High Concentrations Of Divalent Cations Research Articles

Mechanistic insights into allosteric regulation of the A2A adenosine G protein-coupled receptor by physiological cations

Cations play key roles in regulating G-protein-coupled receptors (GPCRs), although their mechanisms are poorly understood. Here, 19F NMR is used to delineate the effects of cations on functional states of the adenosine A2A GPCR. While Na+ reinforces an inactive ensemble and a partial-agonist stabilized state, Ca2+ and Mg2+ shift the equilibrium toward active states. Positive allosteric effects of divalent cations are more pronounced with agonist and a G-protein-derived peptide. In cell membranes, divalent cations enhance both the affinity and fraction of the high affinity agonist-bound state. Molecular dynamics simulations suggest high concentrations of divalent cations bridge specific extracellular acidic residues, bringing TM5 and TM6 together at the extracellular surface and allosterically driving open the G-protein-binding cleft as shown by rigidity-transmission allostery theory. An understanding of cation allostery should enable the design of allosteric agents and enhance our understanding of GPCR regulation in the cellular milieu.

An acid-compatible co-polymer for the solubilization of membranes and proteins into lipid bilayer-containing nanoparticles.

The fundamental importance of membrane proteins in drug discovery has meant that membrane mimetic systems for studying membrane proteins are of increasing interest. One such system has been the amphipathic, negatively charged poly(styrene-co-maleic acid) (SMA) polymer to form "SMA Lipid Particles" (SMALPs) which have been widely adopted to solubilize membrane proteins directly from the cell membrane. However, SMALPs are only soluble under basic conditions and precipitate in the presence of divalent cations required for many downstream applications. Here, we show that the positively charged poly(styrene-co-maleimide) (SMI) forms similar nanoparticles with comparable efficiency to SMA, whilst remaining functional at acidic pH and compatible with high concentrations of divalent cations. We have performed a detailed characterization of the performance of SMI that enables a direct comparison with similar data published for SMA. We also demonstrate that SMI is capable of extracting proteins directly from the cell membrane and can solubilize functional human G-protein coupled receptors (GPCRs) expressed in cultured HEK 293T cells. "SMILPs" thus provide an alternative membrane solubilization method that successfully overcomes some of the limitations of the SMALP method.

Matrix stiffness mechanically conditions EMT and migratory behavior of oral squamous cell carcinoma.

Tumors are composed of heterogeneous phenotypes, each having different sensitivities to the microenvironment. One microenvironment characteristic - matrix stiffness - helps to regulate malignant transformation and invasion in mammary tumors, but its influence on oral squamous cell carcinoma (OSCC) is unclear. We observed that, on stiff matrices, a highly invasive OSCC cell line (SCC25) comprising a low E-cad to N-cad ratio (InvH/E:NL; SCC25) had increased migration velocity and decreased adhesion strength compared to a less invasive OSCC cell line (Cal27) with high E-cad to N-cad ratio (InvL/E:NH; Cal27). However, InvL/E:NH cells acquire a mesenchymal signature and begin to migrate faster when exposed to prolonged time on a stiff niche, suggesting that cells can be mechanically conditioned. Owing to increased focal adhesion assembly, InvL/E:NH cells migrated faster, which could be reduced when increasing integrin affinity with high divalent cation concentrations. Mirroring these data in human patients, we observed that collagen organization, an indicator of matrix stiffness, was increased with advanced disease and correlated with early recurrence. Consistent with epithelial tumors, our data suggest that OSCC cells are mechanically sensitive and that their contribution to tumor progression is mediated in part by this sensitivity.This article has an associated First Person interview with the first author of the paper.

Membrane Order Is a Key Regulator of Divalent Cation-Induced Clustering of PI(3,5)P2 and PI(4,5)P2.

Although the evidence for the presence of functionally important nanosized phosphorylated phosphoinositide (PIP)-rich domains within cellular membranes has accumulated, very limited information is available regarding the structural determinants for compartmentalization of these phospholipids. Here, we used a combination of fluorescence spectroscopy and microscopy techniques to characterize differences in divalent cation-induced clustering of PI(4,5)P2 and PI(3,5)P2. Through these methodologies we were able to detect differences in divalent cation-induced clustering efficiency and cluster size. Ca2+-induced PI(4,5)P2 clusters are shown to be significantly larger than the ones observed for PI(3,5)P2. Clustering of PI(4,5)P2 is also detected at physiological concentrations of Mg2+, suggesting that in cellular membranes, these molecules are constitutively driven to clustering by the high intracellular concentration of divalent cations. Importantly, it is shown that lipid membrane order is a key factor in the regulation of clustering for both PIP isoforms, with a major impact on cluster sizes. Clustered PI(4,5)P2 and PI(3,5)P2 are observed to present considerably higher affinity for more ordered lipid phases than the monomeric species or than PI(4)P, possibly reflecting a more general tendency of clustered lipids for insertion into ordered domains. These results support a model for the description of the lateral organization of PIPs in cellular membranes, where both divalent cation interaction and membrane order are key modulators defining the lateral organization of these lipids.

Coupled Microbial Electrolysis Cells and Struvite Precipitation for Enhanced Nutrient Recovery from Wastewater

Anaerobic digestion is extensively used to recover energy from municipal waste and animal manure. The liquid effluent of anaerobic digestion typically contains high concentrations of ammonium, phosphorus, and divalent cations, which can potentially be recovered by precipitation of sparingly soluble phosphate salts [1-5]. The majority of processes focus on struvite (magnesium ammonium phosphate) crystallization, as both nitrogen and phosphorus are removed and struvite is salable as slow-release plant fertilizer. The potential economics of recovering renewable sources of nitrogen and phosphorus have led to development of various phosphate salt crystallization systems [5-7]. Fluidized bed reactors (FBRs) are commonly used for crystallized phosphate recovery because the design creates an abundance of reactive surface area and solution turbulence [8], enhancing nucleation kinetics and agglomeration. Despite generally high nutrient recovery efficiencies in FBRs, the economic favorability of phosphate salt crystallization systems could be significantly enhanced by increasing the operating pH within the FBR to near the minimum solubility point of struvite (pH = 9 [3]) Sodium hydroxide (NaOH) is often used in research-scale investigation of these technologies because it is highly soluble and can rapidly increase the solution pH [4,7,9,10]. However, such chemical addition approaches can drive up phosphorus recovery costs (as high as $3,500/ton-P) significantly beyond the value of nutrients contained in struvite (~$765/ton-P) [3,11]. Recently, microbial electrochemical technologies (METs), such as microbial fuel cells (MFCs) and microbial electrolysis cells (MECs) have garnered interest as a chemical-free and energy efficient method of enhancing struvite precipitation [12-14]. METs consist of an anode, where anaerobic microbes oxidize organic matter and transfer electrons to an external circuit, and a cathode where the electrons and protons catalytically combine by reducing oxygen to water (MFC) or producing hydrogen gas (MEC). This talk will discuss preliminary results from a bench-scale MEC-FBR system (Figure 1) that demonstrate the potential for an MEC to enable economical pH control in an FBR struvite precipitation system. Net alkali generation rates of approximately 2.3 ± 1.5 mmol m- 2 h-1 were observed in both DC and pulsed-potential MEC tests, which are of reasonable magnitude for typical applications. When only the electrical power input is considered, these experimental data translate to a per-mole cost of alkali generated by an MEC (~7¢ mol-1) that is an order of magnitude lower than the cost of purchased NaOH (~87¢ mol-1). A preliminary net present value (NPV) analysis of the entire MEC unit operation, including both capital and operating costs, suggests a reasonable rate of return can likely be achieved. As well, the data provide preliminary indication of the possibility of further cost advantages from the use of pulsed MEC potentials. The authors acknowledge financial support from USEPA Contract No. EP-D-17-006.

Effects of the bacterial algicide IRI-160AA on cellular morphology of harmful dinoflagellates

The algicide, IRI-160AA, induces mortality in dinoflagellates but not other species of algae, suggesting that a shared characteristic or feature renders this class of phytoplankton vulnerable to the algicide. In contrast to other eukaryotic species, the genome of dinoflagellates is stabilized by high concentrations of divalent cations and transition metals and contains large amounts of DNA with unusual base modifications. These distinctions set dinoflagellates apart from other phytoplankton and suggest that the nucleus may be a dinoflagellate-specific target for IRI-160AA. In this study, morphological and ultrastructural changes in three dinoflagellate species, Prorocentrum minimum, Karlodinium veneficum and Gyrodinium instriatum, were evaluated after short-term exposure to IRI-160AA using super resolution structured illumination microscopy (SR-SIM) and transmission electron microscopy (TEM). Exposure to the algicide resulted in cytoplasmic membrane blebbing, differing chloroplast morphologies, nuclear expansion, and chromosome expulsion and/or destabilization. TEM analysis showed that chromosomes of algicide-treated K. veneficum appeared electron dense with fibrous protrusions. In algicide-treated P. minimum and G. instriatum, chromosome decompaction occurred, while for P. minimum, nuclear expulsion was also observed for several cells. Results of this investigation demonstrate that exposure to the algicide destabilizes dinoflagellate chromosomes, although it was not clear if the nucleus was the primary target of the algicide or if the observed effects on chromosomal structure were due to downstream impacts. In all cases, changes in cellular morphology and ultrastructure were observed within two hours, suggesting that the algicide may be an effective and rapid approach to mitigate dinoflagellate blooms.

Ionotropic Receptor 76b Is Required for Gustatory Aversion to Excessive Na+ in Drosophila.

Avoiding ingestion of excessively salty food is essential for cation homeostasis that underlies various physiological processes in organisms. The molecular and cellular basis of the aversive salt taste, however, remains elusive. Through a behavioral reverse genetic screening, we discover that feeding suppression by Na+-rich food requires Ionotropic Receptor 76b (Ir76b) in Drosophila labellar gustatory receptor neurons (GRNs). Concentrated sodium solutions with various anions caused feeding suppression dependent on Ir76b. Feeding aversion to caffeine and high concentrations of divalent cations and sorbitol was unimpaired in Ir76b-deficient animals, indicating sensory specificity of Ir76b-dependent Na+ detection and the irrelevance of hyperosmolarity-driven mechanosensation to Ir76b-mediated feeding aversion. Ir76b-dependent Na+-sensing GRNs in both L− and s-bristles are required for repulsion as opposed to the previous report where the L-bristle GRNs direct only low-Na+ attraction. Our work extends the physiological implications of Ir76b from low-Na+ attraction to high-Na+ aversion, prompting further investigation of the physiological mechanisms that modulate two competing components of Na+-evoked gustation coded in heterogeneous Ir76b-positive GRNs.

Variation of organic matter quantity and quality in streams at Critical Zone Observatory watersheds

The quantity and chemical composition of dissolved organic matter (DOM) in surface waters influence ecosystem processes and anthropogenic use of freshwater. However, despite the importance of understanding spatial and temporal patterns in DOM, measures of DOM quality are not routinely included as part of large-scale ecosystem monitoring programs and variations in analytical procedures can introduce artifacts. In this study, we used consistent sampling and analytical methods to meet the objective of defining variability in DOM quantity and quality and other measures of water quality in streamflow issuing from small forested watersheds located within five Critical Zone Observatory sites representing contrasting environmental conditions. Results show distinct separations among sites as a function of water quality constituents. Relationships among rates of atmospheric deposition, water quality conditions, and stream DOM quantity and quality are consistent with the notion that areas with relatively high rates of atmospheric nitrogen and sulfur deposition and high concentrations of divalent cations result in selective transport of DOM derived from microbial sources, including in-stream microbial phototrophs. We suggest that the critical zone as a whole strongly influences the origin, composition, and fate of DOM in streams. This study highlights the value of consistent DOM characterization methods included as part of long-term monitoring programs for improving our understanding of interactions among ecosystem processes as controls on DOM biogeochemistry.

Molecular Dynamics Simulation of Sulfobetaine-Type Zwitterionic Surfactants at the Decane/Water Interface: Structure, Interfacial Properties

Molecular dynamics simulations of interfacial properties of new sulfobetaine-type zwitterionic surfactants were performed using a united atomistic model, and a detailed analysis of hydrophilicity, temperature sensitivity, and salt resistance of surfactants was carried out. The new model was parameterized to reproduce the density and showed good agreement with experimental values. The results showed that the surfactant will not cease to be effective with the increase of the temperature from 300 to 373 K but as it rises, the interfacial tension will have a reduction. This kind of surfactant can be proved to remain an activity at high concentration of monovalent and divalent cations. It has commendable salt tolerance.

Cation Coordination Alters the Conformation of a Thrombin-Binding G-Quadruplex DNA Aptamer That Affects Inhibition of Thrombin.

Thrombin-binding aptamers are promising anticoagulants. HD1 is a monomolecular antiparallel G-quadruplex with two G-quartets linked by three loops. Aptamer-thrombin interactions are mediated with two TT-loops that bind thrombin exosite I. Several cations were shown to be coordinated inside the G-quadruplex, including K+, Na+, NH4+, Ba2+, and Sr2+; on the contrary, Mn2+ was coordinated in the grooves, outside the G-quadruplex. K+ or Na+ coordination provides aptamer functional activity. The effect of other cations on aptamer functional activity has not yet been described, because of a lack of relevant tests. Interactions between aptamer HD1 and a series of cations were studied. A previously developed enzymatic method was applied to evaluate aptamer inhibitory activity. The structure-function correlation was studied using the characterization of G-quadruplex conformation by circular dichroism spectroscopy. K+ coordination provided the well-known high inhibitory activity of the aptamer, whereas Na+ coordination supported low activity. Although NH4+ coordination yielded a typical antiparallel G-quadruplex, no inhibitory activity was shown; a similar effect was observed for Ba2+ and Sr2+ coordination. Mn2+ coordination destabilized the G-quadruplex that drastically diminished aptamer inhibitory activity. Therefore, G-quadruplex existence per se is insufficient for aptamer inhibitory activity. To elicit the nature of these effects, we thoroughly analyzed nuclear magnetic resonance (NMR) and X-ray data on the structure of the HD1 G-quadruplex with various cations. The most reasonable explanation is that cation coordination changes the conformation of TT-loops, affecting thrombin binding and inhibition. HD1 counterparts, aptamers 31-TBA and NU172, behaved similarly with some distinctions. In 31-TBA, an additional duplex module stabilized antiparallel G-quadruplex conformation at high concentrations of divalent cations; whereas in NU172, a different sequence of loops in the G-quadruplex module provided an equilibrium of antiparallel and parallel G-quadruplexes that shifted with cation binding. In conclusion, structures of G-quadruplex aptamers are flexible enough and are fine-tuned with different cation coordination.

Ionic mechanisms maintaining action potential conduction velocity at high firing frequencies in an unmyelinated axon.

The descending contralateral movement detector (DCMD) is a high‐performance interneuron in locusts with an axon capable of transmitting action potentials (AP) at more than 500 Hz. We investigated biophysical mechanisms for fidelity of high‐frequency transmission in this axon. We measured conduction velocities (CVs) at room temperature during exposure to 10 mmol/L cadmium, a calcium current antagonist, and found significant reduction in CV with reduction at frequencies >200 Hz of ~10%. Higher temperatures induced greater CV reductions during exposure to cadmium across all frequencies of ~20–30%. Intracellular recordings during 15 min of exposure to cadmium or nickel, also a calcium current antagonist, revealed an increase in the magnitude of the afterhyperpolarization potential (AHP) and the time to recover to baseline after the AHP (Medians for Control: −19.8%; Nickel: 167.2%; Cadmium: 387.2%), that could be due to a T‐type calcium current. However, the removal of extracellular calcium did not mimic divalent cation exposure suggesting calcium currents are not the cause of the AHP increase. Computational modeling showed that the effects of the divalent cations could be modeled with a persistent sodium current which could be blocked by high concentrations of divalent cations. Persistent sodium current shortened the AHP duration in our models and increased CV for high‐frequency APs. We suggest that faithful, high‐frequency axonal conduction in the DCMD is enabled by a mechanism that shortens the AHP duration like a persistent or resurgent sodium current.

Characterization of the FtsZ C-Terminal Variable (CTV) Region in Z-Ring Assembly and Interaction with the Z-Ring Stabilizer ZapD in E. coli Cytokinesis

Polymerization of a ring-like cytoskeletal structure, the Z-ring, at midcell is a highly conserved feature in virtually all bacteria. The Z-ring is composed of short protofilaments of the tubulin homolog FtsZ, randomly arranged and held together through lateral interactions. In vitro, lateral associations between FtsZ protofilaments are stabilized by crowding agents, high concentrations of divalent cations, or in some cases, low pH. In vivo, the last 4–10 amino acid residues at the C-terminus of FtsZ (the C-terminal variable region, CTV) have been implicated in mediating lateral associations between FtsZ protofilaments through charge shielding. Multiple Z-ring associated proteins (Zaps), also promote lateral interactions between FtsZ protofilaments to stabilize the FtsZ ring in vivo. Here we characterize the complementary role/s of the CTV of E. coli FtsZ and the FtsZ-ring stabilizing protein ZapD, in FtsZ assembly. We show that the net charge of the FtsZ CTV not only affects FtsZ protofilament bundling, confirming earlier observations, but likely also the length of the FtsZ protofilaments in vitro. The CTV residues also have important consequences for Z-ring assembly and interaction with ZapD in the cell. ZapD requires the FtsZ CTV region for interaction with FtsZ in vitro and for localization to midcell in vivo. Our data suggest a mechanism in which the CTV residues, particularly K380, facilitate a conformation for the conserved carboxy-terminal residues in FtsZ, that lie immediately N-terminal to the CTV, to enable optimal contact with ZapD. Further, phylogenetic analyses suggest a correlation between the nature of FtsZ CTV residues and the presence of ZapD in the β- γ-proteobacterial species.

Effects of molecular weight-dependent physicochemical heterogeneity of natural organic matter on the aggregation of fullerene nanoparticles in mono- and di-valent electrolyte solutions

Given the wide presence of heterogeneous natural organic matter (NOM) and metal ions (Na+/Ca2+/Mg2+), as well as their significant role in governing nanoparticle stability in aqueous environments, it is of great importance to understand how the molecular weight (MW)-dependent physicochemical properties of NOM impact fundamental transportation processes like the aggregation of engineered nanoparticles (ENPs) in the presence of Na+/Ca2+/Mg2+. Here, we report on the aggregation behavior of a model ENP, fullerene nanoparticles (nC60) in the presence of five MW fractions of Suwannee River NOM (Mf-SRNOMs, separated by ultrafiltration techniques) and three electrolytes (NaCl, CaCl2 and MgCl2). We found that in all NaCl treatments and low concentration CaCl2/MgCl2 treatments, the enhancement of nC60 stability positively correlated with the MW of Mf-SRNOMs. Whereas, the stability efficiency of identical Mf-SRNOM in different electrolytes followed an order of NaCl > MgCl2 > CaCl2, and the enhanced attachment of nC60-SRNOM associations was observed in high MW Mf-SRNOM (SRNOM>100 kD and SRNOM 30–100 kD) at high concentration CaCl2/MgCl2. Our results indicate that although the high MW NOM with large humic-like material is the key component for stabilizing nC60 in monovalent electrolyte, it could play a reversed role in promoting the attachment of nC60, especially in long term aggregations and at high concentrations of divalent cations. Therefore, a detailed understanding of the effects of heterogeneous NOM on the aggregation of ENPs should be highly valued, and properly assessed against different cation species and concentrations.

Kinetics and Equilibrium of Barium and Strontium Sulfate Formation in Marcellus Shale Flowback Water

AbstractFlowback water from natural gas extraction in Marcellus Shale contains very high concentrations of inorganic salts and organic chemicals. Potential reuse of this water in subsequent hydraulic-fracturing operations may be limited by high concentrations of divalent cations (e.g., Ba, Sr, and Ca). Kinetics of barite and celestite precipitation in flowback waters from different well sites was evaluated in this study. Ba reacted rapidly with sulfate and reached equilibrium within 30 min, whereas Sr reacted slowly and took days to reach equilibrium. Equilibrium concentrations of Ba and Sr predicted by thermodynamics models were compared with experimental results. Activity corrections based on the Pitzer equation provided the best agreement with experimental data for both Ba and Sr. Comparison of barite and celestite precipitation kinetics in actual and synthetic flowback water revealed that there was no observable impact of organics and other minor components in actual flowback water on barite precipita...

Improvement on the modified Lowry method against interference of divalent cations in soluble protein measurement

This paper systematically investigated the interference of calcium and magnesium in protein measurement with a modified Lowry method first proposed by Frølund et al. (Appl Microbiol Biotechnol 43:755-761, 1995). This interference has in the past been largely ignored resulting in variable and unreliable results when applied to natural water matrices. We discovered significant formation of calcium and magnesium precipitates that lead to a decline in light absorbance at 750 nm during protein determination. Underestimation of protein concentration (sometimes even yielding negative concentrations) and low experiment reproducibility were demonstrated at high concentrations of divalent cations (e.g., [Ca(2+)] over 1 mmol L(-1)). To eliminate interference from calcium and magnesium, two pretreatment strategies were established based on cation exchange and dialysis. These pretreatments were convenient and were found to be highly effective in removing calcium and magnesium in protein samples. By using the modified Lowry method with these pretreatments, proteins in standard solutions and in wastewater samples were successfully quantified with good reliability and reproducibility. In addition, we demonstrated that simultaneous quantification of humic substances with the modified Lowry method was not affected by the two pretreatments. These approaches are expected to be applicable to protein and humic substance determination in different research fields, in cases where the modified Lowry method is sensitive to divalent cation concentrations.

High concentrations of divalent cations isolate monosynaptic inputs from local circuits in the auditory midbrain

Hierarchical processing of sensory information occurs at multiple levels between the peripheral and central pathway. Different extents of convergence and divergence in top down and bottom up projections makes it difficult to separate the various components activated by a sensory input. In particular, hierarchical processing at sub-cortical levels is little understood. Here we have developed a method to isolate extrinsic inputs to the inferior colliculus (IC), a nucleus in the midbrain region of the auditory system, with extensive ascending and descending convergence. By applying a high concentration of divalent cations (HiDi) locally within the IC, we isolate a HiDi-sensitive from a HiDi-insensitive component of responses evoked by afferent input in brain slices and in vivo during a sound stimulus. Our results suggest that the HiDi-sensitive component is a monosynaptic input to the IC, while the HiDi-insensitive component is a local polysynaptic circuit. Monosynaptic inputs have short latencies, rapid rise times, and underlie first spike latencies. Local inputs have variable delays and evoke long-lasting excitation. In vivo, local circuits have variable onset times and temporal profiles. Our results suggest that high concentrations of divalent cations should prove to be a widely useful method of isolating extrinsic monosynaptic inputs from local circuits in vivo.

Midbrain local circuits shape sound intensity codes

Hierarchical processing of sensory information requires interaction at multiple levels along the peripheral to central pathway. Recent evidence suggests that interaction between driving and modulating components can shape both top down and bottom up processing of sensory information. Here we show that a component inherited from extrinsic sources combines with local components to code sound intensity. By applying high concentrations of divalent cations to neurons in the nucleus of the inferior colliculus in the auditory midbrain, we show that as sound intensity increases, the source of synaptic efficacy changes from inherited inputs to local circuits. In neurons with a wide dynamic range response to intensity, inherited inputs increase firing rates at low sound intensities but saturate at mid-to-high intensities. Local circuits activate at high sound intensities and widen dynamic range by continuously increasing their output gain with intensity. Inherited inputs are necessary and sufficient to evoke tuned responses, however local circuits change peak output. Push–pull driving inhibition and excitation create net excitatory drive to intensity-variant neurons and tune neurons to intensity. Our results reveal that dynamic range and tuning re-emerge in the auditory midbrain through local circuits that are themselves variable or tuned.

Divalent cations modulate membrane binding and pore formation of a potent antibiotic peptide analog of alamethicin

The Ca2+ modulation of pore formation (and disaggregation) kinetics of a synthetic analog of alamethicin F50/5 ([l-Glu(OMe)7,18,19]), a potent antibiotic peptide, was investigated in situ and in vitro. The in situ experiments consisted in whole-cell recording from isolated retinal rod outer segments (OS), because once blocking the only OS endogenous conductance with saturating light, the current flows entirely through the (exogenous) channels formed by the peptide. The kinetics of current change induced by peptide application and removal (in ∼50ms) on the OS extracellular side was measured in the presence of divalent cations at different concentrations. The in vitro experiments consisted on the divalent cations modulation of [l-Glu(OMe)7,18,19] binding to a mimetic OS membrane immobilized on a sensor chip surface, employing surface plasmon resonance spectroscopy (SPR). The presence of even low mM Ca2+ or Mg2+ sufficed to increase the [l-Glu(OMe)7,18,19] apparent affinity for the mimetic OS membrane up to ∼4-fold, which accelerated the activation of the peptide-induced current in OS by ∼10-fold with respect to low Ca2+. In situ and in vitro experiments indicate that high concentrations of divalent cations increased also membrane rigidity, contrasting their effect on increasing the pore formation rate.

Stability of sodium polyphosphate dispersants in mineral processing applications

Polyphosphates are commonly used industrially to provide electrostatic stabilisation to mineral and material suspensions. Polyphosphate solutions however have shown instability under processing conditions such as high temperature and acidic pH which may lead to a reduction in dispersion properties when added to mineral or material suspensions.In this study the influence of pH, temperature and divalent cation concentration pre-treatment on polyphosphate dispersion properties is reported. Dispersion properties are determined using rheological suspension analysis. The suspensions investigated are material-based model systems of titania pigment and high purity boehmite. Infrared spectroscopy studies of polyphosphate solutions were undertaken to directly investigate the polyphosphate structure after pre-treatment. The rheological measurements show reduced polyphosphate dispersion properties with reduced pH, increased temperature and at high calcium concentrations which correlate with directly measured and reported changes in the polyphosphate structure. When pH, temperature and high divalent cation concentrations are combined, the synergistic effect on reduced polyphosphate dispersion performance is particularly prominent.

The Periplasmic Loop Provides Stability to the Open State of the CorA Magnesium Channel

Crystal structures of the CorA Mg(2+) channel have suggested that metal binding in the cytoplasmic domain stabilizes the pentamer in a closed conformation. The open "metal free" state of the channel is, however, still structurally uncharacterized. Here, we have attempted to map conformational states of CorA from Thermotoga maritima by determining which residues support the pentameric structure in the presence or absence of Mg(2+). We find that when Mg(2+) is present, the pentamer is stabilized by the putative gating sites (M1/M2) in the cytoplasmic domain. Strikingly however, we find that the conserved and functionally important periplasmic loop is vital for the integrity of the pentamer when Mg(2+) is absent from the M1/M2 sites. Thus, although the periplasmic loops were largely disordered in the x-ray structures of the closed channel, our data suggests a prominent role for the loops in stabilizing the open conformation of the CorA channels.