared by an in situ thermal-initiated polymerization. The recognition selectivity of a selected monolith strictly relied on the CEC conditions involved. The factors that influence the imprinting selectivity as well as the electroosmotic flow (EOF), including the applied voltage, organic solvent, salt concentration and pH value of the buffer, column temperature, and surfactant modifiers were systematically studied. Once the column was prepared, the experiment results showed that the successful chiral recognition was dependent on CEC variables. For example: the recognition could be observed in acetonitrile and ethanol electrolytes, while methanol and dimethyl sulfoxide (DMSO) electrolytes had no chiral recognition ability. The buffer with pH values of 2.6 or 3.0 at a higher salt concentration had chiral recognition ability. Column temperatures of 25-35 degrees C were optimal. Three surfactants, sodium dodecyl sulfate (SDS), cetyltrimethylammonium bromide (CTAB), and polyoxyethylene sorbitan monolaurate (Tween 20), can improve the recognition. Baseline resolution was obtained under optimized conditions and the column efficiency of the later eluent (S)-naproxen was 90 000 plates/m.

  19. Effects of liquid chromatography mobile phases and buffer salts on phosphorus inductively coupled plasma atomic emission and mass spectrometries utilizing ultrasonic nebulization and membrane desolvation.

    PubMed

    Carr, John E; Kwok, Kaho; Webster, Gregory K; Carnahan, Jon W

    2006-01-23

    Atomic spectrometry, specifically inductively coupled plasma atomic emission spectrometry (ICP-AES) and mass spectrometry (ICP-MS) show promise for heteroatom-based detection of pharmaceutical compounds. The combination of ultrasonic nebulization (USN) with membrane desolvation (MD) greatly enhances detection limits with these approaches. Because pharmaceutical analyses often incorporate liquid chromatography, the study herein was performed to examine the effects of solvent composition on the analytical behaviors of these approaches. The target analyte was phosphorus, introduced as phosphomycin. AES response was examined at the 253.7 nm atom line and mass 31 ions were monitored for the MS experiments. With pure aqueous solutions, detection limits of 5 ppb (0.5 ng in 0.1 mL injection volumes) were obtained with ICP-MS. The ICP-AES detection limit was 150 ppb. Solvent compositions were varied from 0 to 80% organic (acetonitrile and methanol) with nine buffers at concentrations typically used in liquid chromatography. In general, solvents and buffers had statistically significant, albeit small, effects on ICP-AES sensitivities. A few exceptions occurred in cases where typical liquid chromatography buffer concentrations produced higher mass loadings on the plasma. Indications are that isocratic separations can be reliably performed. Within reasonable accuracy tolerances, it appears that gradient chromatography can be performed without the need for signal response normalization. Organic solvent and buffer effects were more significant with ICP-MS. Sensitivities varied significantly with different buffers and organic solvent content. In these cases, gradient chromatography will require careful analytical calibration as solvent and buffer content is varied. However, for most buffer and solvent combinations, signal and detection limits are only moderately affected. Isocratic separations and detection are feasible.

  20. Influence of lime on struvite formation and nitrogen conservation during food waste composting.

    PubMed

    Wang, Xuan; Selvam, Ammaiyappan; Wong, Jonathan W C

    2016-10-01

    This study aimed at investigating the feasibility of supplementing lime with struvite salts to reduce ammonia emission and salinity consequently to accelerate the compost maturity. Composting was performed in 20-L bench-scale reactors for 35days using artificial food waste mixed with sawdust at 1.2:1 (w/w dry basis), and Mg and P salts (MgO and K2HPO4, respectively). Nitrogen loss was significantly reduced from 44.3% to 27.4% during composting through struvite formation even with the addition of lime. Lime addition significantly reduced the salinity to less than 4mS/cm with a positive effect on improving compost maturity. Thus addition of both lime and struvite salts synergistically provide advantages to buffer the pH, reduce ammonia emission and salinity, and accelerate food waste composting. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Development and validation of a stability-indicating high performance liquid chromatographic assay for benoxinate.

    PubMed

    Chorny, Michael; Levy, Daniel; Schumacher, Ilana; Lichaa, Chaim; Gruzman, Boris; Livshits, Oleg; Lomnicky, Yossi

    2003-04-24

    Benoxinate is a local anaesthetic used for ophthalmic applications. The aim of this study was to develop a rapid and simple stability-indicating method for the determination of benoxinate formulated for ophthalmic use, evaluate its long-term stability and identify its major degradation product. Benoxinate was eluted on a 10 microm Spherisorb phenyl column, 250 x 3.2 mm, with a mobile phase consisting of acetonitrile-buffer (pH 3.5) (35:65, v/v), pumped at 0.8 ml min(-1) flow rate. The buffer was composed of sodium dihydrogen phosphate (50 mM), sodium hydrogen sulfate (2.5 mM) and 1-heptanesulfonic acid sodium salt (5 mM). The analyte was quantified spectrophotometrically at 308 nm. The chromatograms of benoxinate formulations obtained by this method showed benoxinate (t = 4.5 min) well resolved from its degradation product (t = 2.3 min), which was separately identified by means of HPLC-MS as 4-amino-3-butoxybenzoic acid. The assay was demonstrated to have high accuracy, precision and linearity. The method was implemented in investigating the long-term stability of benoxinate 0.4% ophthalmic solutions. The method was found to be simple, quick and selective in determining benoxinate concentrations in fresh and aged preparations.

  2. Controllable activation of nanoscale dynamics in a disordered protein alters binding kinetics

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

    Callaway, David J. E.; Matsui, Tsutomu; Weiss, Thomas

    The phosphorylation of specific residues in a flexible disordered activation loop yields precise control of signal transduction. One paradigm is the phosphorylation of S339/S340 in the intrinsically disordered tail of the multi-domain scaffolding protein NHERF1, which affects the intracellular localization and trafficking of NHERF1 assembled signaling complexes. Using neutron spin echo spectroscopy (NSE), we show salt-concentration-dependent excitation of nanoscale motion at the tip of the C-terminal tail in the phosphomimic S339D/S340D mutant. The “tip of the whip” that is unleashed is near the S339/S340 phosphorylation site and flanks the hydrophobic Ezrin-binding motif. The kinetic association rate constant of the bindingmore » of the S339D/S340D mutant to the FERM domain of Ezrin is sensitive to buffer salt concentration, correlating with the excited nanoscale dynamics. The results suggest that electrostatics modulates the activation of nanoscale dynamics of an intrinsically disordered protein, controlling the binding kinetics of signaling partners. Furthermore NSE can pinpoint the nanoscale dynamics changes in a highly specific manner.« less

  3. A Highly Sensitive and Selective Method for the Determination of an Iodate in Table-salt Samples Using Malachite Green-based Spectrophotometry.

    PubMed

    Konkayan, Mongkol; Limchoowong, Nunticha; Sricharoen, Phitchan; Chanthai, Saksit

    2016-01-01

    A simple, rapid, and sensitive malachite green-based spectrophotometric method for the selective trace determination of an iodate has been developed and presented for the first time. The reaction mixture was specifically involved in the liberation of iodine in the presence of an excess of iodide in an acidic condition following an instantaneous reaction between the liberated iodine and malachite green dye. The optimum condition was obtained with a buffer solution pH of 5.2 in the presence of 40 mg L -1 potassium iodide and 1.5 × 10 -5 M malachite green for a 5-min incubation time. The iodate contents in some table-salt samples were in the range of 26 to 45 mg kg -1 , while those of drinking water, tap water, canal water, and seawater samples were not detectable (< 96 ng mL -1 of limits of detection, LOQ) with their satisfied method of recoveries of between 93 and 108%. The results agreed with those obtained using ICP-OES for comparison.

  4. Controllable activation of nanoscale dynamics in a disordered protein alters binding kinetics

    DOE PAGES

    Callaway, David J. E.; Matsui, Tsutomu; Weiss, Thomas; ...

    2017-03-08

    The phosphorylation of specific residues in a flexible disordered activation loop yields precise control of signal transduction. One paradigm is the phosphorylation of S339/S340 in the intrinsically disordered tail of the multi-domain scaffolding protein NHERF1, which affects the intracellular localization and trafficking of NHERF1 assembled signaling complexes. Using neutron spin echo spectroscopy (NSE), we show salt-concentration-dependent excitation of nanoscale motion at the tip of the C-terminal tail in the phosphomimic S339D/S340D mutant. The “tip of the whip” that is unleashed is near the S339/S340 phosphorylation site and flanks the hydrophobic Ezrin-binding motif. The kinetic association rate constant of the bindingmore » of the S339D/S340D mutant to the FERM domain of Ezrin is sensitive to buffer salt concentration, correlating with the excited nanoscale dynamics. The results suggest that electrostatics modulates the activation of nanoscale dynamics of an intrinsically disordered protein, controlling the binding kinetics of signaling partners. Furthermore NSE can pinpoint the nanoscale dynamics changes in a highly specific manner.« less

  5. High-performance liquid-chromatographic separation of subcomponents of antimycin-A

    USGS Publications Warehouse

    Abidi, S.L.

    1988-01-01

    Using a reversed-phase high-performance liquid chromatographic (HPLC) technique, a mixture of antimycins A was separated into eight hitherto unreported subcomponents, Ala, Alb, A2a, A2b, A3a, A3b, A4a, and A4b. Although a base-line resolution of the known four major antimycins Al, A2, A3, and A4 was readily achieved with mobile phases containing acetate buffers, the separation of the new antibiotic subcomponents was highly sensitive to variation in mobile phase conditions. The type and composition of organic modifiers, the nature of buffer salts, and the concentration of added electrolytes had profound effects on capacity factors, separation factors, and peak resolution values. Of the numerous chromatographic systems examined, a mobile phase consisting of methanol-water (70:30) and 0.005 M tetrabutylammonium phosphate at pH 3.0 yielded the most satisfactory results for the separation of the subcomponents. Reversed-phase gradient HPLC separation of the dansylated or methylated antibiotic compounds produced superior chromatographic characteristics and the presence of added electrolytes was not a critical factor for achieving separation. Differences in the chromatographic outcome between homologous and structural isomers were interpretated based on a differential solvophobic interaction rationale. Preparative reversed-phase HPLC under optimal conditions enabled isolation of pure samples of the methylated antimycin subcomponents for use in structural studies.

  6. Arginine-based cationic liposomes for efficient in vitro plasmid DNA delivery with low cytotoxicity.

    PubMed

    Sarker, Satya Ranjan; Aoshima, Yumiko; Hokama, Ryosuke; Inoue, Takafumi; Sou, Keitaro; Takeoka, Shinji

    2013-01-01

    Currently available gene delivery vehicles have many limitations such as low gene delivery efficiency and high cytotoxicity. To overcome these drawbacks, we designed and synthesized two cationic lipids comprised of n-tetradecyl alcohol as the hydrophobic moiety, 3-hydrocarbon chain as the spacer, and different counterions (eg, hydrogen chloride [HCl] salt or trifluoroacetic acid [TFA] salt) in the arginine head group. Cationic lipids were hydrated in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer to prepare cationic liposomes and characterized in terms of their size, zeta potential, phase transition temperature, and morphology. Lipoplexes were then prepared and characterized in terms of their size and zeta potential in the absence or presence of serum. The morphology of the lipoplexes was determined using transmission electron microscopy and atomic force microscopy. The gene delivery efficiency was evaluated in neuronal cells and HeLa cells and compared with that of lysine-based cationic assemblies and Lipofectamine™ 2000. The cytotoxicity level of the cationic lipids was investigated and compared with that of Lipofectamine™ 2000. We synthesized arginine-based cationic lipids having different counterions (ie, HCl-salt or TFA-salt) that formed cationic liposomes of around 100 nm in size. In the absence of serum, lipoplexes prepared from the arginine-based cationic liposomes and plasmid (p) DNA formed large aggregates and attained a positive zeta potential. However, in the presence of serum, the lipoplexes were smaller in size and negative in zeta potential. The morphology of the lipoplexes was vesicular. Arginine-based cationic liposomes with HCl-salt showed the highest transfection efficiency in PC-12 cells. However, arginine-based cationic liposomes with TFA salt showed the highest transfection efficiency in HeLa cells, regardless of the presence of serum, with very low associated cytotoxicity. The gene delivery efficiency of amino acid-based cationic assemblies is influenced by the amino acids (ie, arginine or lysine) present as the hydrophilic head group and their associated counterions.

  7. Arginine-based cationic liposomes for efficient in vitro plasmid DNA delivery with low cytotoxicity

    PubMed Central

    Sarker, Satya Ranjan; Aoshima, Yumiko; Hokama, Ryosuke; Inoue, Takafumi; Sou, Keitaro; Takeoka, Shinji

    2013-01-01

    Background Currently available gene delivery vehicles have many limitations such as low gene delivery efficiency and high cytotoxicity. To overcome these drawbacks, we designed and synthesized two cationic lipids comprised of n-tetradecyl alcohol as the hydrophobic moiety, 3-hydrocarbon chain as the spacer, and different counterions (eg, hydrogen chloride [HCl] salt or trifluoroacetic acid [TFA] salt) in the arginine head group. Methods Cationic lipids were hydrated in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer to prepare cationic liposomes and characterized in terms of their size, zeta potential, phase transition temperature, and morphology. Lipoplexes were then prepared and characterized in terms of their size and zeta potential in the absence or presence of serum. The morphology of the lipoplexes was determined using transmission electron microscopy and atomic force microscopy. The gene delivery efficiency was evaluated in neuronal cells and HeLa cells and compared with that of lysine-based cationic assemblies and Lipofectamine™ 2000. The cytotoxicity level of the cationic lipids was investigated and compared with that of Lipofectamine™ 2000. Results We synthesized arginine-based cationic lipids having different counterions (ie, HCl-salt or TFA-salt) that formed cationic liposomes of around 100 nm in size. In the absence of serum, lipoplexes prepared from the arginine-based cationic liposomes and plasmid (p) DNA formed large aggregates and attained a positive zeta potential. However, in the presence of serum, the lipoplexes were smaller in size and negative in zeta potential. The morphology of the lipoplexes was vesicular. Arginine-based cationic liposomes with HCl-salt showed the highest transfection efficiency in PC-12 cells. However, arginine-based cationic liposomes with TFA salt showed the highest transfection efficiency in HeLa cells, regardless of the presence of serum, with very low associated cytotoxicity. Conclusion The gene delivery efficiency of amino acid-based cationic assemblies is influenced by the amino acids (ie, arginine or lysine) present as the hydrophilic head group and their associated counterions. PMID:23630419

  8. Impact of high pressure freezing on DH5alpha Escherichia coli and red blood cells.

    PubMed

    Suppes, Galen J; Egan, Susan; Casillan, Alfred J; Wei Chan, Kok; Seckar, Bill

    2003-10-01

    The impact of high pressure and freezing on survivability of Escherichia coli and human red blood cells was evaluated to determine the utility of high-pressure transitions for preserving living cells. Based on microscopy and survivability, high pressures did not directly impact physical damage to living cells. E. coli studies showed that increased cell death is due to indirect phenomena with decreasing survivability at increasingly high pressures and exposure times. Pressurization rates up to 1.4kbar/min had negligible effects relative to exposures of >5min at high pressures.Both glycine and control of pH near 7.0 were successful in reducing the adverse impacts of high pressure. Survivability increased from <1% at 5min exposure to 2.1kbar of pressure to typical values >20%. The combination of glycine and the buffer salt led to even further improvements in survivability. Pressure changes were used to traverse temperature and pressures consistent with Ice I and Ice III phase boundaries of pure water.

  9. Gulf of Mexico Integrated Science - Tampa Bay Study - Characterization of Tidal Wetlands

    USGS Publications Warehouse

    McIvor, Carole

    2005-01-01

    Tidal wetlands in Tampa Bay, Florida, consist of mangrove forests and salt marshes. Wetlands buffer storm surges, provide fish and wildlife habitat, and enhance water quality through the removal of water-borne nutrients and contaminants. Substantial areas of both mangroves and salt marshes have been lost to agricultural, residential, and industrial development in this urban estuary. Wetlands researchers are characterizing the biological components of tidal wetlands and examining the physical factors such as salinity, tidal flushing, and sediment deposition that control the composition of tidal wetland habitats. Wetlands restoration is a priority of resource managers in Tampa Bay. Baseline studies such as these are needed for successful restoration planning and evaluation.

  10. Production of Isolated Giant Unilamellar Vesicles under High Salt Concentrations

    PubMed Central

    Stein, Hannah; Spindler, Susann; Bonakdar, Navid; Wang, Chun; Sandoghdar, Vahid

    2017-01-01

    The cell membrane forms a dynamic and complex barrier between the living cell and its environment. However, its in vivo studies are difficult because it consists of a high variety of lipids and proteins and is continuously reorganized by the cell. Therefore, membrane model systems with precisely controlled composition are used to investigate fundamental interactions of membrane components under well-defined conditions. Giant unilamellar vesicles (GUVs) offer a powerful model system for the cell membrane, but many previous studies have been performed in unphysiologically low ionic strength solutions which might lead to altered membrane properties, protein stability and lipid-protein interaction. In the present work, we give an overview of the existing methods for GUV production and present our efforts on forming single, free floating vesicles up to several tens of μm in diameter and at high yield in various buffer solutions with physiological ionic strength and pH. PMID:28243205

  11. Use of ammonium formate in QuEChERS for high-throughput analysis of pesticides in food by fast, low-pressure gas chromatography and liquid chromatography tandem mass spectrometry.

    PubMed

    González-Curbelo, Miguel Ángel; Lehotay, Steven J; Hernández-Borges, Javier; Rodríguez-Delgado, Miguel Ángel

    2014-09-05

    The "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) approach to sample preparation is widely applied in pesticide residue analysis, but the use of magnesium sulfate and other nonvolatile compounds for salting out in the method is not ideal for mass spectrometry. In this study, we developed and evaluated three new different versions of the QuEChERS method using more volatile salts (ammonium chloride and ammonium formate and acetate buffers) to induce phase separation and extraction of 43 representative pesticide analytes of different classes. Fast low-pressure gas chromatography tandem mass spectrometry (LPGC-MS/MS) and liquid chromatography (LC)-MS/MS were used for analysis. The QuEChERS AOAC Official Method 2007.01 was also tested for comparison purposes. Of the studied methods, formate buffering using 7.5g of ammonium formate and 15mL of 5% (v/v) formic acid in acetonitrile for the extraction of 15g of sample (5g for wheat grain) provided the best performance and practical considerations. Method validation was carried out with and without the use of dispersive solid-phase extraction for cleanup, and no significant differences were observed for the majority of pesticides. The method was demonstrated in quantitative analysis for GC- and LC-amenable pesticides in 4 representative food matrices (apple, lemon, lettuce, and wheat grain). With the typical exceptions of certain pH-dependent and labile pesticides, 90-110% recoveries and <10% RSD were obtained. Detection limits were mostly <5ng/g, which met the general need to determine pesticide concentrations as low as 10ng/g for monitoring purposes in food applications. Published by Elsevier B.V.

  12. A revised mechanism for the activation of complement C3 to C3b: a molecular explanation of a disease-associated polymorphism.

    PubMed

    Rodriguez, Elizabeth; Nan, Ruodan; Li, Keying; Gor, Jayesh; Perkins, Stephen J

    2015-01-23

    The solution structure of complement C3b is crucial for the understanding of complement activation and regulation. C3b is generated by the removal of C3a from C3. Hydrolysis of the C3 thioester produces C3u, an analog of C3b. C3b cleavage results in C3c and C3d (thioester-containing domain; TED). To resolve functional questions in relation to C3b and C3u, analytical ultracentrifugation and x-ray and neutron scattering studies were used with C3, C3b, C3u, C3c, and C3d, using the wild-type allotype with Arg(102). In 50 mm NaCl buffer, atomistic scattering modeling showed that both C3b and C3u adopted a compact structure, similar to the C3b crystal structure in which its TED and macroglobulin 1 (MG1) domains were connected through the Arg(102)-Glu(1032) salt bridge. In physiological 137 mm NaCl, scattering modeling showed that C3b and C3u were both extended in structure, with the TED and MG1 domains now separated by up to 6 nm. The importance of the Arg(102)-Glu(1032) salt bridge was determined using surface plasmon resonance to monitor the binding of wild-type C3d(E1032) and mutant C3d(A1032) to immobilized C3c. The mutant did not bind, whereas the wild-type form did. The high conformational variability of TED in C3b in physiological buffer showed that C3b is more reactive than previously thought. Because the Arg(102)-Glu(1032) salt bridge is essential for the C3b-Factor H complex during the regulatory control of C3b, the known clinical associations of the major C3S (Arg(102)) and disease-linked C3F (Gly(102)) allotypes of C3b were experimentally explained for the first time. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. A Revised Mechanism for the Activation of Complement C3 to C3b

    PubMed Central

    Rodriguez, Elizabeth; Nan, Ruodan; Li, Keying; Gor, Jayesh; Perkins, Stephen J.

    2015-01-01

    The solution structure of complement C3b is crucial for the understanding of complement activation and regulation. C3b is generated by the removal of C3a from C3. Hydrolysis of the C3 thioester produces C3u, an analog of C3b. C3b cleavage results in C3c and C3d (thioester-containing domain; TED). To resolve functional questions in relation to C3b and C3u, analytical ultracentrifugation and x-ray and neutron scattering studies were used with C3, C3b, C3u, C3c, and C3d, using the wild-type allotype with Arg102. In 50 mm NaCl buffer, atomistic scattering modeling showed that both C3b and C3u adopted a compact structure, similar to the C3b crystal structure in which its TED and macroglobulin 1 (MG1) domains were connected through the Arg102–Glu1032 salt bridge. In physiological 137 mm NaCl, scattering modeling showed that C3b and C3u were both extended in structure, with the TED and MG1 domains now separated by up to 6 nm. The importance of the Arg102–Glu1032 salt bridge was determined using surface plasmon resonance to monitor the binding of wild-type C3d(E1032) and mutant C3d(A1032) to immobilized C3c. The mutant did not bind, whereas the wild-type form did. The high conformational variability of TED in C3b in physiological buffer showed that C3b is more reactive than previously thought. Because the Arg102-Glu1032 salt bridge is essential for the C3b-Factor H complex during the regulatory control of C3b, the known clinical associations of the major C3S (Arg102) and disease-linked C3F (Gly102) allotypes of C3b were experimentally explained for the first time. PMID:25488663

  14. A Microchip-based Endothelium Mimic Utilizing Open Reservoirs for Cell Immobilization and Integrated Carbon Ink Microelectrodes for Detection

    PubMed Central

    Hulvey, Matthew K; Martin, R. Scott

    2010-01-01

    This paper describes the fabrication and characterization of a microfluidic device that utilizes a reservoir-based approach for endothelial cell immobilization and integrated embedded carbon ink microelectrodes for the amperometric detection of extracellular nitric oxide (NO) release. The design utilizes a buffer channel to continuously introduce buffer or a plug of stimulant to the reservoir as well as a separate sampling channel that constantly withdraws buffer from the reservoir and over the microelectrode. A steel pin is used for both the fluidic connection to the sampling channel and to provide a quasi-reference electrode for the carbon ink microelectrode. Characterization of the device was performed using NO standards produced from a NONOate salt. Finally, NO release from a layer of immobilized endothelial cells was monitored and quantified using the system. This system holds promise as a means to electrochemically detect extracellular NO release from endothelial cells in either an array of reservoirs or concurrently with fluorescence-based intracellular NO measurements. PMID:18989663

  15. Direct analysis of terpenes from biological buffer systems using SESI and IR-MALDESI.

    PubMed

    Nazari, Milad; Malico, Alexandra A; Ekelöf, Måns; Lund, Sean; Williams, Gavin J; Muddiman, David C

    2018-01-01

    Terpenes are the largest class of natural products with a wide range of applications including use as pharmaceuticals, fragrances, flavorings, and agricultural products. Terpenes are biosynthesized by the condensation of a variable number of isoprene units resulting in linear polyisoprene diphosphate units, which can then be cyclized by terpene synthases into a range of complex structures. While these cyclic structures have immense diversity and potential in different applications, their direct analysis in biological buffer systems requires intensive sample preparation steps such as salt cleanup, extraction with organic solvents, and chromatographic separations. Electrospray post-ionization can be used to circumvent many sample cleanup and desalting steps. SESI and IR-MALDESI are two examples of ionization methods that employ electrospray post-ionization at atmospheric pressure and temperature. By coupling the two techniques and doping the electrospray solvent with silver ions, olefinic terpenes of different classes and varying degrees of volatility were directly analyzed from a biological buffer system with no sample workup steps.

  16. Present status and strategy of reel-to-reel TFA-MOD process for coated conductors

    NASA Astrophysics Data System (ADS)

    Izumi, T.; Yoshizumi, M.; Miura, M.; Nakaoka, K.; Ichikawa, Y.; Sutoh, Y.; Miyata, S.; Fukushima, H.; Yamada, Y.; Shiohara, Y.

    2009-10-01

    On the research and development of a reel-to-reel TFA-MOD (Metal Organic Deposition using Trifluoro-acetates) process, a present status is reviewed and its future strategy is addressed. As a base of the study, the 90 m long tape with uniform I c distribution of the 300 A/cm-width level was obtained on the CeO 2 buffered IBAD-Gd 2Zr 2O 7/Hastelloy C276 substrate. The tape has the 56 m region with the end-to-end I c value of 250 A, which corresponds to maximum product of I c × L of 14,000 Am. Based on the results, several directions on R&D have been studied such as “higher I c”, “higher I c- B”, “higher production rate both in coating/calcinations and crystallization steps” and lower cost buffer/substrate”. Then, an extremely high I c value of 735 A/cm-width was achieved in a short tape by the compositional control (e.g. Ba-deficient), in the starting solutions. On the efforts for achieving higher I c- B properties, high I c values of 115 and 35 A/cm-width under the magnetic fields of 1 and 3 T were obtained by the RE mixture of Y and Gd in REBCO, addition of Zr and a growth rate control process. On the other hand, the production rate for the coating/calcinations process was improved by development of new starting solutions, which uses F-free Y salt instead of TFA salt of Y. The high J c value of 1.9 MA/cm 2 was confirmed using the precursor films fabricated at a high traveling rate of 10 m/h. Concerning a higher rate in the crystallization step, the multi-turning system with a vertical gas flow system was developed. The validity of the concept was confirmed using 2-turn parts of the furnace. The high I c value of 250 A/cm-width was realized in the 5 m tape crystallized with a traveling rate of 3 m/h, which is equivalent to 15 m/h for usage of entire area of the furnace of 10-turns. Furthermore, in order to achieve the lower cost, the architecture of the coated conductor with a low cost buffer/substrate system has been developed. An IBAD buffered substrate using IBAD-MgO layer (CeO 2/LMO/IBAD-MgO/Hastelloy C276) was developed and a high production rate of 24 m/h was realized for IBAD-MgO layer using a small ion gun system with the area of 6 × 22 cm 2. The grain texturing of the substrate was reached the Δ ϕ value of 4° in the CeO 2 layer. This substrate was applied to the above mentioned multi-turning crystallization furnace for TFA-MOD process. Then, a 5 m long tape with 260 A/cm-width (@77 K. s.f.) was achieved. According to the TFA-MOD process in the above achievements, the prospects of each issue for the future stage were independently confirmed. Consequently, R&D combining the above-mentioned achievements for longer tapes are expected in the next stage.

  17. Biomedical application of MALDI mass spectrometry for small-molecule analysis.

    PubMed

    van Kampen, Jeroen J A; Burgers, Peter C; de Groot, Ronald; Gruters, Rob A; Luider, Theo M

    2011-01-01

    Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is an emerging analytical tool for the analysis of molecules with molar masses below 1,000 Da; that is, small molecules. This technique offers rapid analysis, high sensitivity, low sample consumption, a relative high tolerance towards salts and buffers, and the possibility to store sample on the target plate. The successful application of the technique is, however, hampered by low molecular weight (LMW) matrix-derived interference signals and by poor reproducibility of signal intensities during quantitative analyses. In this review, we focus on the biomedical application of MALDI-MS for the analysis of small molecules and discuss its favorable properties and its challenges as well as strategies to improve the performance of the technique. Furthermore, practical aspects and applications are presented. © 2010 Wiley Periodicals, Inc.

  18. Petrogenesis of the Pd-rich intrusion at Salt Chuck, Prince of Wales island: an early Paleozoic Alaskan-type ultramafic body

    USGS Publications Warehouse

    Loney, R.A.; Himmelberg, G.R.

    1992-01-01

    The early Paleozoic Salt Chuck intrusion has petrographic and chemical characteristics that are similar to those of Cretaceous Alaskan-type ultramafic-mafic bodies. The intrusion is markedly discordant to the structure of the early Paleozoic Descon Formation, in which it has produced a rather indistinct contact aureole a few meters wide. Mineral assemblages, sequence of crystallization, and mineral chemistry suggest that the intrusion crystallized under low pressures (~2 kbar) with oxidation conditions near those of the NNO buffer, from a hydrous, silica-saturated, orthopyroxene-normative parental magma. The Salt Chuck deposit was probably formed by a two-stage process: 1) a stage of magmatic crystallization in which the sulfides and PGE accumulated in a disseminated manner in cumulus deposits, possibly largely in the gabbro, and 2) a later magmatic-hydrothermal stage during which the sulfides and PGE were remobilized and concentrated in veins and fracture-fillings. In this model, the source of the sulfides and PGE was the magma that produced the Salt Chuck intrusion. -from Authors

  19. Role of the Vascular Wall in Sodium Homeostasis and Salt Sensitivity

    PubMed Central

    Olde Engberink, Rik H.G.; Rorije, Nienke M.G.; Homan van der Heide, Jaap J.; van den Born, Bert-Jan H.

    2015-01-01

    Excessive sodium intake is associated with both hypertension and an increased risk of cardiovascular events, presumably because of an increase in extracellular volume. The extent to which sodium intake affects extracellular volume and BP varies considerably among individuals, discriminating subjects who are salt-sensitive from those who are salt-resistant. Recent experiments have shown that, other than regulation by the kidney, sodium homeostasis is also regulated by negatively charged glycosaminoglycans in the skin interstitium, where sodium is bound to glycosaminoglycans without commensurate effects on extracellular volume. The endothelial surface layer is a dynamic layer on the luminal side of the endothelium that is in continuous exchange with flowing blood. Because negatively charged glycosaminoglycans are abundantly present in this layer, it may act as an intravascular buffer compartment that allows sodium to be transiently stored. This review focuses on the putative role of the endothelial surface layer as a contributor to salt sensitivity, the consequences of a perturbed endothelial surface layer on sodium homeostasis, and the endothelial surface layer as a possible target for the treatment of hypertension and an expanded extracellular volume. PMID:25294232

  20. Identifying the mechanism of Escherichia coli O157:H7 survival by the addition of salt in the treatment with organic acids.

    PubMed

    Bae, Y-M; Yoon, J-H; Kim, J-Y; Lee, S-Y

    2018-01-01

    In this study, the effects of the addition of salt to treatment with acids (one of several organic acids and salt in various solutions including rich or minimal broth, buffer, or distilled water) on the reduction of Escherichia coli O157:H7 were investigated. The protein expression profiles corresponding to acid stress (acetic acid) with or without salt addition were studied using a comparative proteomic analysis of E. coli O157:H7. When acetic, lactic, or propionic acid was combined with 3% NaCl, mutually antagonistic effects of acid and salt on viability of E. coli O157:H7 were observed only in tryptone and yeast extract broth. After exposure to acetic acid alone or in combination with salt, approximately 851 and 916 protein spots were detected, respectively. Analysis of 10 statistically significant differentially expressed proteins revealed that these proteins are mainly related to energy metabolism. When we compared protein expression of E. coli O157:H7 treated with acetic acid and the combination of the acid and salt, the differentially expressed proteins were not related to acid stress- and salt stress-inducible proteins such as stress shock proteins. According to these results, the increased resistance of E. coli O157:H7 to acetic acid after the addition of salt may not be the result of synthesis of proteins related to these phenomena; therefore, further research needs to be conducted to identify the mechanism of the mutually antagonistic effect of some organic acids and salt. © 2017 The Society for Applied Microbiology.

  1. Use of a Phage-Display Method to Identify Peptides that Bind to a Tin Oxide Nanosheets.

    PubMed

    Nakazawa, Hikaru; Seta, Yasuko; Hirose, Tatsuya; Masuda, Yoshitake; Umetsu, Mitsuo

    2018-01-01

    Nanosheets of SnO2 which an n-type semiconductor with a rutile-type crystalline structure are predominantly used as gas sensors. SnO2 nanosheets have a tetragonal crystal structure where growth along the c-axis is suppressed to form a sheet. The major exposed facets of SnO2 nanosheets have {110}, {101} and {211} crystal planes along the a-axis, with the reduced {110} surface having a particularly high surface energy. Identifying peptides that bind to specific crystal planes by using peptide phage-display approach will increase the potential applications of metal oxide nanomaterials by fusing proteins with desirable active sites to peptides that adsorb at high density on the major exposed crystal plane of nanosheets. It may be possible to construct highly sensitive biosensors. The main objective of the present study is to identify peptides that adsorb preferentially to a SnO2 nanosheet by using peptide-phage display approach. Four milligrams of SnO2 nanosheet were mixed with 1011 plaque-forming units of Ph.D.-12 Phage Display Peptide Library. Phage-bound nanosheet particles were washed 10 times with 1 mL of phosphatebuffered saline containing 0.5% Tween 20. Phages bound to the nanosheet were eluted with three different buffers: (1) high-salt buffer containing 2 M NaCl (pH 7.5); (2) acidic buffer containing 200 mM Gly-HCl (pH 2.2); and (3) high-phosphate-ion buffer containing 500 mM NaH2PO4 (pH 7.5). The eluted phages were subjected to four or five rounds of biopanning. At each round, individual plaques were picked from the plates, and the amino acid sequences of the peptides were identified by DNA sequencing. The identified SnO2-binding peptides labeled with fluorescein isothiocyanate were synthesized. Adsorption isotherms were constructed at peptide concentrations ranging from 0.25 to 2.0 µM with 4mg of nanomaterials. We were determined the sequences of 11 clones with the high-salt buffer, 7 with the high-phosphateion buffers, and 6 with the acidic buffer and three peptides (SnO2BPn1, 2, and 3), two peptides (SnO2BPa1 and SnO2BPa2), and one peptide (SnO2BPp1) concentrated under each condition were selected respectively. All six selected peptides contained at least one histidine residue. In addition, the His-Asn-Leu (HNL) sequence was found in two of the peptides (SnO2BPa1 and SnO2BPa2). We constructed adsorption isotherms for the six selected peptides using 4mg of nanosheets. All six peptides were well adsorbed on the SnO2 nanosheet. The adsorption isotherms for SnO2 material with different structure revealed that SnO2BPn1, -2, and -3, and SnO2BPp1, preferentially bound to the spherical SnO2 nanoparticles. SnO2BPa2 preferentially bound to the SnO2 nanosheet, and SnO2BPa1 bound equally to both materials. This result suggested that SnO2BPa2 bound to a specific crystal plane of the nanosheet. The major exposed facet of the SnO2 crystal was the {110} plane, suggesting that SnO2BPa2 likely adsorbed on the {110} plane. SnO2BPn1, SnO2BPn2, SnO2BPn3, SnO2BPa1, and SnO2BPp1 also bound to the other metal oxides, in particular to ZrO2. At pH 7.5, peptides with a negative charge at pH 7.5 (pI 8.5-12) can bind to ZrO2 and SnO2, if the binding is mediated by electrostatic interactions. Thus, it is likely that these five peptides bind to metal oxides via electrostatic interactions. In contrast, SnO2BPa2 had a structurally specific affinity, binding more with the SnO2 nanosheet than with the spherical SnO2 nanoparticles or other metal oxides. We identified six peptides that adsorbed on a SnO2 nanosheet. Five of the selected peptides bound preferentially to spherical SnO2 nanoparticles rather than to the SnO2 nanosheet. Whereas, SnO2BPa2 exhibited specifically binding to the SnO2 nanosheet. Our results suggest that crystal plane recognition and material recognition by these peptides are mediated via different, independent mechanisms. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  2. [Separation of p-aminobenzenearsonic acid and its oxide by ion-pair reversed-phase high performance liquid chromatography].

    PubMed

    Kang, J; Ma, X; Meng, L; Ma, D

    1999-05-01

    To study the separation of p-aminobenzenearsonic acid (PABAA) and its oxide, p-aminophenylarsine oxide (PAPAO), both the absorption spectra were scanned at the wavelengths from 200 nm to 380 nm. PABAA had absorption maximum at 254 nm and PAPAO 258 nm. The effects of salt concentration, column temperature, methanol and ion-pair agent concentrations on the capacity factor were investigated. Compounds of high polarity showed almost no retention on reversed-phase column; as the volume fraction of the methanol decreased from 90% to 10%, the retention time of PABAA gradually increased with broad peak, and partially eluted when methanol volume fraction being below 20%. With temperature rising, the retention time of PABAA was decreased. But PABAA capacity factor can be increased by selecting an appropriate salt concentration for the mobile phase. The cetyltrimethyl and tetrabutyl ammonium ions were separately added as ion-pair agents to the mobile phase containing methanol in phosphate buffer of 10 mmol/L, the changes of retention time were observed. The mechanism of retention based on reversed phase ion-pair model is proposed. Besides, the retention behaviour is also influenced by size exclusion in stationary phase as well as polar interactions with residual silanol group on the silica surface.

  3. Novel hydrophobic interaction chromatography matrix for specific isolation and simple elution of immunoglobulins (A, G, and M) from porcine serum.

    PubMed

    Ramos-Clamont, Gabriela; del Carmen Candia-Plata, Maria; Zamudio, Roberto Guzman; Vazquez-Moreno, Luz

    2006-07-28

    A new, highly acetylated agarose matrix (HA-Sepharose) was synthesized and used as a hydrophobic interaction chromatography (HIC) medium to specifically isolate immunoglobulins (Igs) from porcine serum. Recovery of Igs was in a single step and under mild conditions. HA-Sepharose adsorption was studied in terms of salt, gel acetylation time, flow rate, and protein concentration on the loading buffer. At 0.5 M Na2SO4, control with unmodified Sepharose retained a small fraction (0.70 mg/mL of matrix) of serum albumin. On the contrary HA-Sepharose retained primary Igs (IgA, IgG, and 53% of IgM) as revealed by sodium dodecyl sulphate 10% polyacrylamide gel electrophoresis (SDS-PAGE), quantitative radial immunodiffusion and immunodetection. At a flow rate of 1 mL/min, the HA-Sepharose column capacity (3.9 mg/mL of matrix) was similar to the reported capacity for the commercial thiophilic T-gel. However, HA-Sepharose showed higher recovery of IgA and IgM than the T-gel in the same salt conditions, clearly an advantage in terms of immunoglobulin recovery strategies. Acetylation changed the matrix adsorption from albumin to immunoglobulins; thus, the highly acetylated gel rendered recoveries of Igs from unprocessed porcine serum practically free of albumin.

  4. In-situ Optical Waveguides for Monitoring and Modifying Protein Crystal Growth

    NASA Technical Reports Server (NTRS)

    Gibson, Ursula; Osterberg, Ulf

    2004-01-01

    The use of electric fields in the growth of protein crystals was investigated, both theoretically and experimentally. We used dc, ac and optical fields to change the spatial distribution of proteins. Dc fields had only local effects, due to the conductivity of the growth solution. We found that for low frequency fields, movement of the buffer and salt ions dominated, and that for high frequency ac fields, &electrophoretic effects could be useful for relocating growing protein crystals. The most promising result was that for optical fields, a large gradient in the field could be used to capture a crystal, and observe growth in-situ. This concept could be developed into an experimental setup compatible with automated x-ray diffraction measurements in microgravity.

  5. Root zone salinity and sodicity under seasonal rainfall due to feedback of decreasing hydraulic conductivity

    NASA Astrophysics Data System (ADS)

    van der Zee, S. E. A. T. M.; Shah, S. H. H.; Vervoort, R. W.

    2014-12-01

    Soil sodicity, where the soil cation exchange complex is occupied for a significant fraction by Na+, may lead to vulnerability to soil structure deterioration. With a root zone flow and salt transport model, we modeled the feedback effects of salt concentration (C) and exchangeable sodium percentage (ESP) on saturated hydraulic conductivity Ks(C, ESP) for different groundwater depths and climates, using the functional approach of McNeal (1968). We assume that a decrease of Ks is practically irreversible at a time scale of decades. Representing climate with a Poisson rainfall process, the feedback hardly affects salt and sodium accumulation compared with the case that feedback is ignored. However, if salinity decreases, the much more buffered ESP stays at elevated values, while Ks decreases. This situation may develop if rainfall has a seasonal pattern where drought periods with accumulation of salts in the root zone alternate with wet rainfall periods in which salts are leached. Feedback that affects both drainage/leaching and capillary upward flow from groundwater, or only drainage, leads to opposing effects. If both fluxes are affected by sodicity-induced degradation, this leads to reduced salinity (C) and sodicity (ESP), which suggests that the system dynamics and feedback oppose further degradation. Experiences in the field point in the same direction.

  6. Enhancing recovery of recombinant hepatitis B surface antigen in lab-scale and large-scale anion-exchange chromatography by optimizing the conductivity of buffers.

    PubMed

    Mojarrad Moghanloo, Gol Mohammad; Khatami, Maryam; Javidanbardan, Amin; Hosseini, Seyed Nezamedin

    2018-01-01

    In biopharmaceutical science, ion-exchange chromatography (IEC) is a well-known purification technique to separate the impurities such as host cell proteins from recombinant proteins. However, IEC is one of the limiting steps in the purification process of recombinant hepatitis B surface antigen (rHBsAg), due to its low recovery rate (<50%). In the current study, we hypothesized that ionic strengths of IEC buffers are easy-to-control parameters which can play a major role in optimizing the process and increasing the recovery. Thus, we investigated the effects of ionic strengths of buffers on rHBsAg recovery via adjusting Tris-HCl and NaCl concentrations. Increasing the conductivity of equilibration (Eq.), washing (Wash.) and elution (Elut.) buffers from their initial values of 1.6 mS/cm, 1.6 mS/cm, and 7.0 mS/cm to 1.6 mS/cm, 7 mS/cm and 50 mS/cm, respectively yielded an average recovery rate of 82% in both lab-scale and large-scale weak anion-exchange chromatography without any harsh effect on the purity percentage of rHBsAg. The recovery enhancement via increasing the conductivity of Eq. and Wash. buffers can be explained by their roles in reducing the binding strength and aggregation of retained particles in the column. Moreover, further increase in the salt concentration of Elut. Buffer could substantially promote the ion exchange process and the elution of retained rHBsAg. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. pH tunability and influence of alkali metal basicity on the plasmonic resonance of silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Yadav, Vijay D.; Akhil Krishnan, R.; Borade, Lalit; Shirolikar, Seema; Jain, Ratnesh; Dandekar, Prajakta

    2017-07-01

    Localized surface plasmon resonance has been a unique and intriguing feature of silver nanoparticles (AgNPs) that has attracted immense attention. This has led to an array of applications for AgNPs in optics, sensors, plasmonic imaging etc. Although numerous applications have been reported consistently, the importance of buffer and reaction parameters during the synthesis of AgNPs, is still unclear. In the present study, we have demonstrated the influence of parameters like pH, temperature and buffer conditions (0.1 M citrate buffer) on the plasmonic resonance of AgNPs. We found that neutral and basic pH (from alkali metal) provide optimum interaction conditions for nucleation of plasmon resonant AgNPs. Interestingly, this was not observed in the non-alkali metal base (ammonia). Also, when the nanoparticles synthesized from alkali metal base were incorporated in different buffers, it was observed that the nanoparticles dissolved in the acidic buffer and had reduced plasmonic resonance intensity. This, however, was resolved in the basic buffer, increasing the plasmonic resonance intensity and confirming that nucleation of nanoparticles required basic conditions. The above inference has been supported by characterization of AgNPs using UV-Vis spectrophotometer, Fluorimetry analysis, Infrared spectrometer and TEM analysis. The study concluded that the plasmonic resonance of AgNPs occurs due to the interaction of alkali (Na) and transition metal (Ag) salt in basic/neutral conditions, at a specific temperature range, in presence of a capping agent (citric acid), providing a pH tune to the overall system.

  8. Dynamic interactions between coastal storms and salt marshes: A review

    NASA Astrophysics Data System (ADS)

    Leonardi, Nicoletta; Carnacina, Iacopo; Donatelli, Carmine; Ganju, Neil Kamal; Plater, Andrew James; Schuerch, Mark; Temmerman, Stijn

    2018-01-01

    This manuscript reviews the progresses made in the understanding of the dynamic interactions between coastal storms and salt marshes, including the dissipation of extreme water levels and wind waves across marsh surfaces, the geomorphic impact of storms on salt marshes, the preservation of hurricanes signals and deposits into the sedimentary records, and the importance of storms for the long term survival of salt marshes to sea level rise. A review of weaknesses, and strengths of coastal defences incorporating the use of salt marshes including natural, and hybrid infrastructures in comparison to standard built solutions is then presented. Salt marshes are effective in dissipating wave energy, and storm surges, especially when the marsh is highly elevated, and continuous. This buffering action reduces for storms lasting more than one day. Storm surge attenuation rates range from 1.7 to 25 cm/km depending on marsh and storms characteristics. In terms of vegetation properties, the more flexible stems tend to flatten during powerful storms, and to dissipate less energy but they are also more resilient to structural damage, and their flattening helps to protect the marsh surface from erosion, while stiff plants tend to break, and could increase the turbulence level and the scour. From a morphological point of view, salt marshes are generally able to withstand violent storms without collapsing, and violent storms are responsible for only a small portion of the long term marsh erosion. Our considerations highlight the necessity to focus on the indirect long term impact that large storms exerts on the whole marsh complex rather than on sole after-storm periods. The morphological consequences of storms, even if not dramatic, might in fact influence the response of the system to normal weather conditions during following inter-storm periods. For instance, storms can cause tidal flats deepening which in turn promotes wave energy propagation, and exerts a long term detrimental effect for marsh boundaries even during calm weather. On the other hand, when a violent storm causes substantial erosion but sediments are redistributed across nearby areas, the long term impact might not be as severe as if sediments were permanently lost from the system, and the salt marsh could easily recover to the initial state.

  9. Dynamic interactions between coastal storms and salt marshes: A review

    USGS Publications Warehouse

    Leonardi, Nicoletta; Carnacina, Iacopo; Donatelli, Carmine; Ganju, Neil K.; Plater, Andrew James; Schuerch, Mark; Temmerman, Stijn

    2018-01-01

    This manuscript reviews the progresses made in the understanding of the dynamic interactions between coastal storms and salt marshes, including the dissipation of extreme water levels and wind waves across marsh surfaces, the geomorphic impact of storms on salt marshes, the preservation of hurricanes signals and deposits into the sedimentary records, and the importance of storms for the long term survival of salt marshes to sea level rise. A review of weaknesses, and strengths of coastal defences incorporating the use of salt marshes including natural, and hybrid infrastructures in comparison to standard built solutions is then presented.Salt marshes are effective in dissipating wave energy, and storm surges, especially when the marsh is highly elevated, and continuous. This buffering action reduces for storms lasting more than one day. Storm surge attenuation rates range from 1.7 to 25 cm/km depending on marsh and storms characteristics. In terms of vegetation properties, the more flexible stems tend to flatten during powerful storms, and to dissipate less energy but they are also more resilient to structural damage, and their flattening helps to protect the marsh surface from erosion, while stiff plants tend to break, and could increase the turbulence level and the scour. From a morphological point of view, salt marshes are generally able to withstand violent storms without collapsing, and violent storms are responsible for only a small portion of the long term marsh erosion.Our considerations highlight the necessity to focus on the indirect long term impact that large storms exerts on the whole marsh complex rather than on sole after-storm periods. The morphological consequences of storms, even if not dramatic, might in fact influence the response of the system to normal weather conditions during following inter-storm periods. For instance, storms can cause tidal flats deepening which in turn promotes wave energy propagation, and exerts a long term detrimental effect for marsh boundaries even during calm weather. On the other hand, when a violent storm causes substantial erosion but sediments are redistributed across nearby areas, the long term impact might not be as severe as if sediments were permanently lost from the system, and the salt marsh could easily recover to the initial state.

  10. Photoimageable composition

    DOEpatents

    Simison, Kelby Liv; Dentinger, Paul

    2003-11-11

    The use of selected buffering amines in a photoimageable composition prevents process bias which with conventional photoresists causes designed features to be distorted, especially in corners and high resolution features. It is believed that the amines react with the catalysts, e.g., photoacids, generated to create an inert salt. The presence of the amines also increases resolution. Suitable photoimageable compositions includes: (a) a multifunctional polymeric epoxy resin that is dissolved in an organic solvent wherein the epoxy resin comprises oligomers of bisphenol A that is quantitatively protected by glycidyl ether and wherein the oligomers have an average functionality that ranges from about 3 to 12; (b) a photoactive compound; and (c) an amine that is selected from the group consisting of triisobutylamine, 1,8-bis(dimethylamino)naphthalene (also known as PROTON SPONGET.TM.), 2,2'-diazabicyclo[2.2.2] octane and mixtures thereof. The photoimageable composition is particularly suited for producing high aspect ratio metal microstructures.

  11. Application of enhanced electronegative multimodal chromatography as the primary capture step for immunoglobulin G purification.

    PubMed

    Wang, Yanli; Chen, Quan; Xian, Mo; Nian, Rui; Xu, Fei

    2018-06-01

    In recent studies, electronegative multimodal chromatography with Eshmuno HCX was demonstrated to be a highly promising recovery step for direct immunoglobulin G (IgG) capture from undiluted cell culture fluid. In this study, the binding properties of HCX to IgG at different pH/salt combinations were systematically studied, and its purification performance was significantly enhanced by lowering the washing pH and conductivity after high capacity binding of IgG under its optimal conditions. A single polishing step gave an end-product with non-histone host cell protein (nh-HCP) below 1 ppm, DNA less than 1 ppb, which aggregates less than 0.5% and an overall IgG recovery of 86.2%. The whole non-affinity chromatography based two-column-step process supports direct feed loading without buffer adjustment, thus extraordinarily boosting the overall productivity and cost-savings.

  12. Reversed-phase thin-layer chromatography of homologs of Antimycin-A and related derivatives

    USGS Publications Warehouse

    Abidi, Sharon L.

    1989-01-01

    Using a reversed-phase high-performance liquid chromatographic (HPLC) technique, a mixture of antimycins A was separated into eight hitherto unreported subcomponents, Ala, Alb, A2a, A2b, A3a, A3b, A4a, and A4b. Although a base-line resolution of the known four major antimycins Al, A2, A3, and A4 was readily achieved with mobile phases containing acetate buffers, the separation of the new antibiotic subcomponents was highly sensitive to variation in mobile phase conditions. The type and composition of organic modifiers, the nature of buffer salts, and the concentration of added electrolytes had profound effects on capacity factors, separation factors, and peak resolution values. Of the numerous chromatographic systems examined, a mobile phase consisting of methanol-water (70:30) and 0.005 M tetrabutylammonium phosphate at pH 3.0 yielded the most satisfactory results for the separation of the subcomponents. Reversed-phase gradient HPLC separation of the dansylated or methylated antibiotic compounds produced superior chromatographic characteristics and the presence of added electrolytes was not a critical factor for achieving separation. Differences in the chromatographic outcome between homologous and structural isomers were interpretated based on a differential solvophobic interaction rationale. Preparative reversed-phase HPLC under optimal conditions enabled isolation of pure samples of the methylated antimycin subcomponents for use in structural studies.

  13. Salt disproportionation: A material science perspective.

    PubMed

    Thakral, Naveen K; Kelly, Ron C

    2017-03-30

    While screening the counter-ions for salt selection for an active pharmaceutical substance, there is often an uncertainty about disproportionation of the salt and hence physical stability of the final product formulation to provide adequate shelf life. Several examples of disproportionation reactions are reviewed to explain the concepts of pHmax, microenvironmental pH, and buffering capacity of excipients and APIs to gain mechanistic understanding of disproportionation reaction. Miscellaneous factors responsible for disproportionation are examined. In addition to the dissolution failure due to the formation of less soluble unionized form, various implications of the disproportionation are evaluated with specific examples. During lead optimization and early stages of development, when only a limited amount of material is available, use of predictive tools like mathematical models and model free kinetics to rank order the various counter-ions are discussed in detail. Finally, analytical methods and mitigation strategies are discussed to prevent the disproportionation by detecting it during early stages of drug development. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. (Poly)cation-induced protection of conventional and wireframe DNA origami nanostructures.

    PubMed

    Ahmadi, Yasaman; De Llano, Elisa; Barišić, Ivan

    2018-04-26

    DNA nanostructures hold immense potential to be used for biological and medical applications. However, they are extremely vulnerable towards salt depletion and nucleases, which are common under physiological conditions. In this contribution, we used chitosan and linear polyethyleneimine for coating and long-term stabilization of several three-dimensional DNA origami nanostructures. The impact of the degree of polymerization and the charge density of the polymer together with the N/P charge ratio (ratio of the amines in polycations to the phosphates in DNA) on the stability of encapsulated DNA origami nanostructures in the presence of nucleases and in low-salt media was examined. The polycation shells were compatible with enzyme- and aptamer-based functionalization of the DNA nanostructures. Additionally, we showed that despite being highly vulnerable to salt depletion and nucleolytic digestion, self-assembled DNA nanostructures are stable in cell culture media up to a week. This was contrary to unassembled DNA scaffolds that degraded in one hour, showing that placing DNA strands into a spatially designed configuration crucially affect the structural integrity. The stability of naked DNA nanostructures in cell culture was shown to be mediated by growth media. DNA origami nanostructures kept in growth media were significantly more resistant towards low-salt denaturation, DNase I and serum-mediated digestion than when in a conventional buffer. Moreover, we confirmed that DNA origami nanostructures remain not only structurally intact but also fully functional after exposure to cell media. Agarose gel electrophoresis and negative stain transmission electron microscopy analysis revealed the hybridization of DNA origami nanostructures to their targets in the presence of serum proteins and nucleases. The structural integrity and functionality of DNA nanostructures in physiological fluids validate their use particularly for short-time biological applications in which the shape and structural details of DNA nanodevices are functionally critical.

  15. Low voltage to high voltage level shifter and related methods

    NASA Technical Reports Server (NTRS)

    Mentze, Erik J. (Inventor); Buck, Kevin M. (Inventor); Hess, Herbert L. (Inventor); Cox, David F. (Inventor)

    2006-01-01

    A shifter circuit comprises a high and low voltage buffer stages and an output buffer stage. The high voltage buffer stage comprises multiple transistors arranged in a transistor stack having a plurality of intermediate nodes connecting individual transistors along the stack. The transistor stack is connected between a voltage level being shifted to and an input voltage. An inverter of this stage comprises multiple inputs and an output. Inverter inputs are connected to a respective intermediate node of the transistor stack. The low voltage buffer stage has an input connected to the input voltage and an output, and is operably connected to the high voltage buffer stage. The low voltage buffer stage is connected between a voltage level being shifted away from and a lower voltage. The output buffer stage is driven by the outputs of the high voltage buffer stage inverter and the low voltage buffer stage.

  16. Strategies for the depyrogenation of contaminated immunoglobulin G solutions by histidine-immobilized hollow fiber membrane.

    PubMed

    Legallais, C; Anspach, F B; Bueno, S M; Haupt, K; Vijayalakshmi, M A

    1997-03-28

    The depyrogenation of different IgG solutions using the histidine-linked hollow fiber membrane developed in our laboratory is presented here. Three strategies for endotoxin (ET) removal were investigated according to the immobilized histidine's ability to bind different immunoglobulins: (1) ET removal from 1 mg/ml non histidine-binding mouse monoclonal IgG1 (MabCD4) solution was achieved in the presence of acetate buffer (pH 5.0) without any protein loss. (2) For contaminated human IgG, combined adsorption of ET and IgG in the presence of MOPS of Tris buffer was tested, followed by differential elution using increasing salt concentrations. This attempt was not successful since ET were quantitatively found in the IgG elution fraction. (3) Alternatively, it was proposed to adsorb selectively ET in the presence of acetate buffer (pH 5.0) under non binding conditions for human IgG. Human IgG could then be purified if necessary with the same membrane in the presence of MOPS buffer (pH 6.5). With a 1 m2 histidine-PEVA module under these operating conditions, it is estimated that the depyrogenation of 3 l of 1 mg/ml IgG (human or murine) solution containing 80 EU/ml of ET should be possible.

  17. Protein separation through preliminary experiments concerning pH and salt concentration by tube radial distribution chromatography based on phase separation multiphase flow using a polytetrafluoroethylene capillary tube.

    PubMed

    Kan, Hyo; Tsukagoshi, Kazuhiko

    2017-07-01

    Protein mixtures were separated using tube radial distribution chromatography (TRDC) in a polytetrafluoroethylene (PTFE) capillary (internal diameter=100µm) separation tube. Separation by TRDC is based on the annular flow in phase separation multiphase flow and features an open-tube capillary without the use of specific packing agents or application of high voltages. Preliminary experiments were conducted to examine the effects of pH and salt concentration on the phase diagram of the ternary mixed solvent solution of water-acetonitrile-ethyl acetate (8:2:1 volume ratio) and on the TRDC system using the ternary mixed solvent solution. A model protein mixture containing peroxidase, lysozyme, and bovine serum albumin was analyzed via TRDC with the ternary mixed solvent solution at various pH values, i.e., buffer-acetonitrile-ethyl acetate (8:2:1 volume ratio). Protein was separated on the chromatograms by the TRDC system, where the elution order was determined by the relation between the isoelectric points of protein and the pH values of the solvent solution. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Effect of Hofmeister series salts on Absorptivity of aqueous solutions on Sodium polyacrylate

    NASA Astrophysics Data System (ADS)

    Korrapati, Swathi; Pullela, Phani Kumar; Vijayalakshmi, U.

    2017-11-01

    Sodium polyacrylate (SPA) is a popular super absorbent commonly used in children diapers, sanitary pads, adult diapers etc. The use of SPA is in force from past 30 years and the newer applications like as food preservant are evolving. SPA is recently discovered by our group for improvement of sensitivity of colorimetric agents. Though the discovery of improvement in sensitivity is phenomenal, the mechanism still remains a puzzle. A typical assay reagent contains colorimetric/fluorescent reagents, buffers, salts, stabilizers etc. These chemicals are known to influence the water absorptivity of SPA. If we were to perform chemical/biochemical assays on SPA absorbed reagents effect of salts and other excipients on colorimetric/fluorescence compounds absorbed on SPA is very important. The hofmeister series are standard for studying effect of salts on permeability, stability, aggregation, fluorescence quenching etc. We recently studied affect of urea, sodium chloride, ammonium sulfate, guanidine thiocayanate on fluorescence characteristics of fluorescence compounds and noted that except urea all other reagents have resulted in fluorescence quenching and urea had an opposite effect and increased the fluorescence intensity. This result was attributed to the different water structure around fluorescent in urea solution versus other chaotropic agents.

  19. Experimental and theoretical analysis of the rate of solvent equilibration in the hanging drop method of protein crystal growth

    NASA Technical Reports Server (NTRS)

    Fowlis, William W.; Delucas, Lawrence J.; Twigg, Pamela J.; Howard, Sandra B.; Meehan, Edward J.

    1988-01-01

    The principles of the hanging-drop method of crystal growth are discussed, and the rate of water evaporation in a water droplet (containing protein, buffer, and a precipitating agent) suspended above a well containing a double concentration of precipitating agent is investigated theoretically. It is shown that, on earth, the rate of evaporation may be determined from diffusion theory and the colligative properties of solutions. The parameters affecting the rate of evaporation include the temperature, the vapor pressure of water, the ionization constant of the salt, the volume of the drop, the contact angle between the droplet and the coverslip, the number of moles of salt in the droplet, the number of moles of water and salt in the well, the molar volumes of water and salt, the distance from the droplet to the well, and the coefficient of diffusion of water vapor through air. To test the theoretical equations, hanging-drop experiments were conducted using various reagent concentrations in 25-microliter droplets and measuring the evaporation times at 4 C and 25 C. The results showed good agreement with the theory.

  20. Toward a Molecular Understanding of Protein Solubility: Increased Negative Surface Charge Correlates with Increased Solubility

    PubMed Central

    Kramer, Ryan M.; Shende, Varad R.; Motl, Nicole; Pace, C. Nick; Scholtz, J. Martin

    2012-01-01

    Protein solubility is a problem for many protein chemists, including structural biologists and developers of protein pharmaceuticals. Knowledge about how intrinsic factors influence solubility is limited due to the difficulty of obtaining quantitative solubility measurements. Solubility measurements in buffer alone are difficult to reproduce, because gels or supersaturated solutions often form, making it impossible to determine solubility values for many proteins. Protein precipitants can be used to obtain comparative solubility measurements and, in some cases, estimations of solubility in buffer alone. Protein precipitants fall into three broad classes: salts, long-chain polymers, and organic solvents. Here, we compare the use of representatives from two classes of precipitants, ammonium sulfate and polyethylene glycol 8000, by measuring the solubility of seven proteins. We find that increased negative surface charge correlates strongly with increased protein solubility and may be due to strong binding of water by the acidic amino acids. We also find that the solubility results obtained for the two different precipitants agree closely with each other, suggesting that the two precipitants probe similar properties that are relevant to solubility in buffer alone. PMID:22768947

  1. Supramolecular Assembly of a Biomineralizing Antimicrobial Peptide in Coarse-Grained Monte Carlo Simulations (Postprint)

    DTIC Science & Technology

    2010-07-05

    aqueous solutions at a wide pH range.7 Silica forms rapidly at room temperature in the absence of other ionic catalysts or cofactors (buffers, salts...and other ionic species) that are normally required with in vitro biosilica synthesis. The silaffins are uniquely functionalized; serine residues are...alkaline conditions mimic the properties of the poly- ionic modifications on native silaffins.13,14 The zwitterionic properties facilitate intermolecular

  2. Deposition of zinc sulfide thin films by chemical bath process

    NASA Astrophysics Data System (ADS)

    Oladeji, Isaiah O.; Chow, Lee

    1996-11-01

    Deposition of high quality zinc sulfide (ZnS) thin film over a large area is required if it is to be effectively used in electroluminescent devices, solar cells, and other optoelectronic devices. Of all deposition techniques, chemical bath deposition (CBD) is the least costly technique that meets the above requirements. Recently it is found that the growth of ZnS film, of thickness less than 100 nm in a single dip, by CBD is facilitated by the use of ammonia and hydrazine as complexing agents. Here we report that the thickness of the deposited ZnS film can be increased if ammonium salt is used as a buffer. We also present an analytical study to explain our results and to further understand the ZnS growth process in CBD.

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

    Arieen, A.; Goigoux, C.; Baquey, C.

    Factors affecting liposome transport to the blood compartment after oral administration to rats were evaluated. A high entrapment of calcitonin (CT) was obtained when the vesicles were prepared by sonication and were composed of egg phosphatidylcholine, cholesterol and stearylamine. In vitro tests showed that the liposomes were stable in light acidic or basic buffers, but that they were partly lysed in pH 2.5, 10 mM bile salts and pancreatin. Oral administration of liposomes entrapping calcitonin in fasting rats showed that the vesicles facilitate transport of the hormone to the general circulation and that they increase the lifetime of [sup 125]I-CTmore » in blood. Oral administration of liposomes entrapping radioactive indium in fasting rats did not induce radioactivity in blood. This could be explained by disruption of most of the vesicles in the enterocytes.« less

  4. Salt-induced inhibition of the precipitin reaction of concanavalin A with polysaccharides and glycoprotein.

    PubMed

    Salahuddin, A; Begum, R; Averill, B K

    1984-06-15

    The time course of the precipitin reactions of concanavalin A with glycogen, dextran and ovalbumin was investigated by a light-scattering method near 30 degrees C in 10 mM-Tris/HCl buffer, pH 7.4, containing neutral salts, i.e. NaCl, KCl, NaBr, KI and NaClO4. With 0.8 microM-lectin and 0.36 mg of glycogen/ml, the half-life, t 1/2, of the precipitin reaction was independent of salt concentration between 0.1 M and 1.5 M, and was the same (175s) in the presence of NaCl, KCl, NaBr and KI but was significantly (27%) higher in NaClO4. In contrast, the five salts caused significant to marked enhancement in t 1/2 for the reactions of concanavalin A with dextran and ovalbumin. Likewise, whereas the turbidity produced in 1 h as a result of lectin-glycogen precipitation remained unchanged, those measured for the binding of dextran and ovalbumin were decreased in the presence of three salts. The increase in t 1/2 and decrease in turbidity were found to be higher with NaClO4, followed by KI; NaBr produced moderate and NaCl (or KCl) small but generally significant inhibition of the precipitin reactions with dextran and ovalbumin. The results showed that the lectin-ligand precipitin reactions involve salt-sensitive polar interactions that are less pronounced with compactly folded ligands such as glycogen.

  5. Intrinsic cytosolic calcium buffering properties of single rat cardiac myocytes.

    PubMed Central

    Berlin, J R; Bassani, J W; Bers, D M

    1994-01-01

    Intracellular passive Ca2+, buffering was measured in voltage-clamped rat ventricular myocytes. Cells were loaded with indo-1 (K+ salt) to an estimated cytosolic concentration of 44 +/- 5 microM (Mean +/- SEM, n = 5), and accessible cell volume was estimated to be 24.5 +/- 3.6 pl. Ca2+ transport by the sarcoplasmic reticulum (SR) Ca-ATPase and sarcolemmal Na-Ca exchange was inhibited by treatment with thapsigargin and Na-free solutions, respectively. Extracellular [Ca2+] was maintained at 10 mM and, in some experiments, the mitochondrial uncoupler "1799" was used to assess the degree of mitochondrial Ca2+ uptake. To perform single cell titrations, intracellular Ca2+ ([Ca2+]i) was increased progressively by a train of depolarizing voltage clamp pulses from -40 to +10 mV. The total Ca2+ gain with each pulse was calculated by integration of the Ca current and then analyzed as a function of the rapid change in [Ca2+]i during the pulse. In the range of [Ca2+]i from 0.1 to 2 microM, overall cell buffering was well described as a single lumped Michaelis-Menten type species with an apparent dissociation constant, KD, of of 0.63 +/- 0.07 microM (n = 5) and a binding capacity, Bmax, of 162 +/- 15 mumol/l cell H2O. Correction for buffering attributable to cytosolic indo-1 gives intrinsic cytosolic Ca2+ buffering parameters of KD = 0.96 +/- 0.18 microM and Bmax = 123 +/- 18 mumol/l cell H2O. The fast Ca2+ buffering measured in this manner agrees reasonably with the characteristics of known rapid Ca buffers (e.g., troponin C, calmodulin, and SR Ca-ATPase), but is only about half of the total Ca2+ buffering measured at equilibrium. Inclusion of slow Ca buffers such as the Ca/Mg sites on troponin C and myosin can account for the differences between fast Ca2+ buffering in phase with the Ca current measured in the present experiments and equilibrium Ca2+ buffering. The present data indicate that a rapid rise of [Ca2+]i from 0.1 to 1 microM during a contraction requires approximately 50 microM Ca2+ to be added to the cytosol. PMID:7819510

  6. Interaction between crustal tectonics and salt deformation in the Eastern Sardinian margin, Western Tyrrhenian Sea: seismic data and analogue modelling

    NASA Astrophysics Data System (ADS)

    Vendeville, Bruno; Lymer, Gael; Gaullier, Virginie; Chanier, Frank; Maillard, Agnes; Sage, Françoise; Lofi, Johanna; Thinon, Isabelle

    2014-05-01

    The Tyrrhenian Basin opened by eastward migration of the Apennine subduction system. Rifting along the Eastern Sardinian margin started during the middle to late Miocene times and hence this timing partly overlapped the Messinian Salinity Crisis. The two "METYSS" cruises were conducted to use the deformation of the Messinian salt and its Plio-Quaternary overburden as a proxy for better delineating the tectonic history of the sub-salt basement. Many parts of the study area contain two of the most typical Messinian series of the Western Mediterranean: the Mobile Unit (MU; salt, mainly halite), overlain by the more competent Upper Unit (UU: alternating dolomitic marls and anhydrite). The brittle Plio-Quaternary cover overlies the UU. Usually, the presence of mobile salt is viewed as a nuisance for understanding crustal tectonics because salt's ability to act as a structural buffer between the basement and the cover. However, we illustrate, using examples from the Cornaglia Terrace, how we can use thin-skinned salt tectonics as indicators of vertical movements in the sub-salt, pre-Messinian basement. There, slip along N-S-trending crustal normal faults bounding basement troughs has been recorded by salt and overburden in two different manners: - First, post-salt basement faulting (typically after deposition of the Upper Unit and the early Pliocene), and some crustal-scale southward tilting, triggered along-strike (southward) thin-skinned, gliding of salt and overburden recorded by upslope extension and downslope shortening. - Second, and less obvious at first glance, there was some crustal activity along another basement trough, located East of the Baronie Ridge after deposition of the Messinian salt. This trough is narrow, trends N-S and is bounded by crustal faults. The narrow width of the trough allowed for only minor across-strike (E-W) gliding. The resulting geometry would suggest that nothing happened after Messinian times, but some structural features (confirmed by analogue modelling) show that basement fault slip and tilting (Eastward or Westward) was accommodated by lateral flow of salt, which thinned upslope and inflated downslope, while the overlying sediments remained sub-horizontal.

  7. Stream-Groundwater Interaction Buffers Seasonal Changes in Urban Stream Water Quality

    NASA Astrophysics Data System (ADS)

    Ledford, S. H.; Lautz, L. K.

    2013-12-01

    Urban streams in the northeastern United States have large road salt inputs during winter, increased nonpoint sources of inorganic nitrogen, and decreased short-term and permanent storage of nutrients. Meadowbrook Creek, a first order stream in Syracuse, New York, flows along a negative urbanization gradient, from a channelized and armored stream running through the middle of a roadway to a pool-riffle stream meandering through a broad, vegetated floodplain with a riparian aquifer. In this study we investigated how reconnection to groundwater and introduction of riparian vegetation impacted surface water chemistry by making bi-weekly longitudinal surveys of stream water chemistry in the creek from May 2012 until June 2013. Chloride concentrations in the upstream, urban reach of Meadowbrook Creek were strongly influenced by discharge of road salt to the creek during snow melt events in winter and by the chemistry of water draining an upstream retention basin in summer. Chloride concentrations ranged from 161.2 mg/L in August to 2172 mg/L in February. Chloride concentrations in the downstream, 'connected' reach had less temporal variation, ranging from 252.0 mg/L in August to 1049 mg/L in January, and were buffered by groundwater discharge, as the groundwater chloride concentrations during the sampling period ranged from 84.0 to 655.4 mg/L. Groundwater discharge resulted in higher chloride concentrations in summer and lower concentrations in winter in the connected reach relative to the urban reach, minimizing annual variation. In summer, there was little-to-no nitrate in the urban reach due to a combination of limited sources and high primary productivity. In contrast, during the summer, nitrate concentrations reached over 1 mg N/L in the connected reach due to the presence of riparian vegetation and lower nitrate uptake due to cooler temperatures and shading. During the winter, when temperatures fell below freezing, nitrate concentrations in the urban reach increased to around 0.58 mg N/L, but were still lower than the connected reach, which averaged 0.88 mg N/L. Groundwater discharge rates were measured longitudinally along the creek during a constant rate Rhodamine WT injection and also confirmed qualitatively by longitudinal changes in stream sulfate and δ18O. The buffering capability of groundwater discharge in urban systems has implications for managers trying to mitigate the effects of urbanization on surface water.

  8. Non-monotonic swelling of surface grafted hydrogels induced by pH and/or salt concentration

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

    Longo, Gabriel S.; Department of Biomedical Engineering, Northwestern University, Evanston, Illinois 60208; Chemistry of Life Processes Institute, Northwestern University, Evanston, Illinois 60208

    2014-09-28

    We use a molecular theory to study the thermodynamics of a weak-polyacid hydrogel film that is chemically grafted to a solid surface. We investigate the response of the material to changes in the pH and salt concentration of the buffer solution. Our results show that the pH-triggered swelling of the hydrogel film has a non-monotonic dependence on the acidity of the bath solution. At most salt concentrations, the thickness of the hydrogel film presents a maximum when the pH of the solution is increased from acidic values. The quantitative details of such swelling behavior, which is not observed when themore » film is physically deposited on the surface, depend on the molecular architecture of the polymer network. This swelling-deswelling transition is the consequence of the complex interplay between the chemical free energy (acid-base equilibrium), the electrostatic repulsions between charged monomers, which are both modulated by the absorption of ions, and the ability of the polymer network to regulate charge and control its volume (molecular organization). In the absence of such competition, for example, for high salt concentrations, the film swells monotonically with increasing pH. A deswelling-swelling transition is similarly predicted as a function of the salt concentration at intermediate pH values. This reentrant behavior, which is due to the coupling between charge regulation and the two opposing effects triggered by salt concentration (screening electrostatic interactions and charging/discharging the acid groups), is similar to that found in end-grafted weak polyelectrolyte layers. Understanding how to control the response of the material to different stimuli, in terms of its molecular structure and local chemical composition, can help the targeted design of applications with extended functionality. We describe the response of the material to an applied pressure and an electric potential. We present profiles that outline the local chemical composition of the hydrogel, which can be useful information when designing applications that pursue or require the absorption of biomolecules or pH-sensitive molecules within different regions of the film.« less

  9. Buffers and Oscillations in Intracellular Ca2+ Dynamics

    PubMed Central

    Falcke, Martin

    2003-01-01

    I model the behavior of intracellular Ca2+ release with high buffer concentrations. The model uses a spatially discrete array of channel clusters. The channel subunit dynamics is a stochastic representation of the DeYoung-Keizer model. The calculations show that the concentration profile of fast buffer around an open channel is more localized than that of slow buffers. Slow buffers allow for release of larger amounts of Ca2+ from the endoplasmic reticulum and hence bind more Ca2+ than fast buffers with the same dissociation constant and concentration. I find oscillation-like behavior for high slow buffer concentration and low Ca2+ content of the endoplasmic reticulum. High concentration of slow buffer leads to oscillation-like behavior by repetitive wave nucleation for high Ca2+ content of the endoplasmic reticulum. Localization of Ca2+ release by slow buffer, as used in experiments, can be reproduced by the modeling approach. PMID:12524263

  10. Enhancing longevity of Plasmodium vivax and P. falciparum sporozoites after dissection from mosquito salivary glands.

    PubMed

    Lupton, Emily J; Roth, Alison; Patrapuvich, Rapatbhorn; Maher, Steve P; Singh, Naresh; Sattabongkot, Jetsumon; Adams, John H

    2015-04-01

    The pre-erythrocytic stages of Plasmodium vivax and Plasmodium falciparum remain challenging for experimental research in part due to limited access to sporozoites. An important factor limiting availability is the laboratory support required for producing infected mosquitoes and the ephemeral nature of isolated extracellular sporozoites. This study was undertaken to investigate methods to improve the availability of this limited resource by extending the longevity of the extracellular sporozoites after mosquito dissection. Our goal in this study was to determine whether buffer conditions more closely mimicking the insect microenvironment could prolong longevity of ex vivo P. vivax and P. falciparum sporozoites. The study compared the current standard dissection buffer RPMI1640 to Hank's Balanced Salt Solution with 1g/L glucose (HBSS-1) or 2g/L glucose (HBSS-2) and Grace's Insect Medium for ability to extend longevity of ex vivo P. vivax and P. falciparum sporozoites. The effect of each buffer on sporozoite viability was evaluated by measuring sporozoite gliding motility at 0, 4, 8, and 24h post-dissection from mosquito salivary glands. Comparisons of mean gliding percentages of ex vivo sporozoites in the different buffers and time points found that RPMI and Grace's both showed strong gliding at 0h. In contrast, by 4h post-dissection sporozoites in RPMI consistently had the lowest gliding activity, whereas sporozoites in Grace's had significantly more gliding compared to all other buffers at almost all time points. Our results indicate that P. vivax and P. falciparum sporozoites maintained in insect media rather than the standard dissection buffer RPMI and HBSS retain viability better over time. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  11. Minute synthesis of extremely stable gold nanoparticles.

    PubMed

    Zhou, Min; Wang, Baoxiang; Rozynek, Zbigniew; Xie, Zhaohui; Fossum, Jon Otto; Yu, Xiaofeng; Raaen, Steinar

    2009-12-16

    We describe a rapid environmentally friendly wet-chemical approach to synthesize extremely stable non-toxic, biocompatible, water-soluble monodispersed gold nanoparticles (AuNPs) in one step at room temperature. The particles have been successfully achieved in just a few minutes by merely adding sodium hydroxide (NaOH) acting as an initiator for the reduction of HAuCl(4) in aqueous solution in the presence of polyvinylpyrrolidone (PVP) without the use of any reducing agent. It is also proved to be highly efficient for the preparation of AuNPs with controllable sizes. The AuNPs show remarkable stability in water media with high concentrations of salt, various buffer solutions and physiological conditions in biotechnology and biomedicine. Moreover, the AuNPs are also non-toxic at high concentration (100 microM). Therefore, it provides great opportunities to use these AuNPs for biotechnology and biomedicine. This new approach also involved several green chemistry concepts, such as the selection of environmentally benign reagents and solvents, without energy consumption, and less reaction time.

  12. Investigation of factors affecting in vitro doxorubicin release from PEGylated liposomal doxorubicin for the development of in vitro release testing conditions.

    PubMed

    Shibata, Hiroko; Izutsu, Ken-Ichi; Yomota, Chikako; Okuda, Haruhiro; Goda, Yukihiro

    2015-01-01

    Establishing appropriate drug release testing methods of liposomal products for assuring quality and performance requires the determination of factors affecting in vitro drug release. In this study, we investigated the effects of test conditions (human plasma lot, pH/salt concentration in the test media, dilution factor, temperature, ultrasound irradiation, etc.), and liposomal preparation conditions (pH/concentration of ammonium sulfate solution), on doxorubicin (DXR) release from PEGylated liposomal DXR. Higher temperature and lower pH significantly increased DXR release. The evaluation of DXR solubility indicated that the high DXR release induced by low pH may be attributed to the high solubility of DXR at low pH. Ultrasound irradiation induced rapid DXR release in an amplitude-dependent manner. The salt concentration in the test solution, human plasma lot, and dilution factor had a limited impact on DXR-release. Variations in the ammonium sulfate concentration used in solutions for the formation/hydration of liposomes significantly affected DXR release behavior, whereas differences in pH did not. In addition, heating condition in phosphate-buffered saline at lower pH (<6.5) exhibited higher discriminative ability for the release profiles from various liposomes with different concentrations of ammonium sulfate than did ultrasound irradiation. These results are expected to be helpful in the process of establishing appropriate drug release testing methods for PEGylated liposomal DXR.

  13. Development of a Highly Biocompatible Antituberculosis Nanodelivery Formulation Based on Para-Aminosalicylic Acid—Zinc Layered Hydroxide Nanocomposites

    PubMed Central

    Arulselvan, Palanisamy; El Zowalaty, Mohamed Ezzat; Fakurazi, Sharida; Webster, Thomas J.; Geilich, Benjamin; Hussein, Mohd Zobir

    2014-01-01

    Tuberculosis is a lethal epidemic, difficult to control disease, claiming thousands of lives every year. We have developed a nanodelivery formulation based on para-aminosalicylic acid (PAS) and zinc layered hydroxide using zinc nitrate salt as a precursor. The developed formulation has a fourfold higher efficacy of PAS against mycobacterium tuberculosis with a minimum inhibitory concentration (MIC) found to be at 1.40 μg/mL compared to the free drug PAS with a MIC of 5.0 μg/mL. The newly developed formulation was also found active against Gram-positive bacteria, Gram-negative bacteria, and Candida albicans. The formulation was also found to be biocompatible with human normal lung cells MRC-5 and mouse fibroblast cells-3T3. The in vitro release of PAS from the formulation was found to be sustained in a human body simulated phosphate buffer saline (PBS) solution at pH values of 7.4 and 4.8. Most importantly the nanocomposite prepared using zinc nitrate salt was advantageous in terms of yield and free from toxic zinc oxide contamination and had higher biocompatibility compared to one prepared using a zinc oxide precursor. In summary, these promising in vitro results are highly encouraging for the continued investigation of para-aminosalicylic acid and zinc layered hydroxide nanocomposites in vivo and eventual preclinical studies. PMID:25050392

  14. The adsorption and reaction of adenine nucleotides on montmorillonite

    NASA Technical Reports Server (NTRS)

    Ferris, James P.; Hagan, William J., Jr.

    1986-01-01

    The binding of AMP to Zn(2+)-montmorillonite is investigated in the presence of salts and Good's zwitterion buffers, PIPES and MES. The initial concentrations of nucleotide and the percent adsorbtion are used to calculate the adsorption isotherms, and the Langmuir adsorption equation is used for the analysis of data. The adsorption coefficient was found to be three times greater in the presence of 0.2 M PIPES than in its absence. In addition, basal spacings measured by X-ray diffraction were increased by the buffer. These results are interpreted in terms of a model in which the adsorption of AMP is mediated by a Zn(2+) complex of PIPES in different orientations in the interlamellar region of the montmorillonite. Mixed ligand complexes of this type are reminiscent of the complexes observed between metal ions and biological molecules in living systems.

  15. Force-controlled inorganic crystallization lithography.

    PubMed

    Cheng, Chao-Min; LeDuc, Philip R

    2006-09-20

    Lithography plays a key role in integrated circuits, optics, information technology, biomedical applications, catalysis, and separation technologies. However, inorganic lithography techniques remain of limited utility for applications outside of the typical foci of integrated circuit manufacturing. In this communication, we have developed a novel stamping method that applies pressure on the upper surface of the stamp to regulate the dewetting process of the inorganic buffer and the evaporation rate of the solvent in this buffer between the substrate and the surface of the stamp. We focused on generating inorganic microstructures with specific locations and also on enabling the ability to pattern gradients during the crystallization of the inorganic salts. This approach utilized a combination of lithography with bottom-up growth and assembly of inorganic crystals. This work has potential applications in a variety of fields, including studying inorganic material patterning and small-scale fabrication technology.

  16. Biaxially textured composite substrates

    DOEpatents

    Groves, James R.; Foltyn, Stephen R.; Arendt, Paul N.

    2005-04-26

    An article including a substrate, a layer of a metal phosphate material such as an aluminum phosphate material upon the surface of the substrate, and a layer of an oriented cubic oxide material having a rock-salt-like structure upon the metal phosphate material layer is provided together with additional layers such as a HTS top-layer of YBCO directly upon a layer of a buffer material such as a SrTi.sub.x Ru.sub.1-x O.sub.3 layer.

  17. Acquisition and evaluation of thermodynamic data for morenosite-retgersite equilibria at 0.1 MPa

    USGS Publications Warehouse

    Chou, I.-Ming; Seal, R.R.

    2003-01-01

    Metal-sulfate salts in mine drainage environments commonly occur as solid solutions containing Fe, Cu, Mg, Zn, Al, Mn, Ni, Co, Cd, and other elements. Thermodynamic data for some of the end-member salts containing Fe, Cu, Zn, and Mg have been collected and evaluated previously, and the present study extends to the system containing Ni. Morenosite (NiSO4-7H2O)-retgersite (NiSO4-6H2O) equilibria were determined along five humidity buffer curves at 0.1 MPa and between 5 and 22??C. Reversals along these humidity-buffer curves yield In K = 17.58-6303.35/T, where K is the equilibrium constant, and T is temperature in K. The derived standard Gibbs free energy of reaction is 8.84 kJ/mol, which agrees very well with the values of 8.90, 8.83, and 8.85 kJ/mol based on the vapor pressure measurements of Schumb (1923), Bonnell and Burridge (1935), and Stout et al. (1966). respectively. This value also agrees reasonably well with the values of 8.65 and 9.56 kJ/mol calculated from the data compiled by Wagman et al. (1982) and DeKock (1982), respectively. The temperature-humidity relationships defined by this study for dehydration equilibria between morenosite and retgersite explain the more common occurrence of retgersite relative to morenosite in nature.

  18. Modeling stream-groundwater interactions and associated groundwater salinization in an urban floodplain

    NASA Astrophysics Data System (ADS)

    Ledford, S. H.; Lautz, L.

    2014-12-01

    The salinization of freshwater in the Northeastern United States from road salt application is well documented by the observed long-term increases in chloride concentrations in groundwater over the last fifty years. However, the processes controlling exchange of chloride between surface water and groundwater have not been fully investigated, particularly in urban streams where stream-groundwater interactions can be reduced due to bank armoring and channelization. Our research builds on previous findings that showed the potential for an urban riparian floodplain to buffer seasonal changes in chloride concentrations in an urban stream, resulting in smaller annual ranges of chloride in areas with intact riparian floodplains. A reach of Meadowbrook Creek, in Syracuse, New York, that is disconnected from the groundwater had large seasonal shifts in chloride concentration, varying from 2173 mg/L Cl- in the winter to 161.2 mg/L Cl- in the summer. This is in contrast to a downstream reach of the stream that receives groundwater discharge from a riparian floodplain, where chloride concentrations ranged from 657.0 mg/L in the winter to 252.0 mg/L in the summer. We originally hypothesized that winter snowmelt events caused overbank flooding of saline surface water, which recharged the floodplain groundwater, causing salinization. This saline water was then slowly discharged as baseflow throughout the year and was replaced with freshwater overbank events in the summer. However, a three dimensional model of the floodplain created using Visual MODFLOW indicates that surface water-groundwater interactions, such as hyporheic exchange, may have a greater control on winter salt input than overbank events, while summer flooding recharges the aquifer with freshwater. The model was compared to riparian aquifer samples collected from May 2013 until June 2014 to qualitatively study the impact of different types of surface water-groundwater interactions (e.g. groundwater recharge and discharge, hyporheic interaction) on salt storage and to identify the mechanisms by which urban riparian floodplains buffer seasonal variability of stream chloride concentrations in urban systems impacted by road salt.

  19. Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry

    PubMed Central

    Song, Yong-Ak; Chan, Michael; Celio, Chris; Tannenbaum, Steven R.; Wishnok, John S.; Han, Jongyoon

    2010-01-01

    In this paper, we are evaluating the strategy of sorting peptides / proteins based on the charge to mass without resorting to ampholytes and / or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems, and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration, and isolated peptides within specific pI ranges from sample mixtures, where the pI range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a high-throughput sample fractionation tool for targeted proteomics using LC/MS. PMID:20163146

  20. Nutrition or Detoxification: Why Bats Visit Mineral Licks of the Amazonian Rainforest

    PubMed Central

    Voigt, Christian C.; Capps, Krista A.; Dechmann, Dina K. N.; Michener, Robert H.; Kunz, Thomas H.

    2008-01-01

    Many animals in the tropics of Africa, Asia and South America regularly visit so-called salt or mineral licks to consume clay or drink clay-saturated water. Whether this behavior is used to supplement diets with locally limited nutrients or to buffer the effects of toxic secondary plant compounds remains unclear. In the Amazonian rainforest, pregnant and lactating bats are frequently observed and captured at mineral licks. We measured the nitrogen isotope ratio in wing tissue of omnivorous short-tailed fruit bats, Carollia perspicillata, and in an obligate fruit-eating bat, Artibeus obscurus, captured at mineral licks and at control sites in the rainforest. Carollia perspicillata with a plant-dominated diet were more often captured at mineral licks than individuals with an insect-dominated diet, although insects were more mineral depleted than fruits. In contrast, nitrogen isotope ratios of A. obscurus did not differ between individuals captured at mineral lick versus control sites. We conclude that pregnant and lactating fruit-eating bats do not visit mineral licks principally for minerals, but instead to buffer the effects of secondary plant compounds that they ingest in large quantities during periods of high energy demand. These findings have potential implications for the role of mineral licks for mammals in general, including humans. PMID:18431492

  1. Performance optimization in electric field gradient focusing.

    PubMed

    Sun, Xuefei; Farnsworth, Paul B; Tolley, H Dennis; Warnick, Karl F; Woolley, Adam T; Lee, Milton L

    2009-01-02

    Electric field gradient focusing (EFGF) is a technique used to simultaneously separate and concentrate biomacromolecules, such as proteins, based on the opposing forces of an electric field gradient and a hydrodynamic flow. Recently, we reported EFGF devices fabricated completely from copolymers functionalized with poly(ethylene glycol), which display excellent resistance to protein adsorption. However, the previous devices did not provide the predicted linear electric field gradient and stable current. To improve performance, Tris-HCl buffer that was previously doped in the hydrogel was replaced with a phosphate buffer containing a salt (i.e., potassium chloride, KCl) with high mobility ions. The new devices exhibited stable current, good reproducibility, and a linear electric field distribution in agreement with the shaped gradient region design due to improved ion transport in the hydrogel. The field gradient was calculated based on theory to be approximately 5.76 V/cm(2) for R-phycoerythrin when the applied voltage was 500 V. The effect of EFGF separation channel dimensions was also investigated; a narrower focused band was achieved in a smaller diameter channel. The relationship between the bandwidth and channel diameter is consistent with theory. Three model proteins were resolved in an EFGF channel of this design. The improved device demonstrated 14,000-fold concentration of a protein sample (from 2 ng/mL to 27 microg/mL).

  2. Mechanism of molecular recognition on molecular imprinted monolith by capillary electrochromatography.

    PubMed

    Liu, Zhao-Sheng; Xu, Yan-Li; Yan, Chao; Gao, Ru-Yu

    2005-09-16

    The recognition mechanism of molecularly imprinted polymer (MIP) in capillary electrochromatography (CEC) is complicated since it possesses a hybrid process, which comprises the features of chromatographic retention, electrophoretic migration and molecular imprinting. For an understanding of the molecular recognition of MIP in CEC, a monolithic MIP in a capillary with 1,1'-binaphthyl-2,2'-diamine (BNA) imprinting was prepared by in situ copolymerization of imprinted molecule, methacrylic acid and ethylene glycol dimethacrylate in porogenic solvent, a mixture of toluene-isooctane. Strong recognition ability and high column performance (theory plates was 43,000 plates/m) of BNA were achieved on this monolithic MIP in CEC mode. In addition, BNA and its structural analogue, 1,1'-bi-2, 2'-naphthol, differing in functional groups, were used as model compounds to study imprinting effect on the resultant BNA-imprinted monolithic column, a reference column without imprinting of BNA and a open capillary. The effects of organic modifier concentration, pH value of buffer, salt concentration of buffer and column temperature on the retention and recognition of two compounds were investigated. The results showed that the molecular recognition on MIP monolith in CEC mode mainly derived from imprinting cavities on BNA-imprinted polymer other than chromatographic retention and electrophoretic migration.

  3. Diclofenac Salts, VIII. Effect of the Counterions on the Permeation through Porcine Membrane from Aqueous Saturated Solutions.

    PubMed

    Fini, Adamo; Bassini, Glenda; Monastero, Annamaria; Cavallari, Cristina

    2012-09-12

    The following bases: monoethylamine (EtA), diethylamine (DEtA), triethylamine (TEtA), monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), pyrrolidine (Py), piperidine (Pp), morpholine (M), piperazine (Pz) and their N-2-hydroxyethyl (HE) analogs were employed to prepare 14 diclofenac salts. The salts were re-crystallized from water in order to obtain forms that are stable in the presence of water. Vertical Franz-type cells with a diffusional surface area of 9.62 cm2 were used to study the permeation of these diclofenac salts from their saturated solutions through an internal pig ear membrane. The receptor compartments of the cells contained 100 mL of phosphate buffer (pH 7.4); a saturated solution (5 mL) of each salt was placed in the donor compartment, thermostated at 37 °C. Aliquots were withdrawn at predetermined time intervals over 8 h and then immediately analyzed by HPLC. Fluxes were determined by plotting the permeated amount, normalized for the membrane surface area versus time. Permeation coefficients were obtained dividing the flux values J by the concentration of the releasing phase-that is, water solubility of each salt. Experimental results show that fluxes could be measured when diclofenac salts with aliphatic amines are released from a saturated aqueous solution. Different chemical species (acid, anion, ion pairs) contribute to permeation of the anti-inflammatory agent even though ion-pairs could be hypothesized to operate to a greater extent. Permeation coefficients were found higher when the counterion contains a ring; while hydroxy groups alone do not appear to play an important role, the ring could sustain permeation, disrupting the organized domains of the membrane.

  4. Diclofenac Salts, VIII. Effect of the Counterions on the Permeation through Porcine Membrane from Aqueous Saturated Solutions

    PubMed Central

    Fini, Adamo; Bassini, Glenda; Monastero, Annamaria; Cavallari, Cristina

    2012-01-01

    The following bases: monoethylamine (EtA), diethylamine (DEtA), triethylamine (TEtA), monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), pyrrolidine (Py), piperidine (Pp), morpholine (M), piperazine (Pz) and their N-2-hydroxyethyl (HE) analogs were employed to prepare 14 diclofenac salts. The salts were re-crystallized from water in order to obtain forms that are stable in the presence of water. Vertical Franz-type cells with a diffusional surface area of 9.62 cm2 were used to study the permeation of these diclofenac salts from their saturated solutions through an internal pig ear membrane. The receptor compartments of the cells contained 100 mL of phosphate buffer (pH 7.4); a saturated solution (5 mL) of each salt was placed in the donor compartment, thermostated at 37 °C. Aliquots were withdrawn at predetermined time intervals over 8 h and then immediately analyzed by HPLC. Fluxes were determined by plotting the permeated amount, normalized for the membrane surface area versus time. Permeation coefficients were obtained dividing the flux values J by the concentration of the releasing phase—that is, water solubility of each salt. Experimental results show that fluxes could be measured when diclofenac salts with aliphatic amines are released from a saturated aqueous solution. Different chemical species (acid, anion, ion pairs) contribute to permeation of the anti-inflammatory agent even though ion-pairs could be hypothesized to operate to a greater extent. Permeation coefficients were found higher when the counterion contains a ring; while hydroxy groups alone do not appear to play an important role, the ring could sustain permeation, disrupting the organized domains of the membrane. PMID:24300300

  5. Mercury intracellular partitioning and chelation in a salt marsh plant, Halimione portulacoides (L.) Aellen: strategies underlying tolerance in environmental exposure.

    PubMed

    Válega, M; Lima, A I G; Figueira, E M A P; Pereira, E; Pardal, M A; Duarte, A C

    2009-01-01

    In the presence of metal stress, plants can resort to a series of tolerance mechanisms. Therefore field studies should be undertaken in order to evaluate the real role of these mechanisms in stress coping. The aim of this paper was to clarify the biochemical processes behind mercury tolerance in Halimione portulacoides (L.) Aellen (Caryophyllales: Chenopodiaceae) collected in a mercury contaminated salt marsh. Different fractions of mercury were separated: buffer-soluble (mainly cytosolic) and insoluble mercury (mainly associated with membranes and cell walls). The amounts in each fraction of metal were compared and related to metal distribution within plant organs. Protein-mercury complexes were isolated and analysed for their thiol content in order to assess wether the tolerance of this salt marsh plant was associated with the induction of metal chelation by phytochelatins. Overall, the mercury tolerance strategies of the plant are likely to involve root cell wall immobilization as a major mechanism of metal resistance, rather than metal chelation in the cytosolic fraction. Nevertheless, phytochelatins were demonstrated to chelate mercury under environmental exposure.

  6. Effects of cationic species on visual color formation in model Maillard reactions of pentose sugars and amino acids.

    PubMed

    Rizzi, George P

    2008-08-27

    Effects of cationic species on Maillard browning were examined after heating (ca. 100 degrees C) aqueous pH 7.2 buffered solutions of amino acids and pentose sugars. Metallic ions of Group I metals (Li, Na, K, Rb and Cs) produced a small increase in browning (A420), but somewhat greater effects were observed with ions of Group II metals Ca and Mg. Browning was suppressed by triethylammonium ion, but unaffected by a salt of the stronger base, guanidine. The quaternary amine salt choline chloride produced enhanced browning and served as a model for phospholipid involvement in Maillard reactions. With alpha,omega-diamino acids increases in browning were observed which related to lowered pK2 values resulting from positively charged omega-substituents in these molecules.

  7. Formulation and synthesis of hydrogels having lower critical solution temperature near body temperature

    NASA Astrophysics Data System (ADS)

    Abidin, A. Z.; Graha, H. P. R.; Trirahayu, D. A.

    2017-07-01

    Copolymerization between bacterial cellulose nanocrystal (CN) and methyl cellulose (MC) was carried out using UV light to produce a biocompatible hydrogel at body temperature and liquid at room temperature. Viscosity and salt effect of the MC and copolymer solution at room temperature and its Lower Critical Solution Temperature (LCST) were evaluated. The analysis showed that the higher concentration of methyl cellulose and salt content in the solution produced lower LCST and higher solution viscosity. All samples of polymer solution with MC concentrations of 1 and 2% have a viscosity less than 5000 cP at room temperature. The solutions with MC concentration of 1, 2, and 3% have respectively LCST of 59, 58, and 57°C, while its copolymer solutions with CN concentration of 0.1, 0.3, and 0.5% have respectively LCST of 55, 51, and 41°C. The salt addition to the solution of MC-CN copolymer with concentrations of 1x and 1.5x Phosphat Buffered Saline (PBS) produces respectively LCST of 47 and 38°C. The results suggest that the copolymer solution of MC-CN could produce a lower LCST and the addition of salt could amplify the effect of LCST decrease that can be used to produce a biocompatible hydrogel with LCST as close as body temperature.

  8. Extra Double-stranded RNA Binding Domain (dsRBD) in a Squid RNA Editing Enzyme Confers Resistance to High Salt Environment*

    PubMed Central

    Palavicini, Juan Pablo; Correa-Rojas, Rodrigo A.; Rosenthal, Joshua J. C.

    2012-01-01

    A-to-I RNA editing is particularly common in coding regions of squid mRNAs. Previously, we isolated a squid editing enzyme (sqADAR2) that shows a unique structural feature when compared with other ADAR2 family members: an additional double-stranded RNA (dsRNA) binding domain (dsRBD). Alternative splicing includes or excludes this motif, generating a novel or a conventional variant termed sqADAR2a and sqADAR2b, respectively. The extra dsRBD of sqADAR2a increases its editing activity in vitro. We hypothesized that the high activity is due to an increase in the affinity of the enzyme for dsRNA. This may be important because protein-RNA interactions can be influenced by physical factors. We became particularly interested in analyzing the effects of salt on interactions between sqADAR2 and RNA because squid cells have a ∼3-fold higher ionic strength and proportionally more Cl− than vertebrate cells. To date, in vitro biochemical analyses of adenosine deamination have been conducted using vertebrate-like ionic strength buffers containing chloride as the major anion, although the vast majority of cellular anions are known to be organic. We found that squid-like salt conditions severely impair the binding affinity of conventional ADAR2s for dsRNA, leading to a decrease in nonspecific and site-specific editing activity. Inhibition of editing was mostly due to high Cl− levels and not to the high concentrations of K+, Na+, and organic anions like glutamate. Interestingly, the extra dsRBD in sqADAR2a conferred resistance to the high Cl− levels found in squid neurons. It does so by increasing the affinity of sqADAR2 for dsRNA by 30- or 100-fold in vertebrate-like or squid-like conditions, respectively. Site-directed mutagenesis of squid ADAR2a showed that its increased affinity and editing activity are directly attributable to the RNA binding activity of the extra dsRBD. PMID:22457361

  9. Preparation and characterization of electrodeposited cobalt nanowires

    NASA Astrophysics Data System (ADS)

    Irshad, M. I.; Ahmad, F.; Mohamed, N. M.; Abdullah, M. Z.

    2014-10-01

    Electrochemical deposition technique has been used to deposit cobalt nanowires into the nano sized channels of Anodized Aluminium Oxide (AAO) templates. CoCl2˙6H 2 O salt solution was used, which was buffered with H3BO3 and acidified by dilute H2SO4 to increase the plating life and control pH of the solution. Thin film of copper around 150 nm thick on one side of AAO template coated by e-beam evaporation system served as cathode to create electrical contact. FESEM analysis shows that the as-deposited nanowires are highly aligned, parallel to one another and have high aspect ratio with a reasonably high pore-filing factor. The TEM results show that electrodeposited cobalt nanowires are crystalline in nature. The Hysteresis loop shows the magnetization properties for in and out of plane configuration. The in plane saturation magnetization (Ms) is lower than out of plane configuration because of the easy axis of magnetization is perpendicular to nanowire axis. These magnetic nanowires could be utilized for applications such as spintronic devices, high density magnetic storage, and magnetic sensor applications.

  10. Towards the elucidation of molecular determinants of cooperativity in the liver bile acid binding protein.

    PubMed

    Pedò, Massimo; D'Onofrio, Mariapina; Ferranti, Pasquale; Molinari, Henriette; Assfalg, Michael

    2009-11-15

    Bile acid binding proteins (BABPs) are cytosolic lipid chaperones contributing to the maintenance of bile acid homeostasis and functional distribution within the cell. Liver BABPs act in parallel with ileal transporters to ensure vectorial transport of bile salts in hepatocytes and enterocytes, respectively. We describe the investigation of ligand binding to liver BABP, an essential step in the understanding of intracellular bile salt transport. Binding site occupancies were monitored in NMR titration experiments using (15)N-labelled ligand, while the relative populations of differently bound BABP forms were assessed by mass spectrometry. This site-specific information allowed the determination of intrinsic thermodynamic parameters and the identification of an extremely high cooperativity between two binding sites. Protein-observed NMR experiments revealed a global structural rearrangement which suggests an allosteric mechanism at the basis of the observed cooperativity. The view of a molecular tool capable of buffering against significant concentrations of free bile salts in a large range of solution conditions emerges from the observed pH-dependence of binding. We set to determine the molecular determinants of cooperativity by analysing the binding properties of a protein containing a mutated internal histidine. Both mass spectrometry and NMR experiments are consistent with an overall decreased binding affinity of the mutant, while the measured diffusion coefficients of ligand species reveal that the affinity loss concerns essentially one of the two binding sites. We therefore identified a mutation able to disrupt energetic communication functional to efficient binding and conclude that the buried histidine establishes contacts that stabilize the ternary complex. 2009 Wiley-Liss, Inc.

  11. Effects of middle-term land reclamation on nickel soil-water interaction: a case study from reclaimed salt marshes of Po River Delta, Italy.

    PubMed

    Di Giuseppe, Dario; Melchiorre, Massimiliano; Faccini, Barbara; Ferretti, Giacomo; Coltorti, Massimo

    2017-09-26

    Reclaimed salt marshes are fragile environments where water salinization and accumulation of heavy metals can easily occur. This type of environment constitutes a large part of the Po River Delta (Italy), where intensive agricultural activities take place. Given the higher Ni background of Po River Delta soils and its water-soluble nature, the main aim of this contribution is to understand if reclamation can influence the Ni behavior over time. In this study, we investigated the geochemical features of 40 soils sampled in two different localities from the Po River Delta with different reclamation ages. Samples of salt marsh soils reclaimed in 1964 were taken from Valle del Mezzano while soils reclaimed in 1872 were taken nearby Codigoro town. Batch solubility tests and consecutive determination of Ni in pore-water were compared to bulk physicochemical compositions of soils. Bulk Ni content of the studied soils is naturally high, since these soils originated from Po River sediments derived from the erosion of ultramafic rocks. Moreover, it seems that Ni concentration increases during soil evolution, being probably related to the degradation of serpentine. Instead, the water-soluble Ni measured in the leaching tests is greater in soils recently reclaimed compared to the oldest soils. Soil properties of two soil profiles from a reclaimed wetland area were examined to determine soil evolution over one century. Following reclamation, pedogenic processes of the superficial horizons resulted in organic matter mineralization, pH buffer, and a decrease of Ni water solubility from recently to evolved reclaimed soil.

  12. The effect of sodium chloride on the dissolution of calcium silicate hydrate gels.

    PubMed

    Hill, J; Harris, A W; Manning, M; Chambers, A; Swanton, S W

    2006-01-01

    The use of cement based materials will be widespread in the long-term management of radioactive materials in the United Kingdom. One of the applications could be the Nirex reference vault backfill (NRVB) as an engineered barrier within a deep geological repository. NRVB confers alkaline conditions, which would provide a robust chemical barrier through the control of the solubility of some key radionuclides, enhanced sorption and minimised corrosion of steel containers. An understanding of the dissolution of C-S-H gels in cement under the appropriate conditions (e.g., saline groundwaters) is necessary to demonstrate the expected evolution of the chemistry over time and to provide sufficient cement to buffer the porewater conditions for the required time. A programme of experimental work has been undertaken to investigate C-S-H gel dissolution behaviour in sodium chloride solutions and the effect of calcium/silicon ratio (C/S), temperature and cation type on this behaviour. Reductions in calcium concentration and pH values were observed with samples equilibrated at 45 degrees C compared to those prepared at 25 degrees C. The effect of salt cation type on salt-concentration dependence of the dissolution of C-S-H gels was investigated by the addition of lithium or potassium chloride in place of sodium chloride for gels with a C/S of 1.0 and 1.8. With a C/S of 1.0, similar increases in dissolved calcium concentration with increasing ionic strength were recorded for the different salts. However, at a C/S of 1.8, anomalously high calcium concentrations were observed in the presence of lithium.

  13. Modeling wave attenuation by salt marshes in Jamaica Bay, New York, using a new rapid wave model

    NASA Astrophysics Data System (ADS)

    Marsooli, Reza; Orton, Philip M.; Mellor, George

    2017-07-01

    Using a new rapid-computation wave model, improved and validated in the present study, we quantify the value of salt marshes in Jamaica Bay—a highly urbanized estuary located in New York City—as natural buffers against storm waves. We augment the MDO phase-averaged wave model by incorporating a vegetation-drag-induced energy dissipation term into its wave energy balance equation. We adopt an empirical formula from literature to determine the vegetation drag coefficient as a function of environmental conditions. Model evaluation using data from laboratory-scale experiments show that the improved MDO model accurately captures wave height attenuation due to submerged and emergent vegetation. We apply the validated model to Jamaica Bay to quantify the influence of coastal-scale salt marshes on storm waves. It is found that the impact of marsh islands is largest for storms with lower flood levels, due to wave breaking on the marsh island substrate. However, the role of the actual marsh plants, Spartina alterniflora, grows larger for storms with higher flood levels, when wave breaking does not occur and the vegetative drag becomes the main source of energy dissipation. For the latter case, seasonality of marsh height is important; at its maximum height in early fall, S. alterniflora causes twice the reduction as when it is at a shorter height in early summer. The model results also indicate that the vegetation drag coefficient varies 1 order of magnitude in the study area, and suggest exercising extra caution in using a constant drag coefficient in coastal wetlands.

  14. Films based on neutralized chitosan citrate as innovative composition for cosmetic application.

    PubMed

    Libio, Illen C; Demori, Renan; Ferrão, Marco F; Lionzo, Maria I Z; da Silveira, Nádya P

    2016-10-01

    In this work, citrate and acetate buffers, were investigated as neutralizers to chitosan salts in order to provide biocompatible and stable films. To choose the appropriate film composition for this study, neutralized chitosan citrate and acetate films, with and without the plasticizer glycerol, were prepared and characterized by thickness, moisture content, degree of swelling, total soluble matter in acid medium, simultaneous thermal analysis and differential scanning calorimetry. Chitosan films neutralized in citrate buffer showed greater physical integrity resulted from greater thicknesses, lower moisture absorbance, lower tendency to solubility in the acid medium, and better swelling capacities. According to thermal analyses, these films had higher interaction with water which is considered an important feature for cosmetic application. Since the composition prepared in citrate buffer without glycerol was considered to present better physical integrity, it was applied to investigate hyaluronic acid release in a skin model. Skins treated with those films, with or without hyaluronic acid, show stratum corneum desquamation and hydration within 10min. The results suggest that the neutralized chitosan citrate film prepared without glycerol promotes a cosmetic effect for skin exfoliation in the presence or absence of hyaluronic acid. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Impact of reaction conditions on the laccase-catalyzed conversion of bisphenol A.

    PubMed

    Kim, Young-Jin; Nicell, James A

    2006-08-01

    The oxidative conversion of aqueous BPA catalyzed by laccase from Trametes versicolor was conducted in a closed, temperature-controlled system containing buffer for pH control. The effects of medium pH, buffer concentration, temperature and mediators and the impacts of dissolved wastewater constituents on BPA conversion were investigated. The optimal pH for BPA conversion was approximately 5, with greater than half maximal conversion and good enzyme stability in the range of 4-7. The stability of the enzyme was not impacted by buffer concentration, nor was BPA conversion. Despite the observation that the enzyme tended to be inactivated at elevated temperatures, enhanced conversion of BPA was observed up until a reaction temperature of 45 degrees C. Of the mediators studied, ABTS was most successful at enhancing the conversion of BPA. Dissolved wastewater constituents that were studied included various inorganic salts, organic compounds and heavy metal ions. BPA conversion was inhibited in the presence of anions such as sulfite, thiosulfate, sulfide, nitrite and cyanide. The metal ions Fe(III) and Cu(II) and the halogens chloride and fluoride substantially suppressed BPA conversion, but the presence of selected organic compounds did not significantly reduce the conversion of BPA.

  16. Purification of peroxidase from Horseradish (Armoracia rusticana) roots.

    PubMed

    Lavery, Christopher B; Macinnis, Morgan C; Macdonald, M Jason; Williams, Joanna Bassey; Spencer, Colin A; Burke, Alicia A; Irwin, David J G; D'Cunha, Godwin B

    2010-08-11

    Peroxidase (EC 1.11.1.7) from horseradish ( Armoracia rusticana ) roots was purified using a simple, rapid, three-step procedure: ultrasonication, ammonium sulfate salt precipitation, and hydrophobic interaction chromatography on phenyl Sepharose CL-4B. The preparation gave an overall yield of 71%, 291-fold purification, and a high specific activity of 772 U mg(-1) protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the purified enzyme was homogeneous and had a molecular weight of approximately 40 kDa. The isolated enzyme had an isoelectric point of 8.8 and a Reinheitszahl value of 3.39 and was stable when stored in the presence of glycerol at -20 degrees C, with >95% retention of original enzyme activity for at least 6 months. Maximal activity of purified horseradish peroxidase (HRP) was obtained under different optimized conditions: substrate (guaiacol and H(2)O(2)) concentrations (0.5 and 0.3 mM, respectively), type of buffer (50 mM phosphate buffer), pH (7.0), time (1.0 min), and temperature of incubation (30 degrees C). In addition, the effect of HRP and H(2)O(2) in a neutral-buffered aqueous solution for the oxidation of phenol and 2-chlorophenol substrates was also studied. Different conditions including concentrations of phenol/2-chlorophenol, H(2)O(2), and enzyme, time, pH, and temperature were standardized for the maximal activity of HRP with these substrates; under these optimal conditions 89.6 and 91.4% oxidations of phenol and 2-chlorophenol were obtained, respectively. The data generated from this work could have direct implications in studies on the commercial production of this biotechnologically important enzyme and its stability in different media.

  17. Hypothermic preservation of corneas in a hyperkalaemic solution (CPTES): II. Extended storage in the presence of chondroitin sulphate.

    PubMed Central

    Taylor, M J; Hunt, C J; Madden, P W

    1989-01-01

    Periods of preservation for donor corneas, even for short times, are necessary to facilitate optimum conditions in penetrating keratoplasty. However, current techniques for corneal storage at low temperatures may not provide optimal conditions for maintaining tissue integrity. In particular, the ionic composition of the storage medium has received little attention since it has been assumed throughout that the normal complement of ions in tissue culture media will also be suitable for preservation at reduced temperatures. This study extends our previous investigations on the merits of using CPTES (corneal-potassium-TES), a potassium-rich balanced salt solution containing an impermeant anionic pH buffer (TES), as a storage solution specifically designed to prevent the loss of intracellular potassium and minimise endothelial cell swelling during the time that the normal regulatory processes are switched off. The effect of adding the natural polymer chondroitin sulphate (CS) as a colloid osmotic agent to the hyperkalaemic storage medium is now examined. Corneas stored in CPTES containing 2.5% chondroitin sulphate retained a very high level of structural and functional integrity after three, five, and seven days storage at 0 degrees C; furthermore, stromal swelling was restricted to only 21%. All corneas stored in CPTES + 2.5% CS showed active endothelial function by thinning efficiently at rates that were greater than those previously reported for rabbit corneas stored for similar lengths of time in either M-K medium or K-sol. The zwitterionic buffers TES and HEPES were interchangeable in the hyperkalaemic solution and were non-toxic to corneal endothelium at a concentration of 100 mM. These compounds offer excellent pH buffering in bicarbonate-free medium. Images PMID:2510816

  18. Resistance of Pseudomonas to Quaternary Ammonium Compounds. I. Growth in Benzalkonium Chloride Solution

    PubMed Central

    Adair, Frank W.; Geftic, Sam G.; Gelzer, Justus

    1969-01-01

    Resistant cells of Pseudomonas aeruginosa and a waterborne Pseudomonas sp. (strain Z-R) were able to multiply in nitrogen-free minimal salts solution containing various concentrations of commercially prepared, ammonium acetate-buffered benzalkonium chloride (CBC), a potent antimicrobial agent. As the CBC concentration increased, growth increased until a point was reached at which the extent of growth leveled off or was completely depressed. Minimal salts solutions of pure benzalkonium chloride (PBC) containing no ammonium acetate did not support bacterial growth. When ammonium acetate was added to PBC solutions in the same concentrations found in CBC solutions, growth patterns developed that were comparable to those found with CBC. Likewise, (NH4)2SO4 added to PBC solutions supported growth of both organisms. P. aeruginosa was initially resistant to CBC levels of 0.02% and it was adapted to tolerate levels as high as 0.36%. Strain Z-R was naturally resistant to 0.4% CBC. Since ammonium acetate, carried over by the CBC used in drug formulations and disinfectant solutions, has the potential to support the growth of resistant bacteria and thus make possible the risk of serious infection, it is suggested that regulations allowing the presence of ammonium acetate in CBC solution be reconsidered. PMID:4984761

  19. Efficient chemical and visible-light-driven water oxidation using nickel complexes and salts as precatalysts.

    PubMed

    Chen, Gui; Chen, Lingjing; Ng, Siu-Mui; Lau, Tai-Chu

    2014-01-01

    Chemical and visible-light-driven water oxidation catalyzed by a number of Ni complexes and salts have been investigated at pH 7-9 in borate buffer. For chemical oxidation, [Ru(bpy)3](3+) (bpy = 2,2'-bipyridine) was used as the oxidant, with turnover numbers (TONs) >65 and a maximum turnover frequency (TOFmax) >0.9 s(-1). Notably, simple Ni salts such as Ni(NO3 )2 are more active than Ni complexes that bear multidentate N-donor ligands. The Ni complexes and salts are also active catalysts for visible-light-driven water oxidation that uses [Ru(bpy)3](2+) as the photosensitizer and S2 O8 (2-) as the sacrificial oxidant; a TON>1200 was obtained at pH 8.5 by using Ni(NO3)2 as the catalyst. Dynamic light scattering measurements revealed the formation of nanoparticles in chemical and visible-light-driven water oxidation by the Ni catalysts. These nanoparticles aggregated during water oxidation to form submicron particles that were isolated and shown to be partially reduced β-NiOOH by various techniques, which include SEM, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, XRD, and IR spectroscopy. These results suggest that the Ni complexes and salts act as precatalysts that decompose under oxidative conditions to form an active nickel oxide catalyst. The nature of this active oxide catalyst is discussed. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. A High Salt Diet Inhibits Obesity and Delays Puberty in the Female Rat

    PubMed Central

    Pitynski-Miller, Dori; Ross, Micah; Schmill, Margaret; Schambow, Rachel; Fuller, Teresa; Flynn, Francis W.; Skinner, Donal C.

    2017-01-01

    Background/Objectives Processed foods are considered major contributors to the worldwide obesity epidemic. In addition to high sugar and fat contents, processed foods contain large amounts of salt. Due to correlations with rising adiposity, salt has recently been proposed to be obesogenic. This study investigated three hypotheses: i) high salt contributes to weight gain and adiposity in juvenile female rats, ii) puberty onset would be altered because salt is known to affect neuronal systems involved in activating the reproductive system, and iii) enhanced adiposity will act synergistically with salt to drive early puberty onset. Design Female weanling rats (post-natal day 21, n=105) were fed a low fat/low salt diet, low fat/high salt diet, high fat/low salt diet, or a high salt/high fat diet for 24 days. Metabolic measures, including weight gain, food intake, fecal output, activity, and temperature were recorded in subsets of animals. Results Body weight, retroperitoneal and perirenal fat pad weight, and adipocyte size were all lower in animals fed high fat/high salt compared to animals fed high fat alone. Leptin levels were reduced in high fat/high salt fed animals compared to high fat/low salt fed animals. Daily calorie intake was higher initially but declined with adjusted food intake and was not different among groups after 5 days. Osmolality and corticosterone were not different among groups. Fecal analysis showed excess fat excretion and a decreased digestive efficiency in animals fed high fat/low salt but not in animals fed high fat/high salt. Although respiratory exchange ratio was reduced by high dietary fat or salt, aerobic resting metabolic rate was not affected by diet. High salt delayed puberty onset, regardless of dietary fat content. Conclusions Salt delays puberty and prevents the obesogenic effect of a high fat diet. The reduced weight gain evident in high salt fed animals is not due to differences in food intake or digestive efficiency. PMID:28674441

  1. An Ephemeral Burst-Buffer File System for Scientific Applications

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

    Wang, Teng; Moody, Adam; Yu, Weikuan

    BurstFS is a distributed file system for node-local burst buffers on high performance computing systems. BurstFS presents a shared file system space across the burst buffers so that applications that use shared files can access the highly-scalable burst buffers without changing their applications.

  2. Improvement of cooking quality and gel formation capacity of Bombay duck (Harpodon nehereus) fish meat.

    PubMed

    Rupsankar, Chakrabarti

    2010-10-01

    High moisture content (89%) along with high enzymatic and bacteriological activity in Bombay duck (Harpodon nehereus) meat are responsible for short shelf life and disintegration of meat in cooking. Minimum solubility was at pH 5 (iso-electric point) of muscle protein. Citric acid- sodium citrate buffer (pH 5) with 0.2% potassium sorbate was very effective in reducing moisture in dressed fish and in increasing shelf life up to 4 days at ambient temperature (25-28 °C). Reduction in moisture in meat improved its cooking quality and gel formation capacity with increased protein content. Fish meat contained 1.0-1.5% NaCl and produced stronger gel by using 2% NaCl than conventionally prepared gel with 4% NaCl. Washing fish mince with cold water followed by pressing at pH 5, gave fish cake with more salt soluble protein and better gel strength (>500 gcm) than the same operation done at ambient temperature.

  3. Simultaneous quantification of poly-dispersed anionic, amphoteric and nonionic surfactants in simulated wastewater samples using C18 high-performance liquid chromatography-quadrupole ion-trap mass spectrometry

    NASA Technical Reports Server (NTRS)

    Levine, Lanfang H.; Garland, Jay L.; Johnson, Jodie V.

    2005-01-01

    This paper describes the development of a guantitative method for direct and simultaneous determination of three frequently encountered surfactants, amphoteric (cocoamphoacetate, CAA), anionic (sodium laureth sulfate, SLES), and nonionic (alcohol ethoxylate, AE) using a reversed-phase C18 HPLC coupled with an ESI ion-trap mass spectrometer (MS). Chemical composition, ionization characteristics and fragmentation pathways of the surfactants are presented. Positive ESI was effective for all three surfactants in agueous methanol buffered with ammonium acetate. The method enables rapid determinations in small sample volumes containing inorganic salts (up to 3.5 g L(-1)) and multiple classes of surfactants with high specificity by applying surfactant specific tandem mass spectrometric strategies. It has dynamic linear ranges of 2-60, 1.5-40, 0.8-56 mg L(-1) with R2 egual or greater than 0.999, 0.98 and 0.999 (10 microL injection) for CAA, SLES, and AE, respectively.

  4. Red Light-enhanced Phytochrome Pelletability

    PubMed Central

    Pratt, Lee H.; Marmé, Dieter

    1976-01-01

    Red light-enhanced pelletability of phytochrome was observed in extracts of all 11 plants tested: Avena sativa L., Secale cereale L., Zea mays L., Cucurbita pepo L., Sinapis alba L., Pisum sativum L., Helianthus anuus L., Raphanus sativus L., Glycine max (L.) Merr., Phaseolus vulgaris L., and Lupinus albus L. This enhanced pelletability was observed in all 11 plants following in situ irradiation (in vivo binding) but only in Sinapis and Cucurbita after irradiation of crude extracts (in vitro binding). In vivo binding was not strongly dependent upon pH and, with few exceptions, was not markedly sensitive to high salt concentration, whereas in vitro binding was completely reversed by both high pH and high salt concentration. However, both binding phenomena were observed only with a divalent cation in the extract buffer. In vivo binding was further characterized using Avena which showed an increase in pelletability from less than 10% in dark control extracts to more than 60% in extracts of red light-irradiated shoots. The half-life for binding was 40 seconds at 0.5 C and was strongly temperature-dependent, binding being complete within 5 to 10 sec at 22 C. If pelletable phytochrome in the far red-absorbing form was photoconverted back to the red-absorbing form in situ, phytochrome was released from the pelletable condition with a half-life of 25 minutes at 25 C and 100 minutes at both 13 C and 3 C. No cooperativity in red light-enhanced pelletability with respect to phytochrome-far red-absorbing form was observed. PMID:16659745

  5. Enantioselective determination of (R)-zopiclone and (S)-zopiclone (eszopiclone) in human hair by micropulverized extraction and chiral liquid chromatography/high resolution mass spectrometry.

    PubMed

    Miyaguchi, Hajime; Kuwayama, Kenji

    2017-10-13

    Zopiclone and its (S)-enantiomer (eszopiclone) are commonly prescribed for insomnia. Despite the high demand for enantioselective differentiation, the chiral analysis of zopiclone in hair has not been reported. In this study, a method for the enantioselective quantification of zopiclone in human hair was developed. The extraction medium and duration were optimized using real eszopiclone-positive hair samples. Specifically, micropulverized extraction with 3.0M ammonium phosphate buffer (pH 8.4) involving salting-out assisted liquid-liquid extraction with acetonitrile was utilized to minimize the degradation of zopiclone and for rapid and facile operation. On the other hand, recovery of the conventional solid-liquid extraction involved overnight soaking in 3.0M ammonium phosphate buffer (pH 8.4) was only 0.58±0.12% of the maximum recovery achieved by the present method due to the decomposition in the phosphate buffer. An excellent chiral separation (Rs=5.0) was achieved using a chiral stationary phase comprising cellulose tris(3,5-dichlorophenylcarbamate) and a volatile mobile phase of 10mM ammonium carbonate (pH 8.0)-acetonitrile (25:75, v/v). Detection was carried out using liquid chromatography/high resolution mass spectrometry (LC/HRMS) with electrospray ionization. A Q Exactive mass spectrometer equipped with a quadrupole-Orbitrap analyzer was used for detection. The concentration of 0.50pg/mg was defined as the lowest limit of quantification using 5mg of hair sample. Using the developed approach, the concentration of eszopiclone in hair after a single 2-mg dose was found to be 441pg/mg, which was higher than all the reported values regarding a single administration of zopiclone. After daily administration of racemic zopiclone (3.75mg/day), the concentrations of (R)-enantiomer and (S)-enantiomer in the black hair were 5.30-8.31ng/mg and 7.96-12.8ng/mg, respectively, and the concentration of the (S)-enantiomer was always higher than that of the (R)-enantiomer due to the enantioselective difference in the pharmacokinetics. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Interactions of GRF(1-29)NH2 with plasma proteins and their effects on the release of the peptide from a PLAGA matrix.

    PubMed

    Mariette, B; Coudane, J; Vert, M

    2005-09-02

    The administration of the GRF(1-29)NH2 Growth Hormone Releasing Hormone analog is known as relevant of the concept of drug delivery system using a bioresorbable matrix. However, the release of this peptide from poly(dl-lactic acid-co-glycolic acid) matrices is affected by its insolubility at neutral in salted media and in plasma as well. In order to investigate the origin and the nature of the insolubility in these media in more details, the precipitates collected when the peptide was set in contact with saline, isotonic pH=7.4 phosphate buffer and plasma were analyzed by various techniques, namely weighting, gel chromatography, 1D- and 2D-immunoelectrophoresis, and dialysis to discern the soluble from the insoluble or aggregated fractions. It is shown that precipitation in protein-free salted media is due to a salting out phenomenon complemented by the neutralization of the solubilizing electrostatic charges in the isotonic buffer. In contrast, the precipitation in plasma is due to inter polyelectrolyte-type complexation that involved polyanionic proteins having a rather low isoelectric point like albumin, transferin, haptoglobulin and IgG immunoglobulins. When a rather large quantity of GRF(1-29)NH2 was entrapped in bioresorbable pellets working at a percolating regime after subcutaneous implantation in rats, the peptide was slowly released despite the complexation with plasma proteins. However only a very small part of the peptide was found in blood, this small part being still large enough to cause a detectable increase of the circulating growth hormone concentration. Attempts made to increase the solubility of the peptide in plasma were successful when the peptide was combined with arginine, an amino acid known to promote the poor hormonal activity of injected GRF(1-29)NH2 solutions under clinical conditions.

  7. Effect of Bronsted Acids and Bases, and Lewis Acid (Sn(2+)) on the Regiochemistry of the Reaction of Amines with Trifluoromethyl-β-diketones: Reaction of 3-Aminopyrrole to Selectively Produce Regioisomeric 1H-Pyrrolo[3,2-b]pyridines.

    PubMed

    De Rosa, Michael; Arnold, David; Hartline, Douglas; Truong, Linda; Verner, Roman; Wang, Tianwei; Westin, Christian

    2015-12-18

    Reaction of 3-aminopyrrole (as its salt) with trifluoromethyl-β-diketones gave γ-1H-pyrrolo[3,2-b]pyridines via reaction at the less reactive carbonyl group. The trifluoromethyl group increased the electrophilicity of the adjacent carbonyl group and decreased the basicity of the hydroxyl group of the CF3 amino alcohol formed. This amino alcohol was formed faster, but its subsequent dehydration to the β-enaminone was slow resulting in the preferential formation of the γ-regioisomer. Reaction of 4,4,4-trifluoro-1-phenyl-1,3-butadione with 3-aminopyrrole was carried out using a series of 6 amine buffers. Yields of the α-1H-pyrrolo[3,2-b]pyridine increased as the pKa of the amine buffer decreased. Surprisingly the yield went down at higher pKas. There was a change in mechanism as the reaction mixture became more basic. With strong amines trifluoromethyl-β-diketones were present mainly or completely as the enolate. Under reductive conditions (3-nitropyrrole/Sn/AcOH/trifluoromethyl-β-diketone) the α-1H-pyrrolo[3,2-b]pyridine was the major product as a result of Lewis acid catalysis by Sn(2+). Similar α-regiochemistry was observed when the reaction of the 3-aminopyrrole salt with trifluoromethyl-β-diketones was carried out in the presence of base and tin(II) acetate.

  8. Capacitive deionization on-chip as a method for microfluidic sample preparation.

    PubMed

    Roelofs, Susan H; Kim, Bumjoo; Eijkel, Jan C T; Han, Jongyoon; van den Berg, Albert; Odijk, Mathieu

    2015-03-21

    Desalination as a sample preparation step is essential for noise reduction and reproducibility of mass spectrometry measurements. A specific example is the analysis of proteins for medical research and clinical applications. Salts and buffers that are present in samples need to be removed before analysis to improve the signal-to-noise ratio. Capacitive deionization is an electrostatic desalination (CDI) technique which uses two porous electrodes facing each other to remove ions from a solution. Upon the application of a potential of 0.5 V ions migrate to the electrodes and are stored in the electrical double layer. In this article we demonstrate CDI on a chip, and desalinate a solution by the removal of 23% of Na(+) and Cl(-) ions, while the concentration of a larger molecule (FITC-dextran) remains unchanged. For the first time impedance spectroscopy is introduced to monitor the salt concentration in situ in real-time in between the two desalination electrodes.

  9. Micro-apparatus for rapid determinations of protein solubilities

    NASA Technical Reports Server (NTRS)

    Pusey, Marc L.; Munson, Sibyl

    1991-01-01

    We have developed a column-based micro-technique for rapid determinations of protein solubilities. While retaining a large crystal surface area, the column dead volume has been reduced to equal to or less than 5 micro liters. The technique was tested with tetragonal lysozyme (pH 4.5, 0.1 M acetate, 3.0 percent NaCl, 5-25 C) and column volumes of about 60, 300, and 900 micro liters. Identical solubility data were obtained, indicating that equilibration was obtained even in the smallest columns. In addition, solubility data for Br- and I- salts of lysozyme (pH 4.5, 0.1 M acetate buffer, 0.5 M salt concentrations) were obtained. It appears that the technique can be further miniaturized. The limit in further reducing the crystalline column volume is determined by the minimum solution sample size needed to determine the protein concentration.

  10. The interactions between ionic surfactants and phosphatidylcholine vesicles: Conductometry

    NASA Astrophysics Data System (ADS)

    Tsao, Heng-Kwong; Tseng, Wen Liang

    2001-11-01

    The interaction between ionic surfactants and phosphatidylcholine vesicles, which are prepared without addition of buffer and salt, is investigated by conductivity measurements. On the basis of the vesicle acting as a trap of charge carriers, the bilayer/aqueous phase partition coefficient K and the surfactant/lipid molar ratio Re of nine surfactants are determined. The thermodynamic consistency is satisfied by the measured parameters. The effects of the alkyl chain length (C10-C16) and ionic head group are then studied. The inverse partition coefficient K-1 is linearly related to the critical micelle concentration. The solubilizing ability Reb is a consequence of the competition between the surfactant incorporation into the bilayer and the formation of micelles. Consequently, the K parameter rises whereas the Reb parameter declines as the chain length is increased. The influence due to addition of salt is also discussed.

  11. Tampa Bay Integrated Science Pilot Study: wetland characterization

    USGS Publications Warehouse

    McIvor, Carole C.; Raabe, Ellen; Yates, Kimberly; Carter, Bill; Crane, Mike; Fernandez, Mario; Henningsen, Brandt; Kruse, Sara; Oches, Rich; Proffitt, Ed; Runnels, Randy; Shrestha, Ramesh; Smith, Tom; Travis, Steve

    2001-01-01

    Coastal wetlands in Tampa Bay consist of mangrove forest and tidal salt marsh. Wetlands buffer storm surges, provide fish and wildlife habitat, and enhance water quality through the removal of water-borne nutrients and contaminants. Substantial areas of both mangrove and salt marsh have been lost to agricultural, residential and industrial development in this urban estuary. Wetlands restoration has been initiated in Tampa Bay. Baseline studies on the current condition of wetlands and historical and prehistorical information is needed for successful restoration planning and evaluation.A major objective of this component of the Tampa Bay pilot program was to characterize wetlands in Tampa Bay beginning with areas that differ in their degree of human-induced disturbance (Fig. 1). The Alafia River area is urbanized, industrialized and dredged, whereas the Terra Ceia area has a history of agricultural use with associated soil berms and mosquito ditches, but has not been farmed for at least 20 years (Fig. 2).

  12. High-performance porous polylactide stereocomplex crystallite scaffolds prepared by solution blending and salt leaching.

    PubMed

    Xie, Yan; Lan, Xiao-Rong; Bao, Rui-Ying; Lei, Yang; Cao, Zhi-Qiang; Yang, Ming-Bo; Yang, Wei; Wang, Yun-Bing

    2018-09-01

    Biodegradable stereocomplex crystallite polylactide (SC-PLA) porous scaffolds with well-defined pore structures, high heat resistance, mechanical strength, and solvent resistance together with good biocompatibility, were obtained through solution casting of mixed poly(l-lactide) and poly(d-lactide) solution and subsequent leaching of sodium chloride particles. The pore structure of the SC-PLA scaffolds can be perfectly maintained after a high-pressure sterilization treatment at 121 °C, owing to the extensive formation of stereocomplex crystallites in the scaffolds. In vivo pilot study demonstrates that the fibroblasts of rats can infiltrate into the SC-PLA scaffolds well through the open pores. Degradation tests in phosphate-buffered saline solution reveal that the structure of SC-PLA scaffolds was quite stable due to the enhanced hydrolysis-resistance and improved mechanical properties of the scaffolds. These results reveal that SC-PLA scaffolds with good biocompatibility are potentially to be used as implanted biomaterials for the regeneration and restoration of tissues or organs. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. EURELIOS, the world's first thermomechanical helioelectric power plant

    NASA Astrophysics Data System (ADS)

    Gretz, J.

    Characteristics of the power source, design, costs and operating mechanisms and performance of the EURELIOS central receiver solar electric power plant are described. Noting that the solar input at the earth's surface is about 1 kW/sq m, 6200 sq m of float glass mirrors mounted on 182 heliostats were fabricated to focus the incoming radiation onto a receiver aperture atop a 55 m high tower. The curved mirrors permit the focus of 80% of the energy input to be deposited on a 2.2 m diam aperture which is equipped with heat exchangers imbedded in pyrex and darkened and finned to maximize absorption. Feedwater is superheated in the receiver and is transferred to a buffer tank of hot water at 19 bar and molten salt at 410 C, and then on to turbines for actual power production. The grid serves as back-up power system. Total costs are calculated at $1600/kWe.

  14. SAD phasing with in-house cu Ka radiation using barium as anomalous scatterer.

    PubMed

    Dhanasekaran, V; Velmurugan, D

    2011-12-01

    Phasing of lysozyme crystals using co-crystallized barium ions was performed using single-wavelength anomalous diffraction (SAD) method using Cu Ka radiation with in-house source of data collection. As the ion binding sites vary with respect to the pH of the buffer during crystallization, the highly isomorphic forms of lysozyme crystals grown at acidic and alkaline pH were used for the study. Intrinsic sulphur anomalous signal was also utilized with anomalous signal from lower occupancy ions for phasing. The study showed that to solve the structure by SAD technique, 2.8-fold data redundancy was sufficient when barium was used as an anomalous marker in the in-house copper X-ray radiation source for data collection. Therefore, co-crystallization of proteins with barium containing salt can be a powerful tool for structure determination using lab source.

  15. Competitive counterion complexation allows the true host : guest binding constants from a single titration by ionic receptors.

    PubMed

    Pessêgo, Márcia; Basílio, Nuno; Muñiz, M Carmen; García-Río, Luis

    2016-07-06

    Counterion competitive complexation is a background process currently ignored by using ionic hosts. Consequently, guest binding constants are strongly affected by the design of the titration experiments in such a way that the results are dependent on the guest concentration and on the presence of added salts, usually buffers. In the present manuscript we show that these experimental difficulties can be overcome by just considering the counterion competitive complexation. Moreover a single titration allows us to obtain not only the true binding constants but also the stoichiometry of the complex showing the formation of 1 : 1 : 1 (host : guest : counterion) complexes. The detection of high stoichiometry complexes is not restricted to a single titration experiment but also to a displacement assay where both competitive and competitive-cooperative complexation models are taken into consideration.

  16. Stability of buffer-free freeze-dried formulations: A feasibility study of a monoclonal antibody at high protein concentrations.

    PubMed

    Garidel, Patrick; Pevestorf, Benjamin; Bahrenburg, Sven

    2015-11-01

    We studied the stability of freeze-dried therapeutic protein formulations over a range of initial concentrations (from 40 to 160 mg/mL) and employed a variety of formulation strategies (including buffer-free freeze dried formulations, or BF-FDF). Highly concentrated, buffer-free liquid formulations of therapeutic monoclonal antibodies (mAbs) have been shown to be a viable alternative to conventionally buffered preparations. We considered whether it is feasible to use the buffer-free strategy in freeze-dried formulations, as an answer to some of the known drawbacks of conventional buffers. We therefore conducted an accelerated stability study (24 weeks at 40 °C) to assess the feasibility of stabilizing freeze-dried formulations without "classical" buffer components. Factors monitored included pH stability, protein integrity, and protein aggregation. Because the protein solutions are inherently self-buffering, and the system's buffer capacity scales with protein concentration, we included highly concentrated buffer-free freeze-dried formulations in the study. The tested formulations ranged from "fully formulated" (containing both conventional buffer and disaccharide stabilizers) to "buffer-free" (including formulations with only disaccharide lyoprotectant stabilizers) to "excipient-free" (with neither added buffers nor stabilizers). We evaluated the impacts of varying concentrations, buffering schemes, pHs, and lyoprotectant additives. At the end of 24 weeks, no change in pH was observed in any of the buffer-free formulations. Unbuffered formulations were found to have shorter reconstitution times and lower opalescence than buffered formulations. Protein stability was assessed by visual inspection, sub-visible particle analysis, protein monomer content, charge variants analysis, and hydrophobic interaction chromatography. All of these measures found the stability of buffer-free formulations that included a disaccharide stabilizer comparable to buffer-based formulations, especially at protein concentrations up to and including 115 mg/mL. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Effects of buffer composition and processing conditions on aggregation of bovine IgG during freeze-drying.

    PubMed

    Sarciaux, J M; Mansour, S; Hageman, M J; Nail, S L

    1999-12-01

    The objective of this study was to identify critical formulation and processing variables affecting aggregation of bovine IgG during freeze-drying when no lyoprotective solute is used. Parameters examined were phosphate buffer concentration and counterion (Na versus K phosphate), added salts, cooling rate, IgG concentration, residual moisture level, and presence of a surfactant. No soluble aggregates were detected in any formulation after either freezing/thawing or freeze-drying. No insoluble aggregates were detected in any formulation after freezing, but insoluble aggregate levels were always detectable after freeze-drying. The data are consistent with a mechanism of aggregate formation involving denaturation of IgG at the ice/freeze-concentrate interface which is reversible upon freeze-thawing, but becomes irreversible after freeze-drying and reconstitution. Rapid cooling (by quenching in liquid nitrogen) results in more and larger aggregates than slow cooling on the shelf of the freeze-dryer. This observation is consistent with surface area measurements and environmental electron microscopic data showing a higher surface area of freeze-dried solids after fast cooling. Annealing of rapidly cooled solutions results in significantly less aggregation in reconstituted freeze-dried solids than in nonannealed controls, with a corresponding decrease in specific surface area of the freeze-dried, annealed system. Increasing the concentration of IgG significantly improves the stability of IgG against freeze-drying-induced aggregation, which may be explained by a smaller percentage of the protein residing at the ice/freeze-concentrate interface as IgG concentration is increased. A sodium phosphate buffer system consistently results in more turbid reconstituted solids than a potassium phosphate buffer system at the same concentration, but this effect is not attributable to a pH shift during freezing. Added salts such as NaCl or KCl contribute markedly to insoluble aggregate formation. Both sodium and potassium chloride contribute more to turbidity of the reconstituted solid than either sodium or potassium phosphate buffers at similar ionic strength, with sodium chloride resulting in a substantially higher level of aggregates than potassium chloride. At a given cooling rate, the specific surface area of dried solids is approximately a factor of 2 higher for the formulation containing sodium chloride than the formulation containing potassium chloride. Turbidity is also influenced by the extent of secondary drying, which underscores the importance of minimizing secondary drying of this system. Including a surfactant such as polysorbate 80, either in the formulation or in the water used for reconstitution, decreased, but did not eliminate, insoluble aggregates. There was no correlation between pharmaceutically acceptability of the freeze-dried cake and insoluble aggregate levels in the reconstituted product.

  18. Effect of halide salts on development of surface browning on fresh-cut 'Granny Smith' (Malus × domestica Borkh) apple slices during storage at low temperature.

    PubMed

    Li, Yongxin; Wills, Ron B H; Golding, John B; Huque, Roksana

    2015-03-30

    The postharvest life of fresh-cut apple slices is limited by browning on cut surfaces. Dipping in halide salt solutions was examined for their inhibition of surface browning on 'Granny Smith' apple slices and the effects on biochemical factors associated with browning. Delay in browning by salts was greatest with chloride = phosphate > sulfate > nitrate with no difference between sodium, potassium and calcium ions. The effectiveness of sodium halides on browning was fluoride > chloride = bromide > iodide = control. Polyphenol oxidase (PPO) activity of tissue extracted from chloride- and fluoride-treated slices was not different to control but when added into the assay solution, NaF > NaCl both showed lower PPO activity at pH 3-5 compared to control buffer. The level of polyphenols in treated slices was NaF > NaCl > control. Addition of chlorogenic acid to slices enhanced browning but NaCl and NaF counteracted this effect. There was no effect of either halide salt on respiration, ethylene production, ion leakage, and antioxidant activity. Dipping apple slices in NaCl is a low cost treatment with few impediments to commercial use and could be a replacement for other anti-browning additives. The mode of action of NaCl and NaF is through decreasing PPO activity resulting in reduced oxidation of polyphenols. © 2014 Society of Chemical Industry.

  19. Discovery of potent 2,4-difluoro-linker poly(ADP-ribose) polymerase 1 inhibitors with enhanced water solubility and in vivo anticancer efficacy.

    PubMed

    Chen, Wen-Hua; Song, Shan-Shan; Qi, Ming-Hui; Huan, Xia-Juan; Wang, Ying-Qing; Jiang, Hualiang; Ding, Jian; Ren, Guo-Bin; Miao, Ze-Hong; Li, Jian

    2017-11-01

    Poly (ADP-ribose) polymerase 1 (PARP1) is overexpressed in a variety of cancers, especially in breast and ovarian cancers; tumor cells that are deficient in breast cancer gene 1/2 (BRCA1/2) are highly sensitive to PARP1 inhibition. In this study, we identified a series of 2,4-difluorophenyl-linker analogs (15-55) derived from olaparib as novel PARP1 inhibitors. Four potent analogs 17, 43, 47, and 50 (IC 50 =2.2-4.4 nmol/L) effectively inhibited the proliferation of Chinese hamster lung fibroblast V-C8 cells (IC 50 =3.2-37.6 nmol/L) in vitro, and showed specificity toward BRCA-deficient cells (SI=40-510). The corresponding hydrochloride salts 56 and 57 (based on 43 and 47) were highly water soluble in pH=1.0 buffered salt solutions (1628.2 μg/mL, 2652.5 μg/mL). In a BRCA1-mutated xenograft model, oral administration of compound 56 (30 mg·kg -1 ·d -1 , for 21 d) exhibited more prominent tumor growth inhibition (96.6%) compared with the same dose of olaparib (56.3%); in a BRCA2-mutated xenograft model, oral administration of analog 43 (10 mg·kg -1 ·d -1 , for 28 d) significantly inhibited tumor growth (69.0%) and had no negative effects on the body weights. Additionally, compound 56 exhibited good oral bioavailability (F=32.2%), similar to that of olaparib (F=45.4%). Furthermore, the free base 43 of the hydrochloride salt 56 exhibited minimal hERG inhibition activity (IC 50 =6.64 μmol/L). Collectively, these data demonstrate that compound 56 may be an excellent drug candidate for the treatment of cancer, particularly BRCA-deficient tumors.

  20. Plasma 24,25-dihydroxyvitamin D concentration of Dahl salt-sensitive rats decreases during high salt intake

    NASA Technical Reports Server (NTRS)

    Thierry-Palmer, Myrtle; Tewolde, Teclemicael K.; Forte, Camille; Wang, Min; Bayorh, Mohamed A.; Emmett, Nerimiah L.; White, Jolanda; Griffin, Keri

    2002-01-01

    Dahl salt-sensitive rats, but not salt-resistant rats, develop hypertension in response to high salt intake. We have previously shown an inverse relationship between plasma 25-hydroxyvitamin D (25-OHD) concentration and blood pressure of Dahl salt-sensitive rats during high salt intake. In this study, we report on the relationship between high salt intake and plasma 24,25-dihydroxyvitamin D (24,25-(OH)(2)D) concentration of Dahl salt-sensitive and salt-resistant rats. Rats were fed a high salt diet (8%) and sacrificed at day 2, 7, 14, 21, and 28. Plasma 24,25-(OH)(2)D concentrations of salt-sensitive rats were reduced to 50% of that at baseline at day 2-when blood pressure and plasma 25-OHD concentration were unchanged, but 25-OHD content in the kidney was 81% of that at baseline. Plasma 24,25-(OH)(2)D concentration was reduced further to 10% of that at baseline from day 7 to 14 of high salt intake, a reduction that was prevented in rats switched to a low salt (0.3%) diet at day 7. Exogenous 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D(3)), administered at a level that increased plasma 24,25-(OH)(2)D concentration to five times normal, did not attenuate the salt-induced hypertension of salt-sensitive rats. Plasma 24,25-(OH)(2)D concentration of salt-resistant rats was gradually reduced to 50% of that at baseline at day 14 and returned to baseline value at day 28 of high salt intake. We conclude that the decrease in plasma 24,25-(OH)(2)D concentration in salt-sensitive rats during high salt intake is caused by decreased 25-OHD content in the kidney and also by another unidentified mechanism.

  1. Detection beyond the Debye screening length in a high-frequency nanoelectronic biosensor.

    PubMed

    Kulkarni, Girish S; Zhong, Zhaohui

    2012-02-08

    Nanosensors based on the unique electronic properties of nanotubes and nanowires offer high sensitivity and have the potential to revolutionize the field of Point-of-Care (POC) medical diagnosis. The direct current (dc) detection of a wide array of organic and inorganic molecules has been demonstrated on these devices. However, sensing mechanism based on measuring changes in dc conductance fails at high background salt concentrations, where the sensitivity of the devices suffers from the ionic screening due to mobile ions present in the solution. Here, we successfully demonstrate that the fundamental ionic screening effect can be mitigated by operating single-walled carbon nanotube field effect transistor as a high-frequency biosensor. The nonlinear mixing between the alternating current excitation field and the molecular dipole field can generate mixing current sensitive to the surface-bound biomolecules. Electrical detection of monolayer streptavidin binding to biotin in 100 mM buffer solution is achieved at a frequency beyond 1 MHz. Theoretical modeling confirms improved sensitivity at high frequency through mitigation of the ionic screening effect. The results should promise a new biosensing platform for POC detection, where biosensors functioning directly in physiologically relevant condition are desired. © 2012 American Chemical Society

  2. Use of bicarbonate buffer systems for dissolution characterization of enteric-coated proton pump inhibitor tablets.

    PubMed

    Shibata, Hiroko; Yoshida, Hiroyuki; Izutsu, Ken-Ichi; Goda, Yukihiro

    2016-04-01

    The aim of this study was to assess the effects of buffer systems (bicarbonate or phosphate at different concentrations) on the in vitro dissolution profiles of commercially available enteric-coated tablets. In vitro dissolution tests were conducted using an USP apparatus II on 12 enteric-coated omeprazole and rabeprazole tablets, including innovator and generic formulations in phosphate buffers, bicarbonate buffers and a media modified Hanks (mHanks) buffer. Both omeprazole and rabeprazole tablets showed similar dissolution profiles among products in the compendial phosphate buffer system. However, there were large differences between products in dissolution lag time in mHanks buffer and bicarbonate buffers. All formulations showed longer dissolution lag times at lower concentrations of bicarbonate or phosphate buffers. The dissolution rank order of each formulation differed between mHanks buffer and bicarbonate buffers. A rabeprazole formulation coated with a methacrylic acid copolymer showed the shortest lag time in the high concentration bicarbonate buffer, suggesting varied responses depending on the coating layer and buffer components. Use of multiple dissolution media during in vitro testing, including high concentration bicarbonate buffer, would contribute to the efficient design of enteric-coated drug formulations. © 2016 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology.

  3. High performance liquid chromatography for the simultaneous analysis of penicillin residues in beef and milk using ion-paired extraction and binary water-acetonitrile mixture.

    PubMed

    Kukusamude, Chunyapuk; Burakham, Rodjana; Chailapakul, Orawon; Srijaranai, Supalax

    2012-04-15

    An ion-paired extraction (IPE) has been developed for the analysis of penicillin antibiotics (penicillin G, oxacillin and cloxacillin) in beef and milk samples using tetrabutylammonium bromide (TBABr) as ion-pairing agent and binary water-acetonitrile as extractant. The factors affecting the IPE efficiency were optimized including solution pH, volume of acetonitrile (ACN), concentration of TBABr and electrolyte salt (NH(4))(2)SO(4). The optimum IPE conditions were 10 mmol L(-1) phosphate buffer pH 8, 2 mL of ACN, 6 mmol L(-1) of TBABr and 2.5 mL of saturated ammonium sulfate. Under the HPLC condition: an Xbridge™ C18 reversed-phase column, isocratic elution of 5 mmol L(-1) phosphate buffer (pH 6.6) and acetonitrile (75:25, v/v) and a flow rate of 1 mL min(-1), with UV detection at 215 nm, the separation of three penicillins was achieved within 10 min. Under the selected optimum conditions, the enhancement of 21-53 folds compared to that without preconcentration and limits of detection (LODs) of 1-2 ng mL(-1) were obtained. Good reproducibility was achieved with RSD<2% for retention time and <5% for slope of calibration curves. The average recoveries higher than 85% were obtained. The proposed IPE-HPLC method has shown to be high efficient preconcentration and analysis method for penicillin residues in beef and milk with LOD lower than the maximum residue limits. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Moderate (20%) fructose-enriched diet stimulates salt-sensitive hypertension with increased salt retention and decreased renal nitric oxide.

    PubMed

    Gordish, Kevin L; Kassem, Kamal M; Ortiz, Pablo A; Beierwaltes, William H

    2017-04-01

    Previously, we reported that 20% fructose diet causes salt-sensitive hypertension. In this study, we hypothesized that a high salt diet supplemented with 20% fructose (in drinking water) stimulates salt-sensitive hypertension by increasing salt retention through decreasing renal nitric oxide. Rats in metabolic cages consumed normal rat chow for 5 days (baseline), then either: (1) normal salt for 2 weeks, (2) 20% fructose in drinking water for 2 weeks, (3) 20% fructose for 1 week, then fructose + high salt (4% NaCl) for 1 week, (4) normal chow for 1 week, then high salt for 1 week, (5) 20% glucose for 1 week, then glucose + high salt for 1 week. Blood pressure, sodium excretion, and cumulative sodium balance were measured. Systolic blood pressure was unchanged by 20% fructose or high salt diet. 20% fructose + high salt increased systolic blood pressure from 125 ± 1 to 140 ± 2 mmHg ( P  < 0.001). Cumulative sodium balance was greater in rats consuming fructose + high salt than either high salt, or glucose + high salt (114.2 ± 4.4 vs. 103.6 ± 2.2 and 98.6 ± 5.6 mEq/Day19; P  < 0.05). Sodium excretion was lower in fructose + high salt group compared to high salt only: 5.33 ± 0.21 versus 7.67 ± 0.31 mmol/24 h; P  < 0.001). Nitric oxide excretion was 2935 ± 256  μ mol/24 h in high salt-fed rats, but reduced by 40% in the 20% fructose + high salt group (2139 ± 178  μ mol /24 hrs P  < 0.01). Our results suggest that fructose predisposes rats to salt-sensitivity and, combined with a high salt diet, leads to sodium retention, increased blood pressure, and impaired renal nitric oxide availability. © 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  5. Single molecule force measurements delineate salt, pH and surface effects on biopolymer adhesion

    NASA Astrophysics Data System (ADS)

    Pirzer, T.; Geisler, M.; Scheibel, T.; Hugel, T.

    2009-06-01

    In this paper we probe the influence of surface properties, pH and salt on the adhesion of recombinant spider silk proteins onto solid substrates with single molecule force spectroscopy. A single engineered spider silk protein (monomeric C16 or dimeric (QAQ)8NR3) is covalently bound with one end to an AFM tip, which assures long-time measurements for hours with one and the same protein. The tip with the protein is brought into contact with various substrates at various buffer conditions and then retracted to desorb the protein. We observe a linear dependence of the adhesion force on the concentration of three selected salts (NaCl, NaH2PO4 and NaI) and a Hofmeister series both for anions and cations. As expected, the more hydrophobic C16 shows a higher adhesion force than (QAQ)8NR3, and the adhesion force rises with the hydrophobicity of the substrate. Unexpected is the magnitude of the dependences—we never observe a change of more than 30%, suggesting a surprisingly well-regulated balance between dispersive forces, water-structure-induced forces as well as co-solute-induced forces in biopolymer adhesion.

  6. Quinoa starch granules as stabilizing particles for production of Pickering emulsions.

    PubMed

    Rayner, Marilyn; Sjöö, Malin; Timgren, Anna; Dejmek, Petr

    2012-01-01

    Intact starch granules isolated from quinoa (Chenopodium quinoa Willd.) were used to stabilize emulsion drops in so-called Pickering emulsions. Miglyol 812 was used as dispersed phase and a phosphate buffer (pH7) with different salt (NaCl) concentrations was used as the continuous phase. The starch granules were hydrophobically modified to different degrees by octenyl succinic anhydride (OSA) or by dry heat treatment at 120 degrees C in order to study the effect on the resulting emulsion drop size. The degree of OSA-modification had a low to moderate impact on drop size. The highest level of modification (4.66%) showed the largest mean drop size, and lowest amount of free starch, which could be an effect of a higher degree of aggregation of the starch granules and, thereby, also the emulsion drops stabilized by them. The heat treated starch granules had a poor stabilizing ability and only the starch heated for the longest time (150 min at 120 degrees C) had a better emulsifying capacity than the un-modified native starch granules. The effect of salt concentration was rather limited. However, an increased concentration of salt slightly increased the mean drop size and the elastic modulus.

  7. Radiation sensitivity of poliovirus, a model for norovirus, inoculated in oyster ( Crassostrea gigas) and culture broth under different conditions

    NASA Astrophysics Data System (ADS)

    Jung, Pil-Mun; Park, Jae Seok; Park, Jin-Gyu; Park, Jae-Nam; Han, In-Jun; Song, Beom-Seok; Choi, Jong-il; Kim, Jae-Hun; Byun, Myung-Woo; Baek, Min; Chung, Young-Jin; Lee, Ju-Woon

    2009-07-01

    Poliovirus is a recognized surrogate for norovirus, pathogen in water and food, due to the structural and genetic similarity. Although radiation sensitivity of poliovirus in water or media had been reported, there has been no research in food model such as shellfish. In this study, oyster ( Crassostrea gigas) was incubated in artificial seawater contaminated with poliovirus, and thus radiation sensitivity of poliovirus was determined in inoculated oyster. The effects of ionizing radiation on the sensitivity of poliovirus were also evaluated under different conditions such as pH (4-7) and salt concentration (1-15%) in culture broth, and temperature during irradiation. The D10 value of poliovirus in PBS buffer, virus culture broth and oyster was determined to 0.46, 2.84 and 2.94 kGy, respectively. The initial plaque forming unit (PFU) of poliovirus in culture broth was slightly decreased as the decrease of pH and the increase of salt concentration, but radiation sensitivity was not affected by pH and salt contents. However, radiation resistance of poliovirus was increased at frozen state. These results provide the basic information for the inactivation of pathogenic virus in foods by using irradiation.

  8. The Survival of Burkholderia pseudomallei in Liquid Media

    PubMed Central

    Robertson, Jeannie; Levy, Avram; Sagripanti, Jose-Luis; Inglis, Timothy J. J.

    2010-01-01

    We studied the effect of environmental parameters on the survival of Burkholderia pseudomallei. There was a small increase in bacterial count for up to 28 days in sterilized distilled water or rain water, in water at 20°C or 40°C, and in buffered solutions of pH 4 or higher. Counts of culturable B. pseudomallei declined at pH 3, in the presence of seawater or water with concentrations of 4% salt or higher, and under refrigeration. The morphological appearances of B. pseudomallei changed under conditions that maintained culturable numbers from bacilli to coccoid cells and spiral forms under pH or salt stress. These observations indicate that B. pseudomallei can endure nutrient-depleted environments as well as a wide range of pH, salt concentrations, and temperatures for periods of up to 28 days. The relative stability of B. pseudomallei under these conditions underlines the tenacity of this species and its potential for natural dispersal in water: in surface water collections, in managed water distribution systems, and through rainfall. These survival properties help explain the recent expansion of the known melioidosis endemic zone in Australia and may have played a part in recent melioidosis outbreaks. PMID:20065001

  9. Nitrogen-Polar (000 1 ¯ ) GaN Grown on c-Plane Sapphire with a High-Temperature AlN Buffer.

    PubMed

    Song, Jie; Han, Jung

    2017-03-02

    We demonstrate growing nitrogen-polar (N-polar) GaN epilayer on c-plane sapphire using a thin AlN buffer layer by metalorganic chemical vapor deposition. We have studied the influence of the AlN buffer layer on the polarity, crystalline quality, and surface morphology of the GaN epilayer and found that the growth temperature of the AlN buffer layer played a critical role in the growth of the GaN epilayer. The low growth temperature of the AlN buffer results in gallium-polar GaN. Even a nitridation process has been conducted. High growth temperature for an AlN buffer layer is required to achieve pure N-polarity, high crystalline quality, and smooth surface morphology for a GaN epilayer.

  10. Status of CdS/CdTe solar cell research at NREL

    NASA Astrophysics Data System (ADS)

    Ramanathan, K.; Dhere, R. G.; Coutts, T. J.; Chu, T.; Chu, S.

    1992-12-01

    We report on the deposition of thin cadmium sulfide (CdS) layers from aqueous solutions and their optical properties. CdS layers have been deposited on soda lime glass, tin oxide coated glass and copper indium diselenide (CuInSe2) thin films. A systematic increase in the absorption is found to occur with increasing concentration of the buffer salt used in the bath. CdS/CdTe thin film solar cells have been fabricated by close spaced sublimation of CdTe, yielding 11.3% devices.

  11. The Role of Newly Discovered Exotoxin (S Toxin) in Pseudomonas aeruginosa Infections

    DTIC Science & Technology

    1979-08-01

    sodium or potassium phosphate 6.0-8.0 N-2-hydroxyethylpiperazine-N’-2-ethanesulfonic acid (HEPES) 6.5-8.5 tris 7.0-9.5 sodium borate 7.5-9.5 sodium...was found to be variable with respect to whether sodium or potassium phosphate buffer was used. With sodium phosphate, virtually all the enzyme...activity bound was eluted between 20-100.2M phosphate at pH 6.8. With the potassium salt, elution occurs at 400-?00mM KP04. Since very little protein was

  12. Measuring the Role of Ecological Shift and Environmental Change on Organic Carbon Stocks in Salt Marshes and Mangrove Dominated Wetlands from the Texas Gulf Coast

    NASA Astrophysics Data System (ADS)

    Norwood, M. J.; Louchouarn, P.; Armitage, A. R.; HighField, W.; Brody, S.; White, N.

    2014-12-01

    Texas coastal wetlands are dynamic marsh-mangrove ecotones that play an important role in fishery recruitment, storm buffering, and carbon storage. Historically, C4 salt marsh plants, such as Spartina alterniflora, have dominated the Texas Gulf Coast. For the past 2-3 decades, some of these ecosystems have experienced community shifts with woody tropical plants (Avicennia germinans) competing for resources. This study presents new results on the carbon sequestration potential following such ecological shifts as well as coastal development and wetland loss along the coast of Texas. The recorded change from native grass-dominated C4 salt marshes to wood-dominated C3 mangroves over the last 20 years (1990-2010: 4,660 km2) leads to a non-significant loss in aboveground organic carbon (OC) stocks (-6.5.106 g OC). The most substantial loss of aboveground OC in Texas coastal salt marshes is due to the transformation of these wetlands into tidal flats and open water (-7.53.108 g OC). Similarly, the largest losses in aboveground OC stocks from mangrove ecosystems (-1.57.107 g OC) are due to replacement by open water. Along with the decrease in aboveground OC stocks, we identified a significant decrease in sedimentary OC inventories due to the loss of salt marsh and mangrove coverage (-3.69.109 g OC and 5.71.107 g OC, respectively). In contrast, mangrove expansion into mudflat and salt marsh environments led to a positive addition in aboveground OC stocks (2.78.108 g OC) and increased OC sedimentary inventories (2.32.109 g OC). Mangrove expansion offsets only 70% of the total calculated OC loss (-4.51.109 g OC) in coastal wetlands along the Texas gulf coast over the 20-year study period. This deficit loss is primarily attributed to environmental pressures on coastal salt marshes (i.e., sea level rise, urban and coastal development, erosion).

  13. Characterization of the major proteins of tubers of yam bean (Pachyrhizus ahipa).

    PubMed

    Forsyth, Jane L; Shewry, Peter R

    2002-03-27

    Tubers of six accessions of ahipa (Pachyrhizus ahipa) contained between 0.77 and 1.34% nitrogen on a dry weight basis. This corresponds to 4.8 to 8.4% crude protein based on a nitrogen to protein conversion factor of 6.25; but detailed analysis of AC230 showed that although 93% of the total N was extracted with buffer containing 1.0 M NaCl, about a third of this was lost on dialysis. It was calculated, therefore, that salt-soluble proteins comprise about 60% of the total tuber nitrogen, with low-molecular-mass nitrogenous components comprising a further 30%. Electophoretic analysis of the salt-soluble proteins showed similar patterns of components in the six accessions, with none being present in amounts sufficiently high to suggest a role as storage proteins. Furthermore, light microscopy failed to show significant deposits of protein within the tuber cells. Five "major" protein bands, which together accounted for about 19% of the total salt-soluble protein fraction were purified and subjected to N-terminal amino acid sequencing. Comparison of these with sequences in protein databases revealed similarities to alpha-amylases, chitinases and chitin binding proteins, cysteine proteinases (including major components from P. erosus tubers), a tuberization-specific protein from potato, and proteins induced in soybean and pea by stress or the plant hormone abscisic acid, respectively. It was concluded that the primary roles of these proteins are probably in aspects of tuber metabolism and development and/or conferring protection to pests and pathogens, and that true storage proteins are not present. The absence of storage proteins is consistent with the biological role of the tubers as storage organs for carbohydrates (cf cassava tuberous roots) rather than as propagules (cf yam and potato tubers).

  14. High pressure effects on the iron iron oxide and nickel nickel oxide oxygen fugacity buffers

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

    Campbell, Andrew J; Danielson, Lisa; Righter, Kevin

    The chemical potential of oxygen in natural and experimental samples is commonly reported relative to a specific oxygen fugacity (fO{sub 2}) buffer. These buffers are precisely known at 1 bar, but under high pressures corresponding to the conditions of the deep Earth, oxygen fugacity buffers are poorly calibrated. Reference (1 bar) fO{sub 2} buffers can be integrated to high pressure conditions by integrating the difference in volume between the solid phases, provided that their equations of state are known. In this work, the equations of state and volume difference between the metal-oxide pairs Fe-FeO and Ni-NiO were measured using synchrotronmore » X-ray diffraction in a multi-anvil press and laser heated diamond anvil cells. The results were used to construct high pressure fO{sub 2} buffer curves for these systems. The difference between the Fe-FeO and Ni-NiO buffers is observed to decrease significantly, by several log units, over 80 GPa. The results can be used to improve interpretation of high pressure experiments, specifically Fe-Ni exchange between metallic and oxide phases.« less

  15. Methane fluxes along a salinity gradient on a restored salt marsh, Harpswell, ME

    NASA Astrophysics Data System (ADS)

    Gunn, Cailene; Johnson, Beverly, ,, Dr.; Dostie, Phil; Bohlen, Curtis; Craig, Matthew

    2016-04-01

    This study functions as a pilot project to understand the relationship between salinity and methane emissions on a recently restored salt marsh in Casco Bay, Maine. Salt marshes are dynamic and highly productive ecosystems that provide a multitude of ecosystem services including nutrient filtration, storm-water buffering and carbon sequestration. These ecosystems are highly susceptible to anthropogenic alteration. The emplacement of causeways and narrow culverts, restricts tidal flow and leads to loss of healthy salinity gradients. Consequently, numerous salt marshes have experienced increases in freshwater vegetation growth as a result of coastal population expansion. Recent restoration efforts on Long Marsh, Harpswell, ME replaced a severely undersized culvert with a larger one in February, 2014. The salinity gradient has since been restored along much of the marsh, and freshwater vegetation that encroached on the marsh platform has died back. Vegetation and salinity are key indicators and drivers of CH4 emissions on salt marshes. Using static gas chambers, we quantified CH4 fluxes along two transects at five diverse sites ranging from healthy marsh (salinity of 27 to 31 psu) with Spartina vegetation, to regions invaded by Typha and other freshwater vegetation (salinity of 0 to 4 psu). Sampling was executed in the months of July, August and October. CH4 concentrations were determined using a gas chromatograph with a flame-ionization detector. Preliminary findings suggest reintroduction of healthy tidal flows into the marsh inhibits CH4 production, where the lowest fluxes with least variability were observed at the most saline sites with Spartina vegetation. The largest range of CH4 fluxes exhibited emissions from 0.75 μmol CH4/m2/hr to 518.4 μmol CH4/m2/hr at the Typha dominated sites from July to October. Fluxes at the saltwater and brackish regions were far less variable with ranges from 0.94 μmol CH4/m2/hr to 8.2 μmol CH4/m2/hr and 2.6 to 9.5 μmol CH4/m2/hr, respectively. The transitional sites exhibited ranges from 1.2 μmol CH4/m2/hr to 16.8 μmol CH4/m2/hr. For all sites, lowest fluxes were observed during the month of October, suggesting seasonal influence on CH4 emissions. These data will be complimented by sediment analyses at each site providing δC and % organic carbon using isotope-ratio mass spectrometry, as well as bulk density and rates of decomposition using a tea bag index.

  16. Final Report: Rational Design of Wide Band Gap Buffer Layers for High-Efficiency Thin-Film Photovoltaics

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

    Lordi, Vincenzo

    The main objective of this project is to enable rational design of wide band gap buffer layer materials for CIGS thin-film PV by building understanding of the correlation of atomic-scale defects in the buffer layer and at the buffer/absorber interface with device electrical properties. Optimized wide band gap buffers are needed to reduce efficiency loss from parasitic absorption in the buffer. The approach uses first-principles materials simulations coupled with nanoscale analytical electron microscopy as well as device electrical characterization. Materials and devices are produced by an industrial partner in a manufacturing line to maximize relevance, with the goal of enablingmore » R&D of new buffer layer compositions or deposition processes to push device efficiencies above 21%. Cadmium sulfide (CdS) is the reference material for analysis, as the prototypical high-performing buffer material.« less

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

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

    Melnik, Yurity; Chen, Lu; Kojiri, Hidehiro

    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 morphologymore » 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).« less

  18. Application of hanging drop technique to optimize human IgG formulations.

    PubMed

    Li, Guohua; Kasha, Purna C; Late, Sameer; Banga, Ajay K

    2010-01-01

    The purpose of this work is to assess the hanging drop technique in screening excipients to develop optimal formulations for human immunoglobulin G (IgG). A microdrop of human IgG and test solution hanging from a cover slide and undergoing vapour diffusion was monitored by a stereomicroscope. Aqueous solutions of IgG in the presence of different pH, salt concentrations and excipients were prepared and characterized. Low concentration of either sodium/potassium phosphate or McIlvaine buffer favoured the solubility of IgG. Addition of sucrose favoured the stability of this antibody while addition of NaCl caused more aggregation. Antimicrobial preservatives were also screened and a complex effect at different buffer conditions was observed. Dynamic light scattering, differential scanning calorimetry and size exclusion chromatography studies were performed to further validate the results. In conclusion, hanging drop is a very easy and effective approach to screen protein formulations in the early stage of formulation development.

  19. Determination of residual fluoroquinolones in honey by liquid chromatography using metal chelate affinity chromatography.

    PubMed

    Yatsukawa, Yoh-Ichi; Ito, Hironobu; Matsuda, Takahiro; Nakamura, Munetomo; Watai, Masatoshi; Fujita, Kazuhiro

    2011-01-01

    A new analytical method for the simultaneous determination of seven fluoroquinolones, namely, norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, orbifloxacin, sarafloxacin, and difloxacin, especially in dark-colored honey, has been developed. Fluoroquinolone antibiotics were extracted from samples with MacIlvaine buffer solution (pH 4.0) containing EDTA disodium salt dihydrate. The extracts were treated with both a polymeric cartridge and a metal chelate affinity column preloaded with ferric ion (Fe3+). LC separation with fluorescence detection was performed at 40 degrees C using an Inertsil ODS-4 analytical column (150 x 4.6 mm, 3 microm). The mobile phase was composed of 20 mM/L citrate buffer solution (pH 3.1)-acetonitrile mixture (70 + 30, v/v) containing 1 mM/L sodium dodecyl sulfate. Lomefloxacin was used as an internal standard. The developed method was validated according to the criteria of European Commission Decision 2002/657/EC. Decision limits and detection capabilities were below 2.9 and 4.4 microg/kg, respectively.

  20. A Janus cobalt-based catalytic material for electro-splitting of water

    NASA Astrophysics Data System (ADS)

    Cobo, Saioa; Heidkamp, Jonathan; Jacques, Pierre-André; Fize, Jennifer; Fourmond, Vincent; Guetaz, Laure; Jousselme, Bruno; Ivanova, Valentina; Dau, Holger; Palacin, Serge; Fontecave, Marc; Artero, Vincent

    2012-09-01

    The future of energy supply depends on innovative breakthroughs regarding the design of cheap, sustainable and efficient systems for the conversion and storage of renewable energy sources. The production of hydrogen through water splitting seems a promising and appealing solution. We found that a robust nanoparticulate electrocatalytic material, H2-CoCat, can be electrochemically prepared from cobalt salts in a phosphate buffer. This material consists of metallic cobalt coated with a cobalt-oxo/hydroxo-phosphate layer in contact with the electrolyte and mediates H2 evolution from neutral aqueous buffer at modest overpotentials. Remarkably, it can be converted on anodic equilibration into the previously described amorphous cobalt oxide film (O2-CoCat or CoPi) catalysing O2 evolution. The switch between the two catalytic forms is fully reversible and corresponds to a local interconversion between two morphologies and compositions at the surface of the electrode. After deposition, the noble-metal-free coating thus functions as a robust, bifunctional and switchable catalyst.

  1. The impact of high-salt exposure on cardiovascular development in the early chick embryo.

    PubMed

    Wang, Guang; Zhang, Nuan; Wei, Yi-Fan; Jin, Yi-Mei; Zhang, Shi-Yao; Cheng, Xin; Ma, Zheng-Lai; Zhao, Shu-Zhu; Chen, You-Peng; Chuai, Manli; Hocher, Berthold; Yang, Xuesong

    2015-11-01

    In this study, we show that high-salt exposure dramatically increases chick mortality during embryo development. As embryonic mortality at early stages mainly results from defects in cardiovascular development, we focused on heart formation and angiogenesis. We found that high-salt exposure enhanced the risk of abnormal heart tube looping and blood congestion in the heart chamber. In the presence of high salt, both ventricular cell proliferation and apoptosis increased. The high osmolarity induced by high salt in the ventricular cardiomyocytes resulted in incomplete differentiation, which might be due to reduced expression of Nkx2.5 and GATA4. Blood vessel density and diameter were suppressed by exposure to high salt in both the yolk sac membrane (YSM) and chorioallantoic membrane models. In addition, high-salt-induced suppression of angiogenesis occurred even at the vasculogenesis stage, as blood island formation was also inhibited by high-salt exposure. At the same time, cell proliferation was repressed and cell apoptosis was enhanced by high-salt exposure in YSM tissue. Moreover, the reduction in expression of HIF2 and FGF2 genes might cause high-salt-suppressed angiogenesis. Interestingly, we show that high-salt exposure causes excess generation of reactive oxygen species (ROS) in the heart and YSM tissues, which could be partially rescued through the addition of antioxidants. In total, our study suggests that excess generation of ROS might play an important role in high-salt-induced defects in heart and angiogenesis. © 2015. Published by The Company of Biologists Ltd.

  2. GaAs buffer layer technique for vertical nanowire growth on Si substrate

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

    Xu, Xiaoqing, E-mail: steelxu@stanford.edu; Parizi, Kokab B.; Huo, Yijie

    2014-02-24

    Gold catalyzed vapor-liquid-solid method is widely applied to III–V nanowire (NW) growth on Si substrate. However, the easy oxidation of Si, possible Si contamination in the NWs, high defect density in the NWs, and high sensitivity of the NW morphology to growth conditions largely limit its controllability. In this work, we developed a buffer layer technique by introducing a GaAs thin film with predefined polarity as a template. It is found that samples grown on these buffer layers all have high vertical NW yields in general, due to the single-orientation of the buffer layers. Low temperature buffer with smoother surfacemore » leads to highest yield of vertical NWs, while high temperature (HT) buffer with better crystallinity results in perfect NW quality. The defect-free property we observed here is very promising for optoelectronic device applications based on GaAs NW. Moreover, the buffer layers can eliminate Si contamination by preventing Si-Au alloy formation and by increasing the thickness of the Si diffusion barrier, thus providing more flexibility to vertical NW growth. The buffer layer technique we demonstrated here could be easily extended to other III-V on Si system for electronic and photonic applications.« less

  3. Purification, crystallization and preliminary crystallographic study of haemoglobin from camel (Camelus dromedarius): a high oxygen-affinity lowland species.

    PubMed

    Balasubramanian, M; Moorthy, Pon Sathya; Neelagandan, K; Ponnuswamy, M N

    2009-08-01

    Haemoglobin is a prototypical allosteric protein that is mainly involved in the transportation of oxygen from the lungs to tissues and of carbon dioxide back to the lungs in an intrinsically coordinated manner to maintain the viability of cells. Haemoglobin from Camelus dromedarius provides an interesting case study of adaptation to life in deserts at extremely high temperatures. An ambition to unravel the integrated structural and functional aspects of the casual survival of this animal at high temperatures led us to specifically work on this problem. The present work reports the preliminary crystallographic study of camel haemoglobin. Camel blood was collected and the haemoglobin was purified by anion-exchange chromatography and crystallized using the hanging-drop vapour-diffusion method under buffered high salt concentration using PEG 3350 as a precipitant. Intensity data were collected using a MAR 345 dtb image-plate detector system. Camel haemoglobin crystallized in the monoclinic space group P2(1), with one whole biological molecule (alpha(2)beta(2)) in the asymmetric unit and unit-cell parameters a = 52.759, b = 116.782, c = 52.807 A, beta = 120.07 degrees .

  4. A thiotrophic microbial community in an acidic brine lake in Northern Chile.

    PubMed

    Escudero, Lorena; Oetiker, Nia; Gallardo, Karem; Tebes-Cayo, Cinthya; Guajardo, Mariela; Nuñez, Claudia; Davis-Belmar, Carol; Pueyo, J J; Chong Díaz, Guillermo; Demergasso, Cecilia

    2018-05-10

    The endorheic basins of the Northern Chilean Altiplano contain saline lakes and salt flats. Two of the salt flats, Gorbea and Ignorado, have high acidic brines. The causes of the local acidity have been attributed to the occurrence of volcanic native sulfur, the release of sulfuric acid by oxidation, and the low buffering capacity of the rocks in the area. Understanding the microbial community composition and available energy in this pristine ecosystem is relevant in determining the origin of the acidity and in supporting the rationale of conservation policies. Besides, a comparison between similar systems in Australia highlights key microbial components and specific ones associated with geological settings and environmental conditions. Sediment and water samples from the Salar de Gorbea were collected, physicochemical parameters measured and geochemical and molecular biological analyses performed. A low diversity microbial community was observed in brines and sediments dominated by Actinobacteria, Algae, Firmicutes and Proteobacteria. Most of the constituent genera have been reported to be either sulfur oxidizing microorganisms or ones having the potential for sulfur oxidation given available genomic data and information drawn from the literature on cultured relatives. In addition, a link between sulfur oxidation and carbon fixation was observed. In contrast, to acid mine drainage communities, Gorbea microbial diversity is mainly supported by chemolithoheterotrophic, facultative chemolithoautotrophic and oligotrophic sulfur oxidizing populations indicating that microbial activity should also be considered as a causative agent of local acidity.

  5. DNA recovery from soils of diverse composition.

    PubMed

    Zhou, J; Bruns, M A; Tiedje, J M

    1996-02-01

    A simple, rapid method for bacterial lysis and direct extraction of DNA from soils with minimal shearing was developed to address the risk of chimera formation from small template DNA during subsequent PCR. The method was based on lysis with a high-salt extraction buffer (1.5 M NaCl) and extended heating (2 to 3 h) of the soil suspension in the presence of sodium dodecyl sulfate (SDS), hexadecyltrimethylammonium bromide, and proteinase K. The extraction method required 6 h and was tested on eight soils differing in organic carbon, clay content, and pH, including ones from which DNA extraction is difficult. The DNA fragment size in crude extracts from all soils was > 23 kb. Preliminary trials indicated that DNA recovery from two soils seeded with gram-negative bacteria was 92 to 99%. When the method was tested on all eight unseeded soils, microscopic examination of indigenous bacteria in soil pellets before and after extraction showed variable cell lysis efficiency (26 to 92%). Crude DNA yields from the eight soils ranged from 2.5 to 26.9 micrograms of DNA g-1, and these were positively correlated with the organic carbon content in the soil (r = 0.73). DNA yields from gram-positive bacteria from pure cultures were two to six times higher when the high-salt-SDS-heat method was combined with mortar-and-pestle grinding and freeze-thawing, and most DNA recovered was of high molecular weight. Four methods for purifying crude DNA were also evaluated for percent recovery, fragment size, speed, enzyme restriction, PCR amplification, and DNA-DNA hybridization. In general, all methods produced DNA pure enough for PCR amplification. Since soil type and microbial community characteristics will influence DNA recovery, this study provides guidance for choosing appropriate extraction and purification methods on the basis of experimental goals.

  6. Effects of hypertonic buffer composition on lymph node uptake and bioavailability of rituximab, after subcutaneous administration.

    PubMed

    Fathallah, Anas M; Turner, Michael R; Mager, Donald E; Balu-Iyer, Sathy V

    2015-03-01

    The subcutaneous administration of biologics is highly desirable; however, incomplete bioavailability after s.c. administration remains a major challenge. In this work we investigated the effects of excipient dependent hyperosmolarity on lymphatic uptake and plasma exposure of rituximab as a model protein. Using Swiss Webster (SW) mice as the animal model, we compared the effects of NaCl, mannitol and O-phospho-L-serine (OPLS) on the plasma concentration of rituximab over 5 days after s.c. administration. An increase was observed in plasma concentrations in animals administered rituximab in hypertonic buffer solutions, compared with isotonic buffer. Bioavailability, as estimated by our pharmacokinetic model, increased from 29% in isotonic buffer to 54% in hypertonic buffer containing NaCl, to almost complete bioavailability in hypertonic buffers containing high dose OPLS or mannitol. This improvement in plasma exposure is due to the improved lymphatic trafficking as evident from the increase in the fraction of dose trafficked through the lymph nodes in the presence of hypertonic buffers. The fraction of the dose trafficked through the lymphatics, as estimated by the model, increased from 0.05% in isotonic buffer to 13% in hypertonic buffer containing NaCl to about 30% for hypertonic buffers containing high dose OPLS and mannitol. The data suggest that hypertonic solutions may be a viable option for improving s.c. bioavailability. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Effects of hypertonic buffer composition on lymph node uptake and bioavailability of rituximab, after subcutaneous administration

    PubMed Central

    Fathallah, Anas M.; Turner, Michael R.; Balu-Iyer, Sathy V.

    2015-01-01

    Subcutaneous administration of biologics is highly desirable; however, incomplete bioavailability after sc administration remains a major challenge. In this work we investigated the effects of excipient dependent hyper-osmolarity on lymphatic uptake and plasma exposure of rituximab as a model protein. Using Swiss Webster (SW) mice as our animal model, we compared the effects of NaCl, mannitol and, O-Phospho-L-Serine (OPLS) on plasma concentration of rituximab over 5 days after sc administration. We observed an increase in plasma concentrations in animals administered rituximab in hypertonic buffer solutions, as compared to isotonic buffer. Bioavailability, as estimated by our pharmacokinetic model, increased from 29% in isotonic buffer to 54% in hypertonic buffer containing NaCl, to almost complete bioavailability in hypertonic buffers containing high dose OPLS or mannitol. This improvement in plasma exposure is due to improved lymphatic trafficking as evident from the increase in the fraction of dose trafficked through the lymph node in the presence of hypertonic buffers. The fraction of the dose trafficked through the lymphatic, as estimated by the model, increased from 0.05 % in isotonic buffer to 13% in hyper-tonic buffer containing NaCl to about 30% for hypertonic buffers containing high dose OPLS and mannitol. Our data suggests that hypertonic solutions may be a viable option to improve sc bioavailability. PMID:25377184

  8. Low-Salt Intake during Mating or Gestation in Rats Is Associated with Low Birth and Survival Rates of Babies.

    PubMed

    Chou, Ranna; Hara, Anna; Du, DongDong; Shimizu, Namiko; Sakuyama, Hiroe; Uehara, Yoshio

    2014-01-01

    We investigated the influence of maternal salt restriction during mating or gestation on birth rate and offspring growth in Dahl salt-sensitive rats (DS). DS were divided into 5 groups: DS fed a low-salt (0.3% NaCl, w/w) (DS-low) or high-salt (4% NaCl, w/w) diet (DS-high) during mating and DS-high or DS-low during gestation, and DS fed regular chow (0.75% NaCl, w/w) (DS-regular) throughout mating and gestation. During the unspecified periods, the rats were given regular chow. DS-low during mating delivered fewer infants than high-salt mothers (P < 0.05). The birth rate on regular chow was 87%. Six out of 11 DS-low rats during pregnancy produced pups while the rats fed a high-salt diet all delivered pups (P < 0.025). The pup survival rate was 67% for high-salt mothers during mating and 54% for mothers on a low-salt diet. The pup survival rate was 95% for mothers on a high-salt diet during pregnancy and 64% for mothers on a low-salt diet (P < 0.0001). Seven out of 8 DS-regular rats during mating delivered 59 neonates. However, 66% of the neonates survived. A low-salt diet during mating or pregnancy lowers birth rate and the neonates from low-salt mothers during pregnancy were more likely to die than those from high-salt mothers.

  9. Variation of power generation at different buffer types and conductivities in single chamber microbial fuel cells.

    PubMed

    Nam, Joo-Youn; Kim, Hyun-Woo; Lim, Kyeong-Ho; Shin, Hang-Sik; Logan, Bruce E

    2010-01-15

    Microbial fuel cells (MFCs) are operated with solutions containing various chemical species required for the growth of electrochemically active microorganisms including nutrients and vitamins, substrates, and chemical buffers. Many different buffers are used in laboratory media, but the effects of these buffers and their inherent electrolyte conductivities have not been examined relative to current generation in MFCs. We investigated the effect of several common buffers (phosphate, MES, HEPES, and PIPES) on power production in single chambered MFCs compared to a non-buffered control. At the same concentrations the buffers produced different solution conductivities which resulted in different ohmic resistances and power densities. Increasing the solution conductivities to the same values using NaCl produced comparable power densities for all buffers. Very large increases in conductivity resulted in a rapid voltage drop at high current densities. Our results suggest that solution conductivity at a specific pH for each buffer is more important in MFC studies than the buffer itself given relatively constant pH conditions. Based on our analysis of internal resistance and a set neutral pH, phosphate and PIPES are the most useful buffers of those examined here because pH was maintained close to the pK(a) of the buffer, maximizing the ability of the buffer to contribute to increase current generation at high power densities. Copyright 2009 Elsevier B.V. All rights reserved.

  10. Highly efficient treatment of aerobic vaginitis with simple acidic buffered gels: The importance of pH and buffers on the microenvironment of vaginas.

    PubMed

    Sun, Xiaodong; Qiu, Haiying; Jin, Yiguang

    2017-06-15

    Aerobic vaginitis (AV) leads to uterus deep infection or preterm birth. Antibacterial agents are not optimal therapeutics of AV. Here, we report a series of temperature-sensitive in situ forming acidic buffered gels for topical treatment of AV, involving lactate, acetate, and citrate gels at pH 3.5, 5.0, and 6.5. AV rat models were prepared following vaginal infection with Staphylococcus aureus and Escherichia coli. In vitro/in vivo studies of the buffered gels were performed compared with ofloxacin gels and blank gels. All the buffered gels showed the lower in vitro antibacterial activities than ofloxacin gels but the better in vivo anti-S. aureus effects and similar anti-E. coli effects. The buffered gels improved Lactobacillus growth in the vaginas. Both the healthy rat vaginal pH and the pH of rat vaginas treated with the buffered gels were about 6.5 though the AV rat models or ones treated with ofloxacin gels still remained at the high pH more than 7.0. After treatments with the buffered gels, the vaginal smears changed to a clean state nearly without aerobic bacteria, the vaginal tissues were refreshed, and the immunoreactions were downregulated. The acidic buffered gels bring rapid decrease of local vaginal pH, high antibacterial activities, improvement of probiotics, and alleviation of inflammation. They are simple, highly efficient, and safe anti-AV formulations. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Predictive model for Clostridium perfringens growth in roast beef during cooling and inhibition of spore germination and outgrowth by organic acid salts.

    PubMed

    Sánchez-Plata, Marcos X; Amézquita, Alejandro; Blankenship, Erin; Burson, Dennis E; Juneja, Vijay; Thippareddi, Harshavardhan

    2005-12-01

    Spores of foodborne pathogens can survive traditional thermal processing schedules used in the manufacturing of processed meat products. Heat-activated spores can germinate and grow to hazardous levels when these products are improperly chilled. Germination and outgrowth of Clostridium perfringens spores in roast beef during chilling was studied following simulated cooling schedules normally used in the processed-meat industry. Inhibitory effects of organic acid salts on germination and outgrowth of C. perfringens spores during chilling and the survival of vegetative cells and spores under abusive refrigerated storage was also evaluated. Beef top rounds were formulated to contain a marinade (finished product concentrations: 1% salt, 0.2% potassium tetrapyrophosphate, and 0.2% starch) and then ground and mixed with antimicrobials (sodium lactate and sodium lactate plus 2.5% sodium diacetate and buffered sodium citrate and buffered sodium citrate plus 1.3% sodium diacetate). The ground product was inoculated with a three-strain cocktail of C. perfringens spores (NCTC 8238, NCTC 8239, and ATCC 10388), mixed, vacuum packaged, heat shocked for 20 min at 75 degrees C, and chilled exponentially from 54.5 to 7.2 degrees C in 9, 12, 15, 18, or 21 h. C. perfringens populations (total and spore) were enumerated after heat shock, during chilling, and during storage for up to 60 days at 10 degrees C using tryptose-sulfite-cycloserine agar. C. perfringens spores were able to germinate and grow in roast beef (control, without any antimicrobials) from an initial population of ca. 3.1 log CFU/g by 2.00, 3.44, 4.04, 4.86, and 5.72 log CFU/g after 9, 12, 15, 18, and 21 h of exponential chilling. A predictive model was developed to describe sigmoidal C. perfringens growth curves during cooling of roast beef from 54.5 to 7.2 degrees C within 9, 12, 15, 18, and 21 h. Addition of antimicrobials prevented germination and outgrowth of C. perfringens regardless of the chill times. C. perfringens spores could be recovered from samples containing organic acid salts that were stored up to 60 days at 10 degrees C. Extension of chilling time to > or =9 h resulted in >1 log CFU/g growth of C. perfringens under anaerobic conditions in roast beef. Organic acid salts inhibited outgrowth of C. perfringens spores during chilling of roast beef when extended chill rates were followed. Although C. perfringens spore germination is inhibited by the antimicrobials, this inhibition may represent a hazard when such products are incorporated into new products, such as soups and chili, that do not contain these antimicrobials, thus allowing spore germination and outgrowth under conditions of temperature abuse.

  12. Method of deforming a biaxially textured buffer layer on a textured metallic substrate and articles therefrom

    DOEpatents

    Lee, Dominic F.; Kroeger, Donald M.; Goyal, Amit

    2000-01-01

    The present invention provides methods and biaxially textured articles having a deformed epitaxial layer formed therefrom for use with high temperature superconductors, photovoltaic, ferroelectric, or optical devices. A buffer layer is epitaxially deposited onto biaxially-textured substrates and then mechanically deformed. The deformation process minimizes or eliminates grooves, or other irregularities, formed on the buffer layer while maintaining the biaxial texture of the buffer layer. Advantageously, the biaxial texture of the buffer layer is not altered during subsequent heat treatments of the deformed buffer. The present invention provides mechanical densification procedures which can be incorporated into the processing of superconducting films through the powder deposit or precursor approaches without incurring unfavorable high-angle grain boundaries.

  13. Eco-buffers: A high density agroforestry design using native species

    Treesearch

    William Schroeder

    2012-01-01

    This study showed that Eco-Buffers are characterized by rapid establishment and superior survival when compared to single species buffers. Height of green ash (Fraxinus pennsylvanica Marsh. var. subintegerrima (Vahl.) Fern.) after eight growing seasons averaged 415 cm when growing in an Eco-Buffer compared to 333cm in the single species buffer. Site capture in the Eco-...

  14. Composition of Ragusano cheese during aging.

    PubMed

    Licitra, G; Campo, P; Manenti, M; Portelli, G; Scuderi, S; Carpino, S; Barbano, D M

    2000-03-01

    Ragusano cheese is a brine-salted pasta filata cheese. Composition changes during 12 mo of aging were determined. Historically, Ragusano cheese has been aged in caves at 14 to 16 degrees C with about 80 to 90% relative humidity. Cheeses (n = 132) included in our study of block-to-block variation were produced by 20 farmhouse cheese makers in the Hyblean plain region of the Province of Ragusa in Sicily. Mean initial cheese block weight was about 14 kg. The freshly formed blocks of cheese before brine salting contained about 45.35% moisture, 25.3% protein, and 25.4% fat, with a pH of 5.25. As result of the brining and aging process, a natural rind forms. After 12 mo of aging, the cheese contained about 33.6% moisture, 29.2% protein, 30.0% fat, and 4.4% salt with a pH of 5.54, but block-to-block variation was large. Both soluble nitrogen content and free fatty acid (FFA) content increased with age. The pH 4.6 acetate buffer and 12% TCA-soluble nitrogen as a percentage of total nitrogen were 16 and 10.7%, respectively, whereas the FFA content was about 643 mg/100 g of cheese at 180 d. Five blocks of cheese were selected at 180 d for a study of variation within block. Composition variation within block was large; the center had higher moisture and lower salt in moisture content than did the outside. Composition variation within blocks favored more proteolysis and softer texture in the center.

  15. Effect of yeast extract addition to a mineral salts medium containing hydrolyzed plant xylan on fungal pullulan production.

    PubMed

    Kennedy Ii, Daniel E; West, Thomas P

    2018-05-16

    The ability of the fungus Aureobasidium pullulans ATCC 42023 to produce pullulan from yeast extract-supplemented xylan hydrolysates of the prairie grass prairie cordgrass was examined relative to polysaccharide and cell biomass production, yield, and pullulan content of the polysaccharide. A pullulan concentration of 11.2 g L-1 and yield of 0.79 g g-1 was produced by ATCC 42023 when grown for 168 h at 30°C on the phosphate-buffered hydrolysate supplemented with yeast extract. The highest biomass level being 8.8 g L-1 was produced by ATCC 42023 after 168 h on a yeast extract-supplemented, hydrolysate-containing complete medium lacking sodium chloride. The highest pullulan content of the polysaccharide produced by ATCC 42023 after 168 h on the hydrolysate medium supplemented with yeast extract and ammonium sulfate was 70%. The findings indicate that a polysaccharide with a high pullulan content can be produced at a relatively high yield by the fungus grown on a yeast extract-supplemented xylan hydrolysate, suggesting that pullulan could be produced using a biomass-based process.

  16. Purification of bone morphogenetic protein-2 from refolding mixtures using mixed-mode membrane chromatography.

    PubMed

    Gieseler, Gesa; Pepelanova, Iliyana; Stuckenberg, Lena; Villain, Louis; Nölle, Volker; Odenthal, Uwe; Beutel, Sascha; Rinas, Ursula; Scheper, Thomas

    2017-01-01

    In this study, we present the development of a process for the purification of recombinant human bone morphogenetic protein-2 (rhBMP-2) using mixed-mode membrane chromatography. RhBMP-2 was produced as inclusion bodies in Escherichia coli. In vitro refolding using rapid dilution was carried out according to a previously established protocol. Different membrane chromatography phases were analyzed for their ability to purify BMP-2. A membrane phase with salt-tolerant properties resulting from mixed-mode ligand chemistry was able to selectively purify BMP-2 dimer from refolding mixtures. No further purification or polishing steps were necessary and high product purity was obtained. The produced BMP-2 exhibited a biological activity of 7.4 × 10 5  U/mg, comparable to commercial preparations. Mixed-mode membrane chromatography can be a valuable tool for the direct purification of proteins from solutions with high-conductivity, for example refolding buffers. In addition, in this particular case, it allowed us to circumvent the use of heparin-affinity chromatography, thus allowing the design of an animal-component-free process.

  17. Role of histidine-related compounds to intracellular buffering in fish skeletal muscle.

    PubMed

    Abe, H; Dobson, G P; Hoeger, U; Parkhouse, W S

    1985-10-01

    Histidine-related compounds (HRC) were analyzed in fish skeletal muscle as a means of identifying their precise role in intracellular buffering. Fish muscle was used because it contains two functionally and spatially distinct fiber types, red and white. Two fish species, rainbow trout (Salmo gairdneri) and the Pacific blue marlin (Makaira nigricans), were studied because these species demonstrate widely different activity patterns. Marlin red and white muscle buffer capacity was two times higher than trout with white muscle, buffering being two times greater than red in both species. Buffer capacity was highest in the 6.5-7.5 pH range for all tissues, which corresponded to their high anserine levels. The titrated HRC buffering was greater than the observed HRC buffering, which suggested that not all HRC were available to absorb protons. The HRC contribution to total cellular buffering varied from a high of 62% for marlin white to a low of 7% for trout red. The other principal buffers were found to be phosphate and protein with taurine contributing within red muscle in the 7.0-8.0 pH range. HRC were found to be dominant in skeletal muscle buffering by principally accounting for the buffering capacity differences found between the species and fiber types.

  18. ATP-stabilized amorphous calcium carbonate nanospheres and their application in protein adsorption.

    PubMed

    Qi, Chao; Zhu, Ying-Jie; Lu, Bing-Qiang; Zhao, Xin-Yu; Zhao, Jing; Chen, Feng; Wu, Jin

    2014-05-28

    Calcium carbonate is a common substance found in rocks worldwide, and is the main biomineral formed in shells of marine organisms and snails, pearls and eggshells. Amorphous calcium carbonate (ACC) is the least stable polymorph of calcium carbonate, which is so unstable under normal conditions that it is difficult to be prepared in vitro because it rapidly crystallizes to form one of the more stable polymorphs in aqueous solution. Herein, we report the successful synthesis of highly stable ACC nanospheres in vitro using adenosine 5'-triphosphate disodium salt (ATP) as a stabilizer. The effect of ATP on the stability of ACC nanospheres is investigated. Our experiments show that ATP plays an unique role in the stabilization of ACC nanospheres in aqueous solution. Moreover, the as-prepared ACC nanospheres are highly stable in phosphate buffered saline for a relatively long period of time (12 days) even under relatively high concentrations of calcium and phosphate ions. The cytotoxicity tests show that the as-prepared highly stable ACC nanospheres have excellent biocompatibility. The highly stable ACC nanospheres have high protein adsorption capacity, implying that they are promising for applications in biomedical fields such as drug delivery and protein adsorption. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Mixed-bed ion exchange chromatography employing a salt-free pH gradient for improved sensitivity and compatibility in MudPIT.

    PubMed

    Mommen, Geert P M; Meiring, Hugo D; Heck, Albert J R; de Jong, Ad P J M

    2013-07-16

    In proteomics, comprehensive analysis of peptides mixtures necessitates multiple dimensions of separation prior to mass spectrometry analysis to reduce sample complexity and increase the dynamic range of analysis. The main goal of this work was to improve the performance of (online) multidimensional protein identification technology (MudPIT) in terms of sensitivity, compatibility and recovery. The method employs weak anion and strong cation mixed-bed ion exchange chromatography (ACE) in the first separation dimension and reversed phase chromatography (RP) in the second separation dimension (Motoyama et.al. Anal. Chem 2007, 79, 3623-34.). We demonstrated that the chromatographic behavior of peptides in ACE chromatography depends on both the WAX/SCX mixing ratio as the ionic strength of the mobile phase system. This property allowed us to replace the conventional salt gradient by a (discontinuous) salt-free, pH gradient. First dimensional separation of peptides was accomplished with mixtures of aqueous formic acid and dimethylsulfoxide with increasing concentrations. The overall performance of this mobile phase system was found comparable to ammonium acetate buffers in application to ACE chromatography, but clearly outperformed strong cation exchange for use in first dimensional peptide separation. The dramatically improved compatibility between (salt-free) ion exchange chromatography and reversed phase chromatography-mass spectrometry allowed us to downscale the dimensions of the RP analytical column down to 25 μm i.d. for an additional 2- to 3-fold improvement in performance compared to current technology. The achieved levels of sensitivity, orthogonality, and compatibility demonstrates the potential of salt-free ACE MudPIT for the ultrasensitive, multidimensional analysis of very modest amounts of sample material.

  20. Historical assessments and comparisons of benthic communities and physical habitat in two agricultural streams in California's San Joaquin watershed.

    PubMed

    Hall, Lenwood W; Killen, William D

    2006-01-01

    This study was designed to assess trends in physical habitat and benthic communities (macroinvertebrates) annually in two agricultural streams (Del Puerto Creek and Salt Slough) in California's San Joaquin Valley from 2001 to 2005, determine the relationship between benthic communities and both water quality and physical habitat from both streams over the 5-year period, and compare benthic communities and physical habitat in both streams from 2001 to 2005. Physical habitat, measured with 10 metrics and a total score, was reported to be fairly stable over 5 years in Del Puerto Creek but somewhat variable in Salt Slough. Benthic communities, measured with 18 metrics, were reported to be marginally variable over time in Del Puerto Creek but fairly stable in Salt Slough. Rank correlation analysis for both water bodies combined showed that channel alteration, embeddedness, riparian buffer, and velocity/depth/diversity were the most important physical habitat metrics influencing the various benthic metrics. Correlations of water quality parameters and benthic community metrics for both water bodies combined showed that turbidity, dissolved oxygen, and conductivity were the most important water quality parameters influencing the different benthic metrics. A comparison of physical habitat metrics (including total score) for both water bodies over the 5-year period showed that habitat metrics were more positive in Del Puerto Creek when compared to Salt Slough. A comparison of benthic metrics in both water bodies showed that approximately one-third of the metrics were significantly different between the two water bodies. Generally, the more positive benthic metric scores were reported in Del Puerto Creek, which suggests that the communities in this creek are more robust than Salt Slough.

  1. Salt-responsive gut commensal modulates TH17 axis and disease.

    PubMed

    Wilck, Nicola; Matus, Mariana G; Kearney, Sean M; Olesen, Scott W; Forslund, Kristoffer; Bartolomaeus, Hendrik; Haase, Stefanie; Mähler, Anja; Balogh, András; Markó, Lajos; Vvedenskaya, Olga; Kleiner, Friedrich H; Tsvetkov, Dmitry; Klug, Lars; Costea, Paul I; Sunagawa, Shinichi; Maier, Lisa; Rakova, Natalia; Schatz, Valentin; Neubert, Patrick; Frätzer, Christian; Krannich, Alexander; Gollasch, Maik; Grohme, Diana A; Côrte-Real, Beatriz F; Gerlach, Roman G; Basic, Marijana; Typas, Athanasios; Wu, Chuan; Titze, Jens M; Jantsch, Jonathan; Boschmann, Michael; Dechend, Ralf; Kleinewietfeld, Markus; Kempa, Stefan; Bork, Peer; Linker, Ralf A; Alm, Eric J; Müller, Dominik N

    2017-11-30

    A Western lifestyle with high salt consumption can lead to hypertension and cardiovascular disease. High salt may additionally drive autoimmunity by inducing T helper 17 (T H 17) cells, which can also contribute to hypertension. Induction of T H 17 cells depends on gut microbiota; however, the effect of salt on the gut microbiome is unknown. Here we show that high salt intake affects the gut microbiome in mice, particularly by depleting Lactobacillus murinus. Consequently, treatment of mice with L. murinus prevented salt-induced aggravation of actively induced experimental autoimmune encephalomyelitis and salt-sensitive hypertension by modulating T H 17 cells. In line with these findings, a moderate high-salt challenge in a pilot study in humans reduced intestinal survival of Lactobacillus spp., increased T H 17 cells and increased blood pressure. Our results connect high salt intake to the gut-immune axis and highlight the gut microbiome as a potential therapeutic target to counteract salt-sensitive conditions.

  2. SODR Memory Control Buffer Control ASIC

    NASA Technical Reports Server (NTRS)

    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.

  3. Cost and performance of thermal storage concepts in solar thermal systems, Phase 2-liquid metal receivers

    NASA Astrophysics Data System (ADS)

    McKenzie, A. W.

    Cost and performance of various thermal storage concepts in a liquid metal receiver solar thermal power system application have been evaluated. The objectives of this study are to provide consistently calculated cost and performance data for thermal storage concepts integrated into solar thermal systems. Five alternative storage concepts are evaluated for a 100-MW(e) liquid metal-cooled receiver solar thermal power system for 1, 6, and 15 hours of storage: sodium 2-tank (reference system), molten draw salt 2-tank, sand moving bed, air/rock, and latent heat (phase change) with tube-intensive heat exchange (HX). The results indicate that the all sodium 2-tank thermal storage concept is not cost-effective for storage in excess of 3 or 4 hours; the molten draw salt 2-tank storage concept provides significant cost savings over the reference sodium 2-tank concept; and the air/rock storage concept with pressurized sodium buffer tanks provides the lowest evaluated cost of all storage concepts considered above 6 hours of storage.

  4. Consequences of ions and pH on the supramolecular organization of sphingomyelin and sphingomyelin/cholesterol bilayers.

    PubMed

    Chemin, Caroline; Bourgaux, Claudie; Péan, Jean-Manuel; Pabst, Georg; Wüthrich, Patrick; Couvreur, Patrick; Ollivon, Michel

    2008-06-01

    For drug delivery purpose the anticancer drug S12363 was loaded into ESM/Chol-liposomes using either a pH or an ammonium gradient. Association between the drug and the liposome depends markedly on the liposome membrane structure. Thus, ESM and ESM/Chol bilayer organization had been characterized by coupled DSC and XRDT as a function of both cholesterol concentration and aqueous medium composition. ESM bilayers exhibited a ripple lamellar gel phase P(beta') below the melting temperature and adopted a L(beta)-like gel phase upon Chol insertion. Supramolecular organization of ESM and ESM/Chol bilayers was not modified by citrate buffer or ammonium sulfate solution whatever the pH (3< or = pH < or =7). Nevertheless, in ESM bilayer, ammonium sulfate salt induced a peculiar organization of head groups, leading to irregular d-spacing and weakly correlated bilayers. Moreover, in the presence of salts, a weakening of van der Waals attraction forces was seen and led to a swelling of the water layer.

  5. Ion-specific weak adsorption of salts and water/octanol transfer free energy of a model amphiphilic hexapeptide.

    PubMed

    Déjugnat, Christophe; Dufrêche, Jean-François; Zemb, Thomas

    2011-04-21

    An amphiphilic hexapeptide has been used as a model to quantify how specific ion effects induced by addition of four salts tune the hydrophilic/hydrophobic balance and induce temperature-dependant coacervate formation from aqueous solution. The hexapeptide chosen is present as a dimer with low transfer energy from water to octanol. Taking sodium chloride as the reference state in the Hofmeister scale, we identify water activity effects and therefore measure the free energy of transfer from water to octanol and separately the free energy associated to the adsorption of chaotropic ions or the desorption of kosmotropic ions for the same amphiphilic peptide. These effects have the same order of magnitude: therefore, both energies of solvation as well as transfer into octanol strongly depend on the nature of the electrolytes used to formulate any buffer. Model peptides could be used on separation processes based on criteria linked to "Hofmeister" but different from volume and valency.

  6. Modeling coliform-bacteria concentrations and pH in the salt-wedge reach of the Duwamish River Estuary, King County, Washington

    USGS Publications Warehouse

    Haushild, W.L.; Prych, Edmund A.

    1976-01-01

    Total- and fecal-coliform bacteria, plus pH, alkalinity, and dissolved inorganic carbon are water-quality parameters that have been added to an existing numerical model of water quality in the salt-wedge reach of the Duwamish River estuary in Washington. The coliform bacteria are modeled using a first-order decay (death) rate, which is a function of the local salinity, temperature, and daily solar radiation. The pH is computed by solving a set of chemical-equilibrium equations for carbonate-bicarbonate buffered aqueous solutions. Concentrations of total- and fecal-coliform bacteria computed by the model for the Duwamish River estuary during June-September 1971 generally agreed with observed concentrations within about 40 and 60 percent, respectively. The computed pH generally agreed with observed pH within about a 0.2 pH unit; however, for one 3-week period the computed pH was about a 0.4 unit lower than the observed pH. (Woodard-USGS)

  7. Alteration of the Helicobacter pylori membrane proteome in response to changes in environmental salt concentration

    PubMed Central

    Voss, Bradley J.; Loh, John T.; Hill, Salisha; Rose, Kristie L.; McDonald, W. Hayes; Cover, Timothy L.

    2015-01-01

    Purpose Helicobacter pylori infection and a high dietary salt intake are each risk factors for the development of gastric cancer. We hypothesize that changes in environmental salt concentrations lead to alterations in the H. pylori membrane proteome. Experimental Design Label-free and iTRAQ methods were used to identify H. pylori proteins that change in abundance in response to alterations in environmental salt concentrations. In addition, we biotinylated intact bacteria that were grown under high- or low-salt conditions, and thereby analyzed salt-induced changes in the abundance of surface-exposed proteins. Results Proteins with increased abundance in response to high salt conditions included CagA, the outer membrane protein HopQ, and fibronectin domain-containing protein HP0746. Proteins with increased abundance in response to low salt conditions included VacA, two VacA-like proteins (ImaA and FaaA), outer-membrane iron transporter FecA3, and several proteins involved in flagellar activity. Consistent with the proteomic data, bacteria grown in high salt conditions exhibited decreased motility compared to bacteria grown in lower salt conditions. Conclusions and clinical relevance Alterations in the H. pylori membrane proteome in response to high salt conditions may contribute to the increased risk of gastric cancer associated with a high salt diet. PMID:26109032

  8. An Efficacy and Pharmacokinetic Evaluation of a Dose of Diazepam That Will Reduce the Incidence of Convulsions in Indian Rhesus Monkeys Pretreated with Pyridostigmine Bromide, Challenged with Soman, and Treated with Atropine and Pralidoxime Chloride with the Diazepam

    DTIC Science & Technology

    1990-12-01

    benzophenone (Aldrich 23:985-2), tetrabutylammonium nitrate (Kodak 9664), sodium lauryl sulfate (dodecyl sulfide, sodium salt) (Aldrich 86-201-0), helium gas...phase buffer for the initial identity confirmation using a Supelco LC-I column by dissolving 6.0 g of sodium lauryl sulfate and 1.0 g of...water, glacial acetic acid (Baker Reagent’Grade), tetrabutylammonium chloride (Aldrich g8. percent), sodium lauryl sulfate (Aldrich 98 percent), sodium

  9. Laboratory and Field Evaluation of a Waterless Food Service Sanitation System Used by Military Mobile Kitchen Trailer Crews

    DTIC Science & Technology

    1993-12-01

    Stopper the flask and shake vigorously. The mixture was titrated with 0.003 N sodium lauryl sulfate drcpwise. The endpoint was the first definite...100 g sodium sulfate and 1000 mL distilled water, pill0) and three drops of 0.1% bracu 1henol blue indicator to 50 ML of sample in a 250 mL flask...Product QDS (Syncide Plus) (1) was determined by a bromcphenol blue nethod (7). Add 25 mL of chloroform, 25 mL salt buffer solution (7 g sodium carbonate

  10. Direct Reaction of Amides with Nitric Oxide To Form Diazeniumdiolates

    PubMed Central

    2015-01-01

    We report the apparently unprecedented direct reaction of nitric oxide (NO) with amides to generate ions of structure R(C=O)NH–N(O)=NO–, with examples including R = Me (1a) or 3-pyridyl (1b). The sodium salts of both released NO in pH 7.4 buffer, with 37 °C half-lives of 1–3 min. As NO-releasing drug candidates, diazeniumdiolated amides would have the advantage of generating only 1 equiv of base on hydrolyzing exhaustively to NO, in contrast to their amine counterparts, which generate 2 equiv of base. PMID:25210948

  11. Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis

    PubMed Central

    1978-01-01

    Complexes of plasma membrane segments with desmosomes and attached tonofilaments were separated from the stratum spinosum cells of calf muzzle by means of moderately alkaline buffers of low ionic strength and mechanical homogenization. These structures were further fractionated by the use of various treatments including sonication, sucrose gradient centrifugation, and extraction with buffers containing high concentrations of salt, urea, citric acid, or detergents. Subfractions enriched in desmosome-tonofilament-complexes and tonofilament fragments were studied in detail. The desmosome structures such as the midline, the trilaminar membrane profile, and the desmosomal plaque appeared well preserved and were notably resistant to the various treatments employed. Fractions containing desmosome- tonofilament complexes were invariably dominated by the nonmembranous proteins of the tonofilaments which appeared as five major polypeptide bands (apparent molecular weights: 48,000; 51,000; 58,000; 60,000; 68,000) present in molar ratios of approx. 2:1:1:2:2. Four of these polypeptide bands showed electrophoretic mobilities similar to those of prekeratin polypeptides from bovine hoof. However, the largest polypeptide (68,000 mol wt) migrated significantly less in polyacrylamide gels than the largest component of the hoof prekeratin (approximately 63,000 mol wt). In addition, a series of minor bands, including carbohydrate-containing proteins, were identified and concluded to represent constituents of the desmosomal membrane. The analysis of protein-bound carbohydrates (total 270 microgram/mg phospholipid in desmosome-enriched subfractions) showed the presence of relatively high amounts of glucosamine, mannose, galactose, and sialic acids. These data as well as the lipid composition (e.g., high ratio of cholesterol to phospholipids, relatively high contents of sphingomyelin and gangliosides, and fatty acid pattern) indicate that the desmosomal membrane is complex in protein and lipid composition and has a typical plasma membrane character. The similarity of the desmosome-associated tonofilaments to prekeratin filaments and other forms of intermediate- sized filaments is discussed. PMID:569157

  12. Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis.

    PubMed

    Drochmans, P; Freudenstein, C; Wanson, J C; Laurent, L; Keenan, T W; Stadler, J; Leloup, R; Franke, W W

    1978-11-01

    Complexes of plasma membrane segments with desmosomes and attached tonofilaments were separated from the stratum spinosum cells of calf muzzle by means of moderately alkaline buffers of low ionic strength and mechanical homogenization. These structures were further fractionated by the use of various treatments including sonication, sucrose gradient centrifugation, and extraction with buffers containing high concentrations of salt, urea, citric acid, or detergents. Subfractions enriched in desmosome-tonofilament-complexes and tonofilament fragments were studied in detail. The desmosome structures such as the midline, the trilaminar membrane profile, and the desmosomal plaque appeared well preserved and were notably resistant to the various treatments employed. Fractions containing desmosome-tonofilament complexes were invariably dominated by the nonmembranous proteins of the tonofilaments which appeared as five major polypeptide bands (apparent molecular weights: 48,000; 51,000; 58,000; 60,000; 68,000) present in molar ratios of approx. 2:1:1:2:2. Four of these polypeptide bands showed electrophoretic mobilities similar to those of prekeratin polypeptides from bovine hoof. However, the largest polypeptide (68,000 mol wt) migrated significantly less in polyacrylamide gels than the largest component of the hoof prekeratin (approximately 63,000 mol wt). In addition, a series of minor bands, including carbohydrate-containing proteins, were identified and concluded to represent constituents of the desmosomal membrane. The analysis of protein-bound carbohydrates (total 270 microgram/mg phospholipid in desmosome-enriched subfractions) showed the presence of relatively high amounts of glucosamine, mannose, galactose, and sialic acids. These data as well as the lipid composition (e.g., high ratio of cholesterol to phospholipids, relatively high contents of sphingomyelin and gangliosides, and fatty acid pattern) indicate that the desmosomal membrane is complex in protein and lipid composition and has a typical plasma membrane character. The similarity of the desmosome-associated tonofilaments to prekeratin filaments and other forms of intermediate-sized filaments is discussed.

  13. Current isolating epitaxial buffer layers for high voltage photodiode array

    DOEpatents

    Morse, Jeffrey D.; Cooper, Gregory A.

    2002-01-01

    An array of photodiodes in series on a common semi-insulating substrate has a non-conductive buffer layer between the photodiodes and the semi-insulating substrate. The buffer layer reduces current injection leakage between the photodiodes of the array and allows optical energy to be converted to high voltage electrical energy.

  14. Development of a multi-matrix LC-MS/MS method for urea quantitation and its application in human respiratory disease studies.

    PubMed

    Wang, Jianshuang; Gao, Yang; Dorshorst, Drew W; Cai, Fang; Bremer, Meire; Milanowski, Dennis; Staton, Tracy L; Cape, Stephanie S; Dean, Brian; Ding, Xiao

    2017-01-30

    In human respiratory disease studies, liquid samples such as nasal secretion (NS), lung epithelial lining fluid (ELF), or upper airway mucosal lining fluid (MLF) are frequently collected, but their volumes often remain unknown. The lack of volume information makes it hard to estimate the actual concentration of recovered active pharmaceutical ingredient or biomarkers. Urea has been proposed to serve as a sample volume marker because it can freely diffuse through most body compartments and is less affected by disease states. Here, we report an easy and reliable LC-MS/MS method for cross-matrix measurement of urea in serum, plasma, universal transfer medium (UTM), synthetic absorptive matrix elution buffer 1 (SAMe1) and synthetic absorptive matrix elution buffer 2 (SAMe2) which are commonly sampled in human respiratory disease studies. The method uses two stable-isotope-labeled urea isotopologues, [ 15 N 2 ]-urea and [ 13 C, 15 N 2 ]-urea, as the surrogate analyte and the internal standard, respectively. This approach provides the best measurement consistency across different matrices. The analyte extraction was individually optimized in each matrix. Specifically in UTM, SAMe1 and SAMe2, the unique salting-out assisted liquid-liquid extraction (SALLE) not only dramatically reduces the matrix interferences but also improves the assay recovery. The use of an HILIC column largely increases the analyte retention. The typical run time is 3.6min which allows for high throughput analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Efficient osmolyte-based procedure to increase expression level and solubility of infectious hematopoietic necrosis virus (IHNV) nucleoprotein in E. coli.

    PubMed

    Mohammadinezhad, Rezvan; Farahmand, Hamid; Jalali, Seyed Amir Hossein; Mirvaghefi, Alireza

    2018-05-01

    The nucleoprotein of infectious hematopoietic necrosis virus (IHNV) is considered as the main target antigen for detection of IHNV infection in salmonid fish. This study aimed at improving the expression and solubility of IHNV nucleoprotein (IHNV-NP) in E. coli expression system. The effects of several expression strategies including host strain type, protein expression temperature, heat-shock treatment prior to protein induction, and additives in the growth medium and in the cell lysis buffer were examined. Results showed that bacterial strain type had a great impact on protein expression level, whereas it was not effective in preventing protein aggregation. Production of soluble IHNV-NP was proportionally increased with decreased incubation temperature. Heat-shock treatment prior to protein induction did not change the percent of solubility. For cells grown at low temperature, the presence of additives in the lysis buffer enhanced the solubility of IHNV-NP up to 24%. The highest yield of soluble protein was obtained via incorporation of osmolytes in the growth medium of cells exposed to a mild salt stress, in the following order: sucrose > sorbitol > glycerol > glycine. Soluble protein obtained by the optimized condition was efficiently purified in high yield and successfully detected by two monoclonal antibodies in a sandwich ELISA. Taken together, a combination of proper host strain, low-temperature expression, and timely application of osmolytes in the growth medium provided sufficient quantities of soluble recombinant IHNV-NP that has the potential to be used for diagnostic purposes.

  16. Enjoyment of Spicy Flavor Enhances Central Salty-Taste Perception and Reduces Salt Intake and Blood Pressure.

    PubMed

    Li, Qiang; Cui, Yuanting; Jin, Rongbing; Lang, Hongmei; Yu, Hao; Sun, Fang; He, Chengkang; Ma, Tianyi; Li, Yingsha; Zhou, Xunmei; Liu, Daoyan; Jia, Hongbo; Chen, Xiaowei; Zhu, Zhiming

    2017-12-01

    High salt intake is a major risk factor for hypertension and is associated with cardiovascular events. Most countries exhibit a traditionally high salt intake; thus, identification of an optimal strategy for salt reduction at the population level may have a major impact on public health. In this multicenter, random-order, double-blind observational and interventional study, subjects with a high spice preference had a lower salt intake and blood pressure than subjects who disliked spicy food. The enjoyment of spicy flavor enhanced salt sensitivity and reduced salt preference. Salt intake and salt preference were related to the regional metabolic activity in the insula and orbitofrontal cortex (OFC) of participants. Administration of capsaicin-the major spicy component of chili pepper-enhanced the insula and OFC metabolic activity in response to high-salt stimuli, which reversed the salt intensity-dependent differences in the metabolism of the insula and OFC. In animal study, OFC activity was closely associated with salt preference, and salty-taste information processed in the OFC was affected in the presence of capsaicin. Thus, interventions related to this region may alter the salt preference in mice through fiber fluorometry and optogenetic techniques. In conclusion, enjoyment of spicy foods may significantly reduce individual salt preference, daily salt intake, and blood pressure by modifying the neural processing of salty taste in the brain. Application of spicy flavor may be a promising behavioral intervention for reducing high salt intake and blood pressure. © 2017 American Heart Association, Inc.

  17. Phosphorus sorption and buffering mechanisms in suspended sediments from the Yangtze Estuary and Hangzhou Bay, China

    NASA Astrophysics Data System (ADS)

    Li, M.; Whelan, M. J.; Wang, G.; White, S. M.

    2012-12-01

    The adsorption isotherm and the mechanism of the buffering effect are important controls on phosphorus behaviors in estuaries and are important for estimating phosphate concentrations in aquatic environments. In this paper, we derive phosphate adsorption isotherms in order to investigate sediment adsorption and buffering capacity for phosphorus discharged from sewage outfalls in the Yangtze Estuary and Hangzhou Bay near Shanghai, China. Experiments were also carried out at different temperatures in order to explore the buffering effects for phosphate. The results show that P sorption in sediments with low fine particle fractions was best described using exponential equations. Some P interactions between water and sediment may be caused by the precipitation of CaHPO4 from Ca2+ and HPO42- when the phosphate concentration in the liquid phase is high. Results from the buffering experiments suggest that the Zero Equilibrium Phosphate Concentrations (EPC0) vary from 0.014 mg l-1 to 0.061 mg l-1, which are consistent with measured phosphate concentrations in water samples collected at the same time as sediment sampling. Values of EPC0 and linear sorption coefficients (K) in sediments with high fine particle and organic matter contents are relatively high, which implies that they have high buffering capacity. Both EPC0 and K increase with increasing temperature, indicating a higher P buffering capacity at high temperatures.

  18. Phosphorus sorption and buffering mechanisms in suspended sediments from the Yangtze Estuary and Hangzhou Bay, China

    NASA Astrophysics Data System (ADS)

    Li, M.; Whelan, M. J.; Wang, G. Q.; White, S. M.

    2013-05-01

    The adsorption isotherm and the mechanism of the buffering effect are important controls on phosphorus (P) behaviors in estuaries and are important for estimating phosphate concentrations in aquatic environments. In this paper, we derive phosphate adsorption isotherms in order to investigate sediment adsorption and buffering capacity for phosphorus discharged from sewage outfalls in the Yangtze Estuary and Hangzhou Bay near Shanghai, China. Experiments were also carried out at different temperatures in order to explore the buffering effects for phosphate. The results show that P sorption in sediments with low fine particle fractions was best described using exponential equations. Some P interactions between water and sediment may be caused by the precipitation of CaHPO4 from Ca2+ and HPO42- when the phosphate concentration in the liquid phase is high. Results from the buffering experiments suggest that the Zero Equilibrium Phosphate Concentrations (EPC0) vary from 0.014 mg L-1 to 0.061 mg L-1, which are consistent with measured phosphate concentrations in water samples collected at the same time as sediment sampling. Values of EPC0 and linear sorption coefficients (K) in sediments with high fine particle and organic matter contents are relatively high, which implies that they have high buffering capacity. Both EPC0 and K increase with increasing temperature, indicating a higher P buffering capacity at high temperatures.

  19. Feasibility and application of an HPLC/UVD to determine dinotefuran and its shorter wavelength metabolites residues in melon with tandem mass confirmation.

    PubMed

    Rahman, Md Musfiqur; Park, Jong-Hyouk; Abd El-Aty, A M; Choi, Jeong-Heui; Yang, Angel; Park, Ki Hun; Nashir Uddin Al Mahmud, Md; Im, Geon-Jae; Shim, Jae-Han

    2013-01-15

    A new analytical method was developed for dinotefuran and its metabolites, MNG, UF, and DN, in melon using high-performance liquid chromatography (HPLC) coupled with an ultraviolet detector (UVD). Due to shorter wavelength, lower sensitivity to UV detection, and high water miscibility of some metabolites, QuEChERs acetate-buffered version was modified for extraction and purification. Mobile phases with different ion pairing or ionisation agents were tested in different reverse phase columns, and ammonium bicarbonate buffer was found as the best choice to increase the sensitivity of target analytes to the UV detector. After failure of dispersive SPE clean-up with primary secondary amine, different solid phase extraction cartridges (SPE) were used to check the protecting capability of analytes against matrix interference. Finally, samples were extracted with a simple and rapid method using acetonitrile and salts, and purified through C(18)SPE. The method was validated at two spiking levels (three replicates for each) in the matrix. Good recoveries were observed for all of the analytes and ranged between 70.6% and 93.5%, with relative standard deviations of less than 10%. Calibration curves were linear over the calibration ranges for all the analytes with r(2)≥ 0.998. Limits of detection ranged from 0.02 to 0.05 mg kg(-1), whereas limits of quantitation ranged from 0.06 to 0.16 mg kg(-1) for dinotefuran and its metabolites. The method was successfully applied to real samples, where dinotefuran and UF residues were found in the field-incurred melon samples. Residues were confirmed via LC-tandem mass spectrometry (LC-MS/MS) in positive-ion electrospray ionisation (ESI(+)) mode. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Efficient cell-free production of olfactory receptors: detergent optimization, structure, and ligand binding analyses.

    PubMed

    Kaiser, Liselotte; Graveland-Bikker, Johanna; Steuerwald, Dirk; Vanberghem, Mélanie; Herlihy, Kara; Zhang, Shuguang

    2008-10-14

    High-level production of membrane proteins, particularly of G protein-coupled receptors (GPCRs) in heterologous cell systems encounters a number of difficulties from their inherent hydrophobicity in their transmembrane domains, which frequently cause protein aggregation and cytotoxicity and thus reduce the protein yield. Recent advances in cell-free protein synthesis circumvent those problems to produce membrane proteins with a yield sometimes exceeding the cell-based approach. Here, we report cell-free production of a human olfactory receptor 17-4 (hOR17-4) using the wheat germ extract. Using the simple method, we also successful produced two additional olfactory receptors. To obtain soluble olfactory receptors and to increase yield, we directly added different detergents in varying concentrations to the cell-free reaction. To identify a purification buffer system that maintained the receptor in a nonaggregated form, we developed a method that uses small-volume size-exclusion column chromatography combined with rapid and sensitive dot-blot detection. Different buffer components including salt concentration, various detergents and detergent concentration, and reducing agent and its concentrations were evaluated for their ability to maintain the cell-free produced protein stable and nonaggregated. The purified olfactory receptor displays a typical a alpha-helical CD spectrum. Surface plasmon resonance measurements were used to show binding of a known ligand undecanal to hOR17-4. Our approach to produce a high yield of purified olfactory receptor is a milestone toward obtaining a large quantity of olfactory receptors for designing bionic sensors. Furthermore, this simple approach may be broadly useful not only for other classes of GPCRs but also for other membrane proteins.

  1. Efficient cell-free production of olfactory receptors: Detergent optimization, structure, and ligand binding analyses

    PubMed Central

    Kaiser, Liselotte; Graveland-Bikker, Johanna; Steuerwald, Dirk; Vanberghem, Mélanie; Herlihy, Kara; Zhang, Shuguang

    2008-01-01

    High-level production of membrane proteins, particularly of G protein-coupled receptors (GPCRs) in heterologous cell systems encounters a number of difficulties from their inherent hydrophobicity in their transmembrane domains, which frequently cause protein aggregation and cytotoxicity and thus reduce the protein yield. Recent advances in cell-free protein synthesis circumvent those problems to produce membrane proteins with a yield sometimes exceeding the cell-based approach. Here, we report cell-free production of a human olfactory receptor 17-4 (hOR17-4) using the wheat germ extract. Using the simple method, we also successful produced two additional olfactory receptors. To obtain soluble olfactory receptors and to increase yield, we directly added different detergents in varying concentrations to the cell-free reaction. To identify a purification buffer system that maintained the receptor in a nonaggregated form, we developed a method that uses small-volume size-exclusion column chromatography combined with rapid and sensitive dot-blot detection. Different buffer components including salt concentration, various detergents and detergent concentration, and reducing agent and its concentrations were evaluated for their ability to maintain the cell-free produced protein stable and nonaggregated. The purified olfactory receptor displays a typical a α-helical CD spectrum. Surface plasmon resonance measurements were used to show binding of a known ligand undecanal to hOR17-4. Our approach to produce a high yield of purified olfactory receptor is a milestone toward obtaining a large quantity of olfactory receptors for designing bionic sensors. Furthermore, this simple approach may be broadly useful not only for other classes of GPCRs but also for other membrane proteins. PMID:18840687

  2. Preparation and Evaluation of Biodegradable Scleral Plug Containing Curcumin in Rabbit Eye.

    PubMed

    Zhang, Jun; Sun, Haiyan; Zhou, Nalei; Zhang, Bin; Ma, Jingxue

    2017-12-01

    To test whether biodegradable curcumin-loaded scleral plug is a promising choice for treating posterior ocular diseases, the study investigated the in vitro release profile of the scleral plug and its safety in vivo. Scleral plugs containing 0.5 mg, 1.0 mg and 1.5 mg curcumin were synthesized by a compression-sintering method. These scleral plugs were placed in tubes containing balanced salt solution (BSS) buffer, which was replaced by fresh buffer daily. The curcumin concentration in the removed aliquot was tested daily for 14 days using high-performance liquid chromatography (HPLC). In the study, 44 rabbits were randomly divided into four groups: control, 0.5 mg, 1.0 mg and 1.5 mg curcumin groups. The scleral plug was trans-scleral fixed in the right eye of the rabbits in the three curcumin-treated groups. The control rabbits only received sclerotomy. The treated rabbit eyes were examined by a slit-lamp biomicroscope, an indirect ophthalmoscope and electroretinogram (ERG), and subjected to histological analysis. The concentration of the 1.5 mg curcumin-loaded scleral plug was higher than 15 μg/ml for consecutive 14 days in vitro. The in vivo experiments revealed intraocular pressure, a-wave and b-wave amplitudes of ERG, and conjunctival reaction degree were not significantly different between the four groups. Retinal structure was normal in the curcumin-treated groups. The sclerotomy wound healed after the plug was completely degraded. Anterior chamber reaction or complications were not observed. The study suggests that curcumin-loaded scleral plug could sustain high concentration of curcumin in vitro and is safe in vivo. It might be a promising alternative choice for the treatment of posterior ocular diseases.

  3. Inhibition of Clostridium perfringens spore germination and outgrowth by lemon juice and vinegar product in reduced NaCl roast beef.

    PubMed

    Li, Lin; Valenzuela-Martinez, Carol; Redondo, Mauricio; Juneja, Vijay K; Burson, Dennis E; Thippareddi, Harshavardhan

    2012-11-01

    Inhibition of Clostridium perfringens spore germination and outgrowth in reduced sodium roast beef by a blend of buffered lemon juice concentrate and vinegar (MoStatin LV1) during abusive exponential cooling was evaluated. Roast beef containing salt (NaCl; 1%, 1.5%, or 2%, w/w), blend of sodium pyro- and poly-phosphates (0.3%), and MoStatin LV1 (0%, 2%, or 2.5%) was inoculated with a 3-strain C. perfringens spore cocktail to achieve final spore population of 2.5 to 3.0 log CFU/g. The inoculated products were heat treated and cooled exponentially from 54.4 to 4.4 °C within 6.5, 9, 12, 15, 18, or 21 h. Cooling of roast beef (2.0% NaCl) within 6.5 and 9 h resulted in <1.0 log CFU/g increase in C. perfringens spore germination and outgrowth, whereas reducing the salt concentration to 1.5% and 1.0% resulted in >1.0 log CFU/g increase for cooling times longer than 9 h (1.1 and 2.2 log CFU/g, respectively). Incorporation of MoStatin LV1 into the roast beef formulation minimized the C. perfringens spore germination and outgrowth to <1.0 log CFU/g, regardless of the salt concentration and the cooling time. Cooked, ready-to-eat meat products should be cooled rapidly to reduce the risk of Clostridium perfringens spore germination and outgrowth. Meat processors are reducing the sodium chloride content of the processed meats as a consequence of the dietary recommendations. Sodium chloride reduces the risk of C. perfringens spore germination and outgrowth in meat products. Antimicrobials that contribute minimally to the sodium content of the product should be incorporated into processed meats to assure food safety. Buffered lemon juice and vinegar can be incorporated into meat product formulations to reduce the risk of C. perfringens spore germination and outgrowth during abusive cooling. Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.

  4. The Temperature of Intracellular Ice Formation in Mouse Oocytes vs. the Unfrozen Fraction at that Temperature ⋆

    PubMed Central

    Mazur, Peter; Pinn, Irina L.; Kleinhans, F.W.

    2009-01-01

    We have previously reported [11] that intracellular ice formation (IIF) in mouse oocytes suspended in various concentrations of glycerol and ethylene glycol (EG) occurs at temperatures where the percentage of unfrozen water is about 6% and 12% respectively even though the IIF temperatures varied from −14° to −41°C. However, because of the way the solutions were prepared, the concentrations of salt and glycerol or EG in that unfrozen fraction at IIF were also rather tightly grouped. The experiments reported in the present paper were designed to separate the effects of the unfrozen fraction at IIF from that of the solute concentration in the unfrozen fraction. This separation makes use of two facts. One is that the concentration of solutes in the residual liquid at a given subzero temperature is fixed regardless of their concentration in the initial unfrozen solution. However, second, the fraction unfrozen at a given temperature is dependent on the initial solute concentration. Experimentally, oocytes were suspended in solutions of glycerol/buffered saline and EG/buffered saline of varying total solute concentration with the restriction that the mass ratio of glycerol and EG to salts are held constant. The oocytes were then cooled rapidly enough (20°C/min) to avoid significant osmotic shrinkage, and the temperature at which IIF occurred as noted. When this is done, we find, as previously that the fraction of water remaining unfrozen at the temperature of IIF remains nearly constant at 5 to 8% for both glycerol and EG even though the IIF temperatures vary from −14°C to −50°C. But unlike the previous results, the salt and CPA concentrations in the unfrozen fraction vary by a factor of three. The present procedure for preparing the solutions produces a potentially complicating factor; namely, the cell volumes vary substantially prior to freezing: Substantially greater than isotonic in some solution; substantially smaller in others. However, the data in toto demonstrate that cell volume is not a determining factor in the IIF temperature. PMID:17379206

  5. The temperature of intracellular ice formation in mouse oocytes vs. the unfrozen fraction at that temperature.

    PubMed

    Mazur, Peter; Pinn, Irina L; Kleinhans, F W

    2007-04-01

    We have previously reported [Cryobiology 51 (2005) 29-53] that intracellular ice formation (IIF) in mouse oocytes suspended in various concentrations of glycerol and ethylene glycol (EG) occurs at temperatures where the percentage of unfrozen water is about 6% and 12%, respectively, even though the IIF temperatures varied from -14 to -41 degrees C. However, because of the way the solutions were prepared, the concentrations of salt and glycerol or EG in that unfrozen fraction at IIF were also rather tightly grouped. The experiments reported in the present paper were designed to separate the effects of the unfrozen fraction at IIF from that of the solute concentration in the unfrozen fraction. This separation makes use of two facts. One is that the concentration of solutes in the residual liquid at a given subzero temperature is fixed regardless of their concentration in the initial unfrozen solution. However, second, the fraction unfrozen at a given temperature is dependent on the initial solute concentration. Experimentally, oocytes were suspended in solutions of glycerol/buffered saline and EG/buffered saline of varying total solute concentration with the restriction that the mass ratios of glycerol and EG to salts are held constant. The oocytes were then cooled rapidly enough (20 degrees C/min) to avoid significant osmotic shrinkage, and the temperature at which IIF occurred was noted. When this is done, we find, as previously that the fraction of water remaining unfrozen at the temperature of IIF remains nearly constant at 5-8% for both glycerol and EG even though the IIF temperatures vary from -14 to -50 degrees C. But unlike the previous results, the salt and CPA concentrations in the unfrozen fraction vary by a factor of three. The present procedure for preparing the solutions produces a potentially complicating factor; namely, the cell volumes vary substantially prior to freezing: substantially greater than isotonic in some solutions; substantially smaller in others. However, the data in toto demonstrate that cell volume is not a determining factor in the IIF temperature.

  6. Geographic scale matters in detecting the relationship between neighbourhood food environments and obesity risk: an analysis of driver license records in Salt Lake County, Utah.

    PubMed

    Fan, Jessie X; Hanson, Heidi A; Zick, Cathleen D; Brown, Barbara B; Kowaleski-Jones, Lori; Smith, Ken R

    2014-08-19

    Empirical studies of the association between neighbourhood food environments and individual obesity risk have found mixed results. One possible cause of these mixed findings is the variation in neighbourhood geographic scale used. The purpose of this paper was to examine how various neighbourhood geographic scales affected the estimated relationship between food environments and obesity risk. Cross-sectional secondary data analysis. Salt Lake County, Utah, USA. 403,305 Salt Lake County adults 25-64 in the Utah driver license database between 1995 and 2008. Utah driver license data were geo-linked to 2000 US Census data and Dun & Bradstreet business data. Food outlets were classified into the categories of large grocery stores, convenience stores, limited-service restaurants and full-service restaurants, and measured at four neighbourhood geographic scales: Census block group, Census tract, ZIP code and a 1 km buffer around the resident's house. These measures were regressed on individual obesity status using multilevel random intercept regressions. Obesity. Food environment was important for obesity but the scale of the relevant neighbourhood differs for different type of outlets: large grocery stores were not significant at all four geographic scales, limited-service restaurants at the medium-to-large scale (Census tract or larger) and convenience stores and full-service restaurants at the smallest scale (Census tract or smaller). The choice of neighbourhood geographic scale can affect the estimated significance of the association between neighbourhood food environments and individual obesity risk. However, variations in geographic scale alone do not explain the mixed findings in the literature. If researchers are constrained to use one geographic scale with multiple categories of food outlets, using Census tract or 1 km buffer as the neighbourhood geographic unit is likely to allow researchers to detect most significant relationships. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  7. Domain structure of human complement C4b extends with increasing NaCl concentration: implications for its regulatory mechanism.

    PubMed

    Fung, Ka Wai; Wright, David W; Gor, Jayesh; Swann, Marcus J; Perkins, Stephen J

    2016-12-01

    During the activation of complement C4 to C4b, the exposure of its thioester domain (TED) is crucial for the attachment of C4b to activator surfaces. In the C4b crystal structure, TED forms an Arg 104 -Glu 1032 salt bridge to tether its neighbouring macroglobulin (MG1) domain. Here, we examined the C4b domain structure to test whether this salt bridge affects its conformation. Dual polarisation interferometry of C4b immobilised at a sensor surface showed that the maximum thickness of C4b increased by 0.46 nm with an increase in NaCl concentration from 50 to 175 mM NaCl. Analytical ultracentrifugation showed that the sedimentation coefficient s 20,w of monomeric C4b of 8.41 S in 50 mM NaCl buffer decreased to 7.98 S in 137 mM NaCl buffer, indicating that C4b became more extended. Small angle X-ray scattering reported similar R G values of 4.89-4.90 nm for C4b in 137-250 mM NaCl. Atomistic scattering modelling of the C4b conformation showed that TED and the MG1 domain were separated by 4.7 nm in 137-250 mM NaCl and this is greater than that of 4.0 nm in the C4b crystal structure. Our data reveal that in low NaCl concentrations, both at surfaces and in solution, C4b forms compact TED-MG1 structures. In solution, physiologically relevant NaCl concentrations lead to the separation of the TED and MG1 domain, making C4b less capable of binding to its complement regulators. These conformational changes are similar to those seen previously for complement C3b, confirming the importance of this salt bridge for regulating both C4b and C3b. © 2016 The Author(s).

  8. Daily salt intake estimated by overnight urine collections indicates a high cardiovascular disease risk in Thailand.

    PubMed

    Yokokawa, Hirohide; Yuasa, Motoyuki; Nedsuwan, Supalert; Moolphate, Saiyud; Fukuda, Hiroshi; Kitajima, Tsutomu; Minematsu, Kazuo; Tanimura, Susumu; Marui, Eiji

    2016-01-01

    This cross-sectional study (February 2012 to March 2013) was conducted to estimate daily salt intake and basic characteristics among 793 community-dwelling participants at high risk of cardiovascular disease (Framingham risk score >15%), who had visited diabetes or hypertension clinics at health centres in the Muang district, Chiang Rai, Thailand. We performed descriptive analysis of baseline data and used an automated analyser to estimate the average of 24-hour salt intake estimated from 3 days overnight urine collection. Participants were divided into two groups based on median estimated daily salt intake. Mean age and proportion of males were 65.2 years and 37.6% in the higher salt intake group (>=10.0 g/day, n=362), and 67.5 years and 42.7% in the lower salt intake group (<10.0 g/day, n=431), respectively (p=0.01, p<0.01). The higher salt intake group comprised more patients with a family history of hypertension, antihypertensive drug use, less ideal body mass index (18.5-24.9), higher exercise frequency (>=2 times weekly) and lower awareness of high salt intake. Among higher salt intake participants, those with lower awareness of high salt intake were younger and more often had a family history of hypertension, relative to those with more awareness. Our data indicated that families often share lifestyles involving high salt intake, and discrepancies between actual salt intake and awareness of high salt intake may represent a need for salt reduction intervention aiming at family level. Awareness of actual salt intake should be improved for each family.

  9. Hydrology and Salt Balance in a Large, Hypersaline Coastal Lagoon: Lagoa de Araruama, Brazil

    NASA Astrophysics Data System (ADS)

    Kjerfve, Björn; Schettini, C. A. F.; Knoppers, Bastiaan; Lessa, Guilherme; Ferreira, H. O.

    1996-06-01

    Lagoa de Araruama in the state of Rio de Janeiro, Brazil, is a hypersaline coastal lagoon as a result of semi-arid climate conditions, a small drainage basin and a choked entrance channel. The lagoon has been continuously hypersaline for at least 4·5 centuries, but the mean salinity has varied substantially. It has recently decreased from 57 to 52 as indicated by density (salinity) measurements between 1965 and 1990. Analysis of more than 20 years of salinity time series data, in addition to monthly lagoon cruises to measure the spatial salinity distribution, indicate that the lagoon salinity largely fluctuates in response to the difference between evaporation and precipitation. The major factor explaining the long-term trend of decreasing salinity in the lagoon is the constant pumping of 1 m 3s -1of freshwater to the communities surrounding the lagoon from an adjacent watershed, and subsequent discharge of this water into Lagoa de Araruama. The net salt budget is primarily a balance between the advective import of salt from the coastal ocean and eddy diffusive export of salt to the ocean, although the extensive mining of salt from the lagoon during past decades is also a small but significant contribution to the salt budget. The flushing half-life is proposed as a useful time scale of water exchange, is calculated based on a combination of hydrological and tidal processes, and is excellent for comparison of lagoons and assessing water quality changes. The flushing half-life measures 83·5 days for Lagoa de Araruama, considerably longer than for most other coastal lagoons. The proposed dredging of a second ocean channel to Lagoa de Araruama is probably not a good idea. It is likely to accelerate the decrease of lagoon salinity and somewhat improve the lagoon water exchange. At the same time, this will eliminate the apparent buffering capacity provided by the hypersaline environment, and thus may potentially cause water quality problems.

  10. Influence of mixed electrolytes and pH on adsorption of bovine serum albumin in hydrophobic interaction chromatography.

    PubMed

    Hackemann, Eva; Hasse, Hans

    2017-10-27

    Using salt mixtures instead of single salts can be beneficial for hydrophobic interaction chromatography (HIC). The effect of electrolytes on the adsorption of proteins, however, depends on the pH. Little is known on that dependence for mixed electrolytes. Therefore, the effect of the pH on protein adsorption from aqueous solutions containing mixed salts is systematically studied in the present work for a model system: the adsorption of bovine serum albumin (BSA) on the mildly hydrophobic resin Toyopearl PPG-600M. The pH is adjusted to 4.0, 4.7 or 7.0 using 25mM sodium phosphate or sodium citrate buffer. Binary and ternary salt mixtures of sodium chloride, ammonium chloride, sodium sulfate and ammonium sulfate as well as the pure salts are used at overall ionic strengths between 1500 and 4200mM. The temperature is always 25°C. The influence of the mixed electrolytes on the adsorption behavior of BSA changes completely with varying pH. Positive as well as negative cooperative effects of the mixed electrolytes are observed. The results are analyzed using a mathematical model which was recently introduced by our group. In that model the influence of the electrolytes is described by a Taylor series expansion in the individual ion molarities. After suitable parametrization using a subset of the data determined in the present work, the model successfully predicts the influence of mixed electrolytes on the protein adsorption. Furthermore, results for BSA from the present study are compared to literature data for lysozyme, which are available for the same adsorbent, temperature and salts. By calculating the ratio of the loading of the adsorbent for both proteins particularly favorable separation conditions can be selected. Hence, a model-based optimization of solvents for protein separation is possible. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Enrichment and characterization of anaerobic TNT-degrading bacteria

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

    Roberts, D.J.; Pendharkar, S.

    1995-12-31

    Three media constitutions were used to enrich for mixed cultures capable of degrading 2,4,6-trinitrotoluene (TNT) under strictly anaerobic conditions. The media were derived from a mineral salts solution buffered to pH 7 with CO{sub 2}/bicarbonate and all contained TNT. The cultures were enriched in the TNT mineral salts medium or the TNT mineral salts medium supplemented with glucose, yeast extract, or ammonia in various combinations. Inocula were obtained from a treated soil, previously contaminated with dinoseb and then treated using anaerobic procedures, or from a bench-top aqueous culture, maintained with an extract from a munitions-contaminated soil for more than 4more » years. Several cultures reduced TNT, producing 4-amino-2,6-dinitrotoluene and 2,4-diamino-6-nitrotoluene as the major products. The cultures were unable to effectively remove TNT when cross-transferred to the media they were not enriched on, suggesting that different media had enriched different subcultures form the original inoculum. The treated soil provided the most successful inoculum. Two media were chosen for further studies. Medium 1 contained TNT and glucose and produced a culture that might have used TNT as a nitrogen source. Medium 2, containing TNT and yeast extract, enriched cultures that degraded TNT, accumulating small amounts of p-cresol during the degradation.« less

  12. Remote loading of doxorubicin into liposomes driven by a transmembrane phosphate gradient.

    PubMed

    Fritze, Andreas; Hens, Felicitas; Kimpfler, Andrea; Schubert, Rolf; Peschka-Süss, Regine

    2006-10-01

    This study examines a new method for the remote loading of doxorubicin into liposomes. It was shown that doxorubicin can be loaded to a level of up to 98% into large unilamellar vesicles composed of egg phosphatidylcholine/cholesterol (7/3 mol/mol) with a transmembrane phosphate gradient. The different encapsulation efficiencies which were achieved with ammonium salts (citrate 100%, phosphate 98%, sulfate 95%, acetate 77%) were significantly higher as compared to the loading via sodium salts (citrate 54%, phosphate 52%, sulfate 44%, acetate 16%). Various factors, including pH-value, buffer capacity, solubility of doxorubicin in different salt solutions and base counter-flow, which likely has an influence on drug accumulation in the intraliposomal interior are taken into account. In contrast to other methods, the newly developed remote loading method exhibits a pH-dependent drug release property which may be effective in tumor tissues. At physiological pH-value doxorubicin is retained in the liposomes, whereas drug release is achieved by lowering the pH to 5.5 (approximately 25% release at 25 degrees C or 30% at 37 degrees C within two h). The DXR release of liposomes which were loaded via a sulfate gradient showed a maximum of 3% at pH 5.5.

  13. High-speed atomic force microscopy reveals structural dynamics of amyloid β1–42 aggregates

    PubMed Central

    Watanabe-Nakayama, Takahiro; Ono, Kenjiro; Itami, Masahiro; Takahashi, Ryoichi; Teplow, David B.; Yamada, Masahito

    2016-01-01

    Aggregation of amyloidogenic proteins into insoluble amyloid fibrils is implicated in various neurodegenerative diseases. This process involves protein assembly into oligomeric intermediates and fibrils with highly polymorphic molecular structures. These structural differences may be responsible for different disease presentations. For this reason, elucidation of the structural features and assembly kinetics of amyloidogenic proteins has been an area of intense study. We report here the results of high-speed atomic force microscopy (HS-AFM) studies of fibril formation and elongation by the 42-residue form of the amyloid β-protein (Aβ1–42), a key pathogenetic agent of Alzheimer's disease. Our data demonstrate two different growth modes of Aβ1–42, one producing straight fibrils and the other producing spiral fibrils. Each mode depends on initial fibril nucleus structure, but switching from one growth mode to another was occasionally observed, suggesting that fibril end structure fluctuated between the two growth modes. This switching phenomenon was affected by buffer salt composition. Our findings indicate that polymorphism in fibril structure can occur after fibril nucleation and is affected by relatively modest changes in environmental conditions. PMID:27162352

  14. High Salt Intake Attenuates Breast Cancer Metastasis to Lung.

    PubMed

    Xu, Yijuan; Wang, Wenzhe; Wang, Minmin; Liu, Xuejiao; Lee, Mee-Hyun; Wang, Mingfu; Zhang, Hao; Li, Haitao; Chen, Wei

    2018-04-04

    Diet-related factors are thought to modify the risk of cancers, while the influence of high salt intake remains largely uncharacterized. Breast cancer is the most common cancer in women worldwide. In the present study, we examined the effect of salt intake on breast cancer by using a 4T1 mouse mammary tumor model. Unexpectedly, both the fitness and the survival rate of the tumor-bearing mice were improved by high salt intake. Similarly, high salt intake suppressed the primary tumor growth as well as metastasis to lung in mice. Mechanistically, high salt intake greatly reduced food intake and thus might exert antitumor effect through mimicking calorie restriction. Immunoblotting showed the lower proliferation marker Ki-67 and the higher expression of the tumor suppressor gene p53 in tumors of high salt intake mice. Importantly, high salt intake might induce hyperosmotic stress, which sensitized breast cancer cells to p53-dependent anoikis. Collectively, our findings raise the possibility that endogenous salt deposition might act as the first-line defense system against breast cancer progression as well as metastasis.

  15. Effects of excess salt and fat intake on myocardial function and infarct size in rat.

    PubMed

    Mozaffari, Mahmood S; Patel, Champa; Ballas, Claudia; Schaffer, Stephen W

    2006-03-13

    Important risk factors for cardiovascular disease include excess dietary intake of saturated fat and (or) salt. This study tested the hypothesis that excess intakes of saturated fat (e.g., beef tallow) and salt cause greater myocardial cell death following ischemia-reperfusion injury than each risk factor alone. Male rats were divided into four groups: basal fat diet (4.5% as calories; control), high fat diet (40% as calories; FAT), basal fat diet and high salt (1% NaCl solution; SALT) and high fat diet and high salt (FATSALT). The gain in body weight was significantly higher for FAT and FATSALT groups than those of either the control or the SALT group. Five weeks of exposure to the dietary regimens did not significantly affect the coronary flow rate and except for the salt-fed group, had no effect on the rate-pressure-product of the isolated heart perfused in Langendorff mode. Although infarct size was not affected by the high fat diet, it was reduced by the high salt regimen relative to the high fat diet or the control groups. When rats were fed the FAT and SALT combination, the effect of salt feeding on infarct size was not observed. In addition, the FATSALT group displayed a more marked deterioration in contractile function following ischemia-reperfusion injury than the other groups. In conclusion, short-term intake of a high fat diet, which significantly increases body weight, does not worsen ischemia-reperfusion injury although the treatment prevents the reduction of infarct size associated with high salt feeding.

  16. The impact of ions on allosteric functions in human liver pyruvate kinase

    PubMed Central

    Alontaga, Aileen Y.

    2010-01-01

    Experimental designs used to monitor the magnitude of an allosteric response can greatly influence observed values. We report here the impact of buffer, monovalent cation, divalent cation and anion on the magnitude of the allosteric regulation of the affinity of human liver pyruvate kinase (hL-PYK) for substrate, phosphoenolpyruvate (PEP). The magnitudes of the allosteric activation by fructose-1,6-bisphosphate (Fru-1,6-BP) and the allosteric inhibition by alanine are independent of most, but not all buffers tested. However, these magnitudes are dependent on whether Mg2+ or Mn2+ is included as the divalent cation. In the presence of Mn2+, any change in Kapp-PEP caused by Fru-1,6-BP is minimal. hL-PYK activity does not appear to require monovalent cation. Monovalent cation binding in the active site impacts PEP affinity with minimum influence on the magnitude of allosteric coupling. However, Na+ and Li+ reduce the magnitude of the allosteric response to Fru-1,6-BP, likely due to mechanisms outside of the active site. Which anion is used to maintain a constant monovalent cation concentration also influences the magnitude of the allosteric response. The value of determining the impact of ions on allosteric function can be appreciated by considering that representative structures used in comparative studies have often been determined using protein crystals grown in diverse buffer and salt conditions. PMID:21609859

  17. Influence of Temperature on the Colloidal Stability of Polymer-Coated Gold Nanoparticles in Cell Culture Media.

    PubMed

    Zyuzin, Mikhail V; Honold, Tobias; Carregal-Romero, Susana; Kantner, Karsten; Karg, Matthias; Parak, Wolfgang J

    2016-04-06

    The temperature-dependence of the hydrodynamic diameter and colloidal stability of gold-polymer core-shell particles with temperature-sensitive (poly(N-isopropylacrylamide)) and temperature-insensitive shells (polyallylaminine hydrochloride/polystyrensulfonate, poly(isobutylene-alt-maleic anhydride)-graft-dodecyl) are investigated in various aqueous media. The data demonstrate that for all nanoparticle agglomeration, i.e., increase in effective nanoparticle size, the presence of salts or proteins in the dispersion media has to be taken into account. Poly(N-isopropylacrylamide) coated nanoparticles show a reversible temperature-dependent increase in size above the volume phase transition of the polymer shell when they are dispersed in phosphate buffered saline or in media containing protein. In contrast, the nanoparticles coated with temperature-insensitive polymers show a time-dependent increase in size in phosphate buffered saline or in medium containing protein. This is due to time-dependent agglomeration, which is particularly strong in phosphate buffered saline, and induces a time-dependent, irreversible increase in the hydrodynamic diameter of the nanoparticles. This demonstrates that one has to distinguish between temperature- and time-induced agglomerations. Since the size of nanoparticles regulates their uptake by cells, temperature-dependent uptake of thermosensitive and non-thermosensitive nanoparticles by cells lines is compared. No temperature-specific difference between both types of nanoparticles could be observed. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production

    PubMed Central

    Tufail, Soban; Ismat, Fouzia; Imran, Muhammad; Iqbal, Mazhar; Mirza, Osman; Rhaman, Moazur

    2016-01-01

    Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications. PMID:27093053

  19. Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production.

    PubMed

    Ullah, Raheem; Shah, Majid Ali; Tufail, Soban; Ismat, Fouzia; Imran, Muhammad; Iqbal, Mazhar; Mirza, Osman; Rhaman, Moazur

    2016-01-01

    Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications.

  20. Influence of cell culture medium composition on in vitro dissolution behavior of a fluoride-containing bioactive glass.

    PubMed

    Shah, Furqan A; Brauer, Delia S; Wilson, Rory M; Hill, Robert G; Hing, Karin A

    2014-03-01

    Bioactive glasses are used clinically for bone regeneration, and their bioactivity and cell compatibility are often characterized in vitro, using physiologically relevant test solutions. The aim of this study was to show the influence of varying medium characteristics (pH, composition, presence of proteins) on glass dissolution and apatite formation. The dissolution behavior of a fluoride-containing bioactive glass (BG) was investigated over a period of one week in Eagle's Minimal Essential Medium with Earle's Salts (MEM), supplemented with either, (a) acetate buffer, (b) 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, (c) HEPES + carbonate, or (d) HEPES + carbonate + fetal bovine serum. Results show pronounced differences in pH, ion release, and apatite formation over 1 week: Despite its acidic pH (pH 5.8 after BG immersion, as compared to pH 7.4-8.3 for HEPES-containing media), apatite formation was fastest in acetate buffered (HEPES-free) MEM. Presence of carbonate resulted in formation of calcite (calcium carbonate). Presence of serum proteins, on the other hand, delayed apatite formation significantly. These results confirm that the composition and properties of a tissue culture medium are important factors during in vitro experiments and need to be taken into consideration when interpreting results from dissolution or cell culture studies. Copyright © 2013 Wiley Periodicals, Inc.

  1. Sulfide and Oxide Heterostructures For the SrTiO3 Thin Film Growth on Si and Their Structural and Interfacial Stabilities

    NASA Astrophysics Data System (ADS)

    Yoo, Young‑Zo; Song, Jeong‑Hwan; Konishi, Yoshinori; Kawasaki, Masashi; Koinuma, Hideomi; Chikyow, Toyohiro

    2006-03-01

    Epitaxial SrTiO3 (STO) thin films with high electrical properties were grown on Si using ZnS single- and SrS/MnS hetero-buffer layers. STO films on both ZnS-buffered and SrS/MnS-buffered Si showed two growth orientations, (100) and (110). The temperature dependence of the growth orientation for STO films was different for the ZnS single-buffer layer in comparison with the SrS/MnS heterobuffer layers. (100) growth of STO films on SrS/MnS-buffered Si became dominant at high temperatures about 700 °C, while (100) growth of STO films on ZnS-buffered Si became dominant at a relatively low growth temperature of 550 °C. STO(100) films on ZnS-buffered and SrS/MnS-buffered Si showed lattice and domain matches for epitaxial relationships with [001]ZnS\\parallel[011]STO and SrS[001]\\parallel[011]STO, respectively via 45° in-plane rotation of STO films relative to both ZnS and SrS layers. The ZnS buffer layer contained many stacking faults because of the mismatch between ZnS and Si, however, those defects were terminated at the ZnS/STO interface. In contrast, the MnS buffer was very stable against stacking defect formation. Transmission electron microscopy measurements revealed the presence of a disordered region at the ZnS/Si and MnS/Si interfaces. Auger electron spectroscopy and transmission electron microscopy results showed that a good MnS/Si interface at the initial growth stage degraded to a SiS2-x-rich phase during MnS deposition and again into a SiO2-x-rich phase during STO deposition at the high growth temperature of 700 °C. It was also observed that STO on SrS/MnS-buffered Si showed a markedly high dielectric constant compared with that of STO on ZnS-buffered Si.

  2. The Influence of End-Stop Buffer Characteristics on the Severity of Suspension Seat End-Stop Impacts

    NASA Astrophysics Data System (ADS)

    Wu, X.; Griffin, M. J.

    1998-08-01

    Suspension seat end-stop impacts may be a source of increased risk of injury for the drivers of some machines and work vehicles, such as off-road vehicles. Most suspension seats use rubber buffers to reduce the severity of end-stop impacts, but they still result in a high magnitude of acceleration being transmitted to drivers when an end-stop impact occurs. An experimental study has been conducted to investigate the effect of buffer stiffness and buffer damping on the severity of end-stop impacts. The results show that the end-stop impact performance of suspension seats with only bottom buffers can be improved by the use of both top and bottom buffers. The force-deflection characteristics of rubber buffers had a significant influence on the severity of end-stop impacts. The optimum buffer should have medium stiffness which is nearly linear and occurs over a long deflection, without being compressed to its high stiffness stage. It is shown, theoretically, that buffer damping is capable of significantly reducing the severity of end-stop impacts. However, since current rubber material provides only low damping, alternative materials to those in current use, or either passive or active damping devices, are required.

  3. Heart rate and blood pressure variabilities in salt-sensitive hypertension.

    PubMed

    Piccirillo, G; Bucca, C; Durante, M; Santagada, E; Munizzi, M R; Cacciafesta, M; Marigliano, V

    1996-12-01

    In salt-sensitive hypertension, a high sodium intake causes plasma catecholamines to rise and pulmonary baroreceptor plasticity to fall. In salt-sensitive and salt-resistant hypertensive subjects during low and high sodium intakes, we studied autonomic nervous system activity by power spectral analysis of heart rate and arterial pressure variabilities and baroreceptor sensitivity. In all subjects, high sodium intake significantly enhanced the low-frequency power of heart rate and arterial pressures at rest and after sympathetic stress. It also increased heart rate and arterial pressure variabilities. During high sodium intake, salt-sensitive hypertensive subjects had significantly higher low-frequency powers of systolic arterial pressure (7.5 mm Hg2, P < .05) and of heart rate at rest (59.2 +/- 2.4 normalized units [NU], P < .001) than salt-resistant subjects (6.6 +/- 0.3 mm Hg2, 55.0 +/- 3.2 NU) and normotensive control subjects (5.1 +/- 0.5 mm Hg2, 41.6 +/- 2.9 NU). In salt-sensitive subjects, low sodium intake significantly reduced low-frequency normalized units (P < .001) and the ratio of low- to high-power frequency (P < .001). High-sodium intake significantly increased baroreflex sensitivity in control subjects (from 10.0 +/- 0.7 to 17.5 +/- 0.7 ms/mm Hg, P < .001) and salt-resistant subjects (from 6.9 +/- 0.7 to 13.9 +/- 0.9, P < .05) but not in salt-sensitive subjects (7.4 +/- 0.3 to 7.9 +/- 0.4). In conclusion, a high sodium intake markedly enhances cardiac sympathetic activity in salt-sensitive and salt-resistant hypertension. In contrast, although reduced sodium intake lowers arterial pressure and sympathetic activity, it does so only in salt-sensitive subjects. Hence, in salt-resistant subjects, neither arterial pressure nor sympathetic activity depends on salt intake. During a high sodium intake in normotensive subjects and salt-resistant hypertensive subjects, increased sympathetic activity is probably compensated by enhanced baroreflex sensitivity.

  4. Thermal buffering of concrete by seaweeds during a prolonged summer heatwave

    NASA Astrophysics Data System (ADS)

    Naylor, Larissa; Coombes, Martin

    2014-05-01

    Hard coastal infrastructure is subject to aggressive environmental conditions, including a suite of weathering processes in the intertidal zone. These processes, along with waves, lead to costly deterioration of coastal structures. Existing methods (e.g. coatings, less porous concrete) to reduce the risk of concrete deterioration rapidly lose their effectiveness in the intertidal zone. Additionally, a changing climate will lead to increased frequency of storms, higher sea level and higher extreme temperatures - and therefore, pose an increased risk of deterioration. Might there be a biogenic solution? New research (Coombes et al. 2013) has shown that fucoid seaweeds reduce microclimatic extremes and variability under normal summer conditions. The results presented here supplement these findings in two ways. First, they demonstrate that fucoid seaweeds act as a thermal buffer during a prolonged summer heatwave in Britain (July 2013). Over 36 days of continuous monitoring at two sites in Cornwall, UK, 19 of which were during the official heatwave, there were statistically significant differences (p = 0.000) in the maximum temperatures between thick seaweed (7.5 - 9.5 cm thickness) and thin seaweed (2 - 2.5 cm thickness) plots. Maximum temperatures reached 22°C and 33°C, for thick seaweed and thin seaweed plots, respectively. Variations in maximum temperatures between the two sites appear to be related to aspect. Second, the significantly different maximum temperature results between plots also demonstrate that seaweed thickness is an important factor influencing thermal buffering capacity. These data clearly demonstrate that fucoid seaweeds buffer concrete seawalls against extreme temperature fluxes during a heatwave, probably limiting the efficiency of deteriorative processes such as thermal expansion and contraction and salt crystallisation.

  5. New iodide-based molten salt systems for high temperature molten salt batteries

    NASA Astrophysics Data System (ADS)

    Fujiwara, Syozo; Kato, Fumio; Watanabe, Syouichiro; Inaba, Minoru; Tasaka, Akimasa

    Novel multi-component molten salt systems containing iodides, LiF-LiBr-LiI, LiF-NaBr-LiI, and LiF-LiCl-LiBr-LiI, were investigated for use as electrolytes in high temperature molten salt batteries to improve the discharge rate-capability. The iodide-based molten salts showed higher ionic conductivity (∼3 S cm -1 at 500 °C) than conventional LiCl-KCl, and had low enough melting points (below 400 °C) that can be used in practical high temperature molten salt batteries. The iodide-based salts showed instability at temperatures higher than 280 °C in dried air. The decomposition mechanism of iodide-based molten salts was discussed, and it was found that elimination of oxygen from the environment is effective to stabilize the iodide-based molten salts at high temperatures.

  6. Extremely halophilic archaea from ancient salt sediments and their possible survival in halite fluid inclusions

    NASA Astrophysics Data System (ADS)

    Stan-Lotter, H.; Fendrihan, S.; Gerbl, F. W.; Dornmayr-Pfaffenhuemer, M.; Frethem, C.

    2008-09-01

    Halophilic archaebacteria (haloarchaea) thrive in environments with salt concentrations approaching saturation, such as natural brines, marine solar salterns and alkaline salt lakes; they have also been isolated from ancient subsurface salt sediments of great geological age (195-280 million years) and some of those strains were described as novel species (1). The cells survived perhaps while being enclosed within small fluid inclusions in the halite. The characterization of subsurface microbial life is of astrobiological relevance since extraterrestrial halite has been detected and since microbial life on Mars, if existent, may have retreated into the subsurface. We attempted to simulate the embedding process of extremely halophilic archaea and to analyse any cellular changes which might occur. When enclosing haloarchaea in laboratory grown halite, cells accumulated preferentially in fluid inclusions, as could be demonstrated by pre-staining with fluorescent dyes. With increased time of embedding, rod-shaped cells of Halobacterium salinarum strains were found to assume roundish morphologies. Upon dissolution of the salt crystals, these spheres were stable and viable for months when kept in buffers containing 4 M NaCl. Scanning electron microscopy (SEM) following fixation with glutaraldehyde suggested a potentially gradual transformation from rods to spheres. This notion was supported by fluorescence microscopy of Halobacterium cells, following embedding in halite and staining with SYTO 9. One-dimensional protein patterns of rods and spheres, following SDS polyacrylamide gel electrophoresis, were similar except that the S-layer protein appeared reduced by about 15 - 20 % in spheres. The reddish-orange pigmentation of spheres was much lighter compared to that of rod-shaped cells, suggesting lowered concentrations of carotenoids; this was confirmed by extraction and spectrometry of pigments. The data suggested that Halobacterium cells are capable of forming specific cellular structures upon embedding in fluid inclusions of halite. It is tempting to speculate that such structures may be responsible for long term survival in ancient geological materials such as salt sediments, including extraterrestrial salt. (1) Fendrihan S., Legat A., Pfaffenhuemer M., Gruber C., Weidler W., Gerbl F.W., Stan-Lotter H. (2006) Extremely halophilic archaea and the issue of long-term microbial survival. Reviews in Environmental Science and Bio/technology 5, 1569-1605.

  7. Development of high temperature transport technology for LiCl-KCl eutectic salt in pyroprocessing

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

    Lee, Sung Ho; Lee, Hansoo; Kim, In Tae

    The development of high-temperature transport technologies for molten salt is a prerequisite and a key issue in the industrialization of pyro-reprocessing for advanced fuel cycle scenarios. The solution of a molten salt centrifugal pump was discarded because of the high corrosion power of a high temperature molten salt, so the suction pump solution was selected. An apparatus for salt transport experiments by suction was designed and tested using LiC-KCl eutectic salt. The experimental results of lab-scale molten salt transport by suction showed a 99.5% transport rate (ratio of transported salt to total salt) under a vacuum range of 100 mtorrmore » - 10 torr at 500 Celsius degrees. The suction system has been integrated to the PRIDE (pyroprocessing integrated inactive demonstration) facility that is a demonstrator using non-irradiated materials (natural uranium and surrogate materials). The performance of the suction pump for the transport of molten salts has been confirmed.« less

  8. High temperature ultrasonic testing of materials for internal flaws

    DOEpatents

    Kupperman, David S.; Linzer, Melvin

    1990-01-01

    An apparatus is disclosed for nondestructive evaluation of defects in hot terials, such as metals and ceramics, by sonic signals, which includes a zirconia buffer in contact with a hot material being tested, a liquid couplant of borax in contact with the zirconia buffer and the hot material to be tested, a transmitter mounted on the zirconia buffer sending sonic signals through the buffer and couplant into the hot material, and a receiver mounted on the zirconia buffer receiving sonic signals reflected from within the hot material through the couplant and the buffer.

  9. Micellization and Single-Particle Encapsulation with Dimethylammoniopropyl Sulfobetaines

    PubMed Central

    2017-01-01

    Sulfobetaines (SBs) are a class of zwitterionic surfactants with a reputation for enhancing colloidal stability at high salt concentrations. Here, we present a systematic study on the self-assembly of SB amphiphiles (sultaines or hydroxysultaines) in aqueous solutions, as a function of chain length and composition, ionic strength, and in the presence of alkanethiol-coated Au nanoparticles (GNPs). The diameters of the micelles assembled from SB and amidosulfobetaine (ASB) generally increase monotonically with chain length, although ASB micelles are smaller relative to alkyl SB micelles with similarly sized tailgroups, and oleyl sulfobetaine (OSB) micelles are slightly larger. SB amphiphiles can stabilize alkanethiol-coated GNPs in physiologically relevant buffers at concentrations well below their CMC, with size increases corresponding to single-particle encapsulation. SB-encapsulated GNPs were prepared by three different methods with SB:GNP weight ratios of 10:1, followed by dispersion in water or 1 M NaCl. The low hydrodynamic size of the SB micelles and SB-coated NPs is within the range needed for efficient renal clearance. PMID:28474008

  10. Micellization and Single-Particle Encapsulation with Dimethylammoniopropyl Sulfobetaines.

    PubMed

    Wang, Jianxin; Morales-Collazo, Oscar; Wei, Alexander

    2017-04-30

    Sulfobetaines (SBs) are a class of zwitterionic surfactants with a reputation for enhancing colloidal stability at high salt concentrations. Here, we present a systematic study on the self-assembly of SB amphiphiles (sultaines or hydroxysultaines) in aqueous solutions, as a function of chain length and composition, ionic strength, and in the presence of alkanethiol-coated Au nanoparticles (GNPs). The diameters of the micelles assembled from SB and amidosulfobetaine (ASB) generally increase monotonically with chain length, although ASB micelles are smaller relative to alkyl SB micelles with similarly sized tailgroups, and oleyl sulfobetaine (OSB) micelles are slightly larger. SB amphiphiles can stabilize alkanethiol-coated GNPs in physiologically relevant buffers at concentrations well below their CMC, with size increases corresponding to single-particle encapsulation. SB-encapsulated GNPs were prepared by three different methods with SB:GNP weight ratios of 10:1, followed by dispersion in water or 1 M NaCl. The low hydrodynamic size of the SB micelles and SB-coated NPs is within the range needed for efficient renal clearance.

  11. The purification and properties of placental histaminase

    PubMed Central

    Smith, J. K.

    1967-01-01

    1. Histaminase was extracted from desanguinated human placentae and purified by salt fractionation, ion-exchange chromatography and gel filtration. The purest preparation was still contaminated with haptoglobin–methaemoglobin. 2. Histaminase activity was measured by the o-aminobenzaldehyde method of Holmstedt & Tham (1959), Kapeller-Adler's (1951) test and a modified spectrophotometric indigodisulphonate test of greater sensitivity. 3. Unless contaminant metal ions were removed, enzymic activity on cadaverine, but not on histamine, fell during purification. When EDTA was added to the working buffers, a constant ratio between activities towards cadaverine and histamine was maintained throughout the later stages of purification, and activities towards the two substrates could not be separated by any of the highly resolving chromatographic analyses employed. 4. The purest preparation oxidized histamine, agmatine and benzylamine more slowly than the C4–C6 aliphatic diamines, but mixed-substrate experiments suggested that all these amines were substrates of histaminase. 5. The substrate and inhibitor specificities of placental histaminase were compared with those of related enzymes from other sources. PMID:4962162

  12. Plasmid DNA production combining antibiotic-free selection, inducible high yield fermentation, and novel autolytic purification.

    PubMed

    Carnes, Aaron E; Hodgson, Clague P; Luke, Jeremy M; Vincent, Justin M; Williams, James A

    2009-10-15

    DNA vaccines and gene medicines, derived from bacterial plasmids, are emerging as an important new class of pharmaceuticals. However, the challenges of performing cell lysis processes for plasmid DNA purification at an industrial scale are well known. To address downstream purification challenges, we have developed autolytic Escherichia coli host strains that express endolysin (phage lambdaR) in the cytoplasm. Expression of the endolysin is induced during fermentation by a heat inducible promoter. The endolysin remains in the cytoplasm, where it is separated from its peptidoglycan substrate in the cell wall; hence the cells remain alive and intact and can be harvested by the usual methods. The plasmid DNA is then recovered by autolytic extraction under slightly acidic, low salt buffer conditions and treatment with a low concentration of non-ionic detergent. Under these conditions the E. coli genomic DNA remains associated with the insoluble cell debris and is removed by a solid-liquid separation. Here, we report fermentation, lysis methods, and plasmid purification using autolytic hosts.

  13. Long-term survival of bacterial spores in space

    NASA Technical Reports Server (NTRS)

    Horneck, G.; Bucker, H.; Reitz, G.

    1994-01-01

    On board of the NASA Long Duration Exposure Facility (LDEF), spores of Bacillus subtilis in monolayers (10(exp 6)/sample) or multilayers (10(exp 8)/sample) were exposed to the space environment for nearly six years and their survival was analyzed after retrieval. The response to space parameters, such as vacuum (10(exp -6) Pa), solar electromagnetic radiation up to the highly energetic vacuum-ultraviolet range 10(exp 9) J/sq m) and/or cosmic radiation (4.8 Gy), was studied and compared to the results of a simultaneously running ground control experiment. If shielded against solar ultraviolet (UV)-radiation, up to 80% of spores in multilayers survive in space. Solar UV-radiation, being the most deleterious parameter of space, reduces survival by 4 orders of magnitude or more. However, up to 10(exp 4) viable spores were still recovered, even in completely unprotected samples. Substances, such as glucose or buffer salts serve as chemical protectants. With this 6 year study in space, experimental data are provided to the discussion on the likelihood of 'Panspermia'.

  14. Design of a minimal protein oligomerization domain by a structural approach.

    PubMed

    Burkhard, P; Meier, M; Lustig, A

    2000-12-01

    Because of the simplicity and regularity of the alpha-helical coiled coil relative to other structural motifs, it can be conveniently used to clarify the molecular interactions responsible for protein folding and stability. Here we describe the de novo design and characterization of a two heptad-repeat peptide stabilized by a complex network of inter- and intrahelical salt bridges. Circular dichroism spectroscopy and analytical ultracentrifugation show that this peptide is highly alpha-helical and 100% dimeric tinder physiological buffer conditions. Interestingly, the peptide was shown to switch its oligomerization state from a dimer to a trimer upon increasing ionic strength. The correctness of the rational design principles used here is supported by details of the atomic structure of the peptide deduced from X-ray crystallography. The structure of the peptide shows that it is not a molten globule but assumes a unique, native-like conformation. This de novo peptide thus represents an attractive model system for the design of a molecular recognition system.

  15. Design of a minimal protein oligomerization domain by a structural approach.

    PubMed Central

    Burkhard, P.; Meier, M.; Lustig, A.

    2000-01-01

    Because of the simplicity and regularity of the alpha-helical coiled coil relative to other structural motifs, it can be conveniently used to clarify the molecular interactions responsible for protein folding and stability. Here we describe the de novo design and characterization of a two heptad-repeat peptide stabilized by a complex network of inter- and intrahelical salt bridges. Circular dichroism spectroscopy and analytical ultracentrifugation show that this peptide is highly alpha-helical and 100% dimeric tinder physiological buffer conditions. Interestingly, the peptide was shown to switch its oligomerization state from a dimer to a trimer upon increasing ionic strength. The correctness of the rational design principles used here is supported by details of the atomic structure of the peptide deduced from X-ray crystallography. The structure of the peptide shows that it is not a molten globule but assumes a unique, native-like conformation. This de novo peptide thus represents an attractive model system for the design of a molecular recognition system. PMID:11206050

  16. Growth of calcium phosphates on magnesium substrates for corrosion control in biomedical applications via immersion techniques.

    PubMed

    Shadanbaz, Shaylin; Walker, Jemimah; Staiger, Mark P; Dias, George J; Pietak, Alexis

    2013-01-01

    Magnesium (Mg) has been suggested as a revolutionary biodegradable replacement for current permanent metals used in orthopedic applications. Current investigations concentrate on the control of the corrosion rate to match bone healing. Calcium phosphate coatings have been a recent focus of these investigations through various coating protocols. Within this investigation, an in situ crystallization technique was utilized as an inexpensive and relatively simple method to produce a brushite and monetite coating on pure Mg. Coatings were characterized using energy dispersive spectroscopy, glancing angle X-ray diffraction and field emission scanning electron microscopy. Corrosion protection properties of the coatings were assessed in physiological buffers, Earles balanced salt solution, minimum essential media, and minimum essential media containing serum albumin, over a 4-week period. Using this novel coating protocol, our findings indicate brushite and monetite coated Mg to have significant corrosive protective effects when compared with its uncoated counterpart whilst maintaining high coating substrate adhesion, homogeneity, and reproducibility. Copyright © 2012 Wiley Periodicals, Inc.

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

    PubMed

    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.

  18. Cool barnacles: Do common biogenic structures enhance or retard rates of deterioration of intertidal rocks and concrete?

    PubMed

    Coombes, Martin A; Viles, Heather A; Naylor, Larissa A; La Marca, Emanuela Claudia

    2017-02-15

    Sedentary and mobile organisms grow profusely on hard substrates within the coastal zone and contribute to the deterioration of coastal engineering structures and the geomorphic evolution of rocky shores by both enhancing and retarding weathering and erosion. There is a lack of quantitative evidence for the direction and magnitude of these effects. This study assesses the influence of globally-abundant intertidal organisms, barnacles, by measuring the response of limestone, granite and marine-grade concrete colonised with varying percentage covers of Chthamalus spp. under simulated, temperate intertidal conditions. Temperature regimes at 5 and 10mm below the surface of each material demonstrated a consistent and statistically significant negative relationship between barnacle abundance and indicators of thermal breakdown. With a 95% cover of barnacles, subsurface peak temperatures were reduced by 1.59°C for limestone, 5.54°C for concrete and 5.97°C for granite in comparison to no barnacle cover. The amplitudes of short-term (15-30min) thermal fluctuations conducive to breakdown via 'fatigue' effects were also buffered by 0.70°C in limestone, 1.50°C in concrete and 1.63°C in granite. Furthermore, concentrations of potentially damaging salt ions were consistently lower under barnacles in limestone and concrete. These results indicate that barnacles do not enhance, but likely reduce rates of mechanical breakdown on rock and concrete by buffering near-surface thermal cycling and reducing salt ion ingress. In these ways, we highlight the potential role of barnacles as agents of bioprotection. These findings support growing international efforts to enhance the ecological value of hard coastal structures by facilitating their colonisation (where appropriate) through design interventions. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Constitutive expression of human gastric lipase in Pichia pastoris and site-directed mutagenesis of key lid-stabilizing residues.

    PubMed

    Sams, Laura; Amara, Sawsan; Chakroun, Almahdi; Coudre, Sébastien; Paume, Julie; Giallo, Jacqueline; Carrière, Frédéric

    2017-10-01

    The cDNA encoding human gastric lipase (HGL) was integrated into the genome of Pichia pastoris using the pGAPZα A transfer vector. The HGL signal peptide was replaced by the yeast α-factor to achieve an efficient secretion. Active rHGL was produced by the transformed yeast but its levels and stability were dependent on the pH. The highest activity was obtained upon buffering the culture medium at pH5, a condition that allowed preserving enzyme activity over time. A large fraction (72±2%) of secreted rHGL remained however bound to the yeast cells, and was released by washing the cell pellet with an acid glycine-HCl buffer (pH2.2). This procedure allowed establishing a first step of purification that was completed by size exclusion chromatography. N-terminal sequencing and MALDI-ToF mass spectrometry revealed that rHGL was produced in its mature form, with a global mass of 50,837±32Da corresponding to a N-glycosylated form of HGL polypeptide (43,193Da). rHGL activity was characterized as a function of pH, various substrates and in the presence of bile salts and pepsin, and was found similar to native HGL, except for slight changes in pH optima. We then studied by site-directed mutagenesis the role of three key residues (K4, E225, R229) involved in salt bridges stabilizing the lid domain that controls the access to the active site and is part of the interfacial recognition site. Their substitution has an impact on the pH-dependent activity of rHGL and its relative activities on medium and long chain triglycerides. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Novel inclusion complex of ibuprofen tromethamine with cyclodextrins: physico-chemical characterization.

    PubMed

    Al Omari, Mahmoud M; Daraghmeh, Nidal H; El-Barghouthi, Musa I; Zughul, Mohammad B; Chowdhry, Babur Z; Leharne, Stephen A; Badwan, Adnan A

    2009-10-15

    Guest-host interactions of ibuprofen tromethamine salt (Ibu.T) with native and modified cyclodextrins (CyDs) have been investigated using several techniques, namely phase solubility diagrams (PSDs), proton nuclear magnetic resonance ((1)H NMR), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffractometry (XRPD), scanning-electron microscopy (SEM) and molecular mechanics (MM). From the analysis of PSD data (A(L)-type) it is concluded that the anionic tromethamine salt of ibuprofen (pK(a)=4.55) forms 1:1 soluble complexes with all CyDs investigated in buffered water at pH 7.0, while the neutral form of Ibu forms an insoluble complex with beta-CyD (B(S)-type) in buffered water at pH 2.0. Ibu.T has a lower tendency to complex with beta-CyD (K(11)=58 M(-1) at pH 7.0) compared with the neutral Ibu (K(11)=4200 M(-1)) in water. Complex formation of Ibu.T with beta-CyD (DeltaG(o)=-20.4 kJ/mol) is enthalpy driven (DeltaH(o)=-22.9 kJ/mol) and is accompanied by a small unfavorable entropy (DeltaS(o)=-8.4 J/mol K) change. (1)H NMR studies and MM computations revealed that, on complexation, the hydrophobic central benzene ring of Ibu.T and part of the isobutyl group reside within the beta-CyD cavity leaving the peripheral groups (carboxylate, tromethamine and methyl groups) located near the hydroxyl group networks at either rim of beta-CyD. PSD, (1)H NMR, DSC, FT-IR, XRPD, SEM and MM studies confirmed the formation of Ibu.T/beta-CyD inclusion complex in solution and the solid state.

  1. Emotional labour and work engagement among nurses: examining perceived compassion, leadership and work ethic as stress buffers.

    PubMed

    Mauno, Saija; Ruokolainen, Mervi; Kinnunen, Ulla; De Bloom, Jessica

    2016-05-01

    The study examined whether three resources, that is, compassion, transformational leadership and work ethic feasibility, buffer against the negative effects of emotional labour on work engagement. Emotional labour is a common job stressor among nurses, but little is known about whether certain personal and work resources buffer against it in relation to work engagement. Revealing buffers of emotional labour would help organizations to design tailored interventions. Cross-sectional online survey conducted in 2014. Participants were 3466 Finnish nurses. Hypotheses were tested via hierarchical moderated regression analyses. Higher emotional labour related to lower engagement. Two interaction effects were found. First, work ethic feasibility buffered against emotional labour: the nurses who perceived work ethic feasibility as high in a situation of high emotional labour, scored higher on engagement compared with those nurses who in this stress situation perceived work ethic feasibility to be low. Second, high compassion was detrimental to engagement in the presence of high emotional labour. Transformational leadership did not act as a buffer but showed a positive relationship with engagement. Work ethic feasibility (being able to work according to high ethical standards) is an important resource in nursing as it protects an employee against the negative effects of emotional labour and as it also directly promotes engagement. However, compassion may not always be beneficial in nursing, especially if co-occurring with high job stress. Transformational leadership has potential to improve engagement in nursing although it may not operate as a stress buffer. © 2016 John Wiley & Sons Ltd.

  2. Consumer knowledge and attitudes to salt intake and labelled salt information.

    PubMed

    Grimes, Carley A; Riddell, Lynn J; Nowson, Caryl A

    2009-10-01

    The objective of this study was to investigate consumers' knowledge of health risks of high salt intake and frequency of use and understanding of labelled salt information. We conducted a cross-sectional survey in shopping centres within Metropolitan Melbourne. A sample of 493 subjects was recruited. The questionnaire assessed salt related shopping behaviours, attitudes to salt intake and health and their ability to interpret labelled sodium information. Four hundred and seventy four valid surveys were collected (65% female, 64% being the main shopper). Most participants knew of the relationship between salt intake and high blood pressure (88%). Sixty five percent of participants were unable to correctly identify the relationship between salt and sodium. Sixty nine percent reported reading the salt content of food products when shopping. Salt label usage was significantly related to shoppers concern about the amount of salt in their diet and the belief that their health could improve by lowering salt intake. Approximately half of the sample was unable to accurately use labelled sodium information to pick low salt options. Raising consumer awareness of the health risks associated with high salt consumption may increase salt label usage and purchases of low salt foods. However, for food labels to be effective in helping consumers select low salt foods a more 'user friendly' labelling format is needed.

  3. High-salt in addition to high-fat diet may enhance inflammation and fibrosis in liver steatosis induced by oxidative stress and dyslipidemia in mice.

    PubMed

    Uetake, Yuzaburo; Ikeda, Hitoshi; Irie, Rie; Tejima, Kazuaki; Matsui, Hiromitsu; Ogura, Sayoko; Wang, Hong; Mu, ShengYu; Hirohama, Daigoro; Ando, Katsuyuki; Sawamura, Tatsuya; Yatomi, Yutaka; Fujita, Toshiro; Shimosawa, Tatsuo

    2015-02-13

    It is widely known that salt is an accelerating factor for the progression of metabolic syndrome and causes cardiovascular diseases, most likely due to its pro-oxidant properties. We hypothesized that excessive salt intake also facilitates the development of nonalcoholic steatohepatitis (NASH), which is frequently associated with metabolic syndrome. We examined the exacerbating effect of high-salt diet on high-fat diet-induced liver injury in a susceptible model to oxidative stress, apoE knockout and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) transgenic mice. High-salt diet led to NASH in high-fat diet-fed LOX-1 transgenic/apoE knockout mice without affecting high-fat diet-induced dyslipidemia or hepatic triglyceride accumulation. Additionally, a high-salt and high-fat diet stimulated oxidative stress production and inflammatory reaction to a greater extent than did a high-fat diet in the liver of LOX-1 transgenic/apoE knockout mice. We demonstrated that high-salt diet exacerbated NASH in high-fat diet-fed LOX-1 transgenic /apoE knockout mice and that this effect was associated with the stimulation of oxidative and inflammatory processes; this is the first study to suggest the important role of excessive salt intake in the development of NASH.

  4. Electrical properties of n-type GaSb substrates and p-type GaSb buffer layers for InAs/InGaSb superlattice infrared detectors

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

    Mitchel, W. C., E-mail: William.Mitchel.1@us.af.mil; Haugan, H. J.; Mou, Shin

    2015-09-15

    Lightly doped n-type GaSb substrates with p-type GaSb buffer layers are the preferred templates for growth of InAs/InGaSb superlattices used in infrared detector applications because of relatively high infrared transmission and a close lattice match to the superlattices. We report here temperature dependent resistivity and Hall effect measurements of bare substrates and substrate-p-type buffer layer structures grown by molecular beam epitaxy. Multicarrier analysis of the resistivity and Hall coefficient data demonstrate that high temperature transport in the substrates is due to conduction in both the high mobility zone center Γ band and the low mobility off-center L band. High overallmore » mobility values indicate the absence of close compensation and that improved infrared and transport properties were achieved by a reduction in intrinsic acceptor concentration. Standard transport measurements of the undoped buffer layers show p-type conduction up to 300 K indicating electrical isolation of the buffer layer from the lightly n-type GaSb substrate. However, the highest temperature data indicate the early stages of the expected p to n type conversion which leads to apparent anomalously high carrier concentrations and lower than expected mobilities. Data at 77 K indicate very high quality buffer layers.« less

  5. Nutritional Requirements for Synthesis of Heat-Labile Enterotoxin by Enterotoxigenic Strains of Escherichia coli

    PubMed Central

    Gilligan, Peter H.; Robertson, Donald C.

    1979-01-01

    Optimal growth conditions have been established for production of heat-labile enterotoxin (LT) by both porcine and human strains of enterotoxigenic (ENT+) Escherichia coli. There were no unusual growth factor requirements, and some strains produced fairly high levels of LT in a basal salts medium containing 0.5% glucose if the pH was carefully controlled. Several amino acids markedly stimulated LT synthesis when added to the basal salts-glucose medium. Methionine and lysine were the most stimulatory for both human and porcine strains. Either aspartic acid or glutamic acid further enhanced LT synthesis in the presence of methionine and lysine, with aspartic acid being more stimulatory for porcine strains and glutamic acid more stimulatory for human strains. There were no apparent vitamin requirements and no unusual cations needed for toxin synthesis except that Fe3+ was slightly stimulatory for porcine strains. The stimulation by Fe3+ was observed only in the presence of the three amino acids, suggesting that the effect was indirect rather than on toxin synthesis. The carbon source also influenced the yield of LT. Glucose supported maximal synthesis, but other carbon sources which exhibit a high degree of catabolite repression also supported high levels of synthesis. Little or no LT was released below pH 7.0; therefore, because the pH drops during growth from 7.5 to 6.8, even in highly buffered media, it was necessary to adjust the pH to 8.0 to effect complete release of cell-associated toxin. The defined medium containing three amino acids reduced the amount of UV-absorbing material in culture supernatants about fivefold and increased LT activity for various strains from two- to fivefold over a complex Casamino Acids-yeast extract medium. Conditions found to be optimal for synthesis of LT were inhibitory for the heat-stable enterotoxin. PMID:33900

  6. The use of spin desalting columns in DMSO-quenched H/D-exchange NMR experiments

    PubMed Central

    Chandak, Mahesh S; Nakamura, Takashi; Takenaka, Toshio; Chaudhuri, Tapan K; Yagi-Utsumi, Maho; Chen, Jin; Kato, Koichi; Kuwajima, Kunihiro

    2013-01-01

    Dimethylsulfoxide (DMSO)-quenched hydrogen/deuterium (H/D)-exchange is a powerful method to characterize the H/D-exchange behaviors of proteins and protein assemblies, and it is potentially useful for investigating non-protected fast-exchanging amide protons in the unfolded state. However, the method has not been used for studies on fully unfolded proteins in a concentrated denaturant or protein solutions at high salt concentrations. In all of the current DMSO-quenched H/D-exchange studies of proteins so far reported, lyophilization was used to remove D2O from the protein solution, and the lyophilized protein was dissolved in the DMSO solution to quench the H/D exchange reactions and to measure the amide proton signals by two-dimensional nuclear magnetic resonance (2D NMR) spectra. The denaturants or salts remaining after lyophilization thus prevent the measurement of good NMR spectra. In this article, we report that the use of spin desalting columns is a very effective alternative to lyophilization for the medium exchange from the D2O buffer to the DMSO solution. We show that the medium exchange by a spin desalting column takes only about 10 min in contrast to an overnight length of time required for lyophilization, and that the use of spin desalting columns has made it possible to monitor the H/D-exchange behavior of a fully unfolded protein in a concentrated denaturant. We report the results of unfolded ubiquitin in 6.0M guanidinium chloride. PMID:23339068

  7. Elevation of Fasting Ghrelin in Healthy Human Subjects Consuming a High-Salt Diet: A Novel Mechanism of Obesity?

    PubMed Central

    Zhang, Yong; Li, Fenxia; Liu, Fu-Qiang; Chu, Chao; Wang, Yang; Wang, Dan; Guo, Tong-Shuai; Wang, Jun-Kui; Guan, Gong-Chang; Ren, Ke-Yu; Mu, Jian-Jun

    2016-01-01

    Overweight/obesity is a chronic disease that carries an increased risk of hypertension, diabetes mellitus, and premature death. Several epidemiological studies have demonstrated a clear relationship between salt intake and obesity, but the pathophysiologic mechanisms remain unknown. We hypothesized that ghrelin, which regulates appetite, food intake, and fat deposition, becomes elevated when one consumes a high-salt diet, contributing to the progression of obesity. We, therefore, investigated fasting ghrelin concentrations during a high-salt diet. Thirty-eight non-obese and normotensive subjects (aged 25 to 50 years) were selected from a rural community in Northern China. They were sequentially maintained on a normal diet for three days at baseline, a low-salt diet for seven days (3 g/day, NaCl), then a high-salt diet for seven days (18 g/day). The concentration of plasma ghrelin was measured using an immunoenzyme method (ELISA). High-salt intake significantly increased fasting ghrelin levels, which were higher during the high-salt diet (320.7 ± 30.6 pg/mL) than during the low-salt diet (172.9 ± 8.9 pg/mL). The comparison of ghrelin levels between the different salt diets was statistically-significantly different (p < 0.01). A positive correlation between 24-h urinary sodium excretion and fasting ghrelin levels was demonstrated. Our data indicate that a high-salt diet elevates fasting ghrelin in healthy human subjects, which may be a novel underlying mechanism of obesity. PMID:27240398

  8. Elevation of Fasting Ghrelin in Healthy Human Subjects Consuming a High-Salt Diet: A Novel Mechanism of Obesity?

    PubMed

    Zhang, Yong; Li, Fenxia; Liu, Fu-Qiang; Chu, Chao; Wang, Yang; Wang, Dan; Guo, Tong-Shuai; Wang, Jun-Kui; Guan, Gong-Chang; Ren, Ke-Yu; Mu, Jian-Jun

    2016-05-26

    Overweight/obesity is a chronic disease that carries an increased risk of hypertension, diabetes mellitus, and premature death. Several epidemiological studies have demonstrated a clear relationship between salt intake and obesity, but the pathophysiologic mechanisms remain unknown. We hypothesized that ghrelin, which regulates appetite, food intake, and fat deposition, becomes elevated when one consumes a high-salt diet, contributing to the progression of obesity. We, therefore, investigated fasting ghrelin concentrations during a high-salt diet. Thirty-eight non-obese and normotensive subjects (aged 25 to 50 years) were selected from a rural community in Northern China. They were sequentially maintained on a normal diet for three days at baseline, a low-salt diet for seven days (3 g/day, NaCl), then a high-salt diet for seven days (18 g/day). The concentration of plasma ghrelin was measured using an immunoenzyme method (ELISA). High-salt intake significantly increased fasting ghrelin levels, which were higher during the high-salt diet (320.7 ± 30.6 pg/mL) than during the low-salt diet (172.9 ± 8.9 pg/mL). The comparison of ghrelin levels between the different salt diets was statistically-significantly different (p < 0.01). A positive correlation between 24-h urinary sodium excretion and fasting ghrelin levels was demonstrated. Our data indicate that a high-salt diet elevates fasting ghrelin in healthy human subjects, which may be a novel underlying mechanism of obesity.

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

    DOEpatents

    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.

  10. Molten Salts for High Temperature Reactors: University of Wisconsin Molten Salt Corrosion and Flow Loop Experiments -- Issues Identified and Path Forward

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

    Piyush Sabharwall; Matt Ebner; Manohar Sohal

    2010-03-01

    Considerable amount of work is going on regarding the development of high temperature liquid salts technology to meet future process needs of Next Generation Nuclear Plant. This report identifies the important characteristics and concerns of high temperature molten salts (with lesson learned at University of Wisconsin-Madison, Molten Salt Program) and provides some possible recommendation for future work

  11. Polyamine binding to proteins in oat and Petunia protoplasts

    NASA Technical Reports Server (NTRS)

    Mizrahi, Y.; Applewhite, P. B.; Galston, A. W.

    1989-01-01

    Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants. To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa). In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action. In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH. The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS. Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons. Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.

  12. Preparation and physico-chemical properties of hydrogels from carboxymethyl cassava starch crosslinked with citric acid

    NASA Astrophysics Data System (ADS)

    Boonkham, Sasikan; Sangseethong, Kunruedee; Chatakanon, Pathama; Niamnuy, Chalida; Nakasaki, Kiyohiko; Sriroth, Klanarong

    2014-06-01

    Recently, environmentally friendly hydrogels prepared from renewable bio-based resources have drawn significant attention from both industrial and academic sectors. In this study, chemically crosslinked hydrogels have been developed from cassava starch which is a bio-based polymer using a non-toxic citric acid as a crosslinking agent. Cassava starch was first modified by carboxymethylation to improve its water absorbency property. The carboxymethyl cassava starch (CMCS) obtained was then crosslinked with citric acid at different concentrations and reaction times. The gel fraction of hydrogels increased progressively with increasing citric acid concentration. Free swelling capacity of hydrogels in de-ionized water, saline solution and buffers at various pHs as well as absorption under load were investigated. The results revealed that swelling behavior and mechanical characteristic of hydrogels depended on the citric acid concentration used in reaction. Increasing citric acid concentration resulted in hydrogels with stronger network but lower swelling and absorption capacity. The cassava starch hydrogels developed were sensitive to ionic strength and pH of surrounding medium, showing much reduced swelling capacity in saline salt solution and acidic buffers.

  13. Poly(propyleneimine) glycodendrimers non-covalently bind ATP in a pH- and salt-dependent manner - model studies for adenosine analogue drug delivery.

    PubMed

    Gorzkiewicz, Michał; Buczkowski, Adam; Appelhans, Dietmar; Voit, Brigitte; Pułaski, Łukasz; Pałecz, Bartłomiej; Klajnert-Maculewicz, Barbara

    2018-06-10

    Adenosine analogue drugs (such as fludarabine or cladribine) require transporter-mediated uptake into cells and subsequent phosphorylation for anticancer activity. Therefore, application of nanocarrier systems for direct delivery of active triphosphate forms has been proposed. Here, we applied isothermal titration calorimetry and zeta potential titration to determine the stoichiometry and thermodynamic parameters of interactions between 4th generation poly(propyleneimine) dendrimers (unmodified or sugar-modified for increased biocompatibility) and ATP as a model adenosine nucleotide. We showed that glycodendrimers have the ability to efficiently interact with nucleoside triphosphates and to form stable complexes via electrostatic interactions between the ionized phosphate and amino groups on the nucleotide and the dendrimer, respectively. The complexation process is spontaneous, enthalpy-driven and depends on buffer composition (strongest interactions in organic buffer) and pH (more binding sites in acidic pH). These properties allow us to consider maltose-modified dendrimers as especially promising carriers for adenosine analogues. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Refolding of laccase in dilution additive mode with copper-based ionic liquid.

    PubMed

    Bae, Sang-Woo; Ahn, Kihun; Koo, Yoon-Mo; Ha, Sung Ho

    2013-11-01

    Ionic liquids (ILs) are molten salts which do not crystallize at room temperature. Tunable physicochemical properties of ILs including hydrophobicity and polarity facilitate their applications in many biological processes. In this study, a copper-based IL was employed in order to enhance the refolding efficiency of laccase from Trametes versicolor which requires copper as a cofactor. When 1-ethyl-3-methylimidazolium trichlorocuprate ([EMIM][CuCl₃]) was added to refolding buffer instead of urea, the laccase refolding yield was improved more than 2.7 times compared to the conventional refolding buffer which contains urea. When the refolding of laccase was carried out at different temperatures (4, 25, and 37 °C), the highest refolding yield was obtained at 25 °C. At low temperature, two conflicting effects, i.e., suppression of the aggregate formation and decrease of folding rate, influence the protein refolding. In contrast, a copper-based IL did not enhance the refolding of lysozyme, a non-copper-containing protein. From these results, we can conclude that this copper-based IL, [EMIM][CuCl₃], was exclusively effective on the refolding process of a copper-containing protein.

  15. Polyamine binding to proteins in oat and Petunia protoplasts.

    PubMed

    Mizrahi, Y; Applewhite, P B; Galston, A W

    1989-01-01

    Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants. To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa). In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action. In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH. The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS. Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons. Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.

  16. Emulsion stabilizing capacity of intact starch granules modified by heat treatment or octenyl succinic anhydride.

    PubMed

    Timgren, Anna; Rayner, Marilyn; Dejmek, Petr; Marku, Diana; Sjöö, Malin

    2013-03-01

    Starch granules are an interesting stabilizer candidate for food-grade Pickering emulsions. The stabilizing capacity of seven different intact starch granules for making oil-in-water emulsions has been the topic of this screening study. The starches were from quinoa; rice; maize; waxy varieties of rice, maize, and barley; and high-amylose maize. The starches were studied in their native state, heat treated, and modified by octenyl succinic anhydride (OSA). The effect of varying the continuous phase, both with and without salt in a phosphate buffer, was also studied. Quinoa, which had the smallest granule size, had the best capacity to stabilize oil drops, especially when the granules had been hydrophobically modified by heat treatment or by OSA. The average drop diameter (d 32) in these emulsions varied from 270 to 50 μm, where decreasing drop size and less aggregation was promoted by high starch concentration and absence of salt in the system. Of all the starch varieties studied, quinoa had the best overall emulsifying capacity, and OSA modified quinoa starch in particular. Although the size of the drops was relatively large, the drops themselves were in many instances extremely stable. In the cases where the system could stabilize droplets, even when they were so large that they were visible to the naked eye, they remained stable and the measured droplet sizes after 2 years of storage were essentially unchanged from the initial droplet size. This somewhat surprising result has been attributed to the thickness of the adsorbed starch layer providing steric stabilization. The starch particle-stabilized Pickering emulsion systems studied in this work has potential practical application such as being suitable for encapsulation of ingredients in food and pharmaceutical products.

  17. Improved Epitaxy and Surface Morphology in YBa2Cu3Oy Thin Films Grown on Double Buffered Si Wafers

    NASA Astrophysics Data System (ADS)

    Gao, J.; Kang, L.; Wong, H. Y.; Cheung, Y. L.; Yang, J.

    Highly epitaxial thin films of YBCO have been obtained on silicon wafers using a Eu2CuO4/YSZ (yttrium-stabilized ZrO2) double buffer. Our results showed that application of such a double buffer can significantly enhance the epitaxy of grown YBCO. It also leads to an excellent surface morphology. The average surface roughness was found less than 5 nm in a large range. The results of X-ray small angle reflection and positron spectroscpy demonstrate a very clear and flat interface between YBCO and buffer layers. The Eu2CuO4/YSZ double buffer could be promising for coating high-TC superconducting films on various reactive substrates.

  18. INCREASED RENAL OXIDATIVE STRESS IN SALT-SENSITIVE HUMAN GRK4γ486V TRANSGENIC MICE

    PubMed Central

    Diao, Zhenyu; Asico, Laureano D.; Villar, Van Anthony M.; Zheng, Xiaoxu; Cuevas, Santiago; Armando, Ines; Jose, Pedro A.; Wang, Xiaoyan

    2017-01-01

    We tested the hypothesis that salt-sensitive hypertension is caused by renal oxidative stress by measuring the blood pressure and reactive oxygen species-related proteins in the kidneys of human G protein-coupled receptor kinase 4γ (hGRK4γ) 486V transgenic mice and non-transgenic (Non-T) littermates on normal and high salt diets. High salt diet increased the blood pressure, associated with impaired sodium excretion, in hGRK4γ486V mice. Renal expressions of NOX isoforms were similar in both strains on normal salt diet but NOX2 was decreased by high salt diet to a greater extent in Non-T than hGRK4γ486V mice. Renal HO-2, but not HO-1, protein was greater in hGRK4γ486V than Non-T mice on normal salt diet and normalized by high salt diet. On normal salt diet, renal CuZnSOD and ECSOD proteins were similar but renal MnSOD was lower in hGRK4γ486V than Non-T mice and remained low on high salt diet. High salt diet decreased renal CuZnSOD in hGRK4γ486V but not Non-T mice and decreased renal ECSOD to a greater extent in hGRK4γ486V than Non-T mice. Renal SOD activity, superoxide production, and NOS3 protein were similar in two strains on normal salt diet. However, high salt diet decreased SOD activity and NOS3 protein and increased superoxide production in hGRK4γ486V mice but not in Non-T mice. High salt diet also increased urinary 8-isoprostane and 8-hydroxydeoxyguanosine to a greater extent in hGRK4γ486V than Non-T mice. hGRK4γwild-type mice were normotensive and hGRK4γ142V mice were hypertensive but both were salt-resistant and in normal redox balance. Chronic tempol treatment partially prevented the salt-sensitivity of hGRK4γ486V mice. Thus, hGRK4γ486V causes salt-sensitive hypertension due, in part, to defective renal antioxidant mechanisms. PMID:28189851

  19. High salt reduces the activation of IL-4- and IL-13-stimulated macrophages.

    PubMed

    Binger, Katrina J; Gebhardt, Matthias; Heinig, Matthias; Rintisch, Carola; Schroeder, Agnes; Neuhofer, Wolfgang; Hilgers, Karl; Manzel, Arndt; Schwartz, Christian; Kleinewietfeld, Markus; Voelkl, Jakob; Schatz, Valentin; Linker, Ralf A; Lang, Florian; Voehringer, David; Wright, Mark D; Hubner, Norbert; Dechend, Ralf; Jantsch, Jonathan; Titze, Jens; Müller, Dominik N

    2015-11-02

    A high intake of dietary salt (NaCl) has been implicated in the development of hypertension, chronic inflammation, and autoimmune diseases. We have recently shown that salt has a proinflammatory effect and boosts the activation of Th17 cells and the activation of classical, LPS-induced macrophages (M1). Here, we examined how the activation of alternative (M2) macrophages is affected by salt. In stark contrast to Th17 cells and M1 macrophages, high salt blunted the alternative activation of BM-derived mouse macrophages stimulated with IL-4 and IL-13, M(IL-4+IL-13) macrophages. Salt-induced reduction of M(IL-4+IL-13) activation was not associated with increased polarization toward a proinflammatory M1 phenotype. In vitro, high salt decreased the ability of M(IL-4+IL-13) macrophages to suppress effector T cell proliferation. Moreover, mice fed a high salt diet exhibited reduced M2 activation following chitin injection and delayed wound healing compared with control animals. We further identified a high salt-induced reduction in glycolysis and mitochondrial metabolic output, coupled with blunted AKT and mTOR signaling, which indicates a mechanism by which NaCl inhibits full M2 macrophage activation. Collectively, this study provides evidence that high salt reduces noninflammatory innate immune cell activation and may thus lead to an overall imbalance in immune homeostasis.

  20. Does salt have a permissive role in the induction of puberty?

    PubMed

    Pitynski, Dori; Flynn, Francis W; Skinner, Donal C

    2015-10-01

    Puberty is starting earlier than ever before and there are serious physiological and sociological implications as a result of this development. Current research has focused on the potential role of high caloric, and commensurate high adiposity, contributions to early puberty. However, girls with normal BMI also appear to be initiating puberty earlier. Westernized diets, in addition to being high in fat and sugar, are also high in salt. To date, no research has investigated a link between elevated salt and the reproductive axis. We hypothesize that a high salt diet can result in an earlier onset of puberty through three mechanisms that are not mutually exclusive. (1) High salt activates neurokinin B, a hormone that is involved in both the reproductive axis and salt regulation, and this induces kisspeptin release and ultimate activation of the reproductive axis. (2) Vasopressin released in response to high salt acts on vasopressin receptors expressed on kisspeptin neurons in the anteroventral periventricular nucleus, thereby stimulating gonadotropin releasing hormone and subsequently luteinizing hormone secretion. (3) Salt induces metabolic changes that affect the reproductive axis. Specifically, salt acts indirectly to modulate adiposity, ties in with the obesity epidemic, and further compounds the pathologic effects of obesity. Our overall hypothesis offers an additional cause behind the induction of puberty and provides testable postulates to determine the mechanism of potential salt-mediated affects on puberty. Copyright © 2015. Published by Elsevier Ltd.

  1. Does salt have a permissive role in the induction of puberty?

    PubMed Central

    Pitynski, Dori; Flynn, Francis W.; Skinner, Donal C.

    2017-01-01

    Puberty is starting earlier than ever before and there are serious physiological and sociological implications as a result of this development. Current research has focused on the potential role of high caloric, and commensurate high adiposity, contributions to early puberty. However, girls with normal BMI also appear to be initiating puberty earlier. Westernized diets, in addition to being high in fat and sugar, are also high in salt. To date, no research has investigated a link between elevated salt and the reproductive axis. We hypothesize that a high salt diet can result in an earlier onset of puberty through three mechanisms that are not mutually exclusive. (1) High salt activates neurokinin B, a hormone that is involved in both the reproductive axis and salt regulation, and this induces kisspeptin release and ultimate activation of the reproductive axis. (2) Vasopressin released in response to high salt acts on vasopressin receptors expressed on kisspeptin neurons in the anteroventral periventricular nucleus, thereby stimulating gonadotropin releasing hormone and subsequently luteinizing hormone secretion. (3) Salt induces metabolic changes that affect the reproductive axis. Specifically, salt acts indirectly to modulate adiposity, ties in with the obesity epidemic, and further compounds the pathologic effects of obesity. Our overall hypothesis offers an additional cause behind the induction of puberty and provides testable postulates to determine the mechanism of potential salt-mediated affects on puberty. PMID:26190310

  2. Dietary salt loading increases nitric oxide synthesis in transgenic mice overexpressing sodium-proton exchanger.

    PubMed

    Kiraku, J; Nakamura, T; Sugiyama, T; Takahashi, N; Kuro-o, M; Fujii, J; Nagai, R

    1999-06-01

    We studied the role of nitric oxide (NO) synthesis in amelioration of blood pressure elevation during dietary salt loading in transgenic mice overexpressing sodium proton exchanger. Systolic blood pressure rose after starting salt loading only in the high-salt group of transgenic mice. However, this elevation of blood pressure was not continued. Urinary excretion of inorganic nitrite and nitrate in the high-salt group of transgenic mice was significantly higher than in the high-salt group of control mice. These results suggest that increased NO synthesis in response to salt loading is one of the anti-hypertensive mechanisms in transgenic mice overexpressing sodium proton exchanger.

  3. Improved high temperature integration of Al{sub 2}O{sub 3} on MoS{sub 2} by using a metal oxide buffer layer

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

    Son, Seokki; Choi, Moonseok; Kim, Dohyung

    2015-01-12

    We deposited a metal oxide buffer layer before atomic layer deposition (ALD) of Al{sub 2}O{sub 3} onto exfoliated molybdenum disulfide (MoS{sub 2}) in order to accomplish enhanced integration. We demonstrate that even at a high temperature, functionalization of MoS{sub 2} by means of a metal oxide buffer layer can effectively provide nucleation sites for ALD precursors, enabling much better surface coverage of Al{sub 2}O{sub 3}. It is shown that using a metal oxide buffer layer not only allows high temperature ALD process, resulting in highly improved quality of Al{sub 2}O{sub 3}/MoS{sub 2} interface, but also leaves MoS{sub 2} intact.

  4. Protective effect of dietary potassium against vascular injury in salt-sensitive hypertension.

    PubMed

    Kido, Makiko; Ando, Katsuyuki; Onozato, Maristela L; Tojo, Akihiro; Yoshikawa, Masahiro; Ogita, Teruhiko; Fujita, Toshiro

    2008-02-01

    Hypertensive cardiovascular damage is accelerated by salt loading but counteracted by dietary potassium supplementation. We suggested recently that antioxidant actions of potassium contribute to protection against salt-induced cardiac dysfunction. Therefore, we examined whether potassium supplementation ameliorated cuff-induced vascular injury in salt-sensitive hypertension via suppression of oxidative stress. Four-week-old Dahl salt-sensitive rats were fed a normal-salt (0.3% NaCl), high-salt (8% NaCl), or high-salt plus high-potassium (8% KCl) diet for 5 weeks, and some of the rats fed a high-salt diet were also given antioxidants. One week after the start of the treatments, a silicone cuff was implanted around the femoral artery. Examination revealed increased cuff-induced neointimal proliferation with adventitial macrophage infiltration in arteries from salt-loaded Dahl salt-sensitive rats compared with that in arteries from non-salt-loaded animals (intima/media ratio: 0.471+/-0.070 versus 0.302+/-0.037; P<0.05), associated with regional superoxide overproduction and reduced nicotinamide-adenine dinucleotide phosphate oxidase activation and mRNA overexpression. On the other hand, simultaneous potassium supplementation attenuated salt-induced neointimal hyperplasia (intima/media ratio: 0.205+/-0.012; P<0.001), adventitial macrophage infiltration, superoxide overproduction, and reduced nicotinamide-adenine dinucleotide phosphate oxidase activation and overexpression. Antioxidants, which decrease vascular oxidative stress, also reduced neointima formation induced by salt excess. In conclusion, high-potassium diets seems to have a protective effect against the development of vascular damage induced by salt loading mediated, at least in part, through suppression of the production of reactive oxygen species probably generated by reduced nicotinamide-adenine dinucleotide phosphate oxidase.

  5. High-Salt Diet Has a Certain Impact on Protein Digestion and Gut Microbiota: A Sequencing and Proteome Combined Study.

    PubMed

    Wang, Chao; Huang, Zixin; Yu, Kequan; Ding, Ruiling; Ye, Keping; Dai, Chen; Xu, Xinglian; Zhou, Guanghong; Li, Chunbao

    2017-01-01

    High-salt diet has been considered to cause health problems, but it is still less known how high-salt diet affects gut microbiota, protein digestion, and passage in the digestive tract. In this study, C57BL/6J mice were fed low- or high-salt diets (0.25 vs. 3.15% NaCl) for 8 weeks, and then gut contents and feces were collected. Fecal microbiota was identified by sequencing the V4 region of 16S ribosomal RNA gene. Proteins and digested products of duodenal, jejunal, cecal, and colonic contents were identified by LC-MS-MS. The results indicated that the high-salt diet increased Firmicutes/Bacteroidetes ratio, the abundances of genera Lachnospiraceae and Ruminococcus ( P < 0.05), but decreased the abundance of Lactobacillus ( P < 0.05). LC-MS-MS revealed a dynamic change of proteins from the diet, host, and gut microbiota alongside the digestive tract. For dietary proteins, high-salt diet seemed not influence its protein digestion and absorption. For host proteins, 20 proteins of lower abundance were identified in the high-salt diet group in duodenal contents, which were involved in digestive enzymes and pancreatic secretion. However, no significant differentially expressed proteins were detected in jejunal, cecal, and colonic contents. For bacterial proteins, proteins secreted by gut microbiota were involved in energy metabolism, sodium transport, and protein folding. Five proteins (cytidylate kinase, trigger factor, 6-phosphogluconate dehydrogenase, transporter, and undecaprenyl-diphosphatase) had a higher abundance in the high-salt diet group than those in the low-salt group, while two proteins (acetylglutamate kinase and PBSX phage manganese-containing catalase) were over-expressed in the low-salt diet group than in the high-salt group. Consequently, high-salt diet may alter the composition of gut microbiota and has a certain impact on protein digestion.

  6. High-Salt Diet Has a Certain Impact on Protein Digestion and Gut Microbiota: A Sequencing and Proteome Combined Study

    PubMed Central

    Wang, Chao; Huang, Zixin; Yu, Kequan; Ding, Ruiling; Ye, Keping; Dai, Chen; Xu, Xinglian; Zhou, Guanghong; Li, Chunbao

    2017-01-01

    High-salt diet has been considered to cause health problems, but it is still less known how high-salt diet affects gut microbiota, protein digestion, and passage in the digestive tract. In this study, C57BL/6J mice were fed low- or high-salt diets (0.25 vs. 3.15% NaCl) for 8 weeks, and then gut contents and feces were collected. Fecal microbiota was identified by sequencing the V4 region of 16S ribosomal RNA gene. Proteins and digested products of duodenal, jejunal, cecal, and colonic contents were identified by LC-MS-MS. The results indicated that the high-salt diet increased Firmicutes/Bacteroidetes ratio, the abundances of genera Lachnospiraceae and Ruminococcus (P < 0.05), but decreased the abundance of Lactobacillus (P < 0.05). LC-MS-MS revealed a dynamic change of proteins from the diet, host, and gut microbiota alongside the digestive tract. For dietary proteins, high-salt diet seemed not influence its protein digestion and absorption. For host proteins, 20 proteins of lower abundance were identified in the high-salt diet group in duodenal contents, which were involved in digestive enzymes and pancreatic secretion. However, no significant differentially expressed proteins were detected in jejunal, cecal, and colonic contents. For bacterial proteins, proteins secreted by gut microbiota were involved in energy metabolism, sodium transport, and protein folding. Five proteins (cytidylate kinase, trigger factor, 6-phosphogluconate dehydrogenase, transporter, and undecaprenyl-diphosphatase) had a higher abundance in the high-salt diet group than those in the low-salt group, while two proteins (acetylglutamate kinase and PBSX phage manganese-containing catalase) were over-expressed in the low-salt diet group than in the high-salt group. Consequently, high-salt diet may alter the composition of gut microbiota and has a certain impact on protein digestion. PMID:29033907

  7. Characteristic of molten fluoride salt system LiF-BeF2 (Flibe) and LiF-NaF-KF (Flinak) as coolant and fuel carrier in molten salt reactor (MSR)

    NASA Astrophysics Data System (ADS)

    Bahri, Che Nor Aniza Che Zainul; Al-Areqi, Wadee'ah Mohd; Ruf, Mohd'Izzat Fahmi Mohd; Majid, Amran Ab.

    2017-01-01

    Interest of fluoride salts have recently revived due to the high temperature application in nuclear reactors. Molten Salt Reactor (MSR) was designed to operate at high temperature in range 700 - 800°C and its fuel is dissolved in a circulating molten fluoride salt mixture. Molten fluoride salts are stable at high temperature, have good heat transfer properties and can dissolve high concentration of actinides and fission product. The aim of this paper was to discuss the physical properties (melting temperature, density and heat capacity) of two systems fluoride salt mixtures i.e; LiF-BeF2 (Flibe) and LiF-NaF-KF (Flinak) in terms of their application as coolant and fuel solvent in MSR. Both of these salts showed almost same physical properties but different applications in MSR. The advantages and the disadvantages of these fluoride salt systems will be discussed in this paper.

  8. Nitric oxide contributes to high-salt perception in a blood-sucking insect model.

    PubMed

    Cano, Agustina; Pontes, Gina; Sfara, Valeria; Anfossi, Diego; Barrozo, Romina B

    2017-11-14

    In all organisms, salts produce either appetitive or aversive responses depending on the concentration. While low-salt concentration in food elicits positive responses to ingest, high-salt triggers aversion. Still the mechanisms involved in this dual behavior have just started to be uncovered in some organisms. In Rhodnius prolixus, using pharmacological and behavioral assays, we demonstrated that upon high-salt detection in food a nitric oxide (NO) dependent cascade is activated. This activation involves a soluble guanylate cyclase (sGC) and the production of cyclic guanosine monophosphate (cGMP). Thus, appetitive responses to low-salt diets turn to aversion whenever this cascade is activated. Conversely, insects feed over aversive high-salt solutions when it is blocked by reducing NO levels or by affecting the sGC activity. The activation of NO/sGC/cGMP cascade commands the avoidance feeding behavior in R. prolixus. Investigations in other insect species should examine the possibility that high-salt aversion is mediated by NO/sSG/cGMP signaling.

  9. Granite disposal of U.S. high-level radioactive waste.

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

    Freeze, Geoffrey A.; Mariner, Paul E.; Lee, Joon H.

    This report evaluates the feasibility of disposing U.S. high-level radioactive waste in granite several hundred meters below the surface of the earth. The U.S. has many granite formations with positive attributes for permanent disposal. Similar crystalline formations have been extensively studied by international programs, two of which, in Sweden and Finland, are the host rocks of submitted or imminent repository license applications. This report is enabled by the advanced work of the international community to establish functional and operational requirements for disposal of a range of waste forms in granite media. In this report we develop scoping performance analyses, basedmore » on the applicable features, events, and processes (FEPs) identified by international investigators, to support generic conclusions regarding post-closure safety. Unlike the safety analyses for disposal in salt, shale/clay, or deep boreholes, the safety analysis for a mined granite repository depends largely on waste package preservation. In crystalline rock, waste packages are preserved by the high mechanical stability of the excavations, the diffusive barrier of the buffer, and favorable chemical conditions. The buffer is preserved by low groundwater fluxes, favorable chemical conditions, backfill, and the rigid confines of the host rock. An added advantage of a mined granite repository is that waste packages would be fairly easy to retrieve, should retrievability be an important objective. The results of the safety analyses performed in this study are consistent with the results of comprehensive safety assessments performed for sites in Sweden, Finland, and Canada. They indicate that a granite repository would satisfy established safety criteria and suggest that a small number of FEPs would largely control the release and transport of radionuclides. In the event the U.S. decides to pursue a potential repository in granite, a detailed evaluation of these FEPs would be needed to inform site selection and safety assessment.« less

  10. Protein-protein interactions during high-moisture extrusion for fibrous meat analogues and comparison of protein solubility methods using different solvent systems.

    PubMed

    Liu, KeShun; Hsieh, Fu-Hung

    2008-04-23

    Soy protein, mixed with gluten and starch, was extruded into fibrous meat analogues under high-moisture and high-temperature conditions. The protein solubility of samples collected at different extruder zones and extrudates made with different moistures was determined by 11 extraction solutions consisting of 6 selective reagents and their combinations: phosphate salts, urea, DTT, thiourea, Triton X-100, and CHAPS. Protein solubility by most extractants showed decreasing patterns as the material passed through the extruder, but the solution containing all 6 reagents, known as isoelectric focus (IEF) buffer, solubilized the highest levels and equal amounts of proteins in all samples, indicating that there are no other covalent bonds involved besides disulfide bonds. With regard to relative importance between disulfide bonds and non-covalent interactions, different conclusions could be made from protein solubility patterns, depending on the type of extracting systems and a baseline used for comparison. The observation points out pitfalls and limitation of current protein solubility methodology and explains why controversy exists in the literature. Using the IEF buffer system with omission of one or more selective reagents is considered to be the right methodology to conduct protein solubility study and thus recommended. Results obtained with this system indicate that disulfide bonding plays a more important role than non-covalent bonds in not only holding the rigid structure of extrudates but also forming fibrous texture. The sharpest decrease in protein solubility occurred when the mix passed through the intermediate section of the extruder barrel, indicating formation of new disulfide bonds during the stage of dramatic increase in both temperature and moisture. After this stage, although the physical form of the product might undergo change and fiber formation might occur as it passed through the cooling die, the chemical nature of the product did not change significantly.

  11. Effect of salt intake on beat-to-beat blood pressure nonlinear dynamics and entropy in salt-sensitive versus salt-protected rats.

    PubMed

    Fares, Souha A; Habib, Joseph R; Engoren, Milo C; Badr, Kamal F; Habib, Robert H

    2016-06-01

    Blood pressure exhibits substantial short- and long-term variability (BPV). We assessed the hypothesis that the complexity of beat-to-beat BPV will be differentially altered in salt-sensitive hypertensive Dahl rats (SS) versus rats protected from salt-induced hypertension (SSBN13) maintained on high-salt versus low-salt diet. Beat-to-beat systolic and diastolic BP series from nine SS and six SSBN13 rats (http://www.physionet.org) were analyzed following 9 weeks on low salt and repeated after 2 weeks on high salt. BP complexity was quantified by detrended fluctuation analysis (DFA), short- and long-range scaling exponents (αS and αL), sample entropy (SampEn), and traditional standard deviation (SD) and coefficient of variation (CV(%)). Mean systolic and diastolic BP increased on high-salt diet (P < 0.01) particularly for SS rats. SD and CV(%) were similar across groups irrespective of diet. Salt-sensitive and -protected rats exhibited similar complexity indices on low-salt diet. On high salt, (1) SS rats showed increased scaling exponents or smoother, systolic (P = 0.007 [αL]) and diastolic (P = 0.008 [αL]) BP series; (2) salt-protected rats showed lower SampEn (less complex) systolic and diastolic BP (P = 0.046); and (3) compared to protected SSBN13 rats, SS showed higher αL for systolic (P = 0.01) and diastolic (P = 0.005) BP Hypertensive SS rats are more susceptible to high salt with a greater rise in mean BP and reduced complexity. Comparable mean pressures in sensitive and protective rats when on low-salt diet coupled with similar BPV dynamics suggest a protective role of low-salt intake in hypertensive rats. This effect likely reflects better coupling of biologic oscillators. © 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

  12. Leakage effects in n-GaAs MESFET with n-GaAs buffer layer

    NASA Technical Reports Server (NTRS)

    Wang, Y. C.; Bahrami, M.

    1983-01-01

    Whereas improvement of the interface between the active layer and the buffer layer has been demonstrated, the leakage effects can be important if the buffer layer resistivity is not sufficiently high and/or the buffer layer thickness is not sufficiently small. It was found that two buffer leakage currents exist from the channel under the gate to the source and from drain to the channel in addition to the buffer leakage resistance between drain and source. It is shown that for a 1 micron gate-length n-GaAs MESFET, if the buffer layer resistivity is 12 OHM-CM and the buffer layer thickness h is 2 microns, the performance of the device degrades drastically. It is suggested that h should be below 2 microns.

  13. Structural and thermodynamic analysis of the hetero-association of theophylline with aromatic drug molecules

    NASA Astrophysics Data System (ADS)

    Andrejuk, D. D.; Hernandez Santiago, A. A.; Khomich, V. V.; Voronov, V. K.; Davies, D. B.; Evstigneev, M. P.

    2008-10-01

    The hetero-association of theophylline (THP) with other biologically-active aromatic molecules ( e.g. the anti-cancer drugs daunomycin and novantrone, the antibiotic norfloxacin, the vitamin flavin-mononucleotide and two mutagens ethidium bromide and proflavine) has been studied by NMR in aqueous-salt solution (0.1 M Na-phosphate buffer, p D 7.1). It was found that THP shows an essentially similar hetero-association ability as caffeine (CAF) towards aromatic drugs, except for novantrone (NOV), which has much less affinity to THP than CAF as a result of energetically unfavourable orthogonal orientation of the chromophores of THP and NOV in the hetero-complex.

  14. Explaining variability in sodium intake through oral sensory phenotype, salt sensation and liking

    PubMed Central

    Hayes, John E.; Sullivan, Bridget S.; Duffy, Valerie B.

    2010-01-01

    Our sodium-rich food supply compels investigation of how variation in salt sensation influences liking and intake of high-sodium foods. While supertasters (those with heightened propylthiouracil (PROP) bitterness or taste papillae number) report greater saltiness from concentrated salt solutions, the non-taster/supertaster effect on sodium intake is unclear. We assessed taster effects on salt sensation, liking and intake among 87 healthy adults (45 men). PROP bitterness showed stronger associations with perceived saltiness in foods than did papillae number. Supertasters reported: greater saltiness in chips/pretzels and broth at levels comparable to regular-sodium products; greater sensory and/or liking changes to growing sodium concentration in cheeses (where sodium ions mask bitterness) and broths; and less frequently salting foods. PROP effects were attenuated in women. Compared with men, women reported more saltiness from high-sodium foods and greater liking for broth at salt levels comparable to regular-sodium products. Across men and women, Structural Equation Models showed PROP and papillae number independently explained variability in consuming high-sodium foods by impacting salt sensation and/or liking. PROP supertasters reported greater changes in sensation when more salt was added to broth, which then associated with greater changes in broth liking, and finally with more frequent high-sodium food intake. Greater papillae number was associated with less frequent high-sodium food intake via reduced liking for high-fat/high-sodium foods. In summary, variation in sensations from salt was associated with differences in hedonic responses to high-sodium foods and thus sodium intake. Despite adding less salt, PROP supertasters consumed more sodium through food, as salt was more important to preference, both for its salty taste and masking of bitterness. PMID:20380843

  15. Study of the effects of GaN buffer layer quality on the dc characteristics of AlGaN/GaN high electron mobility transistors

    DOE PAGES

    Ahn, Shihyun; Zhu, Weidi; Dong, Chen; ...

    2015-04-21

    Here we studied the effect of buffer layer quality on dc characteristics of AlGaN/GaN high electron mobility (HEMTs). AlGaN/GaN HEMT structures with 2 and 5 μm GaN buffer layers on sapphire substrates from two different vendors with the same Al concentration of AlGaN were used. The defect densities of HEMT structures with 2 and 5 μm GaN buffer layer were 7 × 10 9 and 5 × 10 8 cm ₋2, respectively, as measured by transmission electron microscopy. There was little difference in drain saturation current or in transfer characteristics in HEMTs on these two types of buffer. However, theremore » was no dispersion observed on the nonpassivated HEMTs with 5 μm GaN buffer layer for gate-lag pulsed measurement at 100 kHz, which was in sharp contrast to the 71% drain current reduction for the HEMT with 2 μm GaN buffer layer.« less

  16. New MBE buffer for micron- and quarter-micron-gateGaAs MESFETs

    NASA Technical Reports Server (NTRS)

    1988-01-01

    A new buffer layer has been developed that eliminates backgating in GaAs MESFETs and substantially reduces short-channel effects in GaAs MESFETs with 0.27-micron-long gates. The new buffer is grown by molecular beam epitaxy (MBE) at a substrate temperature of 200 C using Ga and As sub 4 beam fluxes. The buffer is crystalline, highly resistive, optically inactive, and can be overgrown with high quality GaAs. GaAs MESFETs with a gate length of 0.27 microns that incorporate the new buffer show improved dc and RF properties in comparison with a similar MESFET with a thin undoped GaAs buffer. To demonstrate the backgating performance improvement afforded by the new buffer, MESFETs were fabricated using a number of different buffer layers and structures. A schematic cross section of the MESFET structure used in this study is shown. The measured gate length, gate width, and source-drain spacing of this device are 2,98, and 5.5 microns, respectively. An ohmic contact, isolated from the MESFET by mesa etching, served as the sidegate. The MESFETs were fabricated in MBE n-GaAs layers grown on the new buffer and also in MBE n-GaAs layers grown on buffer layers of undoped GaAs, AlGaAs, and GaAs/AlGaAs superlattices. All the buffer layers were grown by MBE and are 2 microns thick. The active layer is doped to approximately 2 x 10 to the 17th/cu cm with silicon and is 0.3 microns thick.

  17. Power plant I - Fused salt

    NASA Astrophysics Data System (ADS)

    Roche, M.

    A solar thermal power plant using fused salt as the heat transfer fluid for steam power generation is analyzed for the feasibility of economic operation. The salt is also stored in a tank reservoir for maintaining the primary heat loop at temperatures high enough for the salts to remain liquid, and also to provide reserve power for the steam generator. Initial studies were with eutectic (hitec) salt comprising Na, KOH, and nitrites melting at 146 C, and further studies were performed employing draw salt, which has no nitrite, is more stable at high temperature, and melts at 225 C. The use of draw salt was found to allow a 5 percent reduction in storage capacity. Further examinations of the effects of the hitec salts on corrosion and composition degradation at high temperatures are indicated. The molten salt system is projected to offer an efficiency of 26 percent.

  18. Buffer layers for high-Tc thin films on sapphire

    NASA Technical Reports Server (NTRS)

    Wu, X. D.; Foltyn, S. R.; Muenchausen, R. E.; Cooke, D. W.; Pique, A.; Kalokitis, D.; Pendrick, V.; Belohoubek, E.

    1992-01-01

    Buffer layers of various oxides including CeO2 and yttrium-stabilized zirconia (YSZ) have been deposited on R-plane sapphire. The orientation and crystallinity of the layers were optimized to promote epitaxial growth of YBa2Cu3O(7-delta) (YBCO) thin films. An ion beam channeling minimum yield of about 3 percent was obtained in the CeO2 layer on sapphire, indicating excellent crystallinity of the buffer layer. Among the buffer materials used, CeO2 was found to be the best one for YBCO thin films on R-plane sapphire. High Tc and Jc were obtained in YBCO thin films on sapphire with buffer layers. Surface resistances of the YBCO films were about 4 mOmega at 77 K and 25 GHz.

  19. Geochemistry of metal-rich brines from central Mississippi Salt Dome basin, U.S.A.

    USGS Publications Warehouse

    Kharaka, Y.K.; Maest, A.S.; Carothers, W.W.; Law, L.M.; Lamothe, P.J.; Fries, T.L.

    1987-01-01

    Oil-field brines are the most favored ore-forming solutions for the sediment-hosted Mississippi Valley-type ore deposits. Detailed inorganic and organic chemical and isotope analyses of water and gas samples from six oil fields in central Mississippi, one of the very few areas with high metal brines, were conducted to study the inorganic and organic complexes responsible for the high concentrations of these metals. The samples were obtained from production zones consisting of sandstone and limestone that range in depth from 1900 to 4000 m (70-120??C) and in age from Late Cretaceous to Late Jurassic. Results show that the waters are dominantly bittern brines related to the Louann Salt. The brines have extremely high salinities that range from 160,000 to 320,000 mg/l total dissolved solids and are NaCaCl-type waters with very high concentrations of Ca (up to 48,000 mg/l) and other alkaline-earth metals, but with low concentrations of aliphatic acid anions. The concentrations of metals in many water samples are very high, reaching values of 70 mg/l for Pb, 245 mg/l for Zn, 465 mg/l for Fe and 210 mg/l for Mn. The samples with high metal contents have extremely low concentrations (<0.02 mg/l) of H2S. Samples obtained from the Smackover Formation (limestone) have low metal contents that are more typical of oil-field waters, but have very high concentrations (up to 85 mg/l) of H2S. Computations with the geochemical code SOLMINEQ.87 give the following results: (1) both Pb and Zn are present predominantly as aqueous chloride complexes (mainly as PbCl42- and ZnCl42-, respectively); (2) the concentrations of metals complexed with short-chained aliphatic acid anions and reduced S species are minor; (3) organic acid anions are important in controlling the concentrations of metals because they affect the pH and buffer capacity of the waters at subsurface conditions; and (4) galena and sphalerite solubilities control the concentrations of Pb and Zn in these waters. ?? 1988.

  20. Should we eat less salt?

    PubMed

    Delahaye, François

    2013-05-01

    High blood pressure is a major cardiovascular risk factor. There is overwhelming evidence that high salt consumption is a major cause of increased blood pressure. There is also a link between high salt consumption and risk of stroke, left ventricular hypertrophy, renal disease, obesity, renal stones and stomach cancer. Reducing salt consumption leads to a decrease in blood pressure and the incidence of cardiovascular disease. There are no deleterious effects associated with reducing salt consumption and it is also very cost-effective. Many organizations and state governments have issued recommendations regarding the suitable amount of salt consumption. In France, the objective is a salt consumption<8g/day in men and<6.5g/day in women and children. As 80% of consumed salt comes from manufactured products in developed countries, reduction of salt consumption requires the participation of the food industry. The other tool is consumer information and education. Salt consumption has already decreased in France in recent years, but efforts must continue. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  1. The effects of temperature and NaCl concentration on tetragonal lysozyme face growth rates

    NASA Technical Reports Server (NTRS)

    Forsythe, Elizabeth; Pusey, Marc Lee

    1994-01-01

    Measurements were made of the (110) and (101) face growth rates of the tetragonal form of hen egg white lysozyme at 0.1M sodium acetate buffer, pH 4.0, from 4 to 22 C and with 3.0%, 5.0%, and 7.0% NaCl used as the precipitating salt. The data were collected at supersaturation ratios ranging from approximately 4 to approximately 63. Both decreasing temperature and increasing salt concentrations shifted plots of the growth rate versus C/C(sat) to the right, i.e. higher supersaturations were required for comparable growth rates. The observed trends in the growth data are counter to those expected from the solubility data. If tetragonal lysozyme crystal growth is by addition of ordered aggregates from the solution, then the observed growth data could be explained as a result of the effects of lowered temperature and increased salt concentration on the kinetics and equilibrium processes governing protein-protein interactions in solution. The data indicate that temperature would be a more tractable means of controlling the growth rate for tetragonal lysozyme crystals contrary to the usual practice in, e.g., vapor diffusion protein crystal growth, where both the precipitant and protein concentrations are simultaneously increased. However, the available range for control is dependent upon the protein concentration, with the greatest growth rate control being at the lower concentration.

  2. Retention of bile salts in micellar electrokinetic chromatography: relation of capacity factor to octanol-water partition coefficient and critical micellar concentration.

    PubMed

    Lucangioli, S E; Carducci, C N; Tripodi, V P; Kenndler, E

    2001-12-25

    The capacity factors of 16 anionic cholates (from six bile salts, including their glyco- and tauro-conjugates) were determined in a micellar electrokinetic chromatography (MEKC) system consisting of buffer, pH 7.5 (phosphate-boric acid; 20 mmol/l) with 50 mmol/l sodium dodecyl sulfate (SDS) as micelle former and 10% acetonitrile as organic modifier. The capacity factors of the fully dissociated, negatively charged analytes (ranging between 0.2 and 60) were calculated from their mobilities, with a reference background electrolyte (BGE) without SDS representing "free" solution. For comparison, the capacity factors were derived for a second reference BGE where the SDS concentration (5 mmol/l) is close to the critical micellar concentration (CMC). The capacity factors are compared with the logarithm of the octanol-water partition coefficient, log Pow, as measure for lipophilicity. Clear disagreement between these two parameters is found especially for epimeric cholates with the hydroxy group in position 7. In contrast, fair relation between the capacity factor of the analytes and their CMC is observed both depending strongly on the orientation of the OH groups, and tauro-conjugation as well. In this respect the retention behaviour of the bile salts in MEKC seems to reflect their role as detergents in living systems, and might serve as model parameter beyond lipophilicity.

  3. Physical chemistry and evolution of salt tolerance in halobacteria

    NASA Technical Reports Server (NTRS)

    Lanyi, J. K.

    1980-01-01

    The cellular constituents of extremely halophilic bacteria not only tolerate high salt concentration, but in many cases require it for optical functioning. The characteristics affected by salt include enzyme activity, stability, allosteric regulation, conformation and subunit association. The salt effects are of two major kinds: electrostatic shielding of negative charges by cations at low salt concentration, and hydrophobic stabilization by salting-out type salts at high salt concentration. The composition of halobacterial proteins shows an excess of acidic amino acids and a deficiency of nonpolar amino acids, which accounts for these effects. Since the cohesive forces are weaker and the repulsing forces are stronger in these proteins, preventing aggregation in salt, these structures are no longer suited for functioning in the absence of high salt concentrations. Unlike these nonspecific effects, ribosomes in halobacteria show marked preference for potassium over sodium ions. To ensure the proper intracellular ionic composition, powerful ion transport systems have evolved in the halobacteria, resulting in the extrusion of sodium ions and their replacement by potassium. It is likely that such membrane transport system for ionic movements is a necessary requisite for salt tolerance.

  4. The hepatitis E virus open reading frame 3 product interacts with microtubules and interferes with their dynamics.

    PubMed

    Kannan, Harilakshmi; Fan, Sumin; Patel, Deendayal; Bossis, Ioannis; Zhang, Yan-Jin

    2009-07-01

    Hepatitis E virus (HEV) is the causative agent of hepatitis E, a major form of viral hepatitis in developing countries. The open reading frame 3 (ORF3) of HEV encodes a phosphoprotein with a molecular mass of approximately 13 kDa (hereinafter called vp13). vp13 is essential for establishing HEV infections in animals, yet its exact functions are still obscure. Our current study found evidence showing interaction between vp13 and microtubules. Live-cell confocal fluorescence microscopy revealed both filamentous and punctate distribution patterns of vp13 in cells transfected with recombinant ORF3 reporter plasmids. The filamentous pattern of vp13 was altered by a microtubule-destabilizing drug. The vp13 expression led to elevation of acetylated alpha-tubulin, indicating increased microtubule stability. Its association with microtubules was further supported by its presence in microtubule-containing pellets in microtubule isolation assays. Exposure of these pellets to a high-salt buffer caused release of the vp13 to the supernatant, suggesting an electrostatic interaction. Inclusion of ATP and GTP in the lysis buffer during microtubule isolation also disrupted the interaction, indicating its sensitivity to the nucleotides. Further assays showed that motor proteins are needed for the vp13 association with the microtubules because disruption of dynein function abolished the vp13 filamentous pattern. Analysis of ORF3 deletion constructs found that both of the N-terminal hydrophobic domains of vp13 are needed for the interaction. Thus, our findings suggest that the vp13 interaction with microtubules might be needed for establishment of an HEV infection.

  5. Joint Geophysical and Hydrologic Constraints on Shallow Groundwater Flow Systems in Clastic Salt Marshes of the South Atlantic Bight

    NASA Astrophysics Data System (ADS)

    Ruppel, C.; Fulton, P.; Schultz, G. M.; Castillo, L.; Bartlett, J.; Sibley, S.

    2005-12-01

    Salt marsh systems play a critical role in buffering upland coastal areas from the influence of open saltwater bodies and in filtering contaminants that originate offshore or are flushed from uplands. For these reasons, it is important to understand the salt marsh hydrologic cycle, especially the interaction of groundwater and surface water across low-lying coastal fringes and the changes in physical, chemical, and ecological parameters across salinity gradients extending from upland to tidal creek to open water. For the past 5 years, we have conducted hydrogeophysical surveys (inductive EM and DC resistivity) and collected limited, coincident groundwater hydrologic data in clastic salt marshes throughout the South Atlantic Bight (SAB), stretching from South Carolina on the north to the Georgia-Florida border on the south. All of the marshes are dominated by Spartina and Juncus grasses and are cut by tidally-influenced creeks, but both the lithology and age of the marshes vary widely. For example, one highly homogeneous marsh study site has formed only within the past century, while most sites have existed for thousands of years and have laterally and vertically heterogeneous lithology. Geophysical images of the marsh subsurface and coincident monitoring of groundwater temperature, water level, and/or chemistry consistently show that marshes in the mixed energy environment of the middle part of the SAB (GCE LTER) tend to be dominated by submarsh discharge of freshwater to adjacent tidal creeks. In the South Carolina part of the SAB, we have greater evidence for seepage, particularly through biologically-created macropore networks and permeable sediment bodies that intersect tidal creeks. It is possible though that the South Carolina results are not so much 'universal' as reflective of local lithology. In a very young marsh near the Florida border, geophysical imaging implies a mixture of seepage and submarsh flow, and hydrologic data provide unequivocal proof that the near-surface marsh muds act as a low permeability barrier to downward penetration of tidal creek surface waters during periodic inundation of the marsh. Taken together, the results imply that subsurface freshwater bodies flowing beneath some salt marshes act as extensions of the classic freshwater lens that develops beneath uplands and help to resist saline intrusion toward uplands. Certain factors allow us to predict the occurrence of seepage, instead of submarsh flow, in SAB salt marshes with some degree of confidence. Where we have acquired time series, both the hydrogeophysical and hydrologic data suggest that groundwater transport processes are at approximate steady-state at the length scales (vertical and horizontal) and over the duration of our measurements.

  6. Fast simulation of packet loss rates in a shared buffer communications switch

    NASA Technical Reports Server (NTRS)

    Chang, Cheng-Shang; Heidelberger, Philip; Shahabuddin, Perwez

    1993-01-01

    This paper describes an efficient technique for estimating, via simulation, the probability of buffer overflows in a queueing model that arises in the analysis of ATM (Asynchronous Transfer Mode) communication switches. There are multiple streams of (autocorrelated) traffic feeding the switch that has a buffer of finite capacity. Each stream is designated as either being of high or low priority. When the queue length reaches a certain threshold, only high priority packets are admitted to the switch's buffer. The problem is to estimate the loss rate of high priority packets. An asymptotically optimal importance sampling approach is developed for this rare event simulation problem. In this approach, the importance sampling is done in two distinct phases. In the first phase, an importance sampling change of measure is used to bring the queue length up to the threshold at which low priority packets get rejected. In the second phase, a different importance sampling change of measure is used to move the queue length from the threshold to the buffer capacity.

  7. The molecular basis for attractive salt-taste coding in Drosophila.

    PubMed

    Zhang, Yali V; Ni, Jinfei; Montell, Craig

    2013-06-14

    Below a certain level, table salt (NaCl) is beneficial for animals, whereas excessive salt is harmful. However, it remains unclear how low- and high-salt taste perceptions are differentially encoded. We identified a salt-taste coding mechanism in Drosophila melanogaster. Flies use distinct types of gustatory receptor neurons (GRNs) to respond to different concentrations of salt. We demonstrated that a member of the newly discovered ionotropic glutamate receptor (IR) family, IR76b, functioned in the detection of low salt and was a Na(+) channel. The loss of IR76b selectively impaired the attractive pathway, leaving salt-aversive GRNs unaffected. Consequently, low salt became aversive. Our work demonstrated that the opposing behavioral responses to low and high salt were determined largely by an elegant bimodal switch system operating in GRNs.

  8. The prophylactic reduction of aluminium intake.

    PubMed

    Lione, A

    1983-02-01

    The use of modern analytical methods has demonstrated that aluminium salts can be absorbed from the gut and concentrated in various human tissues, including bone, the parathyroids and brain. The neurotoxicity of aluminium has been extensively characterized in rabbits and cats, and high concentrations of aluminium have been detected in the brain tissue of patients with Alzheimer's disease. Various reports have suggested that high aluminium intakes may be harmful to some patients with bone disease or renal impairment. Fatal aluminium-induced neuropathies have been reported in patients on renal dialysis. Since there are no demonstrable consequences of aluminium deprivation, the prophylactic reduction of aluminium intake by many patients would appear prudent. In this report, the major sources of aluminium in foods and non-prescription drugs are summarized and alternative products are described. The most common foods that contain substantial amounts of aluminium-containing additives include some processed cheeses, baking powders, cake mixes, frozen doughs, pancake mixes, self-raising flours and pickled vegetables. The aluminium-containing non-prescription drugs include some antacids, buffered aspirins, antidiarrhoeal products, douches and haemorrhoidal medications. The advisability of recommending a low aluminium diet for geriatric patients is discussed in detail.

  9. Integration of Microdialysis Sampling and Microchip Electrophoresis with Electrochemical Detection

    PubMed Central

    Mecker, Laura C.; Martin, R. Scott

    2009-01-01

    Here we describe the fabrication, optimization, and application of a microfluidic device that integrates microdialysis (MD) sampling, microchip electrophoresis (ME), and electrochemical detection (EC). The manner in which the chip is produced is reproducible and enables the fixed alignment of the MD/ME and ME/EC interfaces. Poly(dimethylsiloxane) (PDMS) -based valves were used for the discrete injection of sample from the hydrodynamic MD dialysate stream into a separation channel for analysis with ME. To enable the integration of ME with EC detection, a palladium decoupler was used to isolate the high voltages associated with electrophoresis from micron-sized carbon ink detection electrodes. Optimization of the ME/EC interface was needed to allow the use of biologically appropriate perfusate buffers containing high salt content. This optimization included changes in the fabrication procedure, increases in the decoupler surface area, and a programmed voltage shutoff. The ability of the MD/ME/EC system to sample a biological system was demonstrated by using a linear probe to monitor the stimulated release of dopamine from a confluent layer of PC 12 cells. To our knowledge, this is the first report of a microchip-based system that couples microdialysis sampling with microchip electrophoresis and electrochemical detection. PMID:19551945

  10. How Does Circadian Rhythm Impact Salt Sensitivity of Blood Pressure in Mice? A Study in Two Close C57Bl/6 Substrains.

    PubMed

    Combe, Roy; Mudgett, John; El Fertak, Lahcen; Champy, Marie-France; Ayme-Dietrich, Estelle; Petit-Demoulière, Benoit; Sorg, Tania; Herault, Yann; Madwed, Jeffrey B; Monassier, Laurent

    2016-01-01

    Mouse transgenesis has provided the unique opportunity to investigate mechanisms underlying sodium kidney reabsorption as well as end organ damage. However, understanding mouse background and the experimental conditions effects on phenotypic readouts of engineered mouse lines such as blood pressure presents a challenge. Despite the ability to generate high sodium and chloride plasma levels during high-salt diet, observed changes in blood pressure are not consistent between wild-type background strains and studies. The present work was designed in an attempt to determine guidelines in the field of salt-induced hypertension by recording continuously blood pressure by telemetry in mice submitted to different sodium and potassium loaded diets and changing experimental conditions in both C57BL/6N and C57BL/6J mice strain (Normal salt vs. Low salt vs. High-salt/normal potassium vs. High salt/low potassium, standard vs. modified light cycle, Non-invasive tail cuff blood pressure vs. telemetry). In this study, we have shown that, despite a strong blood pressure (BP) basal difference between C57BL/6N and C57BL/6J mice, High salt/normal potassium diet increases BP and heart rate during the active phase only (dark period) in the same extent in both strains. On the other hand, while potassium level has no effect on salt-induced hypertension in C57BL/6N mice, high-salt/low potassium diet amplifies the effect of the high-salt challenge only in C57BL/6J mice. Indeed, in this condition, salt-induced hypertension can also be detected during light period even though this BP increase is lower compared to the one occurring during the dark period. Finally, from a methodological perspective, light cycle inversion has no effect on this circadian BP phenotype and tail-cuff method is less sensitive than telemetry to detect BP phenotypes due to salt challenges. Therefore, to carry investigations on salt-induced hypertension in mice, chronic telemetry and studies in the active phase are essential prerequisites.

  11. AlGaSb Buffer Layers for Sb-Based Transistors

    DTIC Science & Technology

    2010-01-01

    transistor ( HEMT ), molecular beam epitaxy (MBE), field-effect transistor (FET), buffer layer INTRODUCTION High-electron-mobility transistors ( HEMTs ) with InAs...monolayers/s. The use of thinner buffer layers reduces molecular beam epitaxial growth time and source consumption. The buffer layers also exhibit...source. In addition, some of the flux from an Sb cell in a molecular beam epitaxy (MBE) system will deposit near the mouth of the cell, eventually

  12. Influence of lime and struvite on microbial community succession and odour emission during food waste composting.

    PubMed

    Wang, Xuan; Selvam, Ammaiyappan; Lau, Sam S S; Wong, Jonathan W C

    2018-01-01

    Lime addition as well as formation of struvite through the addition of magnesium and phosphorus salts provide good pH buffering and may reduce odour emission. This study investigated the odour emission during food waste composting under the influence of lime addition, and struvite formation. Composting was performed in 20-L reactors for 56days using artificial food waste mixed with sawdust at 1.2:1 (w/w dry basis). VFA was one of the most important odours during food waste composting. However, during thermophilic phase, ammonia is responsible for max odour index in the exhaust gas. Trapping ammonia through struvite formation significantly reduced the maximum odour unit of ammonia from 3.0×10 4 to 1.8×10 4 . The generation and accumulation of acetic acid and butyric acid led to the acidic conditions. The addition of phosphate salts in treatment with struvite formation improved the variation of total bacteria, which in turn increased the organic decomposition. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Peptide adsorption to cyanine dye aggregates revealed by cryo-transmission electron microscopy.

    PubMed

    von Berlepsch, Hans; Brandenburg, Enrico; Koksch, Beate; Böttcher, Christoph

    2010-07-06

    The binding interaction between aggregates of the 5-chloro-2-[[5-chloro-3-(3-sulfopropyl)-3H-benzothiazol-2-ylidene]methyl]-3-(3-sulfopropyl)benzothiazolium hydroxide inner salt ammonium salt (CD-1) and alpha-helix, as well as beta-sheet forming de novo designed peptides, was investigated by absorption spectroscopy, circular dichroism spectroscopy, and cryogenic transmission electron microscopy. Both pure dye and pure peptides self-assembled into well-defined supramolecular assemblies in acetate buffer at pH = 4. The dye formed sheetlike and tubular H- and J-aggregates and the peptides alpha-helical coiled-coil assemblies or beta-sheet rich fibrils. After mixing dye and peptide solutions, tubular aggregates with an unusual ultrastructure were found, most likely due to the decoration of dye tubes with monolayers of peptide assemblies based on the strong electrostatic attraction between the oppositely charged species. There was neither indication of a transfer of chirality from the peptides to the dye aggregates nor the opposite effect of a structural transfer from dye aggregates onto the peptides secondary structure.

  14. Electrodeposited Organic Layers Formed from Aryl Diazonium Salts for Inhibition of Copper Corrosion

    PubMed Central

    Chira, Ana; Bucur, Bogdan; Radu, Gabriel-Lucian

    2017-01-01

    Copper substrates deposed on a gold screen-printed electrode were covered with different aryl diazonium salts by electrodeposition at 0.25 mA for 30 or 300 s. Seven compounds were investigated: 4-aminophenylacetic acid, 4-aminophenethyl alcohol, 4-fluoroaniline, 4-(heptadecafluorooctyl)aniline, 4-aminoantipyrine, 4-(4-aminophenyl)butyric acid and 3,4,5-trimethoxyaniline. Quantitative monitoring of the electrodeposition process was carried out by electrogravimetry using quartz crystal microbalance (QCM). The electrodeposited mass varies between 26 ng/cm2 for 4-fluoroaniline formed during 30 s to 442 ng/cm2 for 4-phenylbutyric acid formed during 300 s. The corrosion inhibition properties of aryl-modified layers have been studied in buffer citrate with pH = 3 or 3.5% NaCl solutions using electrochemical noise (ECN) and Tafel potentiodynamic polarization measurements. A corrosion inhibiting efficiency up to 90% was found. The highest corrosion inhibition was obtained for 4-(4-aminophenyl)butyric acid and the lowest for 4-fluoroaniline. A relation between the inhibition efficiency and the chemical nature of the substituents in the protective layer was found. PMID:28772600

  15. Electrodeposited Organic Layers Formed from Aryl Diazonium Salts for Inhibition of Copper Corrosion.

    PubMed

    Chira, Ana; Bucur, Bogdan; Radu, Gabriel-Lucian

    2017-02-28

    Copper substrates deposed on a gold screen-printed electrode were covered with different aryl diazonium salts by electrodeposition at 0.25 mA for 30 or 300 s. Seven compounds were investigated: 4-aminophenylacetic acid, 4-aminophenethyl alcohol, 4-fluoroaniline, 4-(heptadecafluorooctyl)aniline, 4-aminoantipyrine, 4-(4-aminophenyl)butyric acid and 3,4,5-trimethoxyaniline. Quantitative monitoring of the electrodeposition process was carried out by electrogravimetry using quartz crystal microbalance (QCM). The electrodeposited mass varies between 26 ng/cm² for 4-fluoroaniline formed during 30 s to 442 ng/cm² for 4-phenylbutyric acid formed during 300 s. The corrosion inhibition properties of aryl-modified layers have been studied in buffer citrate with pH = 3 or 3.5% NaCl solutions using electrochemical noise (ECN) and Tafel potentiodynamic polarization measurements. A corrosion inhibiting efficiency up to 90% was found. The highest corrosion inhibition was obtained for 4-(4-aminophenyl)butyric acid and the lowest for 4-fluoroaniline. A relation between the inhibition efficiency and the chemical nature of the substituents in the protective layer was found.

  16. Improved biocompatibility of bicarbonate/lactate-buffered PDF is not related to pH.

    PubMed

    Zareie, Mohammad; Keuning, Eelco D; ter Wee, Piet M; Schalkwijk, Casper G; Beelen, Robert H J; van den Born, Jacob

    2006-01-01

    Chronic exposure to conventional peritoneal dialysis fluid (PDF) is associated with functional and structural alterations of the peritoneal membrane. The bioincompatibility of conventional PDF can be due to hypertonicity, high glucose concentration, lactate buffering system, presence of glucose degradation products (GDPs) and/or acidic pH. Although various investigators have studied the sole effects of hyperosmolarity, high glucose, GDPs and lactate buffer in experimental PD, less attention has been paid to the chronic impact of low pH in vivo. Rats received daily 10 ml of either conventional lactate-buffered PDF (pH 5.2; n=7), a standard bicarbonate/lactate-buffered PDF with physiological pH (n=8), bicarbonate/lactate-buffered PDF with acidic pH (adjusted to pH 5.2 with 1 N hydrochloride, n=5), or bicarbonate/lactate buffer, without glucose, pH 7.4 (n=7). Fluids were instilled via peritoneal catheters connected to implanted subcutaneous mini vascular access ports for 8 weeks. Control animals with or without peritoneal catheters served as control groups (n=8/group). Various functional (2 h PET) and morphological/cellular parameters were analyzed. Compared with control groups and the buffer group, conventional lactate-buffered PDF induced a number of morphological/cellular changes, including angiogenesis and fibrosis in various peritoneal tissues (all parameters P<0.05), accompanied by increased glucose absorption and reduced ultrafiltration capacity. Daily exposure to standard or acidified bicarbonate/lactate-buffered PDF improved the performance of the peritoneal membrane, evidenced by reduced new vessel formation in omentum (P<0.02) and parietal peritoneum (P<0.008), reduced fibrosis (P<0.02) and improved ultrafiltration capacity. No significant differences were found between standard and acidified bicarbonate/lactate-buffered PDF. During PET, acidic PDF was neutralized within 15 to 20 min. The bicarbonate/lactate-buffered PDF, acidity per se did not contribute substantially to peritoneal worsening in our in vivo model for PD, which might be explained by the buffering capacity of the peritoneum.

  17. The effect of respiration buffer composition on mitochondrial metabolism and function.

    PubMed

    Wollenman, Lucas C; Vander Ploeg, Matthew R; Miller, Mackinzie L; Zhang, Yizhu; Bazil, Jason N

    2017-01-01

    Functional studies on isolated mitochondria critically rely on the right choice of respiration buffer. Differences in buffer composition can lead to dramatically different respiration rates leading to difficulties in comparing prior studies. The ideal buffer facilities high ADP-stimulated respiratory rates and minimizes substrate transport effects so that the ability to distinguish between various treatments and conditions is maximal. In this study, we analyzed a variety of respiration buffers and substrate combinations to determine the optimal conditions to support mitochondrial function through ADP-stimulated respiration and uncoupled respiration using FCCP. The buffers consisted of a standard KCl based buffer (B1) and three modified buffers with chloride replaced by the K-lactobionate, sucrose, and the antioxidant taurine (B2) or K-gluconate (B3). The fourth buffer (B4) was identical to B2 except that K-lactobionate was replaced with K-gluconate. The substrate combinations consisted of metabolites that utilize different pathways of mitochondrial metabolism. To test mitochondrial function, we used isolated cardiac guinea pig mitochondria and measured oxygen consumption for three respiratory states using an Oroboros Oxygraph-2k. These states were the leak state (energized mitochondria in the absence of adenylates), ADP-stimulated state (energized mitochondria in the presence of saturating ADP concentrations), and uncoupled state (energized mitochondria in the presence of FCCP). On average across all substrate combinations, buffers B2, B3, and B4 had an increase of 16%, 26%, and 35% for the leak state, ADP-simulated state, and uncoupled state, respectively, relative to rates using B1. The common feature distinguishing these buffers from B1 is the notable lack of high chloride concentrations. Based on the respiratory rate metrics obtained with the substrate combinations, we conclude that the adenine nucleotide translocase, the dicarboxylate carrier, and the alpha-ketoglutarate exchanger are partially inhibited by chloride. Therefore, when the goal is to maximize ADP-stimulated respiration, buffers containing K-lactobionate or K-gluconate are superior choices compared to the standard KCl-based buffers.

  18. The effect of respiration buffer composition on mitochondrial metabolism and function

    PubMed Central

    Wollenman, Lucas C.; Vander Ploeg, Matthew R.; Miller, Mackinzie L.; Zhang, Yizhu

    2017-01-01

    Functional studies on isolated mitochondria critically rely on the right choice of respiration buffer. Differences in buffer composition can lead to dramatically different respiration rates leading to difficulties in comparing prior studies. The ideal buffer facilities high ADP-stimulated respiratory rates and minimizes substrate transport effects so that the ability to distinguish between various treatments and conditions is maximal. In this study, we analyzed a variety of respiration buffers and substrate combinations to determine the optimal conditions to support mitochondrial function through ADP-stimulated respiration and uncoupled respiration using FCCP. The buffers consisted of a standard KCl based buffer (B1) and three modified buffers with chloride replaced by the K-lactobionate, sucrose, and the antioxidant taurine (B2) or K-gluconate (B3). The fourth buffer (B4) was identical to B2 except that K-lactobionate was replaced with K-gluconate. The substrate combinations consisted of metabolites that utilize different pathways of mitochondrial metabolism. To test mitochondrial function, we used isolated cardiac guinea pig mitochondria and measured oxygen consumption for three respiratory states using an Oroboros Oxygraph-2k. These states were the leak state (energized mitochondria in the absence of adenylates), ADP-stimulated state (energized mitochondria in the presence of saturating ADP concentrations), and uncoupled state (energized mitochondria in the presence of FCCP). On average across all substrate combinations, buffers B2, B3, and B4 had an increase of 16%, 26%, and 35% for the leak state, ADP-simulated state, and uncoupled state, respectively, relative to rates using B1. The common feature distinguishing these buffers from B1 is the notable lack of high chloride concentrations. Based on the respiratory rate metrics obtained with the substrate combinations, we conclude that the adenine nucleotide translocase, the dicarboxylate carrier, and the alpha-ketoglutarate exchanger are partially inhibited by chloride. Therefore, when the goal is to maximize ADP-stimulated respiration, buffers containing K-lactobionate or K-gluconate are superior choices compared to the standard KCl-based buffers. PMID:29091971

  19. Solution dynamics of synthetic and natural polyelectrolytes

    NASA Astrophysics Data System (ADS)

    Krause, Wendy E.

    Polyelectrolytes are abundant in nature and essential to life, and used extensively in industry. This work discussed two polyelectrolytes: sodium poly(2-acrylamido-2-methylpropanesulfonate) (NaPAMS), synthetic polyelectrolyte, and sodium hyaluronate (NaHA), a glycosaminoglycan. Rheological data of NaPAMS solutions of variable chain length and concentration were reported. A strong dependence of viscosity eta on chain length: eta ˜ M2.4 was found. The comparison of the rheological data with two proposed scaling theories (Dobrynin 1995, Witten 1987) forces the conclusion that neither theory is correct. A possible interpretation of the viscosity data falling between the predictions of the two scaling theories is that some chain rigidity may persist beyond the correlation length. A sample model for the conductivity of semidilute polyelectrolytes with no added salt was presented. The model correctly describes the logarithmic decrease of specific conductance observed for many polyelectrolytes at low concentration (below ca. 10-2M), and is in good agreement with data from NaPAMS solutions. NaHA in phosphate buffered saline behaves as a typical polyelectrolyte in the high-salt limit, as Newtonian viscosities are observed over a wide range of shear rates. There is no evidence of intermolecular hydrogen bonding causing gel formation in NaHA solutions without protein present. The viscosity of 3 mg/mL NaHA was measured in the presence of the selected anti-inflammatory agents. Of the seven additives investigated only (D)-penicillamine significantly altered the rheology of HA. (D)-Penicillamine dramatically reduced the viscosity of HA, probably by disrupting intramolecular hydrogen bonding. The plasma proteins albumin and gamma-globulins bind to HA in solution to form a weak reversible gel. The rheology and osmotic pressure of the simple model for synovial fluid, consisting of 3mg/mL NaHA, 11 mg/mL albumin, and 7 mg/mL gamma-globulins in phosphate buffered saline, were studied in the presence and absence of the seven selected anti-inflammatory agents. Only hydroxychloroquine (HCQ) and (D)-penicillamine strongly influence the theology of the synovial fluid model. HCQ reduces the viscosity of the model solution as well as the model's viscoelasticity. (D)-Penicillamine also reduces the viscosity of the synovial fluid model, but has little effect on the viscoelasticity of the solution. None of the additives effected the osmotic pressure of the synovial fluid model.

  20. Fabrication and characterization of La2Zr2O7 films on different buffer architectures for YBa2Cu3O7-δ coated conductors by RF magnetron sputtering.

    PubMed

    Xu, Da; Liu, Linfei; Xiao, Guina; Li, Yijie

    2013-02-27

    La2Zr2O7 (LZO) films were grown on different buffer architectures by radio frequency magnetron sputtering for the large-scale application of YBa2Cu3O7-x (YBCO)-coated conductors. The three different buffer architectures were cerium oxide (CeO2), yttria-stabilized zirconia (YSZ)/CeO2, and CeO2/YSZ/CeO2. The microstructure and surface morphology of the LZO film were studied by X-ray diffraction, optical microscopy, field emission scanning electron microscopy, and atomic force microscopy. The LZO films prepared on the CeO2, YSZ/CeO2, and CeO2/YSZ/CeO2 buffer architectures were preferentially c-axis-oriented and highly textured. The in-plane texture of LZO film on CeO2 single-buffer architecture was ∆ φ = 5.5° and the out-of-plane texture was ∆ ω = 3.4°. All the LZO films had very smooth surfaces, but LZO films grown on YSZ/CeO2 and CeO2/YSZ/CeO2 buffer architectures had cracks. The highly textured LZO film grown on CeO2-seed buffered NiW tape was suitable for the epitaxial growth of YBCO film with high currents.

  1. Salt-Sensitive Hypertension: Perspectives on Intrarenal Mechanisms

    PubMed Central

    Majid, Dewan S.A.; Prieto, Minolfa C.; Navar, L Gabriel

    2015-01-01

    Salt sensitive hypertension is characterized by increases in blood pressure in response to increases in dietary salt intake and is associated with an enhanced risk of cardiovascular and renal morbidity. Although researchers have sought for decades to understand how salt sensitivity develops in humans, the mechanisms responsible for the increases in blood pressure in response to high salt intake are complex and only partially understood. Until now, scientists have been unable to explain why some individuals are salt sensitive and others are salt resistant. Although a central role for the kidneys in the development of salt sensitivity and hypertension has been generally accepted, it is also recognized that hypertension is of multifactorial origin and a variety of factors can induce, or prevent, blood pressure responsiveness to the manipulation of salt intake. Excess salt intake in susceptible persons may also induce inappropriate central and sympathetic nervous system responses and increase the production of intrarenal angiotensin II, catecholamines and other factors such as oxidative stress and inflammatory cytokines. One key factor is the concomitant inappropriate or paradoxical activation of the intrarenal renin-angiotensin system, by high salt intake. This is reflected by the increases in urinary angiotensinogen during high salt intake in salt sensitive models. A complex interaction between neuroendocrine factors and the kidney may underlie the propensity for some individuals to retain salt and develop salt-dependent hypertension. In this review, we focus mainly on the renal contributions that provide the mechanistic link between chronic salt intake and the development of hypertension. PMID:26028244

  2. Salt Intake and Health Risk in Climate Change Vulnerable Coastal Bangladesh: What Role Do Beliefs and Practices Play?

    PubMed Central

    Rasheed, Sabrina; Siddique, A. K.; Sharmin, Tamanna; Hasan, A. M. R.; Hanifi, S. M. A.; Iqbal, M.; Bhuiya, Abbas

    2016-01-01

    Background High salt consumption is an important risk factor of elevated blood pressure. In Bangladesh about 20 million people are at high risk of hypertension due to climate change induced saline intrusion in water. The objective of this study is to assess beliefs, perceptions, and practices associated with salt consumption in coastal Bangladesh. Methods The study was conducted in Chakaria, Bangladesh between April-June 2011. It was a cross sectional mixed method study. For the qualitative study 6 focus group discussions, 8 key informant interviews, 60 free listing exercises, 20 ranking exercises and 10 observations were conducted. 400 adults were randomly selected for quantitative survey. For analysis we used SPSS for quantitative data, and Anthropac and Nvivo for qualitative data. Results Salt was described as an essential component of food with strong cultural and religious roots. People described both health benefits and risks related to salt intake. The overall risk perception regarding excessive salt consumption was low and respondents believed that the cooking process can render the salt harmless. Respondents were aware that salt is added in many foods even if they do not taste salty but did not recognize that salt can occur naturally in both foods and water. Conclusions In the study community people had low awareness of the risks associated with excess salt consumption and salt reduction strategies were not high in their agenda. The easy access to and low cost of salt as well as unrecognised presence of salt in drinking water has created an environment conducive to excess salt consumption. It is important to design general messages related to salt reduction and test tailored strategies especially for those at high risk of hypertension. PMID:27044049

  3. Chemically Deposited CdS Buffer/Kesterite Cu2ZnSnS4 Solar Cells: Relationship between CdS Thickness and Device Performance.

    PubMed

    Hong, Chang Woo; Shin, Seung Wook; Suryawanshi, Mahesh P; Gang, Myeng Gil; Heo, Jaeyeong; Kim, Jin Hyeok

    2017-10-25

    Earth-abundant, copper-zinc-tin-sulfide (CZTS), kesterite, is an attractive absorber material for thin-film solar cells (TFSCs). However, the open-circuit voltage deficit (V oc -deficit) resulting from a high recombination rate at the buffer/absorber interface is one of the major challenges that must be overcome to improve the performance of kesterite-based TFSCs. In this paper, we demonstrate the relationship between device parameters and performances for chemically deposited CdS buffer/CZTS-based heterojunction TFSCs as a function of buffer layer thickness, which could change the CdS/CZTS interface conditions such as conduction band or valence band offsets, to gain deeper insight and understanding about the V oc -deficit behavior from a high recombination rate at the CdS buffer/kesterite interface. Experimental results show that device parameters and performances are strongly dependent on the CdS buffer thickness. We postulate two meaningful consequences: (i) Device parameters were improved up to a CdS buffer thickness of 70 nm, whereas they deteriorated at a thicker CdS buffer layer. The V oc -deficit in the solar cells improved up to a CdS buffer thickness of 92 nm and then deteriorated at a thicker CdS buffer layer. (ii) The minimum values of the device parameters were obtained at 70 nm CdS thickness in the CZTS TFSCs. Finally, the highest conversion efficiency of 8.77% (V oc : 494 mV, J sc : 34.54 mA/cm 2 , and FF: 51%) is obtained by applying a 70 nm thick CdS buffer to the Cu 2 ZnSn(S,Se) 4 absorber layer.

  4. Effect of salt intake and potassium supplementation on urinary renalase and serum dopamine levels in Chinese adults.

    PubMed

    Wang, Yang; Wang, Dan; Chu, Chao; Mu, Jian-Jun; Wang, Man; Liu, Fu-Qiang; Xie, Bing-Qing; Yang, Fan; Dong, Zhen-Zhen; Yuan, Zu-Yi

    2015-01-01

    The aim of our study was to assess the effects of altered salt and potassium intake on urinary renalase and serum dopamine levels in humans. Forty-two subjects (28–65 years of age) were selected from a rural community of northern China. All subjects were sequentially maintained on a low-salt diet for 7 days (3.0 g/day of NaCl), a high-salt diet for an additional 7 days (18.0 g/day of NaCl), and a high-salt diet with potassium supplementation for a final 7 days (18.0 g/day of NaCl + 4.5 g/day of KCl). Urinary renalase excretions were significantly higher during the high-salt diet intervention than during the low-salt diet. During high-potassium intake, urinary renalase excretions were not significantly different from the high-salt diet, whereas they were significantly higher than the low-salt levels. Serum dopamine levels exhibited similar trends across the interventions. Additionally, a significant positive relationship was observed between the urine renalase and serum dopamine among the different dietary interventions. Also, 24-hour urinary sodium excretion positively correlated with urine renalase and serum dopamine in the whole population. The present study indicates that dietary salt intake and potassium supplementation increase urinary renalase and serum dopamine levels in Chinese subjects. © 2015 S. Karger AG, Basel

  5. Field effect transistors improve buffer amplifier

    NASA Technical Reports Server (NTRS)

    1967-01-01

    Unity gain buffer amplifier with a Field Effect Transistor /FET/ differential input stage responds much faster than bipolar transistors when operated at low current levels. The circuit uses a dual FET in a unity gain buffer amplifier having extremely high input impedance, low bias current requirements, and wide bandwidth.

  6. Influence of organic ions on DNA damage induced by 1 eV to 60 keV electrons.

    PubMed

    Zheng, Yi; Sanche, Léon

    2010-10-21

    We report the results of a study on the influence of organic salts on the induction of single strand breaks (SSBs) and double strand breaks (DSBs) in DNA by electrons of 1 eV to 60 keV. Plasmid DNA films are prepared with two different concentrations of organic salts, by varying the amount of the TE buffer (Tris-HCl and EDTA) in the films with ratio of 1:1 and 6:1 Tris ions to DNA nucleotide. The films are bombarded with electrons of 1, 10, 100, and 60 000 eV under vacuum. The damage to the 3197 base-pair plasmid is analyzed ex vacuo by agarose gel electrophoresis. The highest yields are reached at 100 eV and the lowest ones at 60 keV. The ratios of SSB to DSB are surprisingly low at 10 eV (∼4.3) at both salt concentrations, and comparable to the ratios measured with 100 eV electrons. At all characteristic electron energies, the yields of SSB and DSB are found to be higher for the DNA having the lowest salt concentration. However, the organic salts are more efficient at protecting DNA against the damage induced by 1 and 10 eV electrons. DNA damage and protection by organic ions are discussed in terms of mechanisms operative at each electron energy. It is suggested that these ions create additional electric fields within the groove of DNA, which modify the resonance parameter of 1 and 10 eV electrons, namely, by reducing the electron capture cross-section of basic DNA units and the lifetime of corresponding transient anions. An interstrand electron transfer mechanism is proposed to explain the low ratios for the yields of SSB to those of DSB produced by 10 eV electrons.

  7. Influence of organic ions on DNA damage induced by 1 eV to 60 keV electrons

    PubMed Central

    Zheng, Yi; Sanche, Léon

    2011-01-01

    We report the results of a study on the influence of organic salts on the induction of single strand breaks (SSBs) and double strand breaks (DSBs) in DNA by electrons of 1 eV to 60 keV. Plasmid DNA films are prepared with two different concentrations of organic salts, by varying the amount of the TE buffer (Tris-HCl and EDTA) in the films with ratio of 1:1 and 6:1 Tris ions to DNA nucleotide. The films are bombarded with electrons of 1, 10, 100, and 60 000 eV under vacuum. The damage to the 3197 base-pair plasmid is analyzed ex vacuo by agarose gel electrophoresis. The highest yields are reached at 100 eV and the lowest ones at 60 keV. The ratios of SSB to DSB are surprisingly low at 10 eV (~4.3) at both salt concentrations, and comparable to the ratios measured with 100 eV electrons. At all characteristic electron energies, the yields of SSB and DSB are found to be higher for the DNA having the lowest salt concentration. However, the organic salts are more efficient at protecting DNA against the damage induced by 1 and 10 eV electrons. DNA damage and protection by organic ions are discussed in terms of mechanisms operative at each electron energy. It is suggested that these ions create additional electric fields within the groove of DNA, which modify the resonance parameter of 1 and 10 eV electrons, namely, by reducing the electron capture cross-section of basic DNA units and the lifetime of corresponding transient anions. An interstrand electron transfer mechanism is proposed to explain the low ratios for the yields of SSB to those of DSB produced by 10 eV electrons. PMID:20969428

  8. Rheological characterization of hair shampoo in the presence of dead sea salt.

    PubMed

    Abu-Jdayil, B; Mohameed, H A; Sa'id, M; Snobar, T

    2004-02-01

    In Jordan, a growing industry has been established to produce different types of Dead Sea (DS) cosmetics that have DS salt (contains mainly NaCl, KCl, and MgCl(2)) in their formulas. In this work, the effect of DS salt on the rheology of hair shampoo containing the sodium lauryl ether sulfate as a main active matter was studied. The effects of DS salt and active matter concentration, and the temperature and time of salt mixing, on the rheological properties of hair shampoo were investigated. The salt-free shampoo showed a Newtonian behavior at 'low active matter' (LAM) and shear thinning at 'high active matter' (HAM). The presence of DS salt changed the rheological behavior of LAM shampoo from Newtonian (for the salt-free shampoo) to shear thinning. On the other hand, the behavior of HAM shampoo switched from shear thinning to Newtonian behavior in the presence of high concentration of DS salt. The addition of DS salt increased the apparent viscosity of shampoo to reach a maximum value that corresponded to a salt concentration of 1.5 wt.%. Further addition of DS salt led to a decrease in the shampoo viscosity to reach a value less than that of the salt-free sample at high salt concentration. Changing the mixing temperature (25-45 degrees C) and mixing time (15-120 min) of DS salt with shampoo has no significant influence on the rheological behavior. However, the mixing process increased the apparent viscosity of salt-free shampoo. The power law model fitted well the flow curves of hair shampoo with and without DS salt.

  9. A non-multimacrocyclic heteroditopic receptor that cooperatively binds and effectively extracts KAcO salt.

    PubMed

    Zakrzewski, Maciej; Kwietniewska, Natalia; Walczak, Wojciech; Piątek, Piotr

    2018-06-06

    Prepared in only three synthetic steps, a non-multimacrocyclic heteroditopic receptor binds potassium salts of halides and carboxylates with unusually high cooperativity, suggesting salt binding as associated ion-pairs. Unprecedented extraction of highly hydrophilic KAcO salt from water to organic solution is also demonstrated.

  10. Non-ionic detergents facilitate non-specific binding of M13 bacteriophage to polystyrene surfaces.

    PubMed

    Hakami, Abdulrahim R; Ball, Jonathan K; Tarr, Alexander W

    2015-09-01

    Phage-displayed random peptide libraries are widely used for identifying peptide interactions with proteins and other substrates. Selection of peptide ligands involves iterative rounds of affinity enrichment. The binding properties of the selected phage clones are routinely tested using immunoassay after propagation to high titre in a bacterial host and precipitation using polyethylene glycol (PEG) and high salt concentration. These immunoassays can suffer from low sensitivity and high background signals. Polysorbate 20 (Tween(®) 20) is a non-ionic detergent commonly used in immunoassay washing buffers to reduce non-specific binding, and is also used as a blocking reagent. We have observed that Tween 20 enhances non-specific M13 library phage binding in a peptide-independent manner. Other non-ionic detergents were also found to promote significant, dose-dependent non-specific phage binding in ELISA. This effect was not observed for assays using phage concentrated by ultracentrifugation, suggesting that interactions occur between detergents and the PEG-precipitated phage, irrespective of the displayed peptide motif. This artefact may impact on successful affinity selection of peptides from phage-display libraries. We propose alternative methods for screening phage libraries for identifying binding interactions with target ligands. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. A novel, environmentally friendly sodium lauryl ether sulfate-, cocamidopropyl betaine-, cocamide monoethanolamine-containing buffer for MEKC on microfluidic devices.

    PubMed

    Hoeman, Kurt W; Culbertson, Christopher T

    2008-12-01

    A new buffer has been developed for fast, high-efficiency separations of amino acids by MEKC. This buffer was more environmentally friendly than the most commonly used surfactant-containing buffers for MEKC separations. It used a commercially available dishwashing soap by Seventh Generation (Burlington, VT, USA), which contained three micelle-forming agents. The mixed micelles were composed of sodium lauryl ether sulfate (anionic), cocamidopropyl betaine (zwitterionic), and cocamide monoethanolamine (non-ionic). The optimized buffer contained 5.0% w/w Seventh Generation Free & Clear dishwashing soap, 10 mM sodium borate, and was completely void of organics. The lack of organics and the biodegradability of the surfactant molecules made this buffer more environmentally friendly than typical SDS-containing buffers. This new buffer also had a different selectivity and provided faster separations with higher separation efficiencies than SDS-based buffers. Fast separations of BODIPY FL labeled amino acids yielded peaks with separation efficiencies greater than 100,000 in less than 20 s.

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

    NASA Technical Reports Server (NTRS)

    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.

  13. Control of short-channel effects in InAlN/GaN high-electron mobility transistors using graded AlGaN buffer

    NASA Astrophysics Data System (ADS)

    Han, Tiecheng; Zhao, Hongdong; Peng, Xiaocan; Li, Yuhai

    2018-04-01

    A graded AlGaN buffer is designed to realize the p-type buffer by inducing polarization-doping holes. Based on the two-dimensional device simulator, the effect of the graded AlGaN buffer on the direct-current (DC) and radio-frequency (RF) performance of short-gate InAlN/GaN high-electron mobility transistors (HEMTs) are investigated, theoretically. Compared to standard HEMT, an enhancement of electron confinement and a good control of short-channel effect (SCEs) are demonstrated in the graded AlGaN buffer HEMT. Accordingly, the pinched-off behavior and the ability of gate modulation are significantly improved. And, no serious SCEs are observed in the graded AlGaN buffer HEMT with an aspect ratio (LG/tch) of about 6.7, much lower than that of the standard HEMT (LG/tch = 13). In addition, for a 70-nm gate length, a peak current gain cutoff frequency (fT) of 171 GHz and power gain cutoff frequency (fmax) of 191 GHz are obtained in the grade buffer HEMT, which are higher than those of the standard one with the same gate length.

  14. Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species

    PubMed Central

    Loureiro, João; Rodriguez, Eleazar; Doležel, Jaroslav; Santos, Conceição

    2007-01-01

    Background and Aims After the initial boom in the application of flow cytometry in plant sciences in the late 1980s and early 1990s, which was accompanied by development of many nuclear isolation buffers, only a few efforts were made to develop new buffer formulas. In this work, recent data on the performance of nuclear isolation buffers are utilized in order to develop new buffers, general purpose buffer (GPB) and woody plant buffer (WPB), for plant DNA flow cytometry. Methods GPB and WPB were used to prepare samples for flow cytometric analysis of nuclear DNA content in a set of 37 plant species that included herbaceous and woody taxa with leaf tissues differing in structure and chemical composition. The following parameters of isolated nuclei were assessed: forward and side light scatter, propidium iodide fluorescence, coefficient of variation of DNA peaks, quantity of debris background, and the number of particles released from sample tissue. The nuclear genome size of 30 selected species was also estimated using the buffer that performed better for a given species. Key Results In unproblematic species, the use of both buffers resulted in high quality samples. The analysis of samples obtained with GPB usually resulted in histograms of DNA content with higher or similar resolution than those prepared with the WPB. In more recalcitrant tissues, such as those from woody plants, WPB performed better and GPB failed to provide acceptable results in some cases. Improved resolution of DNA content histograms in comparison with previously published buffers was achieved in most of the species analysed. Conclusions WPB is a reliable buffer which is also suitable for the analysis of problematic tissues/species. Although GPB failed with some plant species, it provided high-quality DNA histograms in species from which nuclear suspensions are easy to prepare. The results indicate that even with a broad range of species, either GPB or WPB is suitable for preparation of high-quality suspensions of intact nuclei suitable for DNA flow cytometry. PMID:17684025

  15. Consumption of a high-salt diet by ewes during pregnancy alters nephrogenesis in 5-month-old offspring.

    PubMed

    Tay, S H; Blache, D; Gregg, K; Revell, D K

    2012-11-01

    Maternal nutrition during pregnancy can affect kidney development in the foetus, which may lead to adverse consequences in the mature kidney. It was expected that high-salt intake by pregnant ewes would lead to a reduction in foetal glomerular number but that the ovine kidney would adapt to maintain homoeostasis, in part by increasing the size of each glomerulus. Merino ewes that were fed either a control (1.5% NaCl) or high-salt (10.5% NaCl) diet during pregnancy, as well as their 5-month-old offspring, were subjected to a dietary salt challenge, and glomerular number and size and sodium excretion were measured. The high-salt offspring had 20% fewer glomeruli compared with the control offspring (P < 0.001), but they also had larger glomerular radii compared with the control offspring (P < 0.001). Consequently, the cross-sectional area of glomeruli was 18% larger in the high-salt offspring than in the control offspring (P < 0.05). There was no difference in the daily urinary sodium excretion between the two offspring groups (P > 0.05), although the high-salt offspring produced urine with a higher concentration of sodium. Our results demonstrated that maternal high-salt intake during pregnancy affected foetal nephrogenesis, altering glomerular number at birth. However, the ability to concentrate and excrete salt was not compromised, which indicates that the kidney was able to adapt to the reduction in the number of glomeruli.

  16. Salt-induced epithelial-to-mesenchymal transition in Dahl salt-sensitive rats is dependent on elevated blood pressure.

    PubMed

    Wang, Y; Mu, J J; Liu, F Q; Ren, K Y; Xiao, H Y; Yang, Z; Yuan, Z Y

    2014-02-01

    Dietary salt intake has been linked to hypertension and cardiovascular disease. Accumulating evidence has indicated that salt-sensitive individuals on high salt intake are more likely to develop renal fibrosis. Epithelial-to-mesenchymal transition (EMT) participates in the development and progression of renal fibrosis in humans and animals. The objective of this study was to investigate the impact of a high-salt diet on EMT in Dahl salt-sensitive (SS) rats. Twenty-four male SS and consomic SS-13(BN) rats were randomized to a normal diet or a high-salt diet. After 4 weeks, systolic blood pressure (SBP) and albuminuria were analyzed, and renal fibrosis was histopathologically evaluated. Tubular EMT was evaluated using immunohistochemistry and real-time PCR with E-cadherin and alpha smooth muscle actin (α-SMA). After 4 weeks, SBP and albuminuria were significantly increased in the SS high-salt group compared with the normal diet group. Dietary salt intake induced renal fibrosis and tubular EMT as identified by reduced expression of E-cadherin and enhanced expression of α-SMA in SS rats. Both blood pressure and renal interstitial fibrosis were negatively correlated with E-cadherin but positively correlated with α-SMA. Salt intake induced tubular EMT and renal injury in SS rats, and this relationship might depend on the increase in blood pressure.

  17. Tritium Control and Capture in Salt-Cooled Fission and Fusion Reactors: Status, Challenges, and Path Forward

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

    Forsberg, Charles W.; Lam, Stephen; Carpenter, David M.

    Three advanced nuclear power systems use liquid salt coolants that generate tritium and thus face the common challenges of containing and capturing tritium to prevent its release to the environment. The fluoride salt–cooled high-temperature reactor (FHR) uses clean fluoride salt coolants and the same graphite-matrix coated-particle fuel as high-temperature gas-cooled reactors. Molten salt reactors (MSRs) dissolve the fuel in a fluoride or chloride salt with release of fission product tritium into the salt. In most FHR and MSR systems, the baseline salts contain lithium where isotopically separated 7Li is proposed to minimize tritium production from neutron interactions with the salt.more » The Chinese Academy of Sciences plans to start operation of a 2-MW(thermal) molten salt test reactor by 2020. For high-magnetic-field fusion machines, the use of lithium enriched in 6Li is proposed to maximize tritium generation—the fuel for a fusion machine. Advances in superconductors that enable higher power densities may require the use of molten lithium salts for fusion blankets and as coolants. Recent technical advances in these three reactor classes have resulted in increased government and private interest and the beginning of a coordinated effort to address the tritium control challenges in 700°C liquid salt systems. In this paper, we describe characteristics of salt-cooled fission and fusion machines, the basis for growing interest in these technologies, tritium generation in molten salts, the environment for tritium capture, models for high-temperature tritium transport in salt systems, alternative strategies for tritium control, and ongoing experimental work. Several methods to control tritium appear viable. Finally, limited experimental data are the primary constraint for designing efficient cost-effective methods of tritium control.« less

  18. Tritium Control and Capture in Salt-Cooled Fission and Fusion Reactors: Status, Challenges, and Path Forward

    DOE PAGES

    Forsberg, Charles W.; Lam, Stephen; Carpenter, David M.; ...

    2017-02-26

    Three advanced nuclear power systems use liquid salt coolants that generate tritium and thus face the common challenges of containing and capturing tritium to prevent its release to the environment. The fluoride salt–cooled high-temperature reactor (FHR) uses clean fluoride salt coolants and the same graphite-matrix coated-particle fuel as high-temperature gas-cooled reactors. Molten salt reactors (MSRs) dissolve the fuel in a fluoride or chloride salt with release of fission product tritium into the salt. In most FHR and MSR systems, the baseline salts contain lithium where isotopically separated 7Li is proposed to minimize tritium production from neutron interactions with the salt.more » The Chinese Academy of Sciences plans to start operation of a 2-MW(thermal) molten salt test reactor by 2020. For high-magnetic-field fusion machines, the use of lithium enriched in 6Li is proposed to maximize tritium generation—the fuel for a fusion machine. Advances in superconductors that enable higher power densities may require the use of molten lithium salts for fusion blankets and as coolants. Recent technical advances in these three reactor classes have resulted in increased government and private interest and the beginning of a coordinated effort to address the tritium control challenges in 700°C liquid salt systems. In this paper, we describe characteristics of salt-cooled fission and fusion machines, the basis for growing interest in these technologies, tritium generation in molten salts, the environment for tritium capture, models for high-temperature tritium transport in salt systems, alternative strategies for tritium control, and ongoing experimental work. Several methods to control tritium appear viable. Finally, limited experimental data are the primary constraint for designing efficient cost-effective methods of tritium control.« less

  19. NUTRITION AND METABOLISM OF MARINE BACTERIA1

    PubMed Central

    Takacs, Frank P.; Matula, Tibor I.; MacLeod, Robert A.

    1964-01-01

    Takacs, Frank P. (McGill University, Montreal, Quebec, Canada), Tibor I. Matula, and Robert A. MacLeod. Nutrition and metabolism of marine bacteria. XIII. Intracellular concentrations of sodium and potassium ions in a marine pseudomonad. J. Bacteriol. 87:510–518. 1964.—Washed cells of a marine pseudomonad were suspended in buffered salt solutions containing, in addition to MgSO4, NaCl and KCl at various concentrations. The cells were centrifuged from the medium and analyzed for Na+ and K+. Inulin and C14-carboxypolyglucose were employed to estimate the volume of extracellular fluid associated with the packed cells. Intracellular Na+ and K+ concentrations were determined by correcting for the amount of Na+ and K+ in the extracellular fluid. At all levels of Na+ in the suspending medium (0 to 1 m), the intracellular and extracellular Na+ concentrations within the limits of experimental error were the same. The intracellular K+ concentrations were approximately double the extracellular concentrations at the two levels of K+ tested (0.01 and 0.15 m) and were not influenced by the amount of Na+ present. Intracellular and extracellular Cl− concentrations were the same at the one level of Cl− examined. The intracellular fluid volume varied with the NaCl or KCl concentration of the suspending medium, being greatest in the absence of added salts, decreasing to a minimum at 0.3 m salt, and then increasing slightly at higher salt concentrations. Most of the intracellular Na+ could be removed by washing with solutions of MgSO4 or sucrose, but a small amount [10 to 15 μmoles/g (dry weight)] remained bound to the cells. PMID:14129666

  20. Molten salt oxidation of organic hazardous waste with high salt content.

    PubMed

    Lin, Chengqian; Chi, Yong; Jin, Yuqi; Jiang, Xuguang; Buekens, Alfons; Zhang, Qi; Chen, Jian

    2018-02-01

    Organic hazardous waste often contains some salt, owing to the widespread use of alkali salts during industrial manufacturing processes. These salts cause complications during the treatment of this type of waste. Molten salt oxidation is a flameless, robust thermal process, with inherent capability of destroying the organic constituents of wastes, while retaining the inorganic ingredients in the molten salt. In the present study, molten salt oxidation is employed for treating a typical organic hazardous waste with a high content of alkali salts. The hazardous waste derives from the production of thiotriazinone. Molten salt oxidation experiments have been conducted using a lab-scale molten salt oxidation reactor, and the emissions of CO, NO, SO 2 , HCl and dioxins are studied. Impacts are investigated from the composition of the molten salts, the types of feeding tube, the temperature of molten carbonates and the air factor. Results show that the waste can be oxidised effectively in a molten salt bath. Temperature of molten carbonates plays the most important role. With the temperature rising from 600 °C to 750 °C, the oxidation efficiency increases from 91.1% to 98.3%. Compared with the temperature, air factor has but a minor effect, as well as the composition of the molten salts and the type of feeding tube. The molten carbonates retain chlorine with an efficiency higher than 99.9% and the emissions of dioxins are below 8 pg TEQ g -1 sample. The present study shows that molten salt oxidation is a promising alternative for the disposal of organic hazardous wastes containing a high salt content.

  1. Buffering capability and limitations in low dispersion photonic crystal waveguides with elliptical airholes.

    PubMed

    Long, Fang; Tian, Huiping; Ji, Yuefeng

    2010-09-01

    A low dispersion photonic crystal waveguide with triangular lattice elliptical airholes is proposed for compact, high-performance optical buffering applications. In the proposed structure, we obtain a negligible-dispersion bandwidth with constant group velocity ranging from c/41 to c/256, by optimizing the major and minor axes of bulk elliptical holes and adjusting the position and the hole size of the first row adjacent to the defect. In addition, the limitations of buffer performance in a dispersion engineering waveguide are well studied. The maximum buffer capacity and the maximum data rate can reach as high as 262bits and 515 Gbits/s, respectively. The corresponding delay time is about 255.4ps.

  2. Effect of Salt Intake and Potassium Supplementation on Serum Renalase Levels in Chinese Adults

    PubMed Central

    Wang, Yang; Liu, Fu-Qiang; Wang, Dan; Mu, Jian-Jun; Ren, Ke-Yu; Guo, Tong-Shuai; Chu, Chao; Wang, Lan; Geng, Li-Ke; Yuan, Zu-Yi

    2014-01-01

    Abstract Renalase, a recently discovered enzyme released by the kidneys, breaks down blood-borne catecholamines and may thus regulate blood pressure (BP). Animal studies have suggested that high levels of dietary salt might reduce blood and kidney renalase levels. We conducted a randomized trial to assess the effects of altered salt and potassium intake on serum renalase levels and the relationship between serum renalase levels and BP in humans. Forty-two subjects (28–65 years of age) were selected from a rural community of northern China. All subjects were sequentially maintained on a low-salt diet for 7 days (3.0 g/day of NaCl), a high-salt diet for additional 7 days (18.0 g/day of NaCl), and a high-salt diet with potassium supplementation for final 7 days (18.0 g/day of NaCl + 4.5 g/day of KCl). Serum renalase levels were significantly higher than baseline levels during the low-salt diet intervention period. Renalase levels decreased with the change from the low-salt to high-salt diet, whereas dietary potassium prevented the decrease in serum renalase induced by the high-salt diet. There was a significant inverse correlation between the serum renalase level and 24-h urinary sodium excretion. No significant correlation was found between the renalase level and BP among the different dietary interventions. The present study indicates that variations in dietary salt intake and potassium supplementation affect the serum renalase concentration in Chinese subjects. PMID:25058146

  3. Effect of salt intake and potassium supplementation on serum renalase levels in Chinese adults: a randomized trial.

    PubMed

    Wang, Yang; Liu, Fu-Qiang; Wang, Dan; Mu, Jian-Jun; Ren, Ke-Yu; Guo, Tong-Shuai; Chu, Chao; Wang, Lan; Geng, Li-Ke; Yuan, Zu-Yi

    2014-07-01

    Renalase, a recently discovered enzyme released by the kidneys, breaks down blood-borne catecholamines and may thus regulate blood pressure (BP). Animal studies have suggested that high levels of dietary salt might reduce blood and kidney renalase levels. We conducted a randomized trial to assess the effects of altered salt and potassium intake on serum renalase levels and the relationship between serum renalase levels and BP in humans.Forty-two subjects (28-65 years of age) were selected from a rural community of northern China. All subjects were sequentially maintained on a low-salt diet for 7 days (3.0 g/day of NaCl), a high-salt diet for additional 7 days (18.0 g/day of NaCl), and a high-salt diet with potassium supplementation for final 7 days (18.0 g/day of NaCl + 4.5 g/day of KCl).Serum renalase levels were significantly higher than baseline levels during the low-salt diet intervention period. Renalase levels decreased with the change from the low-salt to high-salt diet, whereas dietary potassium prevented the decrease in serum renalase induced by the high-salt diet. There was a significant inverse correlation between the serum renalase level and 24-h urinary sodium excretion. No significant correlation was found between the renalase level and BP among the different dietary interventions.The present study indicates that variations in dietary salt intake and potassium supplementation affect the serum renalase concentration in Chinese subjects.

  4. Desoxycorticosterone pivalate-salt treatment leads to non-dipping hypertension in Per1 knockout mice.

    PubMed

    Solocinski, K; Holzworth, M; Wen, X; Cheng, K-Y; Lynch, I J; Cain, B D; Wingo, C S; Gumz, M L

    2017-05-01

    Increasing evidence demonstrates that circadian clock proteins are important regulators of physiological functions including blood pressure. An established risk factor for developing cardiovascular disease is the absence of a blood pressure dip during the inactive period. The goal of the present study was to determine the effects of a high salt diet plus mineralocorticoid on PER1-mediated blood pressure regulation in a salt-resistant, normotensive mouse model, C57BL/6J. Blood pressure was measured using radiotelemetry. After control diet, wild-type (WT) and Per1 (KO) knockout mice were given a high salt diet (4% NaCl) and the long-acting mineralocorticoid deoxycorticosterone pivalate. Blood pressure and activity rhythms were analysed to evaluate changes over time. Blood pressure in WT mice was not affected by a high salt diet plus mineralocorticoid. In contrast, Per1 KO mice exhibited significantly increased mean arterial pressure (MAP) in response to a high salt diet plus mineralocorticoid. The inactive/active phase ratio of MAP in WT mice was unchanged by high salt plus mineralocorticoid treatment. Importantly, this treatment caused Per1 KO mice to lose the expected decrease or 'dip' in blood pressure during the inactive compared to the active phase. Loss of PER1 increased sensitivity to the high salt plus mineralocorticoid treatment. It also resulted in a non-dipper phenotype in this model of salt-sensitive hypertension and provides a unique model of non-dipping. Together, these data support an important role for the circadian clock protein PER1 in the modulation of blood pressure in a high salt/mineralocorticoid model of hypertension. © 2016 Scandinavian Physiological Society. Published by John Wiley & Sons Ltd.

  5. Internal acid buffering in San Joaquin Valley fog drops and its influence on aerosol processing

    NASA Astrophysics Data System (ADS)

    Collett, Jeffrey L.; Hoag, Katherine J.; Rao, Xin; Pandis, Spyros N.

    Although several chemical pathways exist for S(IV) oxidation in fogs and clouds, many are self-limiting: as sulfuric acid is produced and the drop pH declines, the rates of these pathways also decline. Some of the acid that is produced can be buffered by uptake of gaseous ammonia. Additional internal buffering can result from protonation of weak and strong bases present in solution. Acid titrations of high pH fog samples (median pH=6.49) collected in California's San Joaquin Valley reveal the presence of considerable internal acid buffering. In samples collected at a rural location, the observed internal buffering could be nearly accounted for based on concentrations of ammonia and bicarbonate present in solution. In samples collected in the cities of Fresno and Bakersfield, however, significant additional, unexplained buffering was present over a pH range extending from approximately four to seven. The additional buffering was found to be associated with dissolved compounds in the fogwater. It could not be accounted for by measured concentrations of low molecular weight ( C1- C3) carboxylic acids, S(IV), phosphate, or nitrophenols. The amount of unexplained buffering in individual fog samples was found to correlate strongly with the sum of sample acetate and formate concentrations, suggesting that unmeasured organic species may be important contributors. Simulation of a Bakersfield fog episode with and without the additional, unexplained buffering revealed a significant impact on the fog chemistry. When the additional buffering was included, the simulated fog pH remained 0.3-0.7 pH units higher and the amount of sulfate present after the fog evaporated was increased by 50%. Including the additional buffering in the model simulation did not affect fogwater nitrate concentrations and was found to slightly decrease ammonium concentrations. The magnitude of the buffering effect on aqueous sulfate production is sensitive to the amount of ozone present to oxidize S(IV) in these high pH fogs.

  6. High quality Ge epilayer on Si (1 0 0) with an ultrathin Si1-x Ge x /Si buffer layer by RPCVD

    NASA Astrophysics Data System (ADS)

    Chen, Da; Guo, Qinglei; Zhang, Nan; Xu, Anli; Wang, Bei; Li, Ya; Wang, Gang

    2017-07-01

    The authors report a method to grow high quality strain-relaxed Ge epilayer on a combination of low temperature Ge seed layer and Si1-x Ge x /Si superlattice buffer layer by reduced pressure chemical vapor deposition system without any subsequent annealing treatment. Prior to the growth of high quality Ge epilayer, an ultrathin Si1-x Ge x /Si superlattice buffer layer with the thickness of 50 nm and a 460 nm Ge seed layer were deposited successively at low temperature. Then an 840 nm Ge epilayer was grown at high deposition rate with the surface root-mean-square roughness of 0.707 nm and threading dislocation density of 2.5  ×  106 cm-2, respectively. Detailed investigations of the influence of ultrathin low-temperature Si1-x Ge x /Si superlattice buffer layer on the quality of Ge epilayer were performed, which indicates that the crystalline quality of Ge epilayer can be significantly improved by enhancing the Ge concentration of Si1-x Ge x /Si superlattice buffer layer.

  7. Salt intake belief, knowledge, and behavior: a cross-sectional study of older rural Chinese adults.

    PubMed

    Zhang, Jing; Wu, Tao; Chu, Hongling; Feng, Xiangxian; Shi, Jingpu; Zhang, Ruijuan; Zhang, Yuhong; Zhang, Jianxin; Li, Nicole; Yan, Lijing; Niu, Wenyi; Wu, Yangfeng

    2016-08-01

    Excess sodium consumption is a major cause of high blood pressure and subsequent vascular disease. However, the factors driving people's salt intake behavior remains largely unknown. This study aims to assess the relationship of salt intake behaviors with knowledge and belief on salt and health among older adults in rural China.A cross-sectional survey was conducted among 4693 older participants (men ≥50 and women ≥60 years old) randomly selected from 120 rural villages in 5 northern provinces in China. Healthy salt intake behavior was defined as either not eating pickled foods or not adding pickles/soy sauce/salt when food was not salty enough in prior 3 months.There were 81% participants having healthy salt intake behavior. Healthy salt intake behavior was more common among women (P < 0.01) and was positively associated with age (P < 0.01) and poorer health status (P < 0.01), but negatively associated with years in school (P < 0.05). After adjusting for age, sex, years in school, and health status, participants who believed in the harm of high salt intake were more likely to have healthy salt intake behavior, compared with those who did not believe (Odds Ratio = 1.6, P < 0.001). Knowledge of salt intake was not significantly related to healthy salt intake behavior.Our study demonstrated that belief in the harm of high salt intake rather than knowledge about salt and health was associated with healthy salt intake behavior, independent of age, sex, years in school, and health status. Future population salt reduction programs should place more emphasis on establishing health beliefs rather than only delivering salt-related knowledge.Clinical trial registration number of the study is NCT01259700.

  8. Altered potassium ATP channel signaling in mesenteric arteries of old high salt-fed rats

    PubMed Central

    Whidden, Melissa A.; Basgut, Bilgen; Kirichenko, Nataliya; Erdos, Benedek; Tümer, Nihal

    2016-01-01

    [Purpose] Both aging and the consumption of a high salt diet are associated with clear changes in the vascular system that can lead to the development of cardiovascular disease; however the mechanisms are not clearly understood. Therefore, we examined whether aging and the consumption of excess salt alters the function of potassium ATP-dependent channel signaling in mesenteric arteries [Methods] Young (7 months) and old (29 months) Fischer 344 x Brown Norway rats were fed a control or a high salt diet (8% NaCl) for 12 days and mesenteric arteries were utilized for vascular reactivity measurements. [Results] Acetylcholine-induced endothelium relaxation was significantly reduced in old arteries (81 ± 4%) when compared with young arteries (92 ± 2%). Pretreatment with the potassium-ATP channel blocker glibenclamide reduced relaxation to acetylcholine in young arteries but did not alter dilation in old arteries. On a high salt diet, endothelium dilation to acetylcholine was significantly reduced in old salt arteries (60 ± 3%) when compared with old control arteries (81 ± 4%). Glibenclamide reduced acetylcholine-induced dilation in young salt arteries but had no effect on old salt arteries. Dilation to cromakalim, a potassium-ATP channel opener, was reduced in old salt arteries when compared with old control arteries. [Conclusion] These findings demonstrate that aging impairs endothelium-dependent relaxation in mesenteric arteries. Furthermore, a high salt diet alters the function of potassium-ATP-dependent channel signaling in old isolated mesenteric arteries and affects the mediation of relaxation stimuli. PMID:27508155

  9. Selectively Enhanced UV-A Photoresponsivity of a GaN MSM UV Photodetector with a Step-Graded AlxGa1-xN Buffer Layer.

    PubMed

    Lee, Chang-Ju; Won, Chul-Ho; Lee, Jung-Hee; Hahm, Sung-Ho; Park, Hongsik

    2017-07-21

    The UV-to-visible rejection ratio is one of the important figure of merits of GaN-based UV photodetectors. For cost-effectiveness and large-scale fabrication of GaN devices, we tried to grow a GaN epitaxial layer on silicon substrate with complicated buffer layers for a stress-release. It is known that the structure of the buffer layers affects the performance of devices fabricated on the GaN epitaxial layers. In this study, we show that the design of a buffer layer structure can make effect on the UV-to-visible rejection ratio of GaN UV photodetectors. The GaN photodetector fabricated on GaN-on-silicon substrate with a step-graded Al x Ga -x N buffer layer has a highly-selective photoresponse at 365-nm wavelength. The UV-to-visible rejection ratio of the GaN UV photodetector with the step-graded Al x Ga 1-x N buffer layer was an order-of-magnitude higher than that of a photodetector with a conventional GaN/AlN multi buffer layer. The maximum photoresponsivity was as high as 5 × 10 - ² A/W. This result implies that the design of buffer layer is important for photoresponse characteristics of GaN UV photodetectors as well as the crystal quality of the GaN epitaxial layers.

  10. Evaluating the effects of buffer conditions and extremolytes on thermostability of granulocyte colony-stimulating factor using high-throughput screening combined with design of experiments.

    PubMed

    Ablinger, Elisabeth; Hellweger, Monika; Leitgeb, Stefan; Zimmer, Andreas

    2012-10-15

    In this study, we combined a high-throughput screening method, differential scanning fluorimetry (DSF), with design of experiments (DoE) methodology to evaluate the effects of several formulation components on the thermostability of granulocyte colony stimulating factor (G-CSF). First we performed a primary buffer screening where we tested thermal stability of G-CSF in different buffers, pH values and buffer concentrations. The significance of each factor and the two-way interactions between them were studied by multivariable regression analysis. pH was identified as most critical factor regarding thermal stability. The most stabilizing buffer, sodium glutamate, and sodium acetate were determined for further investigations. Second we tested the effect of 6 naturally occurring extremolytes (trehalose, sucrose, ectoine, hydroxyectoine, sorbitol, mannitol) on the thermal stability of G-CSF, using a central composite circumscribed design. At low pH (3.8) and low buffer concentration (5 mM) all extremolytes led to a significant increase in thermal stability except the addition of ectoine which resulted in a strong destabilization of G-CSF. Increasing pH and buffer concentration led to an increase in thermal stability with all investigated extremolytes. The described systematic approach allowed to create a ranking of stabilizing extremolytes at different buffer conditions. Copyright © 2012. Published by Elsevier B.V.

  11. Retention of ionisable compounds on high-performance liquid chromatography XVII. Estimation of the pH variation of aqueous buffers with the change of the methanol fraction of the mobile phase.

    PubMed

    Subirats, Xavier; Bosch, Elisabeth; Rosés, Martí

    2007-01-05

    The use of methanol-aqueous buffer mobile phases in HPLC is a common election when performing chromatographic separations of ionisable analytes. The addition of methanol to the aqueous buffer to prepare such a mobile phase changes the buffer capacity and the pH of the solution. In the present work, the variation of these buffer properties is studied for acetic acid-acetate, phosphoric acid-dihydrogenphosphate-hydrogenphosphate, citric acid-dihydrogencitrate-hydrogencitrate-citrate, and ammonium-ammonia buffers. It is well established that the pH change of the buffers depends on the initial concentration and aqueous pH of the buffer, on the percentage of methanol added, and on the particular buffer used. The proposed equations allow the pH estimation of methanol-water buffered mobile phases up to 80% in volume of organic modifier from initial aqueous buffer pH and buffer concentration (before adding methanol) between 0.001 and 0.01 mol L(-1). From both the estimated pH values of the mobile phase and the estimated pKa of the ionisable analytes, it is possible to predict the degree of ionisation of the analytes and therefore, the interpretation of acid-base analytes behaviour in a particular methanol-water buffered mobile phase.

  12. Sensing small molecule interactions with lipid membranes by local pH modulation.

    PubMed

    Huang, Da; Zhao, Tao; Xu, Wei; Yang, Tinglu; Cremer, Paul S

    2013-11-05

    Herein, we utilized a label-free sensing platform based on pH modulation to detect the interactions between tetracaine, a positively charged small molecule used as a local anesthetic, and planar supported lipid bilayers (SLBs). The SLBs were patterned inside a flow cell, allowing for various concentrations of tetracaine to be introduced over the surface in a buffer solution. Studies with membranes containing POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) yielded an equilibrium dissociation constant value of Kd = 180 ± 47 μm for this small molecule-membrane interaction. Adding cholesterol to the SLBs decreased the affinity between tetracaine and the bilayers, while this interaction tightened when POPE (1-hexadecanoyl-2-(9-Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine) was added. Studies were also conducted with three negatively charged membrane lipids, POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (sodium salt)), POPS (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (sodium salt)), and ganglioside GM1. All three measurements gave rise to a similar tightening of the apparent Kd value compared with pure POPC membranes. The lack of chemical specificity with the identity of the negatively charged lipid indicated that the tightening was largely electrostatic. Through a direct comparison with ITC measurements, it was found that the pH modulation sensor platform offers a facile, inexpensive, highly sensitive, and rapid method for the detection of interactions between putative drug candidates and lipid bilayers. As such, this technique may potentially be exploited as a screen for drug development and analysis.

  13. Multi-pack Disposal Concepts for Spent Fuel (Rev. 0)

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

    Hadgu, Teklu; Hardin, Ernest; Matteo, Edward N.

    2015-12-01

    At the initiation of the Used Fuel Disposition (UFD) R&D campaign, international geologic disposal programs and past work in the U.S. were surveyed to identify viable disposal concepts for crystalline, clay/shale, and salt host media (Hardin et al., 2012). Concepts for disposal of commercial spent nuclear fuel (SNF) and high-level waste (HLW) from reprocessing are relatively advanced in countries such as Finland, France, and Sweden. The UFD work quickly showed that these international concepts are all “enclosed,” whereby waste packages are emplaced in direct or close contact with natural or engineered materials . Alternative “open” modes (emplacement tunnels are keptmore » open after emplacement for extended ventilation) have been limited to the Yucca Mountain License Application Design (CRWMS M&O, 1999). Thermal analysis showed that, if “enclosed” concepts are constrained by peak package/buffer temperature, waste package capacity is limited to 4 PWR assemblies (or 9-BWR) in all media except salt. This information motivated separate studies: 1) extend the peak temperature tolerance of backfill materials, which is ongoing; and 2) develop small canisters (up to 4-PWR size) that can be grouped in larger multi-pack units for convenience of storage, transportation, and possibly disposal (should the disposal concept permit larger packages). A recent result from the second line of investigation is the Task Order 18 report: Generic Design for Small Standardized Transportation, Aging and Disposal Canister Systems (EnergySolution, 2015). This report identifies disposal concepts for the small canisters (4-PWR size) drawing heavily on previous work, and for the multi-pack (16-PWR or 36-BWR).« less

  14. Multi-Pack Disposal Concepts for Spent Fuel (Revision 1)

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

    Hardin, Ernest; Matteo, Edward N.; Hadgu, Teklu

    2016-01-01

    At the initiation of the Used Fuel Disposition (UFD) R&D campaign, international geologic disposal programs and past work in the U.S. were surveyed to identify viable disposal concepts for crystalline, clay/shale, and salt host media. Concepts for disposal of commercial spent nuclear fuel (SNF) and high-level waste (HLW) from reprocessing are relatively advanced in countries such as Finland, France, and Sweden. The UFD work quickly showed that these international concepts are all “enclosed,” whereby waste packages are emplaced in direct or close contact with natural or engineered materials . Alternative “open” modes (emplacement tunnels are kept open after emplacement formore » extended ventilation) have been limited to the Yucca Mountain License Application Design. Thermal analysis showed that if “enclosed” concepts are constrained by peak package/buffer temperature, that waste package capacity is limited to 4 PWR assemblies (or 9 BWR) in all media except salt. This information motivated separate studies: 1) extend the peak temperature tolerance of backfill materials, which is ongoing; and 2) develop small canisters (up to 4-PWR size) that can be grouped in larger multi-pack units for convenience of storage, transportation, and possibly disposal (should the disposal concept permit larger packages). A recent result from the second line of investigation is the Task Order 18 report: Generic Design for Small Standardized Transportation, Aging and Disposal Canister Systems. This report identifies disposal concepts for the small canisters (4-PWR size) drawing heavily on previous work, and for the multi-pack (16-PWR or 36-BWR).« less

  15. Resolving the Intricacies of Lateral Exports of Inorganic Carbon and Alkalinity from Coastal Salt Marshes

    NASA Astrophysics Data System (ADS)

    Wang, A. Z.; Chu, S. N.; Kroeger, K. D.; Gonneea, M. E.; Ganju, N. K.

    2017-12-01

    Dynamic lateral exports of dissolved inorganic carbon (DIC) and total alkalinity (Alk) via tidal exchange from highly productive intertidal marshes are an important piece of puzzle in the coastal carbon cycle, challenging our capability of assessing coastal carbon budgets and projecting future changes under anthropogenic pressure. The effects of these exports on seawater chemistry are profound yet complicated to study. This study presents the latest development of assessing lateral DIC and Alk fluxes from tidal marshes and examining their effects on seawater chemistry and coastal carbon budgets. The study evaluates different approaches to quantify these exports in order to obtain insights on the best and efficient way to capture the dynamics of such exports. A state-of-the-art DIC sensor, Channelized Optical System (CHANOS), was deployed to establish the true DIC fluxes. They are compared to the fluxes derived from empirical modeling and traditional bottle measurements. Salt marshes can acidify and alkalize tidal water by injecting CO2 (DIC) and Alk over a same tidal cycle. However, their generation is decoupled as a result of deferential effects of aerobic and anaerobic respirations. This creates complex scenarios of large swings of seawater chemistry and buffering capacity in tidal water over tidal and seasonal cycles. Marsh exports of DIC and Alk may have complex implications for the future, more acidified ocean. The latest estimates of marsh DIC and Alk exports suggest they are a major term in the marsh carbon budget and can be translated into one of the primary components in the coastal carbon cycle.

  16. Common data buffer

    NASA Technical Reports Server (NTRS)

    Byrne, F.

    1981-01-01

    Time-shared interface speeds data processing in distributed computer network. Two-level high-speed scanning approach routes information to buffer, portion of which is reserved for series of "first-in, first-out" memory stacks. Buffer address structure and memory are protected from noise or failed components by error correcting code. System is applicable to any computer or processing language.

  17. Impact of varying buffer thickness generated strain and threading dislocations on the formation of plasma assisted MBE grown ultra-thin AlGaN/GaN heterostructure on silicon

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

    Chowdhury, Subhra, E-mail: subhra1109@gmail.com; Biswas, Dhrubes; Department of E and E C E, Indian Institute of Technology Kharagpur, Kharagpur 721302

    2015-05-15

    Plasma-assisted molecular beam epitaxy (PAMBE) growth of ultra-thin Al{sub 0.2}Ga{sub 0.8}N/GaN heterostructures on Si(111) substrate with three buffer thickness (600 nm/400 nm/200 nm) have been reported. An unique growth process has been developed that supports lower temperature epitaxy of GaN buffer which minimizes thermally generated tensile strain through appropriate nitridation and AlN initiated epitaxy for achieving high quality GaN buffer which supports such ultra-thin heterostructures in the range of 10-15Å. It is followed by investigations of role of buffer thickness on formation of ultra-thin Al{sub 0.2}Ga{sub 0.8}N/GaN heterostructure, in terms of stress-strain and threading dislocation (TD). Structural characterization were performedmore » by High-Resolution X-Ray Diffraction (HRXRD), room-temperature Photoluminescence (RT-PL), High Resolution Transmission Electron Microscopy (HRTEM) and Atomic Force Microscopy (AFM). Analysis revealed increasing biaxial tensile stress of 0.6918 ± 0.04, 1.1084, 1.1814 GPa in heterostructures with decreasing buffer thickness of 600, 400, 200 nm respectively which are summed up with residual tensile strain causing red-shift in RT-PL peak. Also, increasing buffer thickness drastically reduced TD density from the order 10{sup 10} cm{sup −2} to 10{sup 8} cm{sup −2}. Surface morphology through AFM leads to decrease of pits and root mean square value with increasing buffer thickness which are resulted due to reduction of combined effect of strain and TDs.« less

  18. Increased Dietary Sodium Induces COX2 Expression by activating NFκB in Renal Medullary Interstitial Cells

    PubMed Central

    Zhao, Min; Davis, Linda S.; Blackwell, Timothy S.; Yull, Fiona; Breyer, Matthew D.; Hao, Chuan-Ming

    2013-01-01

    High salt diet induces renal medullary COX2 expression. Selective blockade of renal medullary COX2 activity in rats causes salt sensitive hypertension, suggesting a role for renal medullary COX2 in maintaining systemic sodium balance. The present study characterized the cellular location of COX2 induction in the kidney of mice following high salt diet and examined the role of NFκB in mediating this COX2 induction in response to increased dietary salt. High salt diet (8% NaCl) for 3 days markedly increased renal medullary COX2 expression in C57Bl/6J mice. Co-immunofluorescence using a COX2 antibody and antibodies against AQP2, ClC-K, AQP1 and CD31 showed that high salt diet-induced COX2 was selectively expressed in renal medullary interstitial cells. By using NFκB reporter transgenic mice, we observed a 7 fold increase of luciferase activity in the renal medulla of the NFκB-luciferase reporter mice following high salt diet, and a robust induction of EGFP expression mainly in renal medullary interstitial cells of the NFκB-EGFP reporter mice following high salt diet. Treating high salt diet fed C57Bl/6J mice with selective IκB kinase inhibitor IMD-0354 (8mg/kg bw) substantially suppressed COX2 induction in renal medulla, and also significantly reduced urinary PGE2. These data therefore suggest that renal medullary interstitial cell NFκB plays an important role in mediating renal medullary COX2 expression and promoting renal PGE2 synthesis in response to increased dietary sodium. PMID:23900806

  19. Facilitation through Buffer Saturation: Constraints on Endogenous Buffering Properties

    PubMed Central

    Matveev, Victor; Zucker, Robert S.; Sherman, Arthur

    2004-01-01

    Synaptic facilitation (SF) is a ubiquitous form of short-term plasticity, regulating synaptic dynamics on fast timescales. Although SF is known to depend on the presynaptic accumulation of Ca2+, its precise mechanism is still under debate. Recently it has been shown that at certain central synapses SF results at least in part from the progressive saturation of an endogenous Ca2+ buffer (Blatow et al., 2003), as proposed by Klingauf and Neher (1997). Using computer simulations, we study the magnitude of SF that can be achieved by a buffer saturation mechanism (BSM), and explore its dependence on the endogenous buffering properties. We find that a high SF magnitude can be obtained either by a global saturation of a highly mobile buffer in the entire presynaptic terminal, or a local saturation of a completely immobilized buffer. A characteristic feature of BSM in both cases is that SF magnitude depends nonmonotonically on the buffer concentration. In agreement with results of Blatow et al. (2003), we find that SF grows with increasing distance from the Ca2+ channel cluster, and increases with increasing external Ca2+, [Ca2+]ext, for small levels of [Ca2+]ext. We compare our modeling results with the experimental properties of SF at the crayfish neuromuscular junction, and find that the saturation of an endogenous mobile buffer can explain the observed SF magnitude and its supralinear accumulation time course. However, we show that the BSM predicts slowing of the SF decay rate in the presence of exogenous Ca2+ buffers, contrary to experimental observations at the crayfish neuromuscular junction. Further modeling and data are required to resolve this aspect of the BSM. PMID:15111389

  20. Preconcentration of DNA using magnetic ionic liquids that are compatible with real-time PCR for rapid nucleic acid quantification.

    PubMed

    Emaus, Miranda N; Clark, Kevin D; Hinners, Paige; Anderson, Jared L

    2018-04-28

    Nucleic acid extraction and purification represents a major bottleneck in DNA analysis. Traditional methods for DNA purification often require reagents that may inhibit quantitative polymerase chain reaction (qPCR) if not sufficiently removed from the sample. Approaches that employ magnetic beads may exhibit lower extraction efficiencies due to sedimentation and aggregation. In this study, four hydrophobic magnetic ionic liquids (MILs) were investigated as DNA extraction solvents with the goal of improving DNA enrichment factors and compatibility with downstream bioanalytical techniques. By designing custom qPCR buffers, we directly incorporated DNA-enriched MILs including trihexyl(tetradecyl)phosphonium tris(hexafluoroacetylaceto)nickelate(II) ([P 6,6,6,14 + ][Ni(hfacac) 3 - ]), [P 6,6,6,14 + ] tris(hexafluoroacetylaceto)colbaltate(II) ([Co(hfacac) 3 - ]), [P 6,6,6,14 + ] tris(hexafluoroacetylaceto)manganate(II) ([Mn(hfacac) 3 - ]), or [P 6,6,6,14 + ] tetrakis(hexafluoroacetylaceto)dysprosate(III) ([Dy(hfacac) 4 - ]) into reaction systems, thereby circumventing the need for time-consuming DNA recovery steps. Incorporating MILs into the reaction buffer did not significantly impact the amplification efficiency of the reaction (91.1%). High enrichment factors were achieved using the [P 6,6,6,14 + ][Ni(hfacac) 3 - ] MIL for the extraction of single-stranded and double-stranded DNA with extraction times as short as 2 min. When compared to a commercial magnetic bead-based platform, the [P 6,6,6,14 + ][Ni(hfacac) 3 - ] MIL was capable of producing higher enrichment factors for single-stranded DNA and similar enrichment factors for double-stranded DNA. The MIL-based method was applied for the extraction and direct qPCR amplification of mutation prone-KRAS oncogene fragment in plasma samples. Graphical abstract Magnetic ionic liquid solvents are shown to preconcentrate sufficient KRAS DNA template from an aqueous solution in as short as 2 min without using chaotropic salts or toxic organic solvents. By using custom-designed qPCR buffers, DNA can be directly amplified and quantified from four MILs examined in this study.

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