Ca Influx Research Articles

Zhenxin Anshen formula ameliorates atopic der-matitis-like skin dysfunction in mice and regulation of transient receptor potential vanilloid 1 and transient receptor potential ankyrin 1 in Neural pathways.

To investigate the efficacy of Zhenxin Anshen formula (, ZXAS) on atopic dermatitis (AD) by transient receptor potential vanilloid 1 (TRPV1) and transient receptor potential ankyrin 1 (TRPA1) signalling pathway in mice and . AD-like lesions were induced by 1-chloro-2,4-dinitrobenzene (DNCB) to the shaved dorsal skin of BALB/c mice. BALB/c mice were divided into five groups: normal control, model control, cetirizine, low-, medium-, and high-dose of ZXAS. After ZXAS in-tervention, the skin lesions and blood samples were collected for hematoxylin and eosin-stained and measuring the concentrations of inflammatory cytokines. Immun-oglobulin E (IgE), interleukin (IL)-4, IL-5, IL-13, and thymic stromal lymphopoietin (TSLP) were de-tected by Enzyme-linked immunosorbent assay (ELISA). The spinal cords were collected for measuring the expression of gastrin-releasing peptide receptor (GRPR), TRPV1, and TRPA1 by using immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In addition, 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, ELISA, and Western blotting were conducted for analysis of primary dorsal root ganglia (DRG) neurons . ZXAS treatment improved DNCB-induced AD-like lesions through reducing dermatitis score, number of scratching and epidermal thickness, accompanied by the de-creased IgE and Th2 inflammatory cytokines. ZXAS also supressed the mRNA and protein expression of GRPR, TRPV1, and TRPA1 in the spinal cord. The medicated sera of ZXAS decreased capsaicin-induced Ca influx and downregulated the expression of TRPV1, TRPA1, and phospholipase C in DRG neurons. The therapeutic effect of ZXAS on AD may be related to the regulation of TRPV1 and TRPA1 and inhibition of Ca2+ signals in neurons.

In-situ elemental and boron isotopic variations of tourmaline from the Koktokay pegmatitic rare-metal deposit, China: Insights into external contamination and the source of the granitic pegmatite

This study presents petrochemical and boron isotope compositions of tourmalines from the fertile (No. 3) and relatively barren (No. 1, 2b) pegmatite in the Koktokay pegmatitic rare-metal deposit, to better understand the influence of external contamination on pegmatite mineralization and the source of these pegmatites. There are four types of tourmalines that have been studied: (1) tourmaline in the tourmaline-mica nest of pegmatite No. 1 (Tur-N); (2) tourmalines in the comb-like structure of pegmatite No. 1 (Tur-C1), 2b (Tur-C2), and 3 (Tur-C3), respectively. Tur-N tourmaline is heterogeneous in backscattered electron (BSE) images, Tur-C1 and Tur-C2 appear to be homogenous crystals, while Tur-C3 exhibits to have a complicated zonal structure. These tourmalines are members of the alkali group tourmalines representing mostly dravite-schorl solid solutions. Tur-N contains the lowest Ca content. There is a general increase in the contents of Mg and Ca from Tur-C1 to Tur-C3, implying that the pegmatite melt was contaminated by the host rock (metagabbro) during the emplacement. The contamination's diffusive influx of Mg, Ca, and Fe components into the pegmatite melt impelled the crystallization of tourmaline during the emplacement, which subsequently triggered the deposition of rare element minerals. The external contamination implies that the closed system of melt is extremely favorable for the enrichment and mineralization of rare elements in the pegmatitic rare-metal deposit. The boron isotopes vary slightly among the four types of tourmalines. The B-isotope composition of Tur-C3 is the heaviest (–11.63 ± 0.05 ‰ to –13.16 ± 0.05 ‰), followed by Tur-C1 (–12.29 ± 0.06 ‰ to –12.93 ± 0.05 ‰) and Tur-C2 (–12.51 ± 0.05 ‰ to –13.21 ± 0.05 ‰), Tur-N contains the lowest δ11B value (–13.20 ± 0.05 ‰ to –13.76 ± 0.05 ‰). Synthesized with the previous research results, we hold that the boron isotope of tourmaline (Tur-C3) in the comb-like structure of No. 3 pegmatite might represent the isotope composition of its initial magma, indicating that the tourmaline and associated pegmatite may derive from the melting of clastic sedimentary rock from the Habahe Group forming in Mid Ordovician to Early Devonian in the Chinese Altay orogenic belt.

Negative dietary cation-anion difference and amount of calcium in prepartum diets: Effects on urine and serum minerals

Negative dietary cation-anion difference (DCAD) diets fed prepartum induce a compensated metabolic acidosis, which stimulates metabolic Ca flux before calving and decreases clinical and subclinical hypocalcemia after calving. Effects of low or high dietary Ca in these diets are unclear. Our objective was to determine the effects of inducing a prepartum metabolic acidosis and the amount of dietary Ca on urinary mineral excretion and serum mineral concentrations during the transition period in multiparous Holstein cows (n = 81). Treatment diets fed during the last 28 d before calving were (1) positive DCAD, +6 mEq/100 g of dry matter (DM), target urine pH >7.5, low dietary Ca (0.40% DM; CON); (2) negative DCAD, −24 mEq/100 g of DM, target urine pH 5.5 to 6.0, low dietary Ca (0.40% DM; ND); or (3) negative DCAD, −24 mEq/100 g of DM, target urine pH 5.5 to 6.0, high dietary Ca (2.0% DM; NDCA). Urine was sampled on −21, −14, −7, +1, +2, and +7 d relative to calving. Blood samples were collected on d −30, −21, −14, −7, −4, −2, −1, 0, 1, 2, and 4 relative to parturition. Preplanned treatment contrasts were (1) CON versus ND and NDCA, and (2) ND versus NDCA. Cows fed ND or NDCA had increased urinary volume and excretion of Ca, Cl, and SO4−2, along with decreased excretion of K. Supplementation of Ca to the acidogenic diet increased urinary excretion of Ca at d −21, but this difference lessened as cows approached parturition. Additional Ca increased prepartum urinary excretion of K and Cl. Differences in excretion decreased quickly after parturition, although urinary excretion of Cl remained greater for cows previously fed negative DCAD diets. Acidogenic diets increased serum K and Cl concentrations prepartum. Supplemental Ca decreased serum P relative to ND. During d 0 to 4, serum P and K were greater for cows fed ND or NDCA than for cows fed CON; these differences disappeared by d 4. Acidogenic diets increased serum Ca by increasing Ca flux and excretion in urine. Supplemental Ca in the acidogenic diet modestly increased Ca excretion early in the close-up period.

Calcium isotopes reveal shelf acidification on southern Neotethyan margin during the Smithian-Spathian boundary cooling event

The Smithian-Spathian transition of the late Early Triassic was a critical period of environmental and biological upheavals, following the end-Permian mass extinction. Changes in carbonate deposition during this period have been attributed to intensified upwelling along shelf margins, but relevant studies are scarce. Here, we present calcium isotopes of bulk marine carbonate (δ44/40Cacarb) from a Smithian–Spathian boundary (SSB) succession (Guryul Ravine section, Kashmir) on the southern margin of the Neo-Tethys. Our smoothed δ44/40Cacarb curve reveals a ∼ 0.2‰ negative shift (from ∼ − 1.1‰ to ∼ − 1.3‰) across the SSB, concurrent with a ∼ +10‰ shift in δ13Ccarb. While increased Ca isotopic fractionation could play a role, we specifically examine potential impacts due to changes in marine Ca fluxes. Using a Ca-cycle mass balance model, we explore scenarios of decreased carbonate burial flux (Fcarb), decreased riverine flux (Friv), and a combination of these processes. The modeling suggest that a pulse decrease in Fcarb by 40% over ∼0.06 Myr match the negative shift in δ44/40Cacarb at Guryul Ravine. We infer that this decrease was likely related to intensified upwelling of acidic deep seawater due to invigorated global-oceanic circulation during the SSB cooling event. We suggest that the regionally diverse excursions in δ44/40Cacarb in the Tethyan region could be attributed to spatially varied upwellings in the shelf margin. The upwelling of acidic and anoxic deep seawater may have driven the second-order extinction of ammonoids and conodonts at the beginning of the SSB cooling event.

Ca2+ -activated K+ channels regulate cell volume in human glioblastoma cells.

Glioblastoma (GBM), the most lethal form of brain tumors, bases its malignancy on the strong ability of its cells to migrate and invade the narrow spaces of healthy brain parenchyma. Cell migration and invasion are both critically dependent on changes in cell volume and shape driven by the transmembrane transport of osmotically important ions such as K+ and Cl- . However, while the Cl- channels participating in cell volume regulation have been clearly identified, the precise nature of the K+ channels involved is still uncertain. Using a combination of electrophysiological and imaging approaches in GBM U87-MG cells, we found that hypotonic-induced cell swelling triggered the opening of Ca2+ -activated K+ (KCa ) channels of large and intermediate conductance (BKCa and IKCa , respectively), both highly expressed in GBM cells. The influx of Ca2+ mediated by the hypotonic-induced activation of mechanosensitive channels was found to be a key step for opening both the BKCa and the IKCa channels. Finally, the activation of both KCa channels mediated by mechanosensitive channels was found to be essential for the development of the regulatory volume decrease following hypotonic shock. Taken together, these data indicate that KCa channels are the main K+ channels responsible for the volume regulation in U87-MG cells.

Examining Mechanisms for Voltage-Sensitive Calcium Channel-Mediated Secretion Events in Bone Cells.

In addition to their well-described functions in cell excitability, voltage-sensitive calcium channels (VSCCs) serve a critical role in calcium (Ca2+)-mediated secretion of pleiotropic paracrine and endocrine factors, including those produced in bone. Influx of Ca2+ through VSCCs activates intracellular signaling pathways to modulate a variety of cellular processes that include cell proliferation, differentiation, and bone adaptation in response to mechanical stimuli. Less well understood is the role of VSCCs in the control of bone and calcium homeostasis mediated through secreted factors. In this review, we discuss the various functions of VSCCs in skeletal cells as regulators of Ca2+ dynamics and detail how these channels might control the release of bioactive factors from bone cells. Because VSCCs are druggable, a better understanding of the multiple functions of these channels in the skeleton offers the opportunity for developing new therapies to enhance and maintain bone and to improve systemic health.

Visualizing the triheteromeric N-methyl-D-aspartate receptor subunit composition.

N-methyl-D-aspartate receptors (NMDARs) are one of three ligand-gated ionotropic channels that transduce the effects of neurotransmitter glutamate at excitatory synapses within the central nervous system. Their ability to influx Ca2+ into cells, unlike mature AMPA or kainate receptors, implicates them in a variety of processes ranging from synaptic plasticity to cell death. Many of the receptor's capabilities, including binding glutamate and regulating Ca2+ influx, have been attributed to their subunit composition, determined putatively using cell biology, electrophysiology and/or pharmacology. Here, we show that subunit composition of synaptic NMDARs can also be readily visualized in acute brain slices (rat) using highly specific antibodies directed against extracellular epitopes of the subunit proteins and high-resolution confocal microscopy. This has helped confirm the expression of triheteromeric t-NMDARs (containing GluN1, GluN2, and GluN3 subunits) at synapses for the first time and reconcile functional differences with diheteromeric d-NMDARs (containing GluN1 and GluN2 subunits) described previously. Even though structural information about individual receptors is still diffraction limited, fluorescently tagged receptor subunit puncta coalesce with precision at various magnifications and/or with the postsynaptic density (PSD-95) but not the presynaptic active zone marker Bassoon. These data are particularly relevant for identifying GluN3A-containing t-NMDARs that are highly Ca2+ permeable and whose expression at excitatory synapses renders neurons vulnerable to excitotoxicity and cell death. Imaging NMDAR subunit proteins at synapses not only offers firsthand insights into subunit composition to correlate function but may also help identify zones of vulnerability within brain structures underlying neurodegenerative diseases like Temporal Lobe Epilepsy.

Effectiveness of Nonfunctionalized Graphene Oxide Nanolayers as Nanomedicine against Colon, Cervical, and Breast Cancer Cells.

Recent studies in nanomedicine have intensively explored the prospective applications of surface-tailored graphene oxide (GO) as anticancer entity. However, the efficacy of nonfunctionalized graphene oxide nanolayers (GRO-NLs) as an anticancer agent is less explored. In this study, we report the synthesis of GRO-NLs and their in vitro anticancer potential in breast (MCF-7), colon (HT-29), and cervical (HeLa) cancer cells. GRO-NLs-treated HT-29, HeLa, and MCF-7 cells showed cytotoxicity in the MTT and NRU assays via defects in mitochondrial functions and lysosomal activity. HT-29, HeLa, and MCF-7 cells treated with GRO-NLs exhibited substantial elevations in ROS, disturbances of the mitochondrial membrane potential, an influx of Ca2+, and apoptosis. The qPCR quantification showed the upregulation of caspase 3, caspase 9, bax, and SOD1 genes in GRO-NLs-treated cells. Western blotting showed the depletion of P21, P53, and CDC25C proteins in the above cancer cell lines after GRO-NLs treatment, indicating its function as a mutagen to induce mutation in the P53 gene, thereby affecting P53 protein and downstream effectors P21 and CDC25C. In addition, there may be a mechanism other than P53 mutation that controls P53 dysfunction. We conclude that nonfunctionalized GRO-NLs exhibit prospective biomedical application as a putative anticancer entity against colon, cervical, and breast cancers.

Crop rotation significantly influences the composition of soil, rhizosphere, and root microbiota in canola (Brassica napus L.)

BackgroundCrop rotation is an agronomic practice that is known to enhance productivity and yield, and decrease pest and disease pressure. Economic and other factors have increased the frequency of certain crops, including canola, with unknown effects on the below ground microbial communities that impact plant health and performance. This study investigated the effect of 12 years of crop rotation including canola-wheat; canola-pea-barley; and unrotated canola across three geographic sites in Western Canada with diverse soil types and environmental conditions. To provide data on mature, established crop rotation strategies, root exudate profiles, soil nutrient fluxes, and bacterial and fungal microbial community profiles were determined at the flowering stage in the final two (canola) years of the 12-year rotations.ResultsAfter 12 years of rotation, nutrient fluxes were affected in the soil in an inconsistent manner, with K, NO3, Mg, Ca, P, and Fe fluxes variably impacted by rotation depending on the year and site of sampling. As expected, rotation positively influenced yield and oil content, and decreased disease pressure from Leptosphaeria and Alternaria. In two of the three sites, root exudate profiles were significantly influenced by crop rotation. Bacterial soil, root, and rhizosphere communities were less impacted by crop rotation than the fungal communities. Fungal sequences that were associated with specific rotation strategies were identified in the bulk soil, and included known fungal pathogens in the canola-only strategy. Two closely related fungal sequences identified as Olpidium brassicae were extremely abundant at all sites in both years. One of these sequences was observed uniquely at a single site and was significantly associated with monocropped canola; moreover, its abundance correlated negatively with yield in both years.ConclusionsLong-term canola monoculture affected root exudate profiles and soil nutrient fluxes differently in the three geographic locations. Bacterial communities were less impacted by rotation compared to the fungal communities, which consistently exhibited changes in composition in all ecological niches at all sites, in both years. Fungal sequences identified as O. brassicae were highly abundant at all sites, one of which was strongly associated with canola monoculture. Soil management decisions should include consideration of the effects on the microbial ecosystems associated with the plants in order to inform best management practices.

Mechanistic insights into severe pulmonary inflammation caused by silica stimulation: The role of macrophage pyroptosis

Respirable silica dust is a common hazard faced by occupational workers and prolonged exposure to this dust can lead to pulmonary inflammation, fibrosis and, in severe cases, silicosis. However, the underlying mechanism by which silica exposure causes these physical disorders is not yet understood. In this study, we aimed to shed light on this mechanism by establishing in vitro and in vivo silica exposure models from the perspective of macrophages. Our results showed that compared to the control group, silica exposure resulted in an upregulation of the pulmonary expression of P2X7 and Pannexin-1, but this effect was suppressed by treatment with MCC950, a specific inhibitor of NLRP3. Our in vitro studies showed that silica exposure induced mitochondrial depolarization in macrophages, which led to a reduction of intracellular ATP and an influx of Ca2+. Furthermore, we found that creating an extracellular high potassium environment by adding KCl to the macrophage medium inhibited the expression of pyroptotic biomarkers and pro-inflammatory cytokines such as NLRP3 and IL-1β. Treatment with BBG, a P2X7 antagonist, also effectively inhibited the expression of P2X7, NLRP3, and IL-1β. On the other hand, treatment with FCF, a Pannexin-1 inhibitor, suppressed the expression of Pannexin-1 but had no effect on the expression of pyroptotic biomarkers such as P2X7, NLRP3, and IL-1β. In conclusion, our findings suggest that silica exposure triggers the opening of P2X7 ion channels, resulting in intracellular K+ efflux, extracellular Ca2+ influx, and the assembly of the NLRP3 inflammasome, ultimately leading to macrophage pyroptosis and pulmonary inflammation.

Toward ultraclean correlated metal CaVO3

We report the synthesis and electronic properties of the correlated metal CaVO3, grown by hybrid molecular beam epitaxy. Films were grown on (100) LaAlO3 substrates at a temperature of 900 °C by supplying a flux of elemental Ca through a thermal effusion cell and metalorganic precursor, vanadium oxitriisopropoxide, as a source of vanadium. The presence of a self-regulated growth regime was revealed by the appearance of a specific surface reconstruction detected by reflection high-energy electron diffraction. Films grown within the growth window were characterized by atomically flat surfaces. X-ray reciprocal space maps revealed that the films were coherently strained to the substrate and inherited its twinned microstructure. Despite the presence of twin walls, CaVO3 thin films, grown within the stoichiometric growth window, revealed very low electrical resistivities at low temperatures, with residual resistivity ratios exceeding 90, while films grown at either Ca- or V-excess show deteriorated transport properties, attributed to the presence of extrinsic defects arising from the non-stoichiometry present in these films.

The restriction of calcium influx in metaphase and post-metaphase stages of cell division revealed by imaging secondary ion mass spectrometry (SIMS).

A secondary ion mass spectrometry (SIMS)-based isotopic imaging technique of ion microscopy was used for observing calcium influx in single renal epithelial LLC-PK1 cells. The CAMECA IMS-3f SIMS instrument, used in the study, is capable of producing isotopic images of single cells at 500nm spatial resolution. Due to the high-vacuum requirements of the instrument the cells were prepared cryogenically with a freeze-fracture method and frozen freeze-dried cells were used for SIMS analysis. The influx of calcium was imaged directly by exposure of cells to 44 Ca stable isotope in the extracellular buffer for 10min. The 44 Ca influx was measured at mass 44 and the distribution of endogenous calcium at mass 40 (40 Ca) in the same cell. A direct comparison of interphase cells to cells undergoing division revealed that calcium influx is restricted in metaphase and post-metaphase stages of cell division. This restriction is lifted in late cytokinesis. The net influx of 44 Ca in 10min was approximately half under calcium influx restriction in comparison to interphase cells. Under calcium influx restriction the 44 Ca concentration was the same between the mitotic chromosome and the cytoplasm. These observations indicate that the endoplasmic reticulum (ER) calcium uptake is compromised under calcium influx restriction in cells undergoing division.

Molecular Dynamics Simulations of the Proteins Regulating Synaptic Vesicle Fusion

Neuronal transmitters are packaged in synaptic vesicles (SVs) and released by the fusion of SVs with the presynaptic membrane (PM). An inflow of Ca2+ into the nerve terminal triggers fusion, and the SV-associated protein Synaptotagmin 1 (Syt1) serves as a Ca2+ sensor. In preparation for fusion, SVs become attached to the PM by the SNARE protein complex, a coiled-coil bundle that exerts the force overcoming SV-PM repulsion. A cytosolic protein Complexin (Cpx) attaches to the SNARE complex and differentially regulates the evoked and spontaneous release components. It is still debated how the dynamic interactions of Syt1, SNARE proteins and Cpx lead to fusion. This problem is confounded by heterogeneity in the conformational states of the prefusion protein-lipid complex and by the lack of tools to experimentally monitor the rapid conformational transitions of the complex, which occur at a sub-millisecond scale. However, these complications can be overcome employing molecular dynamics (MDs), a computational approach that enables simulating interactions and conformational transitions of proteins and lipids. This review discusses the use of molecular dynamics for the investigation of the pre-fusion protein-lipid complex. We discuss the dynamics of the SNARE complex between lipid bilayers, as well as the interactions of Syt1 with lipids and SNARE proteins, and Cpx regulating the assembly of the SNARE complex.

Segregation of Ca2+ signaling in olfactory signal transduction.

Olfactory signal transduction is conducted through a cAMP-mediated second messenger cascade. The cytoplasmic Ca2+ concentration increases through the opening of CNG channels, a phenomenon that underlies two major functions, namely, signal boosting and olfactory adaptation. Signal boosting is achieved by an additional opening of the Ca2+-activated Cl- channel whereas adaptation is regulated by Ca2+ feedback to the CNG channel. Thus, the influx of Ca2+ and the resultant increase in cytoplasmic Ca2+ levels play seemingly opposing effects: increasing the current while reducing the current through adaptation. The two functions could be interpreted as compensating for each other. However, in real cells, both functions should be segregated. Ca2+ dynamics in olfactory cilia need to be directly measured, but technical difficulties accompanying the thin structure of olfactory cilia have prevented systematic analyses. In this study, using a combination of electrophysiology, local photolysis of caged cAMP, and Ca2+ imaging, we found that free Ca2+ in the local ciliary cytoplasm decreased along with a reduction in the current containing Ca2+-activated Cl- components returning to the basal level, whereas Ca2+-dependent adaptation persisted for a longer period. The activity of Cl- channels is highly likely to be regulated by the free Ca2+ that is present only immediately after the influx through the CNG channel, and an exclusive interaction between Ca2+ and Ca2+-binding proteins that mediate the adaptation may modulate the adaptation lifetime.

Histological Assessment of Endochondral Ossification and Bone Mineralization

Finely tuned cartilage mineralization, endochondral ossification, and normal bone formation are necessary for normal bone growth. Hypertrophic chondrocytes in the epiphyseal cartilage secrete matrix vesicles, which are small extracellular vesicles initiating mineralization, into the intercolumnar septa but not the transverse partitions of the cartilage columns. Bone-specific blood vessels invade the unmineralized transverse septum, exposing the mineralized cartilage cores. Many osteoblast precursors migrate to the cartilage cores, where they synthesize abundant bone matrices, and mineralize them in a process of matrix vesicle-mediated bone mineralization. Matrix vesicle-mediated mineralization concentrates calcium (Ca) and inorganic phosphates (Pi), which are converted into hydroxyapatite crystals. These crystals grow radially and are eventually get out of the vesicles to form spherical mineralized nodules, leading to collagen mineralization. The influx of Ca and Pi into the matrix vesicle is regulated by several enzymes and transporters such as TNAP, ENPP1, PiT1, PHOSPHO1, annexins, and others. Such matrix vesicle-mediated mineralization is regulated by osteoblastic activities, synchronizing the synthesis of organic bone material. However, osteocytes reportedly regulate peripheral mineralization, e.g., osteocytic osteolysis. The interplay between cartilage mineralization and vascular invasion during endochondral ossification, as well as that of osteoblasts and osteocytes for normal mineralization, appears to be crucial for normal bone growth.

Annexin proteins involved in plasma membrane repair reshape a membrane edge: Direct evidence from molecular simulations.

The Annexin (ANX) family of Ca+2-dependent membrane-binding proteins are implicated in plasma membrane repair in animal cells. The inward flux of Ca+2 upon formation of a plasma membrane lesion/pore drives the binding of cytoplasmic ANX proteins to the membrane, close to the pore. Annexins are deemed responsible for plasma membrane repair and are overexpressed in cancer cells where the plasma membrane is in the need for frequent repair. Annexin A5 (ANXA5) trimers form crystalline arrays solid-supported membranes (1), and owing to their concave disc-like shape, induce negative curvature on a flat plasma membrane (2), and are therefore likely to reshape the membrane in the vicinity of the membrane pore to support membrane repair (3). However, no direct evidence of membrane shaping has yet emerged. Here, we simulate ANXA5 binding on a bicelle, which models a membrane pore under periodic boundary conditions, and demonstrate that increasing concentration of ANXA5 trimers induces a monotonically increasing out-of-plane upward curvature in the bicelle. Interestingly, this overall curvature amounts to half the curvature induced by a single ANXA5 trimer on a flat lipid bilayer. The out of plane curvature induced by ANXA5 trimers near a membrane pore is demonstrated for the first time. Our results have important implications in the membrane-reshaping properties of Annexins and their ability to repair membranes.

Geochemistry of Brine and Paleoclimate Reconstruction during Sedimentation of Messinian Salt in the Tuz Gölü Basin (Türkiye): Insights from the Study of Fluid Inclusions

The halogenesis of the Messinian Tuz Gölü Basin corresponds to the sulfate type and the magnesium sulfate subtype. Compared to the Messinian Sea brines, they have a slightly higher [Na+] concentration, which is 96.6–116.4 g/L, and a much lower [K+] concentration, ranging from 0.1 to 2.35 g/L. During salt sedimentation, the [Mg2+] concentration ranged from 6.1 to 14.0 g/L, and the [SO42−] concentration from 18.2 to 4.5 g/L. Physical–chemical reactions in the basin’s near-surface and bottom waters during the suspension of halite deposition had a decisive influence on the significant reduction of [SO42−] sedimentation brines. During these periods, there was an intensive influx of Ca(HCO3)2 into the sedimentation basin and the formation of glauberite layers. The formation of the glauberite resulted from the slow dissolution of pre-deposited finely dispersed metastable minerals—gypsum, sodium syngenite, or mirabilite. In fluid inclusions in the halite, the sulfate minerals being allogenic crystals of calcium sulfate, are represented by gypsum, bassanite, and anhydrite. Additionally, as the other sulfate minerals, glauberite, anhydrite, and thenardite are found within halite crystals. Sharp fluctuations in daytime air temperatures characterized climatic indicators of the summer period in the Tuz Gölü region: 15.6–49.1 °C. In the spring or cool summer–autumn period, the daytime air temperature in the region ranged from 15.7–22.1 °C, and in late spring and early summer, it ranged from 20.6 °C to 35.0 °C. During some periods, the Tuz Gölü halite crystallized at 61.8–73.5 °C. The extreme high-temperature crystallization regime at the bottom of the salt-bearing basin was achieved due to the emergence of a vertical thermohaline structure. The “greenhouse effect” in the Tuz Gölü was established only briefly but was periodically renewed due to the influx of “fresh” waters.

CaCl2 promotes the cross adaptation of Reaumuria trigyna to salt and drought by regulating Na+, ROS accumulation and programmed cell death

Reaumuria trigyna, a salt-secreting xerophytic shrub endemic to arid desert regions of northwest China, is extremely adaptable to salt and aridity. In this study, we used PEG to simulates drought stress and investigated the effect of NaCl and CaCl2 on R. trigyna seedlings exposed to drought stress. Exogenous application moderate NaCl and CaCl2 were found to stimulate the growth and alleviate drought stress in R. trigyna seedlings. Moderate NaCl and CaCl2 combined treatment increased fresh weight and decreased electrolyte leakage, and malondialdehyde (MDA) content in R. trigyna seedlings under drought stress. Simultaneously, leaf senescence and root damage induced by drought stress were alleviated, with programmed cell death (PCD) related genes expression down-regulated. Among them, the application of CaCl2 under drought and salt treatment is the most effective way to increase osmotic regulators content, antioxidant enzymes activities, and related genes expressions of plants under drought stress, which scavenged excess reactive oxygen species (ROS) and alleviated oxidative damage caused by drought stress. Meanwhile, CaCl2 can reduce the content of Na+and the ratio of Na+/K+ by promoting the outflow of Na+ and inflow of Ca2+, as well as the expression of ion transporter gene, and reduce the ionic toxicity caused by drought and salt cross adaptation. The principal component analysis (PCA) showed that the relevant beneficial indicators were positively correlated with the combined treatment. These results indicated that moderate NaCl can positively regulates defense response to drought stress in R. trigyna, while CaCl2 can significantly promote this process.

S-27-4: OXIDIZED LDL POTENTIATES ANGIOTENSIN II-INDUCED GALPHAQ ACTIVATION THROUGH THE AT1-LOX1 RECEPTOR COMPLEX IN KIDNEY CELLS

Objective: We have reported that the binding of oxidized LDL (oxLDL) to its receptor, LOX-1 activates the G protein-coupled angiotensin II(AII) type 1 receptor (AT1) through membrane complex of the receptors. oxLDL-induced activation of AT1 is characterized by the beta-arrestin-biased signaling with no activation of Galphaq, the main effector of AII-induced cellular response (2021 iScience). In this study, we performed cell experiments to investigate how the co-treatment of oxLDL and AII activates AT1 particularly by focusing on Galphaq-dependent pathway. Design and Methods: We used CHO-cells stably transfected with AT1 (CHO-AT1), LOX-1 and AT1 (CHO-LOX-1-AT1) and two rat kidney cells (NRK49F and NRK52E). Activation of Galphaq and the subsequent induction of Ca influx were quantified by accumulation of IP1 and Fura-2 AM assay, respectively. We also utilized Galphaq-BRET biosensor vector to detect the activity of Galphaq. We assessed mRNA expression of genes associated with oxidative stress, inflammation, and fibrosis in NRK49F and NRK52E. We also assessed protein expression of alphaSMA as a molecular marker of epithelial mesanchymal transition (EMT) after 3 days of the indicated stimulation in NRK49F and NRK52E.The proliferative activity was measured using BrdU assay in NRK49F. Results: The activation of Galphaq was not observed by oxLDL treatment alone but was enhanced by the co-treatment of oxLDL and AII compared to AII alone in CHO-LOX-1-AT1 but not in CHO-AT1. Ca influx was induced not by oxLDL alone but by the co-treatment of oxLDL and a low concentration of AII that was insufficient to induce Ca influx when treated alone in CHO-LOX-1-AT1. We also found that the activation of Galphaq and the induction of Ca influx were detected by the co-treatment of oxLDL and AII but not by single treatment in rat kidney cells. These phenomenons were inhibited by ARB, a Gq inhibitor or the siRNA knockdown of AT1 or LOX-1. Furthermore, combination treatment enhanced the expression of the majority of genes associated with renal injury compared to the single treatment, and EMT was induced only by the combination treatment. These phenomenons were inhibited either by ARB or the Gq inhibitor. The combination treatment enhanced the proliferative activity of kidney fibroblasts compared with single treatment, that was inhibited by ARB, the Gq inhibitor or the siRNA knockdown of AT1 or LOX-1. Conclusions: These findings suggest the synergistic role of the AT1-LOX-1 interaction whereby oxLDL potentiates Ang II-induced Galphaq signaling, leading to the enhancement of renal injury-associated cellular response.

PS-B12-6: OXIDIZED LDL ENHANCES AT1-GQ SIGNALING MEDIATED BY ANGIOTENSIN II THROUGH LOX-1-AT1 COMPLEX IN ADRENAL CELLS.

Objectives: Previous clinical studies have implied that dyslipidemia is associated with increase of blood pressure (BP) or the incidence of hypertension. Oxidized LDL (oxLDL), which is increased in dyslipidemia, binds to lectin-like oxLDL receptor (LOX-1), while angiotensin II (AII), which is involved in the development of hypertension, binds to AII type 1 receptor (AT1), a G protein-coupled receptor. Using vascular endothelial cells, we have shown that LOX-1 and AT1 form a complex on the plasma membrane and oxLDL activates AT1. Notably, the activation of G protein by oxLDL is partial as shown by the results that it activates Gai but not Gaq (Gq), the latter of which mediates the AII-induced BP elevation by mechanisms including the promotion of adrenal aldosterone synthesis. However, we recently found that additional treatment with oxLDL enhanced AT1-Gq signaling compared to AII alone in genetically engineered-Chinese hamster ovary cells expressing both AT1 and LOX-1. In the present study, we examined whether oxLDL enhances AII-Gq signaling in aldosterone-producing adrenal cells, in order to strengthen the hypothesis that oxLDL potentiates AII-induced aldosterone production. Design and Methods: H295R, human adrenal gland carcinoma cell line were treated with oxLDL, AII, or both. Gq-dependent activation of phospholipase C was quantified by measurement of inositol monophosphate (IP1) using the IP-One assay kit. When necessary, cells were pretreated with YM-254890, a Gq inhibitor or siRNA of AT1 or LOX-1. Gq-dependent activation of Ca2+ influx was quantified by measurement of intracellular Ca2+ concentrations using the Fura-2 Ca assay kit. Results: The IP1 assay showed that oxLDL additively increases AII-mediated IP1 production compared to AII alone. Pretreatment of Gq inhibitor or siRNA of AT1 completely abolished and siRNA of LOX-1 partially abolished the activation of AT1-Gq signaling by the combination of oxLDL and AII. In the Fura-2 Ca assay, although AII at concentrations as low as 10–11 M and oxLDL alone did not activate Ca influx, the combination of oxLDL and AII vigorously induced the phenomenon. Conclusions: These results suggest that oxLDL potentiates AII-Gq signaling via a pathway depending on the LOX-1-AT1 interaction in adrenal cells. Further investigation will be required to elucidate if this phenomenon contributes to enhanced production of aldosterone both in vitro and in vivo.