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At least 991 records · Page 55

The Impact of Ethylenediaminetetraacetic Acid (EDTA) on the Sorption of Nd(III), Th{sup (IV)}, and U(VI) onto Dolomite in WIPP-Relevant Brines, GWB and ERDA-6 - 20356

Storage of legacy, transuranic waste at the Waste Isolation Pilot Plant (WIPP) requires accurate risk models for performance assessments. These models must predict the long-term fate and transport of contaminants in the WIPP and surrounding environment. Laboratory support is necessary for the development of an accurate model through better understanding of contaminant behavior in the WIPP. The WIPP is a deep geologic repository that is characterized by high ionic strength due to evaporite deposits in the Salado formation (porewaters up to 7.4 M). High concentrations of sodium chloride (NaCl), calcium chloride (CaCl{sub 2}), and magnesium chloride (MgCl{sub 2}) are particularly of concern due to their potential impact on solubility and ultimately transport of contaminants of concern. Additionally, ligands capable of forming strong complexes with metals are found in significant concentrations in the WIPP environment due to their disposal from waste processing and formation during degradation of repository components (e.g. cellulose degradation by calcium hydroxide in cement). Consequently, there is a need to investigate the fate and transport of contaminants in the WIPP environment with a focus on the effects of high ionic strength and strongly coordinating ligands. Contaminants of high concern include the actinides (U(VI), Pu(IV), and Am(III)). Laboratory support of the WIPP modeling efforts was provided by this research. A series of batch experiments were utilized to study the impact of ionic strength, contaminant concentration, and the presence of ethylenediaminetetraacetic acid (EDTA) on the sorption of Nd(III), Th(IV), and U(VI) onto dolomite, a carbonate mineral found within the Culebra formation. The Culebra formation is the most transmissive layer above the WIPP, making transport of contaminants the most likely route in previous risk assessments. Two brines specifically designed to imitate conditions within the WIPP environment were used in these experiments. The U.S. Energy Research and Development Administration Well 6 (ERDA-6, 95%) brine simulates a low Mg environment, while the generic weep brine (GWB, 95%) simulates the WIPP environment with a high Mg concentration. Mg is added to the repository in the form of MgO as an engineered barrier and is naturally present in some formations. It is expected to scavenge carbonate from solutions decreasing formation of the highly soluble and competitive uranium-carbonate complexes. EDTA is used during manipulations of nuclear materials due to its ability to form strong, stable complexes with the actinides and may be found in high concentrations (up to 0.08 mM) within the WIPP as waste canisters degrade. Further, it may provide additional avenues for unwanted transport of contaminants. Nd(III) and Th(IV) were used as stable analogues of Am(III) and Pu(IV), respectively, to represent the most common oxidation states of actinides in the WIPP environment in addition to U(VI). The potential for colloid formation was also investigated with low and high concentrations of contaminants (10 and 1000 μg/L) using three size-dependent particle separation steps. The results of this research provide a better understanding of potential behavior of actinides in the presence of EDTA and within WIPP-relevant brines. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Ca2+ Effects on Fe(II) Interactions with Mn-Binding Sites in Mn-Depleted Oxygen-Evolving Complexes of Photosystem II and on Fe Replacement of Mn in Mn-Containing, Ca-Depleted Complexes

Fe(II) cations bind with high efficiency and specificity at the high-affinity (HA), Mn-binding site (termed the “blocking effect” since Fe blocks further electron donation to the site) of the oxygen-evolving complex (OEC) in Mn-depleted, photosystem II (PSII) membrane fragments (Semin et al. in Biochemistry 41:5854, 2002). Furthermore, Fe(II) cations can substitute for 1 or 2Mn cations (pH dependent) in Ca-depleted PSII membranes (Semin et al. in Journal of Bioenergetics and Biomembranes 48:227, 2016; Semin et al. in Journal of Photochemistry and Photobiology B 178:192, 2018). In the current study, we examined the effect of Ca2+ cations on the interaction of Fe(II) ions with Mn-depleted [PSII(-Mn)] and Ca-depleted [PSII(-Ca)] photosystem II membranes. We found that Ca2+ cations (about 50 mM) inhibit the light-dependent oxidation of Fe(II) (5 µM) by about 25% in PSII(-Mn) membranes, whereas inhibition of the blocking process is greater at about 40%. Blocking of the HA site by Fe cations also decreases the rate of charge recombination between QA- and YZ•+ from t1/2?=?30 ms to 46 ms. However, Ca2+ does not affect the rate during the blocking process. An Fe(II) cation (20 µM) replaces 1Mn cation in the Mn4CaO5 catalytic cluster of PSII(-Ca) membranes at pH 5.7 but 2 Mn cations at pH 6.5. In the presence of Ca2+ (10 mM) during the substitution process, Fe(II) is not able to extract Mn at pH 5.7 and extracts only 1Mn at pH 6.5 (instead of two without Ca2+). Measurements of fluorescence induction kinetics support these observations. Inhibition of Mn substitution with Fe(II) cations in the OEC only occurs with Ca2+ and Sr2+ cations, which are also able to restore oxygen evolution in PSII(-Ca) samples. Nonactive cations like La3+, Ni2+, Cd2+, and Mg2+ have no influence on the replacement of Mn with Fe. These results show that the location and/or ligand composition of one Mn cation in the Mn4CaO5 cluster is strongly affected by calcium depletion or rebinding and that bound calcium affects the redox potential of the extractable Mn4 cation in the OEC, making it resistant to reduction.

calcium↗

Production of α' H -belite-CSA cement at low firing temperatures

A global increase in population creates a need for infrastructure development to stimulate economic growth and improve quality of life. Concrete plays a significant role in this development due to its availability and mechanical strength. Ordinary Portland cement (OPC) has been the most utilized binder in the construction industry since its formulation. However, OPC production has an enormous energy demand and generates large quantities of CO 2 . This research investigates the production of a clinker, containing mainly the hydraulically activated alpha'H-belite polymorph and calcium sulfoaluminate (ye'elimite or C4A3$\acute{S}$), at a lower firing temperature, using a combination of fluoride and boron oxide. This novel clinker addresses the problem of creating high strength cements at extremely low firing temperatures of 1000–1050 °C (a decrease of 450–500 °C as compared to OPC, and a decrease of 200–250 °C as compared to calcium sulfoaluminate (CSA) cement). This clinker reduces CO 2 emissions from energy consumption and by producing phases with lower limestone requirements. The combination of lower firing temperatures and clinker phases that require lower limestone requirements provide a means to reduce CO 2 emissions.

36 MATERIALS SCIENCE↗

A high-efficiency self-healing cementitious material based on supramolecular hydrogels impregnated with phosphate and ammonium

High-efficiency self-healing cementitious materials using hydroxyapatites as healing product were described. Supramolecular hydrogels impregnated with phosphate and ammonium ions were designed. Morphology of the supramolecular hydrogel and its capability to release phosphate in solution, the structure of healing products and the self-healing efficiency were characterized with a range of analytical techniques. Cracks in the cementitious materials with width up to 1 mm are closed and completely healed. The healing product in the cracks without ammonium ions was brushite with a sponge-like morphology. Released ammonium ions reacted with calcium hydroxide in the cement matrix, increasing the concentration of soluble calcium ions up to 300 times that in reference pore solution. Ammonium ions facilitated the formation and aggregation of healing products which were converted into hydroxyapatites, improving the maximum healable crack width.

36 MATERIALS SCIENCE↗

Strategies for control and mitigation of efflorescence in metakaolin-based geopolymers

Efflorescence can be harmful to material performance as a consequence of microstructural changes due to leaching and carbonation of alkalis. This article identifies mechanisms of efflorescence in metakaolin-based geopolymers, and approaches to reduce free/mobile alkali cations and pore network connectivity, and hence reduce efflorescence. Mechanical, mass transport and microstructural properties are assessed in metakaolin-based geopolymer cements produced using different alkali concentrations in the alkali activator and substitution/addition of blast furnace slag, silica fume, calcium aluminate cement, silicone oil and a dispersive admixture. The most effective strategy to reduce efflorescence results from increasing Si/Al in the geopolymer gel by addition of soluble silicate, silica fume or silicone oil. Addition of calcium aluminate cement or a dispersive admixture also reduce efflorescence, but are less effective. These results confirm that efflorescence may be reduced by controlling the amount of soluble silicates and reducing free alkali cations in geopolymers, providing enhanced performance and durability.

36 MATERIALS SCIENCE↗

Continuous optical in-situ pH monitoring during early hydration of cementitious materials

A luminescent sensor technique for pH analyses has been tested in cementitious media. The optical sensors monitor in-situ pH every 5 s during early hydration reactions over a period of 100 h from initial fluid up to consolidated stage. The pH development was analysed in calcium aluminate, calcium sulfoaluminate and OPC/slag cements up to a pH level of 13.3 and directly correlated with the heat of hydration. Thus tracing the dissolution and formation processes of solid phases via pH sensor monitoring is possible. This measurement technique enables the recording of continuous and highly reproducible pH data sets from slurries, pastes, and hardening materials leading to new vistas for individual (trans)formation pathways of solids, hydration and consolidation reactions in high alkaline cementitious systems.

36 MATERIALS SCIENCE↗

Isotopic fractionation accompanying CO 2 hydroxylation and carbonate precipitation from high pH waters at The Cedars, California, USA

The Cedars ultramafic block hosts alkaline springs (pH > 11) in which calcium carbonate forms upon uptake of atmospheric CO 2 and at times via mixing with surface water. Here, these processes lead to distinct carbonate morphologies with “floes” forming at the atmosphere-water interface, “snow” of fine particles accumulating at the bottom of pools and terraced constructions of travertine. Floe material is mainly composed of aragonite needles despite CaCO 3 precipitation occurring in waters with low Mg/Ca (<0.01). Precipitation of aragonite is likely promoted by the high pH (11.5–12.0) of pool waters, in agreement with published experiments illustrating the effect of pH on calcium carbonate polymorph selection.

58 GEOSCIENCES↗

New insights into Mn 2+ and Mg 2+ inhibition of calcite growth

Impurity ion and isotope partitioning into carbonate minerals provide a window into the molecular processes occurring at the fluid-mineral interface during crystal growth. Here, we employ calcium isotope fractionation together with process-based modeling to elucidate the mechanisms by which two divalent cations with starkly contrasting compatibility, magnesium and manganese, inhibit calcite growth and incorporate into the mineral lattice. Calcite growth inhibition by Mg 2+ is log-linear and K Mg is on the order of 0.02–0.03 throughout the range of {Mg 2+ }/{Ca 2+ } studied here (0.01–2.6). Mn 2+ exhibits much stronger log-linear growth rate inhibition at low Mn 2+ concentrations (fluid {Mn 2+ }/{Ca 2+ } = 0.001–0.02). Mn 2+ is readily incorporated into the calcite lattice to form a calcite-rhodochrosite solid solution, with large partition coefficients (K Mn 4.6–15.6) inversely correlated to growth rate. For both Mn 2+ and Mg 2+ , calcium isotope fractionation is found to be invariant with {Me 2+ }/{Ca 2+ } despite more than an order of magnitude decline in growth rate. This invariant Δ 44/40 Ca suggests that the presence of Mn 2 + or Mg 2+ does not significantly change the relative rates of Ca 2+ attachment and detachment at kink sites during growth, indicative of a dominantly kink blocking inhibition mechanism. Because the partitioning behavior dictates that Mn 2+ must attach to the surface significantly faster than Ca 2+ , attachment of Mn 2+ is likely to be as a non-monomer species such as an ion pair or possibly a larger polynuclear cluster. We propose that calcite growth rate inhibition by Mn is determined by the kinetics of carbonate attachment at Mn-occupied kink sites, potentially due to slow re-orientation kinetics of carbonate ions that have formed an inner-sphere complex with Mn 2+ at the surface but must reorient to incorporate into the lattice. We demonstrate that patterns in Mg 2+ partitioning and inhibition behavior are broadly consistent with growth inhibition driven by slow Mg 2+ -aquo complex dehydration relative to Ca 2+ but argue that this mechanism likely represents one endmember scenario, seen in Mg-calcite growth at low supersaturations and net precipitation rates. During growth at faster net precipitation rates, some portion of Mg 2+ is likely incorporated as a partially hydrated or otherwise complexed species, but calcite growth remains significantly inhibited by the kinetics of CO 3 2- attachment at Mg 2+ kink sites. These findings suggest a hybrid classical/nonclassical growth mechanism whereby Ca 2+ incorporates largely as a free ion at kink sites while Mn 2+ and some portion of Mg 2+ are incorporated via non-monomer attachment. This pattern may be generalizable; trace constituent cations with aquo-complex desolvation rates significantly slower than the mineral growth rate preferentially incorporate as a non-monomer species during otherwise classical crystal growth.

58 GEOSCIENCES↗

The Roles of Oil–Water Interfaces in Forming Ultrasmall CaSO 4 Nanoparticles

In natural and engineered environmental systems, calcium sulfate (CaSO 4 ) nucleation commonly occurs at dynamic liquid–liquid interfaces. Although CaSO 4 is one of the most common minerals in oil spills and oil–water separation, the mechanisms driving its nucleation at these liquid–liquid interfaces remain poorly understood. Here, in this study, using in situ small-angle X-ray scattering (SAXS), we examined CaSO 4 nucleation at oil–water interfaces and found that within 60 minutes of reaction, short rod-shaped nanoparticles (with a radius of gyration (R g ) of 17.2 ± 2.7 nm and a length of 38.2 ± 5.8 nm) had formed preferentially at the interfaces. Wide-angle X-ray scattering (WAXS) analysis identified these nanoparticles as gypsum (CaSO 4 ·2H 2 O). In addition, spherial nanoparticles measuring 4.1 nm in diameter were observed at oil–water interfaces, where surface-enhanced Raman spectroscopy (SERS) revealed an elevated pH compared to the bulk solution. The negatively charged oil–water interfaces preferentially adsorb calcium ions, collectively promoting CaSO 4 formation there. CaSO 4 particle formation at the oil–water interface follows a nonclassical nucleation (N-CNT) pathway by forming ultrasmall amorphous spherical particles which then aggregate to form intermediate nanoparticles, subsequently growing into nanorod-shaped gypsum. These findings of this study provide insights into mineral scaling during membrane separation and can inform more efficient oil transport in energy recovery systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Shape-preserving amorphous-to-crystalline transformation of CaCO3 revealed by in situ TEM

Organisms use inorganic ions and macromolecules to regulate crystallization from amorphous- precursors, endowing natural biominerals with complex morphologies and enhanced properties. The mechanisms by which modifiers enable these shape preserving transformations are poorly understood. We used in situ liquid-phase transmission electron microscopy to follow the evolution from amorphous calcium carbonate to calcite in the presence of additives. A combination of contrast analysis and infrared spectroscopy shows that Mg ions, which are widely present in seawater and biological fluids, alter the transformation pathway in a concentration-dependent manner. The ions bring excess (structural) water into the amorphous bulk so that a direct transformation is triggered by dehydration in the absence of morphological changes. Molecular dynamics simulations suggest Mg-incorporated water induces structural fluctuations, allowing transformation without the need to nucleate a separate crystal. Thus, the obtained calcite retains the original morphology of the amorphous state, biomimetically achieving the morphological control of crystals seen in biominerals

crystallization, biomineralization, calcium carbon↗

SPRUCE Precipitation Chemistry and Bulk Atmospheric Deposition Beginning in 2013

This data set reports the chemistry of precipitation water that was collected at the SPRUCE site located in the S1 bog and calculated values of bulk atmospheric deposition of solutes in precipitation. Sample collection and analyses started during December of 2013 and will continue for the duration of the experiment. Precipitation samples were collected year round on an event-basis from up to three individual collectors. Samples were either composited or individually analyzed, depending on the total volume of water available relative to the amount needed to complete all laboratory analyses. Funnel/bottle collectors were used to collect rainfall and buckets were used to collect snowfall and mixed precipitation. Samples were analyzed for pH, specific conductivity, nutrient concentrations (ammonium, nitrate, soluble reactive phosphorus, total nitrogen), cation concentrations (calcium, magnesium), anion concentrations (chloride, sulfate), total organic carbon concentrations, and natural-abundance stable isotopes of water (δ18O-H2O and δD-H2O). Bulk deposition was calculated by multiplying rainfall amount by solute concentration for ammonium, nitrate, soluble reactive phosphorus (SRP), total nitrogen, calcium, magnesium, chloride, sulfate, and total organic carbon. All measurements were made at the 8.1-ha S1 bog forest site in northern Minnesota, 40 km north of Grand Rapids, in the USDA Forest Service Marcell Experimental Forest (MEF).

54 ENVIRONMENTAL SCIENCES↗

Changes in the C-terminal, N-terminal, and histidine regions of amelogenin reveal the role of oligomer quaternary structure on adsorption and hydroxyapatite mineralization

Adsorption interactions between amelogenin and calcium phosphate minerals are believed to be important to amelogenin’s function in enamel formation, however, the role of specific amino acid residues and domains within the protein in controlling adsorption is not well known. We synthesized “mechanistic probes” by systematically removing charged regions of amelogenin in order to elucidate their roles. The probes included amelogenin without the charged residues in the N-terminus (SEKR), without two, three, or eight histidines (H) in the central protein region (H2, H3, H8), or without the C-terminal residues (Delta). In-situ atomic force microscopy (AFM) adsorption studies onto hydroxyapatite (HAP) single crystals confirmed that the C-terminus was the dominant domain in promoting adsorption. We propose that subtle changes in protein-protein interactions for proteins with histidines and N-terminal residues removed resulted in changes in the oligomer quaternary size and structure that also affected protein adsorption. HAP mineralization studies revealed that the oligomer-HAP binding energy and protein layer thickness were factors in controlling the amorphous calcium phosphate (ACP) to HAP induction time. Our studies with mechanistic probes reveal the importance of the oligomer quaternary structure in controlling amelogenin adsorption and HAP mineralization.

59 BASIC BIOLOGICAL SCIENCES↗

Influence of variable rates of neritic carbonate deposition on atmospheric carbon dioxide and pelagic sediments

Short-term imbalances in the global cycle of shallow water calcium carbonate deposition and dissolution may be responsible for much of the observed Pleistocene change in atmospheric carbon dioxide content. However, any proposed changes in the alkalinity balance of the ocean must be reconciled with the sedimentary record of deep-sea carbonates. The possible magnitude of the effect of shallow water carbonate deposition on the dissolution of pelagic carbonate can be tested using numerical simulations of the global carbon cycle. Boundary conditions can be defined by using extant shallow water carbonate accumulation data and pelagic carbonate deposition/dissolution data. On timescales of thousands of years carbonate deposition versus dissolution is rarely out of equilibrium by more than 1.5 x 10(13) mole yr-1. Results indicate that the carbonate chemistry of the ocean is rarely at equilibrium on timescales less than 10 ka. This disequilibrium is probably due to sea level-induced changes in shallow water calcium carbonate deposition/dissolution, an interpretation that does not conflict with pelagic sedimentary data from the central Pacific.

NASA Discipline Exobiology↗

Effect of calmodulin antagonists on the growth and graviresponsiveness of primary roots of maize

We examined the effect of calmodulin (CaM) antagonists applied at the root tip on root growth, gravity-induced root curvature, and the movement of calcium across the root tip and auxin (IAA) across the elongation zone of gravistimulated roots. All of the CaM antagonists used in these studies delayed gravity-induced curvature at a concentration (1 micromole) that did not affect root growth. Calmodulin antagonists (> or = 1 micromole) inhibited downward transport of label from 45Ca2+ across the caps of gravistimulated roots relative to the downward transport of 45Ca2+ in gravistimulated roots which were not treated with CaM antagonists. Application of CaM antagonists at the root tip (> or = 1 micromole) also decreased the relative downward movement of label from 3H-IAA applied to the upper side of the elongation zone of gravistimulated roots. In general, tip application of antagonists inhibited neither the upward transport of 45Ca2+ in the root tip nor the upward movement of label from 3H-IAA in the elongation zone of gravistimulated roots. Thus, roots treated with CaM antagonists > or = 1 micromole become less graviresponsive and exhibit reduced or even a reversal of downward polarity of calcium transport across the root tip and IAA transport across the elongation zone. The results indicate that calmodulin-regulated events play a role in root gravitropism.

NASA Discipline Plant Biology↗

Calbindin-D(28k) controls [Ca(2+)](i) and insulin release. Evidence obtained from calbindin-d(28k) knockout mice and beta cell lines

The role of the calcium-binding protein, calbindin-D(28k) in potassium/depolarization-stimulated increases in the cytosolic free Ca(2+) concentration ([Ca(2+)](i)) and insulin release was investigated in pancreatic islets from calbindin-D(28k) nullmutant mice (knockouts; KO) or wild type mice and beta cell lines stably transfected and overexpressing calbindin. Using single islets from KO mice and stimulation with 45 mM KCl, the peak of [Ca(2+)](i) was 3.5-fold greater in islets from KO mice compared with wild type islets (p < 0.01) and [Ca(2+)](i) remained higher during the plateau phase. In addition to the increase in [Ca(2+)](i) in response to KCl there was also a significant increase in insulin release in islets isolated from KO mice. Evidence for modulation by calbindin of [Ca(2+)](i) and insulin release was also noted using beta cell lines. Rat calbindin was stably expressed in betaTC-3 and betaHC-13 cells. In response to depolarizing concentrations of K(+), insulin release was decreased by 45-47% in calbindin expressing betaTC cells and was decreased by 70-80% in calbindin expressing betaHC cells compared with insulin release from vector transfected betaTC or betaHC cells (p < 0.01). In addition, the K(+)-stimulated intracellular calcium peak was markedly inhibited in calbindin expressing betaHC cells compared with vector transfected cells (225 nM versus 1,100 nM, respectively). Buffering of the depolarization-induced rise in [Ca(2+)](i) was also observed in calbindin expressing betaTC cells. In summary, our findings, using both isolated islets from calbindin-D(28k) KO mice and beta cell lines, establish a role for calbindin in the modulation of depolarization-stimulated insulin release and suggest that calbindin can control the rate of insulin release via regulation of [Ca(2+)](i).

Non-NASA Center↗

Antisense oligodeoxynucleotide inhibition of a swelling-activated cation channel in osteoblast-like osteosarcoma cells

By patch-clamp analysis, we have shown that chronic, intermittent mechanical strain (CMS) increases the activity of stretch-activated cation channels of osteoblast-like UMR-106.01 cells. CMS also produces a swelling-activated whole-cell conductance (Gm) regulated by varying strain levels. We questioned whether the swelling-activated conductance was produced by stretch-activated cation channel activity. We have identified a gene involved in the increase in conductance by using antisense oligodeoxynucleotides (ODN) derived from the alpha 1-subunit genes of calcium channels found in UMR-106.01 cells (alpha1S, alpha1C, and alpha1D). We demonstrate that alpha 1C antisense ODNs abolish the increase in Gm in response to hypotonic swelling following CMS. Antisense ODNs to alpha1S and alpha1D, sense ODNs to alpha1C, and sham permeabilization had no effect on the conductance increase. In addition, during cell-attached patch-clamp studies, antisense ODNs to alpha1c completely blocked the swelling-activated and stretch-activated nonselective cation channel response to strain. Antisense ODNs to alpha1S treatment produced no effect on either swelling-activated or stretch-activated cation channel activity. There were differences in the stretch-activated and swelling-activated cation channel activity, but whether they represent different channels could not be determined from our data. Our data indicate that the alpha1C gene product is involved in the Gm and the activation of the swelling-activated cation channels induced by CMS. The possibility that swelling-activated cation channel genes are members of the calcium channel superfamily exists, but if alpha1c is not the swelling-activated cation channel itself, then its expression is required for induction of swelling-activated cation channel activity by CMS.

NASA Discipline Musculoskeletal↗

Cellular localization of the Ca2+ binding TCH3 protein of Arabidopsis

TCH3 is an Arabidopsis touch (TCH) gene isolated as a result of its strong and rapid upregulation in response to mechanical stimuli, such as touch and wind. TCH3 encodes an unusual calcium ion-binding protein that is closely related to calmodulin but has the potential to bind six calcium ions. Here it is shown that TCH3 shows a restricted pattern of accumulation during Arabidopsis vegetative development. These data provide insight into the endogenous signals that may regulate TCH3 expression and the sites of TCH3 action. TCH3 is abundant in the shoot apical meristem, vascular tissue, the root columella and pericycle cells that give rise to lateral roots. In addition, TCH3 accumulation in cells of developing shoots and roots closely correlates with the process of cellular expansion. Following wind stimulation, TCH3 becomes more abundant in specific regions including the branchpoints of leaf primordia and stipules, pith parenchyma, and the vascular tissue. The consequences of TCH3 upregulation by wind are therefore spatially restricted and TCH3 may function at these sites to modify cell or tissue characteristics following mechanical stimulation. Because TCH3 accumulates specifically in cells and tissues that are thought to be under the influence of auxin, auxin levels may regulate TCH3 expression during development. TCH3 is upregulated in response to low levels of exogenous indole-3-acetic acid (IAA), but not by inactive auxin-related compounds. These results suggest that TCH3 protein may play roles in mediating physiological responses to auxin and mechanical environmental stimuli.

NASA Discipline Plant Biology↗

Identification of a new EF-hand superfamily member from Trypanosoma brucei

We identified several open reading frames between the regions encoding calmodulin and ubiquitin-EP52/1 in the genome of Trypanosoma brucei. One of these, EFH5, encodes a protein 192 amino acids long. The EFH5 transcript is present in poly(A)+ mRNA and is present at similar levels in the mammalian bloodstream form and the insect procyclic form. EFH5 contains four EF-hand homolog domains, two of which are inferred to bind Ca2+ ions. We expressed EFH5 as a fusion protein in Escherichia coli and demonstrated calcium-binding activity of the fusion protein using the 45Ca-overlay technique. The function of EFH5 remains unknown; however, as the fourth EF-hand homolog identified in trypanosomes, it attests to the broad range of functions assumed by calcium functioning as a second messenger. EFH5, which is most closely related to LAV1-2 from Physarum, represents a distinct subfamily among the EF-hand-containing proteins.

Non-NASA Center↗