Low Mg2 Research Articles

An assessment of the influence of sulfidic mine wastes on rainwater quality in a semiarid climate (SE Spain)

This study assessed the influence of the Sierra Minera de Cartagena – La Unión mining wastes on the surrounding areas, due to the sediment transport in the atmosphere. Monitoring of 15 weather stations located in the Campo de Cartagena land (SE Spain) was performed in the period December, 2004–March, 2008. A total of 920 rainfall water samples were collected and analyzed in this study. The network of weather stations covered a wide range of soil use and human activities (industrial, urban, agricultural, and mining sites). The physicochemical characterization involved determination of pH, EC, major ions, and the metals of interest in the soluble fraction of rainwater. Precipitations had an alkaline pH (7.0–7.9) and, compared to World Health Organization guidelines for drinking-water quality, samples were characterized by low EC (76–930 μS/cm), Ni (0.1–8 μg/L), Cu (3–88 μg/L), As (0.04–2 μg/L), Na+ (3–16 mg/L), K+ (0.41–30 mg/L), Ca2+ (6–51 mg/L), Mg2+ (up to 5 mg/L), Cl– (4–23 mg/L), NO3– (1–30 mg/L), and SO42− (4–35 mg/L) values. In certain locations, Zn, Cd, and Pb exceeded the guideline limits. A downward trend of H+, Zn, Cd, and Pb concentrations in total depositions was observed while the distance to the area affected by mining activities increased in the first 20 km. Rainwater quality was found to be just subtly affected by the metal-sulfide abandoned minesites, but without presenting a current threat to neither public health nor natural ecosystem. Nevertheless, greater attention should be paid in the future projections of climate change, where a significant increase in aridity may favor the atmospheric transport of pollutant particles.

Low-Mg(2+) treatment increases sensitivity of voltage-gated Na(+) channels to Ca(2+)/calmodulin-mediated modulation in cultured hippocampal neurons.

Culture of hippocampal neurons in low-Mg(2+) medium (low-Mg(2+) neurons) results in induction of continuous seizure activity. However, the underlying mechanism of the contribution of low Mg(2+) to hyperexcitability of neurons has not been clarified. Our data, obtained using the patch-clamp technique, show that voltage-gated Na(+) channel (VGSC) activity, which is associated with a persistent, noninactivating Na(+) current (INa,P), was modulated by calmodulin (CaM) in a concentration-dependent manner in normal and low-Mg(2+) neurons, but the channel activity was more sensitive to Ca(2+)/CaM regulation in low-Mg(2+) than normal neurons. The increased sensitivity of VGSCs in low-Mg(2+) neurons was partially retained when CaM12 and CaM34, CaM mutants with disabled binding sites in the N or C lobe, were used but was diminished when CaM1234, a CaM mutant in which all four Ca(2+) sites are disabled, was used, indicating that functional Ca(2+)-binding sites from either lobe of CaM are required for modulation of VGSCs in low-Mg(2+) neurons. Furthermore, the number of neurons exhibiting colocalization of CaM with the VGSC subtypes NaV1.1, NaV1.2, and NaV1.3 was significantly higher in low- Mg(2+) than normal neurons, as shown by immunofluorescence. Our main finding is that low-Mg(2+) treatment increases sensitivity of VGSCs to Ca(2+)/CaM-mediated regulation. Our data reveal that CaM, as a core regulating factor, connects the functional roles of the three main intracellular ions, Na(+), Ca(2+), and Mg(2+), by modulating VGSCs and provides a possible explanation for the seizure discharge observed in low-Mg(2+) neurons.

Metabolic responses differentiate between interictal, ictal and persistent epileptiform activity in intact, immature hippocampus in vitro

Interictal spikes, ictal responses, and status epilepticus are characteristic of abnormal neuronal activity in epilepsy. Since these events may involve different energy requirements, we evaluated metabolic function (assessed by simultaneous NADH and FAD+ imaging and tissue O2 recordings) in the immature, intact mouse hippocampus (P5-P7, in vitro) during spontaneous interictal spikes and ictal-like events (ILEs), induced by increased neuronal network excitability with either low Mg2+ media or decreased inhibition with bicuculline. In low Mg2+ medium NADH fluorescence showed a small decrease both during the interictal build-up leading to an ictal event and before ILE occurrences, but a large positive response during and after ILEs (up to 10% net change). Tissue O2 recordings (pO2) showed an oxygen dip (indicating oxygen consumption) coincident with each ILE at P5 and P7, closely matching an NADH fluorescence increase, indicating a large surge in oxidative metabolism. The ILE O2 dip was significantly larger at P7 as compared to P5 suggesting a higher metabolic response at P7. After several ILEs at P7, continuous, low voltage activity (late recurrent discharges: LRDs) occurred. During LRDs, whilst the epileptiform activity was relatively small (low voltage synchronous activity) oxygen levels remained low and NADH fluorescence elevated, indicating persistent oxygen utilization and maintained high metabolic demand. In bicuculline, NADH fluorescence levels decreased prior to the onset of epileptiform activity, followed by a slow positive phase, which persisted during interictal responses. Metabolic responses can thus differentiate between interictal, ictal-like and persistent epileptiform activity resembling status epilepticus, and confirm that spreading depression did not occur. These results demonstrate clear translational value to the understanding of metabolic requirements during epileptic conditions.

Structural and functional consequences of removing the N-terminal domain from the magnesium chelatase ChlH subunit of Thermosynechococcus elongatus.

Magnesium chelatase (MgCH) initiates chlorophyll biosynthesis by catalysing the ATP-dependent insertion of Mg2+ into protoporphyrin. This large enzyme complex comprises ChlH, I and D subunits, with I and D involved in ATP hydrolysis, and H the protein that handles the substrate and product. The 148 kDa ChlH subunit has a globular N-terminal domain attached by a narrow linker to a hollow cage-like structure. Following deletion of this ~18 kDa domain from the Thermosynechoccus elongatus ChlH, we used single particle reconstruction to show that the apo- and porphyrin-bound forms of the mutant subunit consist of a hollow globular protein with three connected lobes; superposition of the mutant and native ChlH structures shows that, despite the clear absence of the N-terminal ‘head’ region, the rest of the protein appears to be correctly folded. Analyses of dissociation constants shows that the ΔN159ChlH mutant retains the ability to bind protoporphyrin and the Gun4 enhancer protein, although the addition of I and D subunits yields an extremely impaired active enzyme complex. Addition of the Gun4 enhancer protein, which stimulates MgCH activity significantly especially at low Mg2+ concentrations, partially reactivates the ΔN159ChlH–I–D mutant enzyme complex, suggesting that the binding site or sites for Gun4 on H do not wholly depend on the N-terminal domain.

Magnesium depletion in patients treated with therapeutic hypothermia after cardiac arrest.

Magnesium (Mg(2+)) depletion can have detrimental effects in postcardiac arrest patients through multiple potential mechanisms. Therapeutic hypothermia (TH) produces a Mg(2+) diuresis, but the effects of postcardiac arrest TH on serum Mg(2+) levels in patients with postcardiac arrest syndrome (PCAS) are yet to be systematically quantified. We conducted a retrospective chart review of 119 consecutive comatose PCAS patients treated with TH between 2005 and 2010 and compared them to 33 matched historic controls (HCs) seen at the same institution between 2002 and 2005 who were not treated with TH. We abstracted data from the first 96 hours postarrest, including date, time, and value of serum Mg(2+) levels and date, time, and amount of Mg(2+) repletion, along with outcomes at discharge. The median Mg(2+) level of TH patients was 2.0 mg/dL [interquartile range (IQR), 1.9-2.2 mg/dL] (0.82 mmol/L [IQR, 0.78-0.90 mmol/L]) versus 2.2 mg/dL [IQR, 1.9-2.4 mg/dL] (0.90 mmol/L [IQR, 0.82-0.99 mmol/L]) (p=0.2) in HCs. In addition, 42.9% (520/1214) of Mg(2+) levels in TH patients versus 31.9% (43/135) (p=0.014) in HC patients were below 2.0 mg/dL [0.82 mmol/L]. The average number of times the Mg(2+) level was checked in TH patients was 10.2 (range 1-18) versus 4.1 (range 1-10) in HCs. The TH patients were more likely to receive supplemental Mg(2+) than HCs (81.5% [97/119] vs. 27.3% [9/33] [p<0.01]). The mean supplemental Mg(2+) dose was 1.9 g for TH patients versus 0.5 g for HC patients. Mortality in patients treated with TH was 53.1% (60/113) versus 78.6% (22/28) (p=0.014) in HCs. Low serum Mg(2+) levels with subsequent Mg(2+) supplementation were more common in comatose patients with PCAS treated with TH compared to normothermic HC patients. The effect of untreated hypomagnesemia on postcardiac arrest outcomes remains to be determined.

Determinants of the Rate of mRNA Translocation in Bacterial Protein Synthesis

Studying the kinetics of translocation of mRNA and tRNAs on the translating ribosome is technically difficult since the rate-limiting steps involve large conformational changes without covalent bond formation or disruption. Here, we have developed a unique assay system for precise estimation of the full translocation cycle time at any position in any type of open reading frame (ORF). Using a buffer system optimized for high accuracy of tRNA selection together with high concentration of elongation factor G, we obtained in vivo compatible translocation rates. We found that translocation was comparatively slow early in the ORF and faster further downstream of the initiation codon. The maximal translocation rate decreased from the in vivo compatible value of 30s−1 at 1mM free Mg2+ concentration to the detrimentally low value of 1s−1 at 6mM free Mg2+ concentration. Thus, high and in vivo compatible accuracy of codon translation, as well as high and in vivo compatible translocation rate, required a remarkably low Mg2+ concentration. Finally, we found that the rate of translocation deep inside an ORF was not significantly affected upon variation of the standard free energy of interaction between a 6-nt upstream Shine-Dalgarno (SD)-like sequence and the anti-SD sequence of 16S rRNA in a range of 0–6kcal/mol. Based on these experiments, we discuss the optimal choice of Mg2+ concentration for maximal fitness of the living cell by taking its effects on the accuracy of translation, the peptide bond formation rate and the translocation rate into account.

Magnesium in medicine and treatment

Magnesium (Mg2+) plays a critical role in numerous metabolic function in our body. Normal levels of both intraand extra cellular magnesium are necessary for correct cellular processes. The specific clinical conditions in which Mg2+ deficiency has been implicated to play a pathophysiological role include depression, ischaemic heart disease, arrhythmias, preeclampsia, asthma, intradialytic hypotension or critical illness. Correct administration of magnesium can eliminate perioperative pain and muscle spasms, keep blood flowing smoothly, and prevent platelet stickiness. Although magnesium sulphate is now widely used to prevent or control eclamptic convulsions, there is no consensus on the optimal dosage and concentration of magnesium sulphate for the management of eclamptic seizures. Owing to its bronchodilating and anti-inflammatory effects, Mg2+ is an encouraging adjuvant therapy for paediatric patients, although its use is often limited to patients who do not respond to conventional treatment. Studies have also suggested that magnesium might be useful as an agent to control the rigidity and spasms of severe tetanus or for the enhancement of neuromuscular blockade during anaesthesia, but its actual efficacy as an anticonvulsant or an adjuvant to analgesics and anaesthetics to induce and maintain anaesthesia remains unclear. Moreover, a negative correlation has been shown between intradialytic changes of the serum magnesium and hypotensive episodes during dialysis sessions. Low Mg2+ haemodialysis solution resulted in both hypomagnesaemia and hypocalcaemia. Mg2+ compounds are now becoming recognized as safe, effective and cost-efficient alternatives to other phosphorus binders, with a significant added benefit of substantially reducing the risk and impact of cardiovascular diseases. It seems that magnesium is underused in clinical conditions, considering its therapeutic capacity. On the other hand, numerous studies have found conflicting results, questioning the implicit efficacy of magnesium in several clinical conditions.

Ail and PagC-Related Proteins in the Entomopathogenic Bacteria of Photorhabdus Genus

Among pathogenic Enterobacteriaceae, the proteins of the Ail/OmpX/PagC family form a steadily growing family of outer membrane proteins with diverse biological properties, potentially involved in virulence such as human serum resistance, adhesion and entry into eukaryotic culture cells. We studied the proteins Ail/OmpX/PagC in the bacterial Photorhabdus genus. The Photorhabdus bacteria form symbiotic complexes with nematodes of Heterorhabditis species, associations which are pathogenic to insect larvae. Our phylogenetic analysis indicated that in Photorhabdus asymbiotica and Photorhabdus luminescens only Ail and PagC proteins are encoded. The genomic analysis revealed that the Photorhabdus ail and pagC genes were present in a unique copy, except two ail paralogs from P. luminescens. These genes, referred to as ail1 Pl and ail2 Pl, probably resulted from a recent tandem duplication. Surprisingly, only ail1 Pl expression was directly controlled by PhoPQ and low external Mg2+ conditions. In P. luminescens, the magnesium-sensing two-component regulatory system PhoPQ regulates the outer membrane barrier and is required for pathogenicity against insects. In order to characterize Ail functions in Photorhabdus, we showed that only ail2 Pl and pagC Pl had the ability, when expressed into Escherichia coli, to confer resistance to complement in human serum. However no effect in resistance to antimicrobial peptides was found. Thus, the role of Ail and PagC proteins in Photorhabdus life cycle is discussed.

Activation of calcineurin underlies altered trafficking of α2 subunit containing GABAA receptors during prolonged epileptiform activity

Fast inhibitory signalling in the mammalian brain is mediated by gamma-aminobutyric acid type A receptors (GABAARs), which are targets for anti-epileptic therapy such as benzodiazepines. GABAARs undergo tightly regulated trafficking processes that are essential for maintenance and physiological modulation of inhibitory strength. The trafficking of GABAARs to and from the membrane is altered during prolonged seizures such as in Status Epilepticus (SE) and has been suggested to contribute to benzodiazepine pharmacoresistance in patients with SE. However, the intracellular signalling mechanisms that cause this modification in GABAAR trafficking remain poorly understood. In this study, we investigate the surface stability of GABAARs during SE utilising the low Mg2+ model in hippocampal rat neurons. Live-cell imaging of super ecliptic pHluorin (SEP)-tagged α2 subunit containing GABAARs during low Mg2+ conditions reveals that the somatic surface receptor pool undergoes down-regulation dependent on N-methyl-d-aspartate receptor (NMDAR) activity. Analysis of the intracellular Ca2+ signal during low Mg2+ using the Ca2+-indicator Fluo4 shows that this reduction of surface GABAARs correlates well with the timeline of intracellular Ca2+ changes. Furthermore, we show that the activation of the phosphatase calcineurin was required for the decrease in surface GABAARs in neurons undergoing epileptiform activity. These results indicate that somatic modulation of GABAAR trafficking during epileptiform activity in vitro is mediated by calcineurin activation which is linked to changes in intracellular Ca2+ concentrations. These mechanisms could account for benzodiazepine pharmacoresistance and the maintenance of recurrent seizure activity, and reveal potential novel targets for the treatment of SE.This article is part of the Special Issue entitled ‘GABAergic Signaling in Health and Disease’.

Physicochemical characteristics of drip waters: Influence on mineralogy and crystal morphology of recent cave carbonate precipitates

Speleothems are one of the most intensively explored continental archives for palaeoclimate variability. The parameters, however, that control speleothem petrography and its changes with time and space, specifically calcite crystal morphology and carbonate mineralogy, are still poorly understood. In order to shed light on processes and their products, precipitation experiments of recent carbonate crystals on watch glasses and glass plates were performed in seven selected caves. Drip water sites were analysed for their fluid Mg/Ca molar ratio, pH, degree of saturation for calcite and aragonite and drip rates. Corresponding precipitates were analysed with respect to their mineralogy, calcite crystal morphology and Mg/Ca molar ratio of calcite. The following results are found: High fluid Mg/Ca ratios are found only for caves situated in dolostone, thus the hostrock lithology indirectly controls the carbonate mineralogy and calcite crystal morphology of speleothems. The precipitation of aragonite in place of calcite occurred only in dolostone caves and is bound to very specific conditions. These are: high fluid Mg/Ca ratios (⩾0.5), high fluid pH (>8.2) and low fluid saturation indices for calcite (<0.8). These specific conditions are induced by slow drip rates of <0.2ml/min as often under more arid conditions, causing the precipitation of calcite/aragonite prior to reaching the stalagmite top. Due to this, fluid chemistry is altered, which in turn leads to changes in carbonate mineralogy and geochemistry on the stalagmite top. Calcite growth is inhibited at high fluid Mg/Ca ratios and hence, aragonite precipitation is kinetically stabilised. An increase of the drip water Mg/Ca ratio leads to an increased incorporation of Mg2+ into the calcite crystal lattice and thus, to a change in calcite crystal morphology. Four distinctive changes occur with increasing Mg2+ incorporation: (i) development of new forms (steeper rhombohedra and base pinacoid) at the edges and corners of the crystal seed, (ii) crystal habit tend to elongate along [001] due to slower growth of faces with high Mg2+ densities, (iii) reconstitution of crystal faces with low Mg2+ densities, and (iv) occurrence of calcite crystals with bended faces and edges due to very high Mg2+ (Mg/Ca ratios of 0.009–0.051) incorporation. Growth rates and possibly also organic compounds, however, may also affect the morphology of calcite crystals. Based on the data shown here, the relation of Mg2+ incorporation and the resulting changes in calcite crystal morphologies as well as the conditions of aragonite precipitation are now clearly better understood. Further work should aim at linking the calcite crystal morphology of watch glass precipitates with calcite crystal fabrics in speleothems in order to exploit the petrographic archive of speleothem deposits.

The importance of the magnesium transporter MgtB for virulence of Yersinia pseudotuberculosis and Yersinia pestis.

Mg(2+) has been shown to be an important signal controlling gene regulation via the PhoPQ two-component regulatory system for a range of Gram-negative bacteria, including Yersinia pestis and Yersinia pseudotuberculosis. The magnesium ion transporter MgtB is part of the complex PhoPQ regulon, being upregulated in response to low Mg(2+). Despite the presence of other Mg(2+) transport systems in Yersinia, inactivation of mgtB had a significant effect on the ability of the bacteria to scavenge this crucial ion. Whereas inactivation of PhoPQ is reported to adversely affect intracellular survival, we show that Y. pestis and Y. pseudotuberculosis ΔmgtB mutants survived equally as well as the respective parent strain within macrophages, although they were more sensitive to killing in the Galleria model of infection. Surprisingly, despite MgtB being only one member of the Mg(2+) stimulon and PhoPQ controlling the expression levels of a range of genes including mgtB, the Yersinia ΔmgtB mutants were more highly attenuated than the equivalent Yersinia ΔphoP mutants in mouse models of infection. MgtB may be a suitable target for development of novel antimicrobials, and investigation of its role may help elucidate the contribution of this component of the PhoPQ regulon to pathogenesis.

Serratia marcescens arn, a PhoP-regulated locus necessary for polymyxin B resistance.

Polymyxins, which are increasingly being used to treat infections caused by multidrug-resistant bacteria, perform poorly against Serratia marcescens. To investigate the underlying mechanisms, Tn5 mutagenesis was performed and two mutants exhibiting increased polymyxin B (PB) susceptibility were isolated. The mutants were found to have Tn5 inserted into the arnB and arnC genes. In other bacteria, arnB and arnC belong to the seven-gene arn operon, which is involved in lipopolysaccharide (LPS) modification. LPSs of arn mutants had greater PB-binding abilities than that of wild-type LPS. Further, we identified PhoP, a bacterial two-component response regulator, as a regulator of PB susceptibility in S. marcescens. By the reporter assay, we found PB- and low-Mg2+-induced expression of phoP and arn in the wild-type strain but not in the phoP mutant. Complementation of the phoP mutant with the full-length phoP gene restored the PB MIC and induction by PB and low Mg2+ levels, as in the wild type. An electrophoretic mobility shift assay (EMSA) further demonstrated that PhoP bound directly to the arn promoter. The PB challenge test confirmed that pretreatment with PB and low Mg2+ levels protected S. marcescens from a PB challenge in the wild-type strain but not in the phoP mutant. Real-time reverse transcriptase-PCR also indicated that PB serves as a signal to regulate expression of ugd, a gene required for LPS modification, in S. marcescens through a PhoP-dependent pathway. Finally, we found that PB-resistant clinical isolates displayed greater expression of arnA upon exposure to PB than did susceptible isolates. This is the first report to describe the role of S. marcescens arn in PB resistance and its modulation by PB and Mg2+ through the PhoP protein.

The inflammatory molecules IL-1β and HMGB1 can rapidly enhance focal seizure generation in a brain slice model of temporal lobe epilepsy.

Epilepsy is a neurological disorder characterized by a hyperexcitable brain tissue and unpredictable seizures, i.e., aberrant firing discharges in large neuronal populations. It is well established that proinflammatory cytokines, in addition to their canonical involvement in the immune response, have a crucial role in the mechanism of seizure generation. The purpose of the present study was to investigate the role of interleukin-1β (IL-1β) and high mobility group B1 (HMGB1) in the generation of seizure-like discharges using two models of focal epilepsy in a rat entorhinal cortex slice preparation. Seizure like-discharges were evoked by either slice perfusion with low Mg2+ and picrotoxin or with a double NMDA local stimulation in the presence of the proconvulsant 4-amino-pyridine. The effects of IL-1β or HMGB1 were evaluated by monitoring seizure discharge generation through laser scanning microscope imaging of Ca2+ signals from neurons and astrocytes. In the picrotoxin model, we revealed that both cytokines increased the mean frequency of spontaneous ictal-like discharges, whereas only IL-1β reduced the latency and prolonged the duration of the first ictal-like event. In the second model, a single NMDA pulse, per se ineffective, became successful when it was performed after IL-β or HMGB1 local applications. These findings demonstrate that both IL-1β and HMGB1 can rapidly lower focal ictal event threshold and strengthen the possibility that targeting these inflammatory pathways may represent an effective therapeutic strategy to prevent seizures.

RpoS integrates CRP, Fis, and PhoP signaling pathways to control Salmonella Typhi hlyE expression.

BackgroundSPI-18 is a pathogenicity island found in some Salmonella enterica serovars, including S. Typhi. SPI-18 harbors two ORFs organized into an operon, hlyE and taiA genes, both implicated in virulence. Regarding the hlyE regulation in S. Typhi, it has been reported that RpoS participates as transcriptional up-regulator under low pH and high osmolarity. In addition, CRP down-regulates hlyE expression during exponential growth. Previously, it has been suggested that there is another factor related to catabolite repression, different from CRP, involved in the down-regulation of hlyE. Moreover, PhoP-dependent hlyE up-regulation has been reported in bacteria cultured simultaneously under low pH and low concentration of Mg2+. Nevertheless, the relative contribution of each environmental signal is not completely clear. In this work we aimed to better understand the regulation of hlyE in S. Typhi and the integration of different environmental signals through global regulators.ResultsWe found that Fis participates as a CRP-independent glucose-dependent down-regulator of hlyE. Also, Fis and CRP seem to exert the repression over hlyE through down-regulating rpoS. Moreover, PhoP up-regulates hlyE expression via rpoS under low pH and low Mg2+ conditions.ConclusionsAll these results together show that, at least under the tested conditions, RpoS is the central regulator in the hlyE regulatory network, integrating multiple environmental signals and global regulators.

MiR-134 blockade prevents status epilepticus like-activity and is neuroprotective in cultured hippocampal neurons

Status epilepticus (SE) is a life-threatening neurological disorder associated with significant morbidity and mortality. MicroRNAs (miRNAs) are small, non-coding RNAs that act post-transcriptionally modulating messenger RNA (mRNA) translation or stability which may have important roles in the pathogenesis of epilepsy. It has been reported that silencing microRNA-134 in vivo has significant neuroprotective and prolonged seizure-suppressive effects. However, the mechanism by which miR-134 inhibition suppressed seizures and whether miR-134 inhibition works in an in vitro model of SE, is unknown. Compared to a complex in vivo system, in vitro models of SE-like electrographic activity can be powerful tools to study this miRNA. Using a cell culture model of low Mg2+ treatment of rat hippocampal neurons, we found SE-like electrographic activity increased expression of miR-134. Inhibiting expression of miR-134 using an inhibitor lentivirus with two miR-134 binding sites reduced SE-like electrographic activity in the hippocampal neurons and reduced neuronal death. This study provides direct evidence that inhibition of miR-134 can block status epilepticus-like discharges and is neuroprotective in hippocampal neuronal cultures and implies that inhibiting miR-134 may be a potential candidate for the clinical treatment of SE.

Arabidopsis Transporter MGT6 Mediates Magnesium Uptake and Is Required for Growth under Magnesium Limitation.

Although magnesium (Mg2+) is the most abundant divalent cation in plant cells, little is known about the mechanism of Mg2+ uptake by plant roots. Here, we report a key function of Magnesium Transport6 (MGT6)/Mitochondrial RNA Splicing2-4 in Mg2+ uptake and low-Mg2+ tolerance in Arabidopsis thaliana. MGT6 is expressed mainly in plant aerial tissues when Mg2+ levels are high in the soil or growth medium. Its expression is highly induced in the roots during Mg2+ deficiency, suggesting a role for MGT6 in response to the low-Mg2+ status in roots. Silencing of MGT6 in transgenic plants by RNA interference (RNAi) resulted in growth retardation under the low-Mg2+ condition, and the phenotype was restored to normal growth after RNAi plants were transferred to Mg2+-sufficient medium. RNAi plants contained lower levels of Mg2+ compared with wild-type plants under low Mg2+ but not under Mg2+-sufficient conditions. Further analysis indicated that MGT6 was localized in the plasma membrane and played a key role in Mg2+ uptake by roots under Mg2+ limitation. We conclude that MGT6 mediates Mg2+ uptake in roots and is required for plant adaptation to a low-Mg2+ environment.

Influence of Mg2+ on CaCO3 precipitation during subsurface reactive transport in a homogeneous silicon-etched pore network

Calcium carbonate (CaCO3) geochemical reactions exert a fundamental control on the evolution of porosity and permeability in shallow-to-deep subsurface siliciclastic and limestone rock reservoirs. As a result, these carbonate water–rock interactions play a critically important role in research on groundwater remediation, geological carbon sequestration, and hydrocarbon exploration. A study was undertaken to determine the effects of Mg2+ concentration on CaCO3 crystal morphology, precipitation rate, and porosity occlusion under flow and mixing conditions similar to those in subsurface aquifers. This was accomplished by promoting CaCO3 precipitation through the mixing of two solutions flowing parallel to each other in a microfluidic pore structure, containing uniform concentrations of dissolved Ca2+ and carbonate (CO32−), and systematic variations in the concentration of Mg2+. Raman spectroscopy indicates that all three polymorphs of CaCO3 (calcite, aragonite, and vaterite) were present under all experimental conditions. Coordinated brightfield imaging results show the morphology of calcite with increasing Mg2+ progressed from blocky and dogtooth approximately 10–80μm in size, to anhedral spheroidal approximately 5–30μm in size. The morphology of aragonite with increasing Mg2+ progressed from shrubs and fuzzy dumbells to spheroidal, and the size increased from approximately 5–60μm to 20–200μm. Recrystallization was observed in all experiments, but more so at low Mg2+, in which many small microcrystals dissolved and re-precipitated as one or a few larger calcite crystals. Analysis of brightfield images indicates calcite is the most abundant polymorph under all conditions. However, the area of pore space with aragonite increased from <5% when no Mg2+ was present to >20% at the highest Mg2+ concentration. The initial apparent precipitation rate of mineral polymorphs with no Mg2+ present was 2.5 times greater than when 40mM Mg2+ was added, and large (20–200μm) aragonite crystals formed primarily near to and below the center mixing zone with increasing Mg2+ concentration. Pore-scale modeling results are consistent with experiments, and indicate that all three polymorphs are thermodynamically favorable, with calcite and aragonite being the most favorable and having similar saturation ratios (SR>100). The influence of Mg2+ on mineral precipitation rates is consistent with previous studies showing that calcite precipitation rates decrease with increasing Mg2+ concentrations. The precipitation of aragonite below the center-mixing zone is not predicted by thermodynamic SRs, but is consistent with the literature and our modeling results showing aragonite precipitation is kinetically more favorable in regions with higher Mg2+/Ca2+ ratios. Hence, both thermodynamic and kinetic constraints affect precipitation rates, the distribution of mineral polymorphs, and the corresponding extent of porosity occlusion. A tracer study demonstrated that mineral precipitation along the center-mixing zone under all experimental conditions led to substantial pore blockage. Imaging results suggest that with increasing Mg2+ concentration, slower crystal growth rates will increase the time period before pore blockage occurs, and the transition to more spherical and larger aragonite crystals below the center mixing line will increase pore occlusion and decrease mixing. Hence, understanding how Mg2+ affects calcium carbonate precipitation is very important for predicting mixing and reactive transport in subsurface reservoirs.

Elucidation of the distal convoluted tubule transcriptome identifies Pcbd1 as new causative gene for hereditary hypomagnesemia (860.4)

The distal convoluted tubule (DCT) is instrumental in fine‐tuning of the renal Mg2+ handling. Here, we aim to discover new Mg2+‐related genes in DCT. Transgenic mice expressing eGFP under a DCT‐specific parvalbumin promoter were subjected to Mg2+‐deficient or Mg2+‐enriched diets. Subsequently, the Complex Object Parametric Analyzer and Sorter (COPAS) allowed for the first time isolation of eGFP‐positive DCT cells. RNA extracts thereof were analyzed by DNA microarrays to identify Mg2+ regulatory genes and 46 genes showed differential expression. Several known magnesiotropic genes, such as Trpm6 and Parvalbumin, were upregulated under low dietary Mg2+. Moreover, new genes were identified that are potentially involved in renal Mg2+ handling. Among others, Pcbd1 was identified to be higher expressed in low Mg2+ conditions. Pcbd1 encodes a transcriptional co‐activator of Hnf1b, which was previously implicated in renal Mg2+ handling and maturity onset diabetes of the young (MODY). Interestingly, when examining three independent patients with Pcbd1 mutations, two individuals were diagnosed with hypomagnesemia and renal Mg2+ loss. Moreover, two patients also developed diabetes with characteristics of MODY regardless of the serum Mg2+ levels. By elucidating the Mg2+‐sensitive DCT transcriptome new candidate genes in renal Mg2+ handling have been identified.

The major-ion composition of Carboniferous seawater

The major-ion chemistry (Na+, Mg2+, Ca2+, K+, SO42−, and Cl−) of Carboniferous seawater was determined from chemical analyses of fluid inclusions in marine halites, using the cryo scanning electron microscopy (Cryo-SEM) X-ray energy-dispersive spectrometry (EDS) technique. Fluid inclusions in halite from the Mississippian Windsor and Mabou Groups, Shubenacadie Basin, Nova Scotia, Canada (Asbian and Pendleian Substages, 335.5–330Ma), and from the Pennsylvanian Paradox Formation, Utah, USA, (Desmoinesian Stage 309–305Ma) contain Na+–Mg2+–K+–Ca2+–Cl− brines, with no measurable SO42−, which shows that the Carboniferous ocean was a “CaCl2 sea”, relatively enriched in Ca2+ and low in SO42− with equivalents Ca2+ >SO42−+HCO3−.δ34S values from anhydrite in the Mississippian Shubenacadie Basin (13.2–14.0 ‰) and the Pennsylvanian Paradox Formation (11.2–12.6 ‰) support seawater sources. Br in halite from the Shubenacadie Basin (53–111ppm) and the Paradox Basin (68–147ppm) also indicate seawater parentages.Carboniferous seawater, modeled from fluid inclusions, contained ∼22mmol Ca2+/kg H2O (Mississippian) and ∼24mmol Ca2+/kg H2O (Pennsylvanian). Estimated sulfate concentrations are ∼14mmol SO42−/kg H2O (Mississippian), and ∼12mmol SO42−/kg H2O (Pennsylvanian). Calculated Mg2+/Ca2+ ratios are 2.5 (Mississippian) and 2.3 (Pennsylvanian), with an estimated range of 2.0–3.2.The fluid inclusion record of seawater chemistry shows a long period of CaCl2 seas in the Paleozoic, from the Early Cambrian through the Carboniferous, when seawater was enriched in Ca2+ and relatively depleted in SO42−. During this ∼200Myr interval, Ca2+ decreased and SO42− increased, but did not cross the Ca2+–SO42− chemical divide to become a MgSO4 sea (when SO42− in seawater became greater than Ca2+) until the latest Pennsylvanian or earliest Permian (∼309–295Ma). Seawater remained a MgSO4 sea during the Permian and Triassic, for ∼100Myr.Fluid inclusions also record a long interval, from the Early Cambrian to the Middle Devonian, when seawater had low Mg2+/Ca2+ ratios (<2) that coincide with calcite seas. The Mg2+/Ca2+ ratio of seawater rose from 0.9 in the Middle Devonian, to 2.5 in the Middle/Late Mississippian, 2.3 in the Middle Pennsylvanian, and 3.5 in the Early Permian. The transition from calcite seas to aragonite seas, established from the mineralogy of oölites and early marine cements, occurred in the Late Mississippian. Fluid inclusions show that seawater Mg2+/Ca2+ ratios rose above 2 by the Middle to Late Mississippian coinciding exactly with the shift to aragonite seas. Aragonite seas existed for ∼100Myr, from the Late Mississippian until the Late Triassic/Early Jurassic.

Intracellular Gold Nanoparticles Increase Neuronal Excitability and Aggravate Seizure Activity in the Mouse Brain

Due to their inert property, gold nanoparticles (AuNPs) have drawn considerable attention; their biological application has recently expanded to include nanomedicine and neuroscience. However, the effect of AuNPs on the bioelectrical properties of a single neuron remains unknown. Here we present the effect of AuNPs on a single neuron under physiological and pathological conditions in vitro. AuNPs were intracellularly applied to hippocampal CA1 neurons from the mouse brain. The electrophysiological property of CA1 neurons treated with 5- or 40-nm AuNPs was assessed using the whole-cell patch-clamp technique. Intracellular application of AuNPs increased both the number of action potentials (APs) and input resistance. The threshold and duration of APs and the after hyperpolarization (AHP) were decreased by the intracellular AuNPs. In addition, intracellular AuNPs elicited paroxysmal depolarizing shift-like firing patterns during sustained repetitive firings (SRF) induced by prolonged depolarization (10 sec). Furthermore, low Mg2+-induced epileptiform activity was aggravated by the intracellular AuNPs. In this study, we demonstrated that intracellular AuNPs alter the intrinsic properties of neurons toward increasing their excitability, and may have deleterious effects on neurons under pathological conditions, such as seizure. These results provide some considerable direction on application of AuNPs into central nervous system (CNS).