Ion Current Research Articles

Abstract WP27: Tissue Kallikrein Attenuates Ischemic Edema by Increasing Water Transport Function of AQP4 in Glymphatic System in pMCAO Mice

Aquaporin-4 (AQP4) is an essential water channel protein in GS (glymphatic system), and is widely distributed in astrocytes at the brain/cerebrospinal fluid interface, where it facilitates water movement. Altered AQP4 polarization and expression are evident in cytotoxic edema in pMCAO mice in our previous study, glymphatic MRI of which also shows reduced CSF tracer entry and clearance. Tissue Kallikrein treatment provides cerebral infarction patients with vasodilatory activity, which may facilitate the dynamics of CSF in GS. We hypothesized that TK could attenuate depolarization of AQP4 to confer benefit in brain edema by boosting the kinetics in GS. In this study, we evaluated the effects of TK on mice acute ischemic edema in both wild and AQP4-/- pMCAO models. A fluorescent tracer was injected into the striatum of both mice, and the infusion pattern of the fluorescent tracer in the brain parenchyma was detected to confirm the function of AQP4 in alleviating edema by TK. Concurrently, immunohistochemistry analysis was performed to explore the altered AQP4 polarization and expression in TK treatment animals. Our results indicated that TK attenuated brain edema was associated with improved transport function of AQP4 to CSF in pMCAO mice, which might relate to the alteration of polarization and expression of AQP4.

Magnetic Trapping of Galactic Cosmic Rays in the Outer Heliosheath and Their Preferential Entry into the Heliosphere

This paper examines the geometry of interstellar magnetic field lines close to the boundary of the heliosphere in the direction of the unperturbed local interstellar magnetic field, where the field lines are spread apart by the heliopause (HP). Such field parting establishes a region of weaker magnetic field of about 300 au in size in the northern hemisphere that acts as a giant magnetic trap affecting the propagation of galactic cosmic rays (GCRs). The choice of an analytic model of the magnetic field in the very local interstellar medium allows us to qualitatively study the resulting magnetic field draping pattern while avoiding unphysical dissipation across the HP-impeding numerical magnetohydrodynamic (MHD) models. We investigate GCR transport in the region exterior to the heliosphere, including the magnetic trap, subject to guiding center drifts, pitch angle scattering, and perpendicular diffusion. The transport coefficients were derived from Voyager 1 observations of magnetic turbulence in the VLISM. Our results predict a ring current of energetic ions drifting around the interior of the magnetic trap. It is also demonstrated that GCRs cross the HP for the first time preferentially through a crescent-shaped region between the magnetic trap and the upwind direction. The paper includes results of MHD modeling of the heliosphere that provide the coordinates of the center of the magnetic trap in ecliptic coordinates. In addition to the heliosphere, we examine several extreme field draping configurations that could describe the astrospheres of other stars.

Mitochondrial Morphology and Function Abnormality in Ovarian Granulosa Cells of Patients with Diminished Ovarian Reserve.

In this study, we examined the changes in the mitochondrial structure and function in cumulus granulosa cells of patients with diminished ovarian reserve (DOR) to explore the causes and mechanisms of decreased mitochondrial quality. The mitochondrial ultrastructure was observed by transmission electron microscope, and the function was determined by detecting the ATP content, reactive oxygen species (ROS) levels, the number of mitochondria, and the mitochondrial membrane potential. The expression of ATP synthases in relation to mitochondrial function was analyzed. Additionally, protein immunoblotting was used to compare the expression levels of mitochondrial kinetic protein, the related channel protein in the two groups. Patients with DOR had abnormal granulosa cell morphology, increased mitochondrial abnormalities, decreased mitochondrial function, and disturbed mitochondrial dynamics. Additionally, the silent information regulator 1 (SIRT1)/phospho-AMP-activated protein kinase (P-AMPK)-peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) pathway expression was decreased, which was speculated to be associated with the decreased mitochondrial mass in the DOR group. The mitochondrial mass was decreased in granulosa cells of patients in the DORgroup. The mitochondrial dysfunction observed in granulosa cells of patients in the DOR group may be associated with dysregulation of the SIRT1/P-AMPK-PGC-1α-mitochondrial transcription factor A (TFAM) pathway.

Ignition Delay and Reaction Time Measurements of Hydrogen–Air Mixtures at High Temperatures

Induction and reaction times of hydrogen–air mixtures (ϕ = 0.5–2) have been measured behind reflected shock waves at temperatures of 1000–1600 K, pressures of 0.1, 0.3, 0.6 MPa in the domain of the extended second explosion limit. The measurements were performed in the shock tube with a completely transparent test section of 0.5 m long, which provides pressure, ion current, OH and high-speed chemiluminescence observations. The experimental induction time plots demonstrate a clear increasing of the global activation energy from high- to low temperature post-shock conditions. This trend is strongly pronounced at higher post-shock pressures. For a high-temperature range of T > 1200 K, induction time measurements show an activation energy for the global reaction rate of hydrogen oxidation of 64–83 kJ/mole. Detected reaction times exhibit a big scatter and a weak temperature dependence. The minimum reaction time value was nearly 2 µs. Obtained induction time data were compared with calculations carried out in accordance with the known kinetic mechanisms. For current and former shock-tube experiments within a pressure range of 0.1–2 MPa, critical temperatures required for strong (1000–1100 K), transient and weak auto-ignition modes behind reflected shock waves were identified by means of the pressure and ion-probe measurements in stoichiometric hydrogen-air mixture. The transfer from the strong volumetric self-ignition near the reflecting wall to the hot spot ignition (transient) was established and visualized below <1200 K with a post-shock temperature decreasing.

Next Generation Sequencing and Electromyography Reveal the Involvement of the P2RX6 Gene in Myopathy.

Ion channelopathies result from impaired ion channel protein function, due to mutations affecting ion transport across cell membranes. Over 40 diseases, including neuropathy, pain, migraine, epilepsy, and ataxia, are associated with ion channelopathies, impacting electrically excitable tissues and significantly affecting skeletal muscle. Gene mutations affecting transmembrane ionic flow are strongly linked to skeletal muscle disorders, particularly myopathies, disrupting muscle excitability and contraction. Electromyography (EMG) analysis performed on a patient who complained of weakness and fatigue revealed the presence of primary muscular damage, suggesting an early-stage myopathy. Whole exome sequencing (WES) did not detect potentially causative variants in known myopathy-associated genes but revealed a novel homozygous deletion of the P2RX6 gene likely disrupting protein function. The P2RX6 gene, predominantly expressed in skeletal muscle, is an ATP-gated ion channel receptor belonging to the purinergic receptors (P2RX) family. In addition, STRING pathways suggested a correlation with more proteins having a plausible role in myopathy. No previous studies have reported the implication of this gene in myopathy. Further studies are needed on patients with a defective ion channel pathway, and the use of in vitro functional assays in suppressing P2RX6 gene expression will be required to validate its functional role.

Multiphysics Modeling of Electrochemical Impedance Spectroscopy Responses of SAM-Modified Screen-Printed Electrodes

In this work, we present a multiphysics modeling approach capable of simulating electrochemical impedance spectroscopy (EIS) responses of screen-printed electrodes (SPEs) modified with self-assembled monolayers of 11-Mercaptoundecanoic acid (MUA). Commercially available gold SPEs are electrochemically characterized through experimental cyclic voltammetry and EIS measurements with 10 mM [Fe(CN)6]3−/4− redox couple in phosphate buffered saline before and after the surface immobilization of MUA at different concentrations. We design the multiphysics model through COMSOL Multiphysics® based on the 3D geometry of the devices under test. The model includes four different physics considering the metal/solution interface electrochemical phenomena, the ion and electron potentials and currents, and the measurement set-up. The model is calibrated through a set of experimental measurements, allowing the tuning of the parameters used by the model. We use the calibrated model to simulate the EIS response of MUA-modified SPEs, comparing the results with experimental data. The simulations fit the experimental curves well, following the variation of MUA concentration on the surface from 1 µM to 100 µM. The EIS parameters, retrieved through a CPE-modified Randles’ circuit, confirm the consistency with the experimental data. Notably, the simulated surface coverage estimates and the variation of charge transfer resistance due to MUA-immobilization are well matched with their experimental counterparts, reporting only a 2% difference and being consistent with the experimental electrochemical behavior of the SPEs.

Transient Receptor Potential Channels: Multiple Modulators of Peripheral Neuropathic Pain in Several Rodent Models.

Neuropathic pain, a prevalent chronic condition in clinical settings, has attracted widespread societal attention. This condition is characterized by a persistent pain state accompanied by affective and cognitive disruptions, significantly impacting patients' quality of life. However, current clinical therapies fall short of addressing its complexity. Thus, exploring the underlying molecular mechanism of neuropathic pain and identifying potential targets for intervention is highly warranted. The transient receptor potential (TRP) receptors, a class of widely distributed channel proteins, in the nervous system, play a crucial role in sensory signaling, cellular calcium regulation, and developmental influences. TRP ion channels are also responsible for various sensory responses including heat, cold, pain, and stress. This review highlights recent advances in understanding TRPs in various rodent models of neuropathic pain, aiming to uncover potential therapeutic targets for clinical management.

Lead inhibits microglial cell migration via suppression of store-operated calcium entry

Lead (Pb) is a non-biodegradable environmental pollutant that can lead to neurotoxicity by inducing neuroinflammation. Microglial activation plays a key role in neuroinflammation, and microglial migration is one of its main features. However, whether Pb affects microglial migration has not yet been elucidated. Herein, the effect of Pb on microglial migration was investigated using BV-2 microglial cells and primary microglial cells. The results showed that cell activation markers (TNF-α and CD206) in BV-2 cells were increased after Pb treatment. The migration ability of microglia was inhibited by Pb. Both store-operated calcium entry (SOCE) and the Ca2+ release-activated Ca2+ (CRAC) current were downregulated by microglia treatment with Pb in a dose-dependent manner. However, there was no statistical difference in the protein levels of stromal interaction molecule (STIM) 1, STIM2, or Ca2+ release-activated Ca2+ channel protein (Orai) 1 in microglia. The external Ca2+ influx and cell migration ability were restored to a certain extent after overexpression of either STIM1 or its CRAC activation domain in microglia. These results indicated that Pb inhibits microglial migration by downregulation of SOCE and impairment of the function of STIM1.

Corrosion behavior of laser powder bed fusion additive manufacturing produced TiNi alloy by micro-arc oxidation

To improve the corrosion resistance of TiNi alloy fabricated by laser powder bed fusion (LPBF), a porous oxidation layer was synthesized by micro-arc oxidation in a sodium aluminate and sodium silicate electrolyte. The influences of the applied voltage and the processing time on the morphology of oxidation layer were investigated, and the corrosion behavior of the oxidation layer in artificial saliva was evaluated and compared with that of the as-fabricated LPBF alloy. The results indicate that, as increasing the applied voltage and the processing time, the oxidation layer becomes uniform and integrated. The optimum parameters are with an applied voltage of 450 V and processing time of 40 min. The oxidation layer primarily contains α-Al2O3 and consists of two layers, i.e., a thin, compact and uniform inner layer and a porous outer layer. The formation of stable α-Al2O3 phase in the coating and its almost non-porous dense structure reduce the channels for corrosion ions to penetrate into the substrate through coating, thereby improving the corrosion resistance of TiNi alloy.

The apical 70-pS potassium K channel in the thick ascending limb of Henle's loop is a large-conductance Na+-and Cl--activated, KNa1.1-like, channel.

Apical potassium channels are crucial for thick ascending limb (TAL) of Henle's loop transport function. The ROMK (KNCJ1) gene encodes a 30-pS K channel whose loss of function causes the reduced NaCl reabsorption in the TAL associated with Type 2 Bartter's syndrome. In contrast, the molecular basis of a functionally ROMK-related 70-pS K channel is still unclear. The aim of this study was to highlight new specific channel properties that may give insights on its molecular identity. Using the patch-clamp technique on the apical membrane of mouse split-open TAL tubules, we observed that 70-pS K channel activity, but not ROMK channel activity, increases with the internal Na+ and Cl- concentrations, with relative 50 % effective concentrations (EC50) and Hill coefficients (nH) of 40.6 mM (SD 1.65) and 2.4 (SD 0.28) for Na+, and of 29.3 mM (SD 2.35) and 2.2 (SD 0.39) for Cl-. Conversely, 70-pS K channel activity was inhibited by internal K+ with a relative EC50 of 64 mM (SD 13.5) and a nH of 3.5 (SD 2.3), and by internal NH4+ and Ca2+. The reevaluation of channel conductive properties revealed an actual inward conductance of ~ 170 pS, with multiple subconductance levels and an inward rectification, and a substantial permeability to NH4+ ( = 0.2). We conclude that the apical 70-pS K channel in TAL cells is a large-conductance Na+- and Cl--activated potassium channel functionally resembling a KNa1.1 channel and propose that ROMK determines its functional expression possibly at the level of channel protein synthesis or trafficking.

A Mechanical–Electrical Model to Describe the Negative Differential Resistance in Membranotronic Devices

Membranotronic devices are artificial neural membranes mimicing the functionality of biological neural networks. These devices rely on the emergence of negative differential resistance (NDR). A minimalistic physical model for membranotronic devices capable of generating NDR is presented. The model features a deformable membrane with holes that facilitate ion currents. The deformation of the membrane, induced by electrostatic pressure from an applied voltage, modulates these currents. The model comprises a well‐established mechanical framework for describing deformable membranes with holes, alongside a model for ionic current that considers temperature‐dependent ion mobilities. It is demonstrated that the model can faithfully reproduce NDR across a wide and physically realistic range of parameter combinations. Furthermore, the simulations reveal that the temperature of the electrolyte can exceed its boiling point, resulting in bubble formation. To mitigate this issue, materials with high heat transfer coefficients and low conductivity are recommended. In essence, the work bridges the gap between artificial membranotronic devices and biological neural networks by providing a robust physical model capable of emulating NDR, a key feature in the operation of such systems. This advancement in membranotronics holds great promise for the development of bioinspired soft artificial neuromimetic systems that closely mimic their biological counterparts.

A MXene heterostructure-based piezoionic sensor for wearable sensing applications

Humans rely on ion transport to sense and process tactile information. Piezoionic sensor can directly convert mechanical energy into electrical energy through similar ion migration, showing applications in wearable sensing. However, affected by the electrode materials properties, the relatively low voltage output and poor stability of the piezoionic sensors restrict their development. Here, we designed a MXene/Ag nanoparticles heterostructure electrode-based piezoionic sensor with stable and high-performance voltage output. During ion mediated mechanoelectric conversion, the layered MXene nanosheets in electrodes provide transport channels for movable ions. While the interlayer insertion of Ag nanoparticles increases the interlayer spacing between MXene nanosheets, improving the ion storage and transport. Under a bending deformation of 0.7 % strain, the sensor outputs 11.1 mV voltage. The voltage can maintain stable under cyclic deformation for about 13000 s. Based on the excellent mechanical-to-electric conversion, this sensor can be applied to monitor human body activities. The feasibility of the sensor for guiding high-quality cardiopulmonary resuscitation (CPR) training is also demonstrated. Moreover, it can be adhered on the finger to generate pulse or square wave electrical signals by rhythmic bending the finger, which can transmit information based on the coding principle of Morse code. These results indicate the potential of the self-powered piezoionic sensor in medical rehabilitation and human–computer interaction.

Hypertrophic cardiomyopathy dysfunction mimicked in human engineered heart tissue and improved by sodium-glucose cotransporter 2 inhibitors.

Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiomyopathy, often caused by pathogenic sarcomere mutations. Early characteristics of HCM are diastolic dysfunction and hypercontractility. Treatment to prevent mutation-induced cardiac dysfunction is lacking. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are a group of antidiabetic drugs that recently showed beneficial cardiovascular outcomes in patients with acquired forms of heart failure. We here studied if SGLT2i represent a potential therapy to correct cardiomyocyte dysfunction induced by an HCM sarcomere mutation. Contractility was measured of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) harbouring an HCM mutation cultured in 2D and in 3D engineered heart tissue (EHT). Mutations in the gene encoding β-myosin heavy chain (MYH7-R403Q) or cardiac troponin T (TNNT2-R92Q) were investigated. In 2D, intracellular [Ca2+], action potential and ion currents were determined. HCM mutations in hiPSC-CMs impaired relaxation or increased force, mimicking early features observed in human HCM. SGLT2i enhance the relaxation of hiPSC-CMs, to a larger extent in HCM compared to control hiPSC-CMs. Moreover, SGLT2i-effects on relaxation in R403Q EHT increased with culture duration, i.e. hiPSC-CMs maturation. Canagliflozin's effects on relaxation were more pronounced than empagliflozin and dapagliflozin. SGLT2i acutely altered Ca2+ handling in HCM hiPSC-CMs. Analyses of SGLT2i-mediated mechanisms that may underlie enhanced relaxation in mutant hiPSC-CMs excluded SGLT2, Na+/H+ exchanger, peak and late Nav1.5 currents, and L-type Ca2+ current, but indicate an important role for the Na+/Ca2+ exchanger. Indeed, electrophysiological measurements in mutant hiPSC-CM indicate that SGLT2i altered Na+/Ca2+ exchange current. SGLT2i (canagliflozin > dapagliflozin > empagliflozin) acutely enhance relaxation in human EHT, especially in HCM and upon prolonged culture. SGLT2i may represent a potential therapy to correct early cardiac dysfunction in HCM.

Transcriptomics-based analysis reveals the nephrotoxic effects of triphenyltin (TPT) on SD rats by affecting RAS, AQPs and lipid metabolism

Triphenyltin (TPT) is a class of organotin compounds that are extensively used in industry and agriculture. They have endocrine-disrupting effects and cause severe environmental contamination. Pollutants may accumulate in the kidneys and cause pathological complications. However, the mechanism of TPT's toxicological effects on the kidney remains unclear. This study aimed to investigate the toxic effects and mechanism of action of TPT exposure on renal impairment in rats. Male SD rats were divided into four groups: the Ctrl group (control group), TPT-L group (0.5 mg/kg/d), TPT-M group (1 mg/kg/d), and TPT-H group (2 mg/kg/d). After 28 days of exposure to TPT, we observed the morphology and structure of kidney tissue using HE, PASM, and Masson staining. We also detected serum biochemical indexes, performed transcriptome sequencing of rat kidney tissue using RNA-seq. Furthermore, protein expression levels were measured through immunohistochemistry and gene expression levels were determined using RT-qPCR. The study results indicated a decrease in kidney weight and relative kidney weight after 28 days of exposure to TPT. Additionally, TPT caused damage to kidney structure and function, as evidenced by HE staining, PASM staining, and serum biochemical tests. Transcriptomics identified 352 DEGs, and enrichment analyses revealed that TPT exposure primarily impacted the renin-angiotensin system (RAS). The expression levels of water channel proteins were reduced, and the expression levels of RAS and lipid metabolism-related genes (Mme, Ace, Fasn, Cyp4a8, Cpt1b and Ppard) were significantly decreased in the TPT-treated group. In summary, exposure to TPT may impair renal structure and function in rats by affecting RAS, AQPs, and lipid metabolism.

Bio-inspired High-Performance Artificial Ion Pump Mediated by Subnanoscale Dehydration Hydration Effects.

The energy conversion in plant chloroplast is carried out by pumping protons into the thylakoid for driving ATP synthesis. Inspired by ion active transport in living organisms, we attempted to design an artificial ion pump induced by subnanoconfinement effects. This ionic device uses two polarity functional nanoporous films as ion-selective valves at both ends and UiO-66 metal-organic framework-filled microchannels as ion storage cavities. In the charging process, ions could be pumped into the central cavities by nanovalves, which produced an ion gradient 10 to 100 times higher than the bulk, and were trapped within the subnanocages by dehydration. In the discharging process, the enriched ions were rehydrated and slowly released by the surface charge of the nanovalves, producing a sustainable ion current. The ion storage efficiency of this nanofluidic device could be improved to 60.3%, and the release time of ion current was also prolonged by 1 order of magnitude. This work combines the active and passive transport of ions to realize fast storage and slow release of ionic current, which provides an ion gradient-mediated novel energy conversion strategy.

Chloride Intracellular Channel Protein 1 (CLIC1) Is a Critical Host Cellular Factor for Influenza A Virus Replication.

(1) Background: Influenza A Virus (IAV) uses host cellular proteins during replication in host cells. IAV infection causes elevated expression of chloride intracellular channel protein 1 (CLIC1) in lung epithelial cells, but the importance of this protein in IAV replication is unknown. (2) In this study, we determined the role of CLIC1 in IAV replication by investigating the effects of CLIC1 knockdown (KD) on IAV viral protein translation, genomic RNA transcription, and host cellular proteome dysregulation. (3) Results: CLIC1 KD in A549 human lung epithelial cells resulted in a significant decrease in progeny supernatant IAV, but virus protein expression was unaffected. However, a significantly larger number of viral RNAs accumulated in CLIC1 KD cells. Treatment with a CLIC1 inhibitor also caused a significant reduction in IAV replication, suggesting that CLIC1 is an important host factor in IAV replication. SomaScan®, which measures 1322 proteins, identified IAV-induced dysregulated proteins in wild-type cells and in CLIC1 KD cells. The expression of 116 and 149 proteins was significantly altered in wild-type and in CLIC1 KD cells, respectively. A large number of the dysregulated proteins in CLIC1 KD cells were associated with cellular transcription and predicted to be inhibited during IAV replication. (4) Conclusions: This study suggests that CLIC1 is involved in later stages of IAV replication. Further investigation should clarify mechanism(s) for the development of anti-IAV drugs targeting CLIC1 protein.

Functional characterization of inactivating ABCC8 variants causing congenital hyperinsulinism.

Congenital hyperinsulinism (CHI; OMIM: 256450) is characterized by persistent insulin secretion despite severe hypoglycemia. The most common causes are variants in the ATP-binding cassette subfamily C member 8(ABCC8) and potassium inwardly-rectifying channel subfamily J member 11(KCNJ11) genes. These encode ATP-sensitive potassium (KATP) channel subunit sulfonylurea receptor 1 (SUR1) and inwardly rectifying potassium channel (Kir6.2) proteins. A 7-day-old male infant presented with frequent hypoglycemic episodes and was clinically diagnosed with CHI, underwent trio-whole-exome sequencing, revealing compound heterozygous ABCC8 variants (c.307C>T, p.His103Tyr; and c.3313_3315del, p.Ile1105del) were identified. In human embryonic kidney 293 (HEK293) and rat insulinoma cells (INS-1) transfected with wild-type and variant plasmids, KATP channels formed by p.His103Tyr were delivered to the plasma membrane, whereas p.Ile1105del or double variants (p.His103Tyr coupled with p.Ile1105del) failed to be transported to the plasma membrane. Compared to wild-type channels, the channels formed by the variants (p.His103Tyr; p.Ile1105del) had elevated basal [Ca2+]i, but did not respond to stimulation by glucose. Our results provide evidence that the two ABCC8 variants may be related to CHI owing to defective trafficking and dysfunction of KATP channels.

Thermodynamics and S-Palmitoylation Dependence of Interactions between Human Aquaporin-4 M1 Tetramers in Model Membranes.

Aquaporin-4 (AQP4) is a water channel protein found primarily in the central nervous system (CNS) that helps to regulate water-ion homeostasis. AQP4 exists in two major isoforms: M1 and M23. While both isoforms have a homotetrameric quaternary structure and are functionally identical when transporting water, the M23 isoform forms large protein aggregates known as orthogonal arrays of particles (OAPs). In contrast, the M1 isoform creates a peripheral layer around the outside of these OAPs, suggesting a thermodynamically stable interaction between the two. Structurally, the M1 isoform has an N-terminal tail that is 22 amino acids longer than the M23 isoform and contains two solvent-accessible cysteines available for S-palmitoylation at cysteine-13 (Cys-13) and cysteine-17 (Cys-17) in the amino acid sequence. Earlier work suggests that the palmitoylation of these cysteines might aid in regulating AQP4 assemblies. This work discusses the thermodynamic driving forces for M1 protein-protein interactions and how the palmitoylation state of M1 affects them. Using temperature-dependent single-particle tracking, the standard state free energies, enthalpies, and entropies were measured for these interactions. Furthermore, we present a binding model based on measured thermodynamics and a structural modeling study. The results of this study demonstrate that the M1 isoform will associate with itself according to the following expressions: 2[AQP4-M1]4 ↔ [[AQP4-M1]4]2 when palmitoylated and 3[AQP4-M1]4 ↔ [AQP4-M1]4 + [[AQP4-M1]4]2 ↔ [[AQP4-M1]4]3 when depalmitoylated. This is primarily due to a conformational change induced by adding the palmitic acid groups at Cys-13 and Cys-17 in the N-terminal tails of the homotetramers. In addition, a statistical mechanical model was developed to estimate the Gibbs free energy, enthalpy, and entropy for forming dimers and trimers. These results were in good agreement with experimental values.

Calcium-sensing receptor and NF-κB pathways in TN breast cancer contribute to cancer-induced cardiomyocyte damage via activating neutrophil extracellular traps formation.

Cardiovascular disorders are commonly prevalent in cancer patients, yet the mechanistic link between them remains poorly understood. Because neutrophil extracellular traps (NETs) have implications not just in cardiovascular diseases (CVD), but also in breast cancer (BC), it was hypothesized to contribute to CVD in the context of oncogenesis. We established a mouse model using nude mice to simulate liver metastasis of triple-negative BC (TNBC) through the injection of MDA-MB-231 cells. Multiple imaging and analysis techniques were employed to assess the cardiac function and structure, including echocardiography, HE staining, Masson staining, and transmission electron microscopy (TEM). MDA-MB-231 cells underwent treatment with a CaSR inhibitor, CaSR agonist, and NF-κB channel blocker. The phosphorylation of NF-κB channel protein p65 and the expression and secretion of IL-8 were assessed using qRT-PCR, Western Blot, and ELISA, respectively. In addition, MDA-MB-231 cells were co-cultured with polymorphonuclear neutrophils (PMN) under varying conditions. The co-localization of PMN extracellular myeloperoxidase (MPO) and DNA were observed by cellular immunofluorescence staining to identify the formation of NETs. Then, the cardiomyocytes were co-cultured with the above medium that contains NETs or not, respectively; the effects of NETs on cardiomyocytes apoptosis were perceived by flow cytometry. The ultrastructural changes of myocardial cells were perceived by TEM, and ELISA detected the levels of myocardial enzyme (LDH, MDA and SOD). Overall, according to our research, CaSR has been found to have a regulatory role in IL-8 secretion in MDA-MB-231 cells, as well as in the formation of NETs by PMN cells. These findings suggest CaSR-mediated stimulation in PMN can lead to increased NETs formation and subsequently to cytotoxicity in cardiomyocytes, which potentially via activation of the NF-κB signaling cascade of BC cell.

The low-affinity calcium channel protein AaFig1 is essential for the growth and development, infection structure differentiation, and pathogenicity of Alternaria alternata

Fig1, as a low-affinity Ca2+ channel, is needed for Ca2+ influx and normal mating found in a few species, but its detailed molecular regulating mechanism in fungi, especially in Alternaria alternata, the causal agent of pear black spot, is still unclear. In this study, Fig1 in A. alternata was cloned, identified and named as AaFig1, and its function was characterized by targeted gene knockout strategy. The results showed that the ΔAaFig1 was severely decreased in vegetative growth, biomass accumulation and sporulation. Fluo-4, AM staining indicated that the Ca2+ content in ΔAaFig1 was decreased. Deletion of AaFig1 reduced the response of A. alternata to exogenous Ca2+ but improved the tolerance to osmotic stresses. Interestingly, the appressorium formation of the ΔAaFig1 strain on hydrophobic and pear wax extract coated surface, and infection hyphae formation on the onion epidermis were significantly reduced, meanwhile, the ΔAaFig1 strain could not penetrate the cellophane. In addition, virulence on wound-inoculated pear fruit and intact pear leaves, melanin production and the activities of cell wall degrading enzymes in AaFig1 strain were reduced. In summary, these results indicate that AaFig1-mediated Ca2+ homeostasis positively regulates the growth and development, infection structure formation and pathogenicity of A. alternata. The findings will provide a basis for developing new strategies to control this pathogen in pear fruit.