Calcium Ion Activity Research Articles

Fish scale gelatin/diatom biosilica composite hemostasis sponge with ultrafast dispersing and in situ gelation for hemorrhage control.

Rapid control of hemorrhage is vital in first-aid and surgery. As representative of emergency hemostatic materials, inorganic porous materials achieve rapid hemostasis through concentrating protein coagulation factors by water adsorption to accelerate the coagulation reaction process, however their efficacy is often limited by the insufficient contact of material with blood and the lack of blood clot strength. Herein, we report an ultrafast dispersing and in situ gelation sponge (SG/DB) based on anchoring interface effect for hemorrhage control using freeze drying method after mixing fish scale gel (SG) and tert-butyl alcohol (TBA) pre-crystallized diatom biosilica (DB). This design retains the hierarchical porous structure of DB in SG matrix, and granting the SG/DB the capability to disperse ultrafast, achieving dissolution in both water and blood within 3 s. The DB and SG released by disintegration of SG/DB can activate intrinsic coagulation pathway and strengthen fibrin clot gelation through the anchoring interface effect, even realizing coagulation of anticoagulant whole blood without calcium ion activation. Animal studies showed 10%T-SG/DB has superior hemostatic properties to various commercially available hemostatic materials (rat liver and artery, 100 s; rabbit liver, artery, and heart, 3.2, 4.6, and 2.9 min, respectively), reducing bleeding by 30 % compared to QuikClot Combat Gauze®, and is easily removable without residue.

3D-Printed Bioceramic Scaffolds Reinforced by the In Situ Oriented Growth of Grains for Supercritical Bone Defect Reconstruction.

Porous calcium phosphate ceramics have attracted widespread attention owing to their excellent bioactivity. However, their poor mechanical properties severely limit their clinical applications. Significantly improving the mechanical strength of porous CaP ceramics while maintaining their bioactivity remains a major challenge. To address this issue, calcium sulfate is used to regulate the directional growth of hydroxyapatite grains during ceramic sintering. The in situ oriented grains can not only alleviate the stress concentration but also strengthen the bonding force between the ceramic grain boundaries. Calcium sulfate improves the release of active calcium ions from calcium phosphate ceramics, further enhancing their bioactivity and osteoinductivity in vivo. Transcriptome and proteome sequencing reveals that the in situ whisker-reinforced ceramics increase the expression of proteins related to calcium ion binding and promote the expression of osteogenesis-related proteins. In the supercritical bone defect repair model, repair of the defect is achieved within 3 months, with mechanical recovery reaching more than 70% of the autologous bone.

Deciphering the SAM- and metal-dependent mechanism of O-methyltransferases in cystargolide and belactosin biosynthesis: A structure–activity relationship study

Cystargolides and belactosins are natural products with a distinct dipeptide structure and an electrophilic β-lactone warhead. They are known to inhibit proteases such as the proteasome or caseinolytic protease P, highlighting their potential in treating cancers and neurodegenerative diseases. Recent genetic analyses have shown homology between the biosynthetic pathways of the two inhibitors. Here, we characterize the O-methyltransferases BelI and CysG, which catalyze the initial step of β-lactone formation. Employing techniques such as crystallography, computational analysis, mutagenesis, and activity assays, we identified a His-His-Asp (HHD) motif in the active sites of the two enzymes, which is crucial for binding a catalytically active calcium ion. Our findings thus elucidate a conserved divalent metal-dependent mechanism in both biosynthetic pathways that distinguishes BelI and CysG from previously characterized O-methyltransferases.

The Influence of Polygonatum King ilium on the Activity of Calcium-Regulating Enzymes and Calcium Ion Concentration in SH-SY5Y Cells Damaged by Aβ25-35 In Vitro

Objective To explore the effects of Polygonatum King ilium and Earthworm Formula (RPEWM) on the activity of calcium-regulating enzymes and free calcium ion (Ca2+) concentration in SH-SY5Y cells with Aβ damage in vitro. Methods SH-SY5Y cells were induced to differentiate with retinoic acid, then damaged by Aβ to mimic Alzheimer's disease (AD) injury. RPEWM extract was added during the AD injury to protect the cells, and its effects on cell proliferation, apoptosis, and the activity of cellular Ca2+-ATPase､Ca2+-Mg2+- ATPase ,as well as free calcium ion concentration were observed. Results RPEWM extract significantly protected cells from AD injury, resisted apoptosis, and promoted cell proliferation; it increased the activity of intracellular Ca2+-ATPase､Ca2+-Mg2+- ATPase, reduced Ca2+ concentration, and showed a dose-dependent relationship. Conclusion RPEWM has a good protective effect on differentiated SH-SY5Y cells with Aβ25-35damage and has a certain therapeutic effect on AD.

Purification and characterization of limonin D-ring lactone hydrolase from sweet orange (Citrus sinensis (L.) Osbeck) seeds.

Citrus products often suffer from delayed bitterness, which is generated from the conversion of non-bitter precursors (limonoate A-ring lactone, LARL) to limonin under the catalysis of limonin D-ring lactone hydrolase (LDLH). In this study, LDLH was isolated and purified from sweet orange seeds, and a rapid and accurate high-performance liquid chromatography method to quantify LARL was developed and applied to analyze the activity and enzymatic properties of purified LDLH. Purified LDLH (25.22 U mg-1) showed bands of 245 kDa and 17.5 kDa molecular weights in native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate PAGE analysis respectively. After a 24 h incubation under strongly acidic (pH 3) or strongly alkaline (pH 9) conditions, LDLH still retained approximately 100% activity. Moreover, LDLH activity was not impaired by thermal treatment at 50 °C for 120 min. Enzyme inhibition assays showed that LDLH was inactivated only after ethylenediaminetetraacetic acid treatment, and other enzyme inhibitors showed no significant effect on its activity. In addition, the LDLH activity of calcium ion (Ca2+) intervention was 108% of that in the blank group, and that of zinc ion (Zn2+) intervention was 71%. LDLH purified in this study was a multimer containing 17.5 kDa monomer with a wide pH tolerance range (pH 3-9) and excellent thermal stability. Moreover, LDLH might be a metallopeptidase, and its activity was stimulated by Ca2+ and significantly inhibited by Zn2+. These findings improve our understanding of LDLH and provide some important implications for reducing the bitterness in citrus products in the future. © 2024 Society of Chemical Industry.

Development of Anisotropic Electrically Conductive GNP-Reinforced PCL-Collagen Scaffold for Enhanced Neurogenic Differentiation under Electrical Stimulation.

The internal electric field of the human body plays a crucial role in regulating various biological processes, such as, cellular interactions, embryonic development and the healing process. Electrical stimulation (ES) modulates cytoskeleton and calcium ion activities to restore nervous system functioning. When exposed to electrical fields, stem cells respond similarly to neurons, muscle cells, blood vessel linings, and connective tissue (fibroblasts), depending on their environment. This study develops cost-effective electroconductive scaffolds for regenerative therapy. This was achieved by incorporating carboxy functionalized graphene nanoplatelets (GNPs) into a Polycaprolactone (PCL)-collagen matrix. ES was used to assess the scaffolds' propensity to boost neuronal differentiation from MSCs. This study reported that aligned GNP-reinforced PCL-Collagen scaffolds demonstrate substantial MSC differentiation with ES. This work effectively develops scaffolds using a simple, cost-effective synthesis approach. The direct coupling approach generated a homogeneous electric field to stimulate cells cultured on GNP-reinforced scaffolds. The scaffolds exhibited improved mechanical and electrical characteristics, as a result of the reinforcement with carbon nanofillers. In vitro results suggest that electrical stimulation helps differentiation of mesenchymal stem-like cells (MSC-like) towards neuronal. This finding holds great potential for the development of effective treatments for tissue injuries related to the nervous system.

Comparing physicochemical properties related to thermal stability of caprine and bovine milk protein concentrate dispersions

Thermal stability and physicochemical properties as a function of heating temperature, pH and salt addition was compared between caprine and bovine milk protein concentrate dispersions. Caprine milk protein concentrate (MPC) had a lower amount of heat coagulation time and a higher percentage of sediments than bovine MPC. Calcium ion activity of caprine MPC was higher than that of bovine. The amount of κ-casein in the serum of caprine MPC dispersions was higher than that of bovine, and increased with increasing temperatures, pH and the addition of NaCl and citrate. The amount of whey protein in serum was also higher in heated caprine MPC compared with bovine. Particle size of caprine MPC was larger than bovine MPC and was more susceptible to induced aggregation. The lower thermal stability of caprine MPC might be associated with the higher calcium ion activity and the higher amount of κ-casein dissociation promoted by whey protein denaturation and complexation in serum.

Insights into the influence of calcium ions on the adsorption behavior of sodium oleate and its response to flotation of quartz: FT-IR, XPS and AMF studies

Assiduous efforts on the high-efficiency flotation of quartz with calcium ion activation have been ongoing, while there is a lack of understanding of its potential mechanism. In this research, insights into the influence of calcium ions on the adsorption behavior of sodium oleate and its response to flotation of quartz were systematically investigated. Micro-flotation experimental results show that addition order of calcium ions influences the quartz flotation. Ca2+/NaOL complexes exhibit higher flotation recovery of quartz (92.18 %) under the recommended conditions. The Zeta potential and Fourier transform infrared spectroscopy results indicate that the adsorption of NaOL onto bare quartz surfaces is extremely difficult, and the addition of Ca2+ promotes the adsorption of NaOL. X-ray photoelectron spectroscopy analysis results confirm that Ca2+/NaOL complexes reinforce the adsorption of NaOL and Ca2+ onto quartz surface by the formation of -Si-O-Ca-OOCR, and the Ca-OOCR content in the total Ca reaches 62.61 %. The adsorption amount results further illustrate that the adsorption amount of Ca2+ and NaOL increase to 18.56 × 10−2 mg/g and 40.83 × 10−2 mg/g, respectively. Additionally, starch slightly decrease the adsorption amount of Ca2+ on quartz surfaces. Atomic force microscope analysis reveal that Ca2+/NaOL components promote the formation of Ca-OOCR colloid precipitates and the transportation of the precipitates to the quartz surface. The main adsorption of Ca-OOCR colloid precipitates contributes to the enveloped bulge image. Notably, the Ca-OOCR colloid precipitates adsorption mechanism exerts more important role in the adsorption of sodium oleate and calcium ion species, as well as its response to flotation of quartz.

FIM: A fatigued-injured muscle model based on the sliding filament theory

Skeletal muscle modeling has a vital role in movement studies and the development of therapeutic approaches. In the current study, a Huxley-based model for skeletal muscle is proposed, which demonstrates the impact of impairments in muscle characteristics. This model focuses on three identified ions: H+, inorganic phosphate Pi, and Ca2+. Modifications are made to actin-myosin attachment and detachment rates to study the effects of H+ and Pi. Additionally, an activation coefficient is included to represent the role of calcium ions interacting with troponin, highlighting the importance of Ca2+. It is found that maximum isometric muscle force decreases by 9.5% due to a reduction in pH from 7.4 to 6.5 and by 47.5% in case of the combination of a reduction in pH and an increase of Pi concentration up to 30 mM, respectively. Then the force decline caused by a fall in the active calcium ions is studied. When only 15% of the total calcium in the myofibrillar space is able to interact with troponin, up to 80% force drop is anticipated by the model. The proposed fatigued-injured muscle model is useful to study the effect of various shortening velocities and initial muscle-tendon lengths on muscle force; in addition, the benefits of the model go beyond predicting the force in different conditions as it can also predict muscle stiffness and power. The power and stiffness decrease by 40% and 6.5%, respectively, due to the pH reduction, and the simultaneous accumulation of H+ and Pi leads to a 50% and 18% drop in power and stiffness.

Identification of a drug binding pocket in TMEM16F calcium-activated ion channel and lipid scramblase

The dual functions of TMEM16F as Ca2+-activated ion channel and lipid scramblase raise intriguing questions regarding their molecular basis. Intrigued by the ability of the FDA-approved drug niclosamide to inhibit TMEM16F-dependent syncytia formation induced by SARS-CoV-2, we examined cryo-EM structures of TMEM16F with or without bound niclosamide or 1PBC, a known blocker of TMEM16A Ca2+-activated Cl- channel. Here, we report evidence for a lipid scrambling pathway along a groove harboring a lipid trail outside the ion permeation pore. This groove contains the binding pocket for niclosamide and 1PBC. Mutations of two residues in this groove specifically affect lipid scrambling. Whereas mutations of some residues in the binding pocket of niclosamide and 1PBC reduce their inhibition of TMEM16F-mediated Ca2+ influx and PS exposure, other mutations preferentially affect the ability of niclosamide and/or 1PBC to inhibit TMEM16F-mediated PS exposure, providing further support for separate pathways for ion permeation and lipid scrambling.

Surface plasmon resonance and microscale thermophoresis approaches for determining the affinity of perforin for calcium ions.

Perforin is a pore-forming protein that plays a crucial role in the immune system by clearing virus-infected or tumor cells. It is released from cytotoxic granules of immune cells and forms pores in targeted lipid membranes to deliver apoptosis-inducing granzymes. It is a very cytotoxic protein and is therefore adapted not to act in producing cells. Its activity is regulated by the requirement for calcium ions for optimal activity. However, the exact affinity of perforin for calcium ions has not yet been determined. We conducted a molecular dynamics simulation in the absence or presence of calcium ions that showed that binding of at least three calcium ions is required for stable perforin binding to the lipid membrane. Biophysical studies using surface plasmon resonance and microscale thermophoresis were then performed to estimate the binding affinities of native human and recombinant mouse perforin for calcium ions. Both approaches showed that mouse perforin has a several fold higher affinity for calcium ions than that of human perforin. This was attributed to a particular residue, tryptophan at position 488 in mouse perforin, which is replaced by arginine in human perforin. This represents an additional mechanism to control the activity of human perforin.

Abstract A010: ANO7 expression changes in prostate cancer progression — Association with an aggressive phenotype?

Abstract Introduction The expression level and single nucleotide polymorphisms of the prostate-specific gene ANO7 have been shown to correlate with prostate cancer (PrCa) outcome. Very little is known about ANO7, apart from its expression profile in tissues and that the protein is either a calcium-activated ion channel or a lipid scramblase. Our results suggest that ANO7 likely has a role in PrCa progression and that loss of ANO7 is associated with malignant PrCa. Materials and methods Sections of a prostate cancer formalin-fixed paraffin-embedded tissue microarray were used for ANO7 mRNA and protein detection. mRNA was detected with fluorescent in situ hybridization (FISH). Protein was detected with immunohistochemistry (IHC) using a polyclonal anti-ANO7 antibody (HPA035730). mRNA from fresh radical prostatectomy primary tumor samples were sequenced and subsequent gene expression levels were analyzed for co-expression with ANO7. The sample set consisted of a matched tumor and benign tissue sample (15+15) from each patient. Results The FISH images indicated explicitly that ANO7 mRNA is exclusively expressed in the luminal cells of the prostatic glandular epithelium. ANO7 mRNA is expressed in all cancerous lumina-forming glandular structures, but not in poorly-formed glands or sheets of cancer, in which ANO7 mRNA expression is very low or non-existent. Morphologically benign glands amidst cancerous tissue showed no expression, in contrast to tissue in which benign glands are predominating. The IHC showed a similar staining pattern to FISH.ANO7 co-expression gene set enrichment analysis resulted in 19 positively correlating and 77 negatively correlating pathways in KEGG (Kyoto Encyclopedia of Genes and Genomes) terms. Most of the positively correlating terms relate to metabolism, whereas among the negatively correlating terms, there were many signal transduction and cell growth and death-related pathways, such as Hippo, Ras, and p53 signaling pathways. Conclusions ANO7 mRNA and protein expression correlate spatially in the tissue, showing that the expression is specific to differentiated luminal cells and highly reduced in high-grade cancer. This notion is supported by the co-expressed gene-set enrichment analysis, which is showing a negative correlation of ANO7 with many pathways that are associated with aggressive PrCa. Mainly metabolism-related processes were positively correlated with ANO7, which could indicate that ANO7 is linked to the reprogramming of metabolism in the cells during cancer progression. Further research aims to identify the pathways in which ANO7 plays a significant role and to elucidate how the ANO7 protein functionally alters signaling related to PrCa progression. Citation Format: Olli Metsälä, Gudrun Wahlström, Pekka Taimen, Johanna Schleutker. ANO7 expression changes in prostate cancer progression — Association with an aggressive phenotype? [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A010.

Evaluation of the Anticancer Activity of Calcium Ions Introduced into Human Breast Adenocarcinoma Cells MCF-7/WT and MCF-7/DOX by Electroporation.

Breast cancer ranks among the top three most common malignant neoplasms in Poland. The use of calcium ion-assisted electroporation is an alternative approach to the classic treatment of this disease. The studies conducted in recent years confirm the effectiveness of electroporation with calcium ions. Electroporation is a method that uses short electrical pulses to create transitional pores in the cell membrane to allow the penetration of certain drugs. The aim of this study was to investigate the antitumor effects of electroporation alone and calcium ion-assisted electroporation on human mammary adenocarcinoma cells that are sensitive (MCF-7/WT) and resistant to doxorubicin (MCF-7/DOX). The cell viability was assessed using independent tests: MTT and SRB. The type of cell death after the applied therapy was determined by TUNEL and flow cytometry (FACS) methods. The expression of Cav3.1 and Cav3.2 proteins of T-type voltage-gated calcium channels was assessed by immunocytochemistry, and changes in the morphology of CaEP-treated cells were visualized using a holotomographic microscope. The obtained results confirmed the effectiveness of the investigated therapeutic method. The results of the work constitute a good basis for planning research at the in vivo level and in the future to develop a more effective and safer method of breast cancer treatment for patients.

PIEZO1 and PECAM1 interact at cell-cell junctions and partner in endothelial force sensing

Two prominent concepts for the sensing of shear stress by endothelium are the PIEZO1 channel as a mediator of mechanically activated calcium ion entry and the PECAM1 cell adhesion molecule as the apex of a triad with CDH5 and VGFR2. Here, we investigated if there is a relationship. By inserting a non-disruptive tag in native PIEZO1 of mice, we reveal in situ overlap of PIEZO1 with PECAM1. Through reconstitution and high resolution microscopy studies we show that PECAM1 interacts with PIEZO1 and directs it to cell-cell junctions. PECAM1 extracellular N-terminus is critical in this, but a C-terminal intracellular domain linked to shear stress also contributes. CDH5 similarly drives PIEZO1 to junctions but unlike PECAM1 its interaction with PIEZO1 is dynamic, increasing with shear stress. PIEZO1 does not interact with VGFR2. PIEZO1 is required in Ca2+-dependent formation of adherens junctions and associated cytoskeleton, consistent with it conferring force-dependent Ca2+ entry for junctional remodelling. The data suggest a pool of PIEZO1 at cell junctions, the coming together of PIEZO1 and PECAM1 mechanisms and intimate cooperation of PIEZO1 and adhesion molecules in tailoring junctional structure to mechanical requirement.

Evaluation of carbonation conversion of recycled concrete fines using high-temperature CO2: Reaction kinetics and statistical method for parameters optimization

Accelerated carbonation of recycled concrete fines (RCF) is an eco-friendly technology and has been considered as an effective way to achieve CO2 fixation and improve the cementitious activity of RCF. However, the slow gas-solid reaction under a standard (20 °C) environment is an issue affecting the carbonation performance. On the other hand, the surface water of alkaline solids plays a crucial role as an ionic transferring medium for high-temperature carbonation. This work, therefore, investigates the influences of operating parameters, i.e., temperature (60–180 °C), water content (W/S ratio; 0.1–0.5 ml/g) and carbonation duration (5–50 min) on the reaction of RCF. After that, multiple kinetics models, namely shrinking-core model and surface water coverage model, are presented to elucidate the carbonation kinetics of RCF under high-temperature environments. The maximum diffusivity of CO2 and carbonation rate of RCF are 1.33 × 10−6 cm2/min and 0.052 min−1, respectively at 100 °C, whereas the carbonated particles are surrounded by polyhedral calcite crystals. Kinetics study shows that the activity of calcium ions in water film decreases with higher temperature due to faster evaporation of water and the development of an electrical double layer in the interface particles. In addition, the non-evaporated surface water at high W/S ratios limits the CO2 penetration and reaction rate. Finally, the response surface methodology is employed for statistical prediction of the maximum conversion of RCF. Accordingly, the highest carbonation conversion of RCF is 56.69% for particles wetted with a W/S ratio of 0.3 ml/g and carbonated at 100 °C for 40 min.

Effect of microbial transglutaminase on thermal stability of milk protein

At present, Transglutaminase (abbreviated as TG, EC2.3.2.13) has been used in meat products, dairy products and baked goods in Japan and other countries. The stability of MP (Milk protein) colloid system is very important to the quality of dairy products. The effect of MTG (Microbial TG) on the thermal stability of MP was studied in this experiment. The change of HCT (Heat Coagulation Time) of MP emulsion before and after adding MTG was measured. The results show that the effect of MTG on MP leads to the decrease of its solubility in the region of pH < 4.0 or pH > 7.0, but it increases in the range near the isoelectric point of MP (pH 4.0 ~ 6.0). In the range of acidic pH value with low stability, the high calcium ion activity, low Zeta potential and micelle hydration make the milk have poor thermal stability and will solidify rapidly at 150℃. The hydrophobic interaction between MTG and MP changes the conformation of MP, which leads to the change of thermal stability.

Single-step genome-wide association analyses of claw horn lesions in Holstein cattle using linear and threshold models

BackgroundLameness in dairy cattle is primarily caused by foot lesions including the claw horn lesions (CHL) of sole haemorrhage (SH), sole ulcers (SU), and white line disease (WL). This study investigated the genetic architecture of the three CHL based on detailed animal phenotypes of CHL susceptibility and severity. Estimation of genetic parameters and breeding values, single-step genome-wide association analyses, and functional enrichment analyses were performed.ResultsThe studied traits were under genetic control with a low to moderate heritability. Heritability estimates of SH and SU susceptibility on the liability scale were 0.29 and 0.35, respectively. Heritability of SH and SU severity were 0.12 and 0.07, respectively. Heritability of WL was relatively lower, indicating stronger environmental influence on the presence and development of WL than the other two CHL. Genetic correlations between SH and SU were high (0.98 for lesion susceptibility and 0.59 for lesion severity), whereas genetic correlations of SH and SU with WL also tended to be positive. Candidate quantitative trait loci (QTL) were identified for all CHL, including some on Bos taurus chromosome (BTA) 3 and 18 with potential pleiotropic effects associated with multiple foot lesion traits. A genomic window of 0.65 Mb on BTA3 explained 0.41, 0.50, 0.38, and 0.49% of the genetic variance for SH susceptibility, SH severity, WL susceptibility, and WL severity, respectively. Another window on BTA18 explained 0.66, 0.41, and 0.70% of the genetic variance for SH susceptibility, SU susceptibility, and SU severity, respectively. The candidate genomic regions associated with CHL harbour annotated genes that are linked to immune system function and inflammation responses, lipid metabolism, calcium ion activities, and neuronal excitability.ConclusionsThe studied CHL are complex traits with a polygenic mode of inheritance. Most traits exhibited genetic variation suggesting that animal resistance to CHL can be improved with breeding. The CHL traits were positively correlated, which will facilitate genetic improvement for resistance to CHL as a whole. Candidate genomic regions associated with lesion susceptibility and severity of SH, SU, and WL provide insights into a global profile of the genetic background underlying CHL and inform genetic improvement programmes aiming at enhancing foot health in dairy cattle.

Rhythmic calcium ion activity related to glioma growth reveals the mechanism of ion transmission

Rhythmic calcium ion activity related to glioma growth reveals the mechanism of ion transmission

Effect of sodium hexametaphosphate on heat-induced changes in micellar casein isolate solutions

Micellar casein isolate (MCI) solutions (9%, w/w, casein; pH 6.7) containing 0, 12 or 24 mEq L−1 sodium hexametaphosphate (SHMP) were subject to three different thermal treatments: in-container sterilisation (121 °C for 8 min) and two different continuous flow sterilisation treatments (124 °C for 5 min and 140 °C for 5.8 s). Samples were analysed after 1–60 d storage at 20 and 40 °C for pH, calcium ion activity, turbidity, particle size, viscosity and protein and mineral distribution. SHMP-induced casein micelle disruption in untreated samples was apparent from reductions in turbidity, particle size and increases in sedimentable caseins and calcium, and from strong increases in viscosity. After heating, the pH and viscosity decreased strongly due to the heat-induced hydrolysis of SHMP. SHMP altered casein micelle structure, but during heat treatment and storage, samples with SHMP showed good stability.

A TMEM16J variant leads to dysregulated cytosolic calcium which may lead to renal disease.

SIGIRR (single immunoglobulin IL-1 related receptor), PKP3 (plakophilin 3), and TMEM16J (anoctamin 9), a putative calcium-activated ion channel and phospholipid scramblase, control the immune response and the extent of inflammation. Variants of SIGIRR/PKP3/TMEM16J lead to severe inflammatory diseases such as pneumonia, enterocolitis, and kidney graft rejection. Meta-analysis of genome-wide association studies identified TMEM16J-T604A as a promotor for chronic kidney disease (CKD), but the disease mechanism and function of TMEM16J remain unknown. Here, we demonstrate TMEM16J as a calcium-activated calcium-permeable channel, which is expressed in the endoplasmic reticulum (ER). TMEM16J controls the intracellular distribution of calcium, and inhibits intracellular receptor-mediated Ca2+ signals and Ca2+ -dependent activation of ion channels, but augments transcription and release of pro-inflammatory cytokines. Renal epithelial cells expressing the variant TMEM16J-T604A show enhanced calcium signals when compared to cells expressing wt-TMEM16J, and demonstrate spontaneous transcription and release of cytokines. This study identifies TMEM16J as an important regulator of intracellular Ca2+ signals, ion channel activity, and cytokine release. TMEM16J may therefore affect immune response in renal tissue and immune cells.