Reconstruction and Simulation of Neocortical Microcircuitry.

We present a first-draft digital reconstruction of the microcircuitry of somatosensory cortex of juvenile rat. The reconstruction uses cellular and synaptic organizing principles to algorithmically reconstruct detailed anatomy and physiology from sparse experimental data. An objective anatomical method defines a neocortical volume of 0.29 ± 0.01 mm(3) containing ~31,000 neurons, and patch-clamp studies identify 55 layer-specific morphological and 207 morpho-electrical neuron subtypes. When digitally reconstructed neurons are positioned in the volume and synapse formation is restricted to biological bouton densities and numbers of synapses per connection, their overlapping arbors form ~8 million connections with ~37 million synapses. Simulations reproduce an array of in vitro and in vivo experiments without parameter tuning. Additionally, we find a spectrum of network states with a sharp transition from synchronous to asynchronous activity, modulated by physiological mechanisms. The spectrum of network states, dynamically reconfigured around this transition, supports diverse information processing strategies. PAPERCLIP: VIDEO ABSTRACT.

Cold and exogenous calcium alter Allium fistulosum cell wall pectin to depress intracellular freezing temperatures.

De-methyl esterification of homogalacturonan and subsequent cross-linking with Ca2+ is hypothesized to enhance the freezing survival of cold acclimated plants by reducing the porosity of primary cell walls. To test this theory, we collected leaf epidermal peels from non- (23/18 °C) and cold acclimated (2 weeks at 12/4 °C) Japanese bunching onion (Allium fistulosum L.). Cold acclimation enhanced the temperature at which half the cells survived freezing injury by 8 °C (LT50 =-20 °C), and reduced tissue permeability by 70-fold compared with non-acclimated epidermal cells. These effects were associated with greater activity of pectin methylesterase (PME) and a reduction in the methyl esterification of homogalacturonan. Non-acclimated plants treated with 50 mM CaCl2 accumulated higher concentrations of galacturonic acid, Ca2+ in the cell wall, and a lower number of visible cell wall pores compared with that observed in cold acclimated plants. Using cryo-microscopy, we observed that 50 mM CaCl2 treatment did not lower the LT50 of non-acclimated cells, but reduced the lethal intracellular ice nucleation to temperatures observed in cold acclimated epidermal cells. We postulate that the PME-homogalacturonan-mediated reduction in cell wall porosity is integral to intracellular freezing avoidance strategies in cold acclimated herbaceous cells.

Examining a synchrotron-based approach for in situ analyses of Al speciation in plant roots.

Aluminium (Al) K- and L-edge X-ray absorption near-edge structure (XANES) has been used to examine Al speciation in minerals but it remains unclear whether it is suitable for in situ analyses of Al speciation within plants. The XANES analyses for nine standard compounds and root tissues from soybean (Glycine max), buckwheat (Fagopyrum tataricum), and Arabidopsis (Arabidopsis thaliana) were conducted in situ. It was found that K-edge XANES is suitable for differentiating between tetrahedral coordination (peak of 1566 eV) and octahedral coordination (peak of 1568 to 1571 eV) Al, but not suitable for separating Al binding to some of the common physiologically relevant compounds in plant tissues. The Al L-edge XANES, which is more sensitive to changes in the chemical environment, was then examined. However, the poorer detection limit for analyses prevented differentiation of the Al forms in the plant tissues because of their comparatively low Al concentration. Where forms of Al differ markedly, K-edge analyses are likely to be of value for the examination of Al speciation in plant tissues. However, the apparent inability of Al K-edge XANES to differentiate between some of the physiologically relevant forms of Al may potentially limit its application within plant tissues, as does the poorer sensitivity at the L-edge.

Uranium(IV) adsorption by natural organic matter in anoxic sediments.

Uranium is an important carbon-free fuel source and environmental contaminant that accumulates in the tetravalent state, U(IV), in anoxic sediments, such as ore deposits, marine basins, and contaminated aquifers. However, little is known about the speciation of U(IV) in low-temperature geochemical environments, inhibiting the development of a conceptual model of U behavior. Until recently, U(IV) was assumed to exist predominantly as the sparingly soluble mineral uraninite (UO2+x) in anoxic sediments; however, studies now show that this is not often the case. Yet a model of U(IV) speciation in the absence of mineral formation under field-relevant conditions has not yet been developed. Uranium(IV) speciation controls its reactivity, particularly its susceptibility to oxidative mobilization, impacting its distribution and toxicity. Here we show adsorption to organic carbon and organic carbon-coated clays dominate U(IV) speciation in an organic-rich natural substrate under field-relevant conditions. Whereas previous research assumed that U(IV) speciation is dictated by the mode of reduction (i.e., whether reduction is mediated by microbes or by inorganic reductants), our results demonstrate that mineral formation can be diminished in favor of adsorption, regardless of reduction pathway. Projections of U transport and bioavailability, and thus its threat to human and ecosystem health, must consider U(IV) adsorption to organic matter within the sediment environment.

A quantum access network.

The theoretically proven security of quantum key distribution (QKD) could revolutionize the way in which information exchange is protected in the future. Several field tests of QKD have proven it to be a reliable technology for cryptographic key exchange and have demonstrated nodal networks of point-to-point links. However, until now no convincing answer has been given to the question of how to extend the scope of QKD beyond niche applications in dedicated high security networks. Here we introduce and experimentally demonstrate the concept of a 'quantum access network': based on simple and cost-effective telecommunication technologies, the scheme can greatly expand the number of users in quantum networks and therefore vastly broaden their appeal. We show that a high-speed single-photon detector positioned at a network node can be shared between up to 64 users for exchanging secret keys with the node, thereby significantly reducing the hardware requirements for each user added to the network. This point-to-multipoint architecture removes one of the main obstacles restricting the widespread application of QKD. It presents a viable method for realizing multi-user QKD networks with efficient use of resources, and brings QKD closer to becoming a widespread technology.

Importance of the RpoE Regulon in Maintaining the Lipid Bilayer during Antimicrobial Treatment with the Polycationic Agent, Chlorhexidine.

The emergence of multidrug resistance in bacteria has reached alarming levels. To solve this growing problem, discovery of novel cellular targets or pathways important for antimicrobial resistance is urgently needed. In this study, we explored how the alternative sigma factor, RpoE, protects Escherichia coli O157 against the toxic effects of the polycationic antimicrobial agent, chlorhexidine (CHX). Susceptibility of this organism to CHX was found to directly correlate to the growth rate, with the faster replicating wild-type being more susceptible to CHX than its more slowly replicating ΔrpoE O157 mutant. Once the wild-type and rpoE mutant strains had undergone growth arrest (entered the stationary growth phase), their resistance to CHX became entirely dependent on the functionality of RpoE. The RpoE regulon plays a critical role in maintaining the integrity of the asymmetric lipid bilayer of E. coli, thereby preventing the intracellular accumulation of CHX. Finally, using a single-cell, high-resolution, synchrotron-based approach, we discovered a subpopulation of the rpoE mutant strain with no detectable intracellular CHX, a predominant characteristic of the wild-type CHX-resistant population. This finding reveals a role of phenotypic heterogeneity in antimicrobial resistance.

Multispecies Biofilms Transform Selenium Oxyanions into Elemental Selenium Particles: Studies Using Combined Synchrotron X-ray Fluorescence Imaging and Scanning Transmission X-ray Microscopy.

Selenium (Se) is an element of growing environmental concern, because low aqueous concentrations can lead to biomagnification through the aquatic food web. Biofilms, naturally occurring microbial consortia, play numerous important roles in the environment, especially in biogeochemical cycling of toxic elements in aquatic systems. The complexity of naturally forming multispecies biofilms presents challenges for characterization because conventional microscopic techniques require chemical and physical modifications of the sample. Here, multispecies biofilms biotransforming selenium oxyanions were characterized using X-ray fluorescence imaging (XFI) and scanning transmission X-ray microscopy (STXM). These complementary synchrotron techniques required minimal sample preparation and were applied correlatively to the same biofilm areas. Sub-micrometer XFI showed distributions of Se and endogenous metals, while Se K-edge X-ray absorption spectroscopy indicated the presence of elemental Se (Se0). Nanoscale carbon K-edge STXM revealed the distributions of microbial cells, extracellular polymeric substances (EPS), and lipids using the protein, saccharide, and lipid signatures, respectively, together with highly localized Se0 using the Se LIII edge. Transmission electron microscopy showed the electron-dense particle diameter to be 50-700 nm, suggesting Se0 nanoparticles. The intimate association of Se0 particles with protein and polysaccharide biofilm components has implications for the bioavailability of selenium in the environment.

Metal-organic framework based upon the synergy of a Brønsted acid framework and Lewis acid centers as a highly efficient heterogeneous catalyst for fixed-bed reactions.

We report a strategy of combining a Brønsted acid metal-organic framework (MOF) with Lewis acid centers to afford a Lewis acid@Brønsted acid MOF with high catalytic activity, as exemplified in the context of MIL-101-Cr-SO3H·Al(III). Because of the synergy between the Brønsted acid framework and the Al(III) Lewis acid centers, MIL-101-Cr-SO3H·Al(III) demonstrates excellent catalytic performance in a series of fixed-bed reactions, outperforming two benchmark zeolite catalysts (H-Beta and HMOR). Our work therefore not only provides a new approach to achieve high catalytic activity in MOFs but also paves a way to develop MOFs as a new type of highly efficient heterogeneous catalysts for fixed-bed reactions.

Validating the scalability of soft X-ray spectromicroscopy for quantitative soil ecology and biogeochemistry research.

Synchrotron-based soft-X-ray scanning transmission X-ray microscopy (STXM) has the potential to provide nanoscale resolution of the associations among biological and geological materials. However, standard methods for how samples should be prepared, measured, and analyzed to allow the results from these nanoscale imaging and spectroscopic tools to be scaled to field scale biogeochemical results are not well established. We utilized a simple sample preparation technique that allows one to assess detailed mineral, metal, and microbe spectroscopic information at the nano- and microscale in soil colloids. We then evaluated three common approaches to collect and process nano- and micronscale information by STXM and the correspondence of these approaches to millimeter scale soil measurements. Finally, we assessed the reproducibility and spatial autocorrelation of nano- and micronscale protein, Fe(II) and Fe(III) densities in a soil sample. We demonstrate that linear combination fitting of entire spectra provides slightly different Fe(II) mineral densities compared to image resonance difference mapping but that difference mapping results are highly reproducible between among sample replicates. Further, STXM results scale to the mm scale in complex systems with an approximate geospatial range of 3 µm in these samples.

Microscopic and spectroscopic analyses of chlorhexidine tolerance in Delftia acidovorans biofilms.

The physicochemical responses of Delftia acidovorans biofilms exposed to the commonly used antimicrobial chlorhexidine (CHX) were examined in this study. A CHX-sensitive mutant (MIC, 1.0 µg ml(-1)) was derived from a CHX-tolerant (MIC, 15.0 µg ml(-1)) D. acidovorans parent strain using transposon mutagenesis. D. acidovorans mutant (MT51) and wild-type (WT15) strain biofilms were cultivated in flow cells and then treated with CHX at sub-MIC and inhibitory concentrations and examined by confocal laser scanning microscopy (CLSM), scanning transmission X-ray microscopy (STXM), and infrared (IR) spectroscopy. Specific morphological, structural, and chemical compositional differences between the CHX-treated and -untreated biofilms of both strains were observed. Apart from architectural differences, CLSM revealed a negative effect of CHX on biofilm thickness in the CHX-sensitive MT51 biofilms relative to those of the WT15 strain. STXM analyses showed that the WT15 biofilms contained two morphochemical cell variants, whereas only one type was detected in the MT51 biofilms. The cells in the MT51 biofilms bioaccumulated CHX to a similar extent as one of the cell types found in the WT15 biofilms, whereas the other cell type in the WT15 biofilms did not bioaccumulate CHX. STXM and IR spectral analyses revealed that CHX-sensitive MT51 cells accumulated the highest levels of CHX. Pretreating biofilms with EDTA promoted the accumulation of CHX in all cells. Thus, it is suggested that a subpopulation of cells that do not accumulate CHX appear to be responsible for greater CHX resistance in D. acidovorans WT15 biofilm in conjunction with the possible involvement of bacterial membrane stability.

Field trial of a quantum secured 10 Gb/s DWDM transmission system over a single installed fiber.

We present results from the first field-trial of a quantum-secured DWDM transmission system, in which quantum key distribution (QKD) is combined with 4 × 10 Gb/s encrypted data and transmitted simultaneously over 26 km of field installed fiber. QKD is used to frequently refresh the key for AES-256 encryption of the 10 Gb/s data traffic. Scalability to over 40 DWDM channels is analyzed.

Phosphatation of zeolite H-ZSM-5: a combined microscopy and spectroscopy study.

A variety of phosphated zeolite H-ZSM-5 samples are investigated by using a combination of Fourier transfer infrared (FTIR) spectroscopy, single pulse (27)Al, (29)Si, (31)P, (1)H-(31)P cross polarization (CP), (27)Al-(31)P CP, and (27)Al 3Q magic angle spinning nuclear magnetic resonance (MAS NMR) spectroscopy, scanning transmission X-ray microscopy (STXM) and N2 physisorption. This approach leads to insights into the physicochemical processes that take place during phosphatation. Direct phosphatation of H-ZSM-5 promotes zeolite aggregation, as phosphorus does not penetrate deep into the zeolite material and is mostly found on and close to the outer surface of the zeolite, acting as a glue. Phosphatation of pre-steamed H-ZSM-5 gives rise to the formation of a crystalline tridymite AlPO4 phase, which is found in the mesopores of dealuminated H-ZSM-5. Framework aluminum species interacting with phosphorus are not affected by hydrothermal treatment. Dealuminated H-ZSM-5, containing AlPO4 , retains relatively more framework Al atoms and acid sites during hydrothermal treatment than directly phosphated H-ZSM-5.

Soft X-ray spectromicroscopy study of mineral-organic matter associations in pasture soil clay fractions.

There is a growing acceptance that associations with soil minerals may be the most important overarching stabilization mechanism for soil organic matter. However, direct investigation of organo-mineral associations has been hampered by a lack of methods that can simultaneously characterize organic matter (OM) and soil minerals. In this study, STXM-NEXAFS spectroscopy at the C 1s, Ca 2p, Fe 2p, Al 1s, and Si 1s edges was used to investigate C associations with Ca, Fe, Al, and Si species in soil clay fractions from an upland pasture hillslope. Bulk techniques including C and N NEXAFS, Fe K-edge EXAFS spectroscopy, and XRD were applied to provide additional information. Results demonstrated that C was associated with Ca, Fe, Al, and Si with no separate phase in soil clay particles. In soil clay particles, the pervasive C forms were aromatic C, carboxyl C, and polysaccharides with the relative abundance of carboxyl C and polysaccharides varying spatially at the submicrometer scale. Only limited regions in the soil clay particles had aliphatic C. Good C-Ca spatial correlations were found for soil clay particles with no CaCO3, suggesting a strong role of Ca in organo-mineral assemblage formation. Fe EXAFS showed that about 50% of the total Fe in soils was contained in Fe oxides, whereas Fe-bearing aluminosilicates (vermiculite and Illite) accounted for another 50%. Fe oxides in the soil were mainly crystalline goethite and hematite, with lesser amounts of poorly crystalline ferrihydrite. XRD revealed that soil clay aluminosilicates were hydroxy-interlayered vermiculite, Illite, and kaolinite. C showed similar correlation with Fe to Al and Si, implying a similar association of Fe oxides and aluminosilicates with organic matter in organo-mineral associations. These direct microscopic determinations can help improve understanding of organo-mineral interactions in soils.

Properties of Fe-organic matter associations via coprecipitation versus adsorption.

The association of organic matter (OM) with minerals is recognized as the most important stabilization mechanism for soil organic matter. This study compared the properties of Fe-OM complexes formed from adsorption (reaction of OM to postsynthesis ferrihydrite) versus coprecipitation (formation of Fe solids in the presence of OM). Coprecipitates and adsorption complexes were synthesized using dissolved organic matter (DOM) extracts from a forest little layer at varying molar C/Fe ratios of 0.3-25.0. Sample properties were studied by N2 gas adsorption, XRD, FTIR, Fe EXAFS, and STXM-NEXAFS techniques. Coprecipitation resulted in much higher maximum C contents (∼130 mg g(-1) C difference) in the solid products than adsorption, which may be related to the formation of precipitated insoluble Fe(III)-organic complexes at high C/Fe ratios in the coprecipitates as revealed by Fe EXAFS analysis. Coprecipitation led to a complete blockage of mineral surface sites and pores with ≥177 mg g(-1) C and molar C/Fe ratios ≥2.8 in the solid products. FTIR and STXM-NEXAFS showed that the coprecipitated OM was similar in composition to the adsorbed OM. An enrichment of aromatic C was observed at low C/Fe ratios. Association of carboxyl functional groups with Fe was shown with FTIR and STXM-NEXAFS analysis. STXM-NEXAFS analysis showed a continuous C distribution on minerals. Desorption of the coprecipitated OM was less than that of the adsorbed OM at comparable C/Fe ratios. These results are helpful to understand C and Fe cycling in the natural environments with periodically fluctuating redox conditions, where coprecipitation can occur.

Synchrotron-based chemical nano-tomography of microbial cell-mineral aggregates in their natural, hydrated state.

Chemical nano-tomography of microbial cells in their natural, hydrated state provides direct evidence of metabolic and chemical processes. Cells of the nitrate-reducing Acidovorax sp. strain BoFeN1 were cultured in the presence of ferrous iron. Bacterial reduction of nitrate causes precipitation of Fe(III)-(oxyhydr)oxides in the periplasm and in direct vicinity of the cells. Nanoliter aliquots of cell-suspension were injected into custom-designed sample holders wherein polyimide membranes collapse around the cells by capillary forces. The immobilized, hydrated cells were analyzed by synchrotron-based scanning transmission X-ray microscopy in combination with angle-scan tomography. This approach provides three-dimensional (3D) maps of the chemical species in the sample by employing their intrinsic near-edge X-ray absorption properties. The cells were scanned through the focus of a monochromatic soft X-ray beam at different, chemically specific X-ray energies to acquire projection images of their corresponding X-ray absorbance. Based on these images, chemical composition maps were then calculated. Acquiring projections at different tilt angles allowed for 3D reconstruction of the chemical composition. Our approach allows for 3D chemical mapping of hydrated samples and thus provides direct evidence for the localization of metabolic and chemical processes in situ.

Embedding Reverse Electron Transfer Between Stably Bare Cu Nanoparticles and Cation-Vacancy CuWO4.

Cu nanoparticles (NPs) have attracted widespread attention in electronics, energy, and catalysis. However, conventionally synthesized Cu NPs face some challenges such as surface passivation and agglomeration in applications, which impairs their functionalities in the physicochemical properties. Here, the issues above by engineering an embedded interface of stably bare Cu NPs on the cation-vacancy CuWO4 support is addressed, which induces the strong metal-support interactions and reverse electron transfer. Various atomic-scale analyses directly demonstrate the unique electronic structure of the embedded Cu NPs with negative charge and anion oxygen protective layer, which mitigates the typical degradation pathways such as oxidation in ambient air, high-temperature agglomeration, and CO poisoning adsorption. Kinetics and in situ spectroscopic studies unveil that the embedded electron-enriched Cu NPs follow the typical Eley-Rideal mechanism in CO oxidation, contrasting the Langmuir-Hinshelwood mechanism on the traditional Cu NPs. This mechanistic shift is driven by the Coulombic repulsion in anion oxygen layer, enabling its direct reaction with gaseous CO to form the easily desorbed monodentate carbonate.

Influence of compost on the mobility of arsenic in soil and its uptake by bean plants (Phaseolus vulgaris L.) irrigated with arsenite-contaminated water.

The influence of compost on the growth of bean plants irrigated with As-contaminated waters and its influence on the mobility of As in the soils and the uptake of As (as NaAs(III)O2) by plant components was studied at various compost application rates (3·10(4) and 6·10(4) kg ha(-1)) and at three As concentrations (1, 2 and 3 mg kg(-1)). The biomass and As and P concentrations of the roots, shoots and beans were determined at harvest time, as well as the chlorophyll content of the leaves and nonspecific and specifically bound As in the soil. The bean plants exposed to As showed typical phytotoxicity symptoms; no plants however died over the study. The biomass of the bean plants increased with the increasing amounts of compost added to the soil, attributed to the phytonutritive capacity of compost. Biomass decreased with increasing As concentrations, however, the reduction in the biomass was significantly lower with the addition of compost, indicating that the As phytotoxicity was alleviated by the compost. For the same As concentration, the As content of the roots, shoots and beans decreased with increasing compost added compared to the Control. This is due to partial immobilization of the As by the organic functional groups on the compost, either directly or through cation bridging. Most of the As adsorbed by the bean plants accumulated in the roots, while a scant allocation of As occurred in the beans. Hence, the addition of compost to soils could be used as an effective means to limit As accumulation in crops from As-contaminated waters.

Proof-of-principle for SERS imaging of Aspergillus nidulans hyphae using in vivo synthesis of gold nanoparticles.

High spatial resolution methods to assess the physiology of growing cells should permit analysis of fungal biochemical composition. Whole colony methods cannot capture the details of physiology and organism-environment interaction, in part because the structure, function and composition of fungal hyphae vary within individual cells depending on their distance from the growing apex. Surface Enhanced Raman Scattering (SERS) can provide chemical information on materials that are in close contact with appropriate metal substrates, such as nanopatterned gold surfaces and gold nanoparticles (AuNPs). Since nanoparticles can be generated by living cells, we have created conditions for AuNP formation within and on the surface of Aspergillus nidulans hyphae in order to explore their potential for SERS analysis. AuNP distribution and composition have been assessed by UV-Vis spectroscopy, fluorescence light microscopy, transmission electron microscopy, and scanning transmission X-ray microscopy. AuNPs were often associated with hyphal walls, both in the peripheral cytoplasm and on the outer wall surface. Interpretation of SERS spectra is challenging, and will require validation for the diversity of organic molecules present. Here, we show proof-of-principle that it is possible to generate SERS spectra from nanoparticles grown in situ by living hyphae.

Soft X-ray induced photoreduction of organic Cu(II) compounds probed by X-ray absorption near-edge (XANES) spectroscopy.

Photoreduction is a major obstacle for using the X-ray absorption near-edge structure (XANES) fingerprint to perform metal speciation at the molecular level in biological and environmental samples, especially for metalloproteins. In this study, soft X-ray induced photoreduction was observed in organic Cu(II) compounds during XANES measurements in a third-generation synchrotron source. Next Cu L(3)-edge, O K-edge, and C K-edge XANES spectroscopy, together with the scanning transmission X-ray microscopy (STXM), were used to probe the specific radiation damage processes of Cu acetate with similar local structures to Cu metalloproteins. Breakup of the Cu-Cu bond was hypothesized for the initial photoreduction of Cu acetate. The following radiation damage of Cu acetate produced CuO and an organic Cu(I) compound with a C═C bond, and the further photoreduction of the resulting CuO to Cu metal was also demonstrated. Our results indicated the importance of consideration of photoreduction during soft XANES measurements for the solid state compounds with high valence metals. Reducing the radiation dose to ~0.1 MGy effectively prevented the photoreduction of organic Cu(II) compounds during these measurements. This proposed radiation damage mechanism in Cu acetate may be generally useful in explaining the photoreduction process in Cu metalloproteins.

Sodium adsorption by reusable zeolite adsorbents: integrated adsorption cycles for salinised groundwater treatment.

Using Canadian (CMZ), Bear River (BRZ), and St. Cloud (SCZ) zeolites, this study investigates the application of natural and pre-treated zeolites for Na+ removal from salinised groundwater. Natural BRZ achieved better Na+ removal for initial concentrations of 250-10,000 mg Na+/L and had the highest maximum adsorption capacity (14.3 ± 0.4 mg/g) compared to natural CMZ (5.8 ± 0.5 mg/g) and SCZ (5.6 ± 0.7 mg/g). Natural BRZ exhibited a higher cation exchange capacity (CEC), mineralogical purity, and natural abundance of exchangeable calcium. The natural abundance of Na+ on CMZ and SCZ may have reduced Na+ adsorption. H-form BRZ and H-form CMZ were also prepared through conventional acidic pre-treatment. Acid treatment improved zeolite properties for adsorption (surface area and CEC). Synchrotron-based X-ray scanning transmission microscopy (STXM) indicated that Na+ adsorption sites in the H-form zeolites were associated with the mineral framework. However, sorption effluents were highly acidic (pH ∼2) and Al3+ leached significantly due to the dealumination induced by acid treatment. Alternatively, hard water softening was cyclically integrated with sodium adsorption as a zeolite treatment to generate Ca/Mg-form CMZ. This integration suggested the feasibility of combining CMZ cycles for water softening and sodium reduction for an extended CMZ lifecycle. Natural CMZ was first used to treat hard water, which enriched the CMZ with Ca2+ and Mg2+ and increased its subsequent Na+ removal rate by over 77%, without producing acidic effluents. The Canadian zeolite adsorbed more sodium when water softening was integrated with sodium removal, which is a repeatable dual-treatment.