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G-protein coupled receptors inhibit Cav2.2 N-type Ca2+ channels by a fast, voltage-dependent pathway mediated by Gαi/Gβγ and a slow, voltage-independent pathway mediated by Gαq-dependent reductions in phosphatidylinositol 4,5-bisphosphate (PIP2) or increases in arachidonic acid. Studies of these forms of regulation generally employ Ba2+ as the permeant ion, despite that Ca2+ -dependent pathways may impinge upon G-protein modulation. To address this possibility, we compared tonic G-protein inhibition of currents carried by Ba2+ (IBa) and Ca2+ (ICa) in HEK293T cells transfected with Cav2.2. Both IBa and ICa exhibited voltage-dependent facilitation (VDF), consistent with Gβγ unbinding from the channel. Compared to that for IBa, VDF of ICa was less sensitive to an inhibitor of Gα proteins (GDP-β-S) and an inhibitor of Gβγ (C-terminal construct of G-protein coupled receptor kinase 2). While insensitive to high intracellular Ca2+ buffering, VDF of ICa that remained in GDP-β-S was blunted by reductions in PIP2. We propose that when G-proteins are inhibited, Ca2+ influx through Cav2.2 promotes a form of VDF that involves PIP2. Our results highlight the complexity whereby Cav2.2 channels integrate G-protein signaling pathways, which may enrich the information encoding potential of chemical synapses in the nervous system., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Thomas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Waterborne infections caused by pathogenic microorganisms represent serious health risks for humans. Ternary zinc-tin oxide nanoparticles have great potential as a cost-effective, environmentally friendly, and efficient candidate for waterborne infections; however, their photocatalytic and antibacterial effects are quite limited due to insufficient visible light absorption and rapid electron-hole recombination. Herein, barium-doped zinc stannate (Ba@ZTO) nanoparticles were synthesized by the hydrothermal method and used for the first time not only as antibacterial agents to prevent the spread of the harmful bacteria S. aureus and E. coli but also as photocatalysts to degrade the organic pollutant rhodamine B. Unexpectedly, Ba 2+ ions exhibited compressive stress behavior instead of the predicted tensile stress when inserted into the ZTO crystal lattice, playing an active role in increasing oxygen vacancies within the crystal lattice and in the formation of hydroxyl radicals in the bulk solution and hydrogen peroxide (H 2 O 2 ) radicals, significantly improving the photocatalytic and antibacterial properties. Strain-induced defects created by the insertion of larger barium ions into the ZTO lattice promote the increase of shallow traps for boosting photocatalytic/disinfection properties while suppressing deep-level traps that encourage nonradiative recombination. In essence, defect and strain engineering opens a promising route to achieve high disinfection efficiency by inducing larger cation ions under visible light in oxide-based materials.

The selectivity filter of K+ channels catalyzes a rapid and highly selective transport of K+ while serving as a gate. To understand the control of this filter gate, we use the pore-only K+ channel KcvNTS in which gating is exclusively determined by the activity of the filter gate. It has been previously shown that a mutation at the C-terminus of the pore-helix (S42T) increases K+ permeability and introduces distinct voltage-dependent and K+-sensitive channel closures at depolarizing voltages. Here, we report that the latter are not generated by intrinsic conformational changes of the filter gate but by a voltage-dependent block caused by nanomolar trace contaminations of Ba2+ in the KCl solution. Channel closures can be alleviated by extreme positive voltages and they can be completely abolished by the high-affinity Ba2+ chelator 18C6TA. By contrast, the same channel closures can be augmented by adding Ba2+ at submicromolar concentrations to the cytosolic buffer. These data suggest that a conservative exchange of Ser for Thr in a crucial position of the filter gate increases the affinity of the filter for Ba2+ by >200-fold at positive voltages. While Ba2+ ions apparently remain only for a short time in the filter-binding sites of the WT channel before passing the pore, they remain much longer in the mutant channel. Our findings suggest that the dwell times of permeating and blocking ions in the filter-binding sites are tightly controlled by interactions between the pore-helix and the selectivity filter., (© 2024 Tewes et al.)

Purpose: To evaluate the effects of H 2 O 2 as an oxidant on the electroretinogram (ERG) in isolated rat retina., Methods: Retinas were isolated from rat eyes and perfused with a nutrient solution. ERGs were recorded every 3 min. Once the signal was at a steady state, H 2 O 2 was added to the perfusion solution., Results: H 2 O 2 caused instantaneous and transient changes in amplitudes and implicit times of the ERG, followed by changes in retinal survival curves. H 2 O 2 0.2 mM produced a rapid increase in b-wave amplitude, followed by a return to the initial value and a survival curve above the control (without H 2 O 2 ). A slight increase in a-wave was observed, followed by a decrease and a recovery above the control. The slow PIII decreased and then recovered to the initial value. H 2 O 2 0.6 mM induced a small increase in b-wave amplitude, followed by a rapid decrease without recovery. The a-wave and slow PIII decreased rapidly without recovery. The implicit times of the a-wave and b-wave increased moderately with a low dose of H 2 O 2 , whereas they significantly increased with a high dose. Whatever the dose, the slow PIII implicit time increased significantly, followed by a return to the initial value. Barium increased the a-wave and b-wave, and then H 2 O 2 reduced the two waves with a stronger effect on the a-wave. Aspartate and barium isolated the fast PIII, which decreased after H 2 O 2 application., Conclusions: H 2 O 2 affects retinal function as shown by ERGs in isolated rat retina. The response differs with the dose of H 2 O 2 , suggesting that mechanisms underlying the action at low doses might be different from those at high doses. Our results also suggest an effect of H 2 O 2 on ionic currents and/or neurotransmitter releases involved in the generation of the ERG and indicate a more pronounced effect on photoreceptors than on postsynaptic cells.

Infection of bone defects remains a challenging issue in clinical practice, resulting in various complications. The current clinical treatments include antibiotic therapy and surgical debridement, which can cause drug-resistance and potential postoperative complications. Therefore, there is an urgent need for an efficient treatment to sterilize and promote bone repair in situ. In this work, an ultrasound responsive selenium modified barium titanate nanoparticle (Se@BTO NP) was fabricated, which exhibited significant antibacterial and bone regeneration effects. Selenium nanoparticle (Se NP) was modified on the surface of barium titanate nanoparticle (BTO NP) to form heterostructure, which facilitated the second distribution of piezo-induced carriers under ultrasound (US) irradiation and improved the separation of electron-hole pairs. The Se@BTO NPs exhibited remarkable antibacterial efficiency with an antibacterial rate of 99.23 % against Staphylococcus aureus (S.aureus) and significantly promoted the osteogenic differentiation under ultrasound irradiation. The in vivo experiments exhibited that Se@BTO NPs successfully repaired the femoral condylar bone defects of rats infected by S.aureus, resulting in significant promotion of bone regeneration. Overall, this work provided an innovative strategy for the utilization of US responsive nanomaterials in efficient bacteria elimination and bone regeneration. STATEMENT OF SIGNIFICANCE: Infectious bone defects remain a challenging issue in clinical practice. Current antibiotic therapy and surgical debridement has numerous limitations such as drug-resistance and potential complications. Herein, we designed an innovative ultrasound responsive selenium modified barium titanate nanoparticle (Se@BTO NP) to achieve efficient non-invasive bacteria elimination and bone regeneration. In this work, Se@BTO nanoparticles can enhance the separation of electrons and holes, facilitate the transfer of free carriers due to the cooperative effect of ultrasound induced piezoelectric field and heterojunction construction, and thus exhibit remarkable antibacterial and osteogenesis effect. Overall, our study provided a promising strategy for the utilization of piezocatalytic nanomaterials in efficient antibacterial and bone regeneration., (Copyright © 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

During videofluoroscopic swallowing study (VFSS), barium sulfate (BaSO 4 ) is commonly added into food samples as a radiopaque contrast media for bolus visualization and examination. Accordingly, the consistency and flow behavior of barium stimuli can differ significantly from their non-barium counterparts. Such differences may have a subsequent impact on the validity of VFSS. Therefore, in this study, effects of barium sulfate on the shear and extensional rheological properties and IDDSI (International Dysphagia Diet Standardization Initiative) flow consistency of liquids prepared using various commercial thickening powders were investigated. Results showed that all barium stimuli exhibited shear thinning behavior but with significantly higher shear viscosity compared to the non-barium counterparts. A shift factor of viscosity at shear rate 50 s -1 with values in range of 1.21-1.73 could be used to describe the increase in the viscosity for samples thickened with gum-based thickeners. However, the change in the viscosity was not invariant for the stimuli prepared starch-based thickener. The addition of BaSO 4 had a negative impact on extensional properties of samples by demonstrating a faster filament rupture. The extent of impact on the decrease in filament breakup time was more pronounced in xanthan > guar gum ≈ tara gum-based thickeners. Based on the IDDSI flow test, no significant effect of BaSO 4 was found on the gum-based thickeners, whereas there was a marked effect in the starch-based sample. These results can be used beneficially to assist clinicians in the dysphagia diagnosis for matching rheological properties of the barium stimuli to enhance effectiveness dysphagia interventions., (© 2023 Wiley Periodicals LLC.)

3D-printed scaffolds are suitable for patient-specific implant preparation for bone regeneration in large-scale critical bone defects. In addition, these scaffolds should have mechanical and biological properties similar to those of natural bone tissue. In this study, 3D-printed barium-doped calcium silicate (BaCS)/poly-ε-caprolactone (PCL) composite scaffolds were fabricated as an alternative strategy for bone tissue engineering to achieve appropriate physicochemical characteristics and stimulate osteogenesis. Scaffolds containing 10% Ba (Ba10) showed optimal mechanical properties, preventing premature scaffold degradation during immersion while enabling ion release in a sustained manner to achieve the desired therapeutic goals. In addition, Wharton's jelly mesenchymal stem cells (WJMSCs) were used to assess biocompatibility and osteogenic differentiation behaviour. WJMSCs were cultured on the scaffold and permeabilised via ICP to analyse the presence of Si and Ba ions in the medium and cell lysates, suggesting that the ions released by the scaffold could effectively enter the cells. The protein expression of CaSR, PI3K, Akt, and JNK confirmed that CaSR could activate cells cultured in Ba10, thereby affecting the subsequent PI3k/Akt and JNK pathways and further promoting osteogenic differentiation. The in vivo performance of the proposed scaffolds was assessed using micro-CT and histological slices, which revealed that the BaCS scaffolds could further enhance bone regeneration, compared with bare scaffolds. These results suggest the potential use of 3D-printed BaCS/PCL scaffolds as next-generation substitutes for bone regeneration.

Ceftriaxone reduces gallbladder and ileal contractility. Many studies have shown that ceftriaxone causes biliary sludge and pseudolithiasis. However, its effect on intestinal transit time has not been investigated. This study aimed to investigate the effect of ceftriaxone on intestinal transit time. Sixteen rats were examined in two groups: The study group (group A, n = 8) was administered with 100 mg/kg ceftriaxone intramuscularly for 7 days. The control group (group B, n = 8) was administered with intramuscular distilled water for 7 days. On the seventh day, a mixture of 2 ml barium and saline was given orally to both groups. Barium transit was evaluated using serial digital X-ray images. The stomach was full and the transition into the small intestine loop was observed in all rats at 45 min in both groups. At the 2nd hour, colonic transition was observed in two rats in group A (2/8, 25%) and in seven rats in group B (7/8, 87.5%). At the 4th hour, five (62.5%) rats in group A had transverse colonic transition, and all rats in group B (8/8, 100%) had transverse and/or left colonic transition. At the 6th hour, no rat in group A had rectal transition, and all rats in group B (8/8, 100%) had complete passage of colonic contrast material. Ceftriaxone significantly prolongs the small intestine transit time, large intestine transit time, and total intestinal transit times., (© 2022 Société Française de Pharmacologie et de Thérapeutique. Published by John Wiley & Sons Ltd.)

Previous epidemiological studies have reported that prenatal exposure to metals might have influence on fetal growth. Most studies assessed the effect of individual metals, while the investigation on the relationship between multiple metal exposure and fetal growth is sparse. The objective of the present study is to assess the joint impact of metal mixtures on fetal growth during pregnancy. A total of 1275 maternal-infant pairs from the Jiangsu Birth Cohort (JBC) Study were included to investigate the effect of maternal metal exposure on fetal biometry measures at 22-24, 30-32, and 34-36 weeks of gestation. Lead (Pb), arsenic (As), cadmium (Cd), mercury (Hg), chromium (Cr), vanadium(V), thallium (Tl) and barium (Ba) were measured by inductively coupled plasma mass spectrometry (ICP-MS) in maternal urine samples collected in the first trimester. We used general linear models and restricted cubic splines to test dose-response relationships between single metals and fetal growth. The weighted quantile sum (WQS) models were then applied to evaluate the overall effect of all these metals. We observed inverse associations of exposure to Pb, V and Cr with estimated fetal weight (EFW) at 34-36 weeks of gestation. Notably, maternal exposure to metal mixtures was significantly associated with reduced EFW at 34-36 weeks of gestation after adjusting for some covariates and confounders (aβ -0.05 [95% CI: 0.09, -0.01], P = 0.023), and this association was mainly driven by Cr (30.41%), Pb (23.92%), and Tl (15.60%). These findings indicated that prenatal exposure to metal mixtures might impose adverse effects on fetal growth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Triple-negative breast cancer (TNBC) is an aggressive BC subtype with a higher metastatic rate and a worse 5-year survival ratio than the other BC. It is an urgent need to develop a noninvasive treatment with high efficiency to resist TNBC cell proliferation and invasion. Internal wireless electric stimulation (ES) based on piezoelectric materials is an emerging noninvasive strategy, with adjustable ES intensity and excellent biosafety. In this study, three different barium titanate nanoparticles (BTNPs) with different crystal phases and piezoelectric properties were studied. Varying intensities of internal ES were generated from the three BTNPs ( i.e ., BTO, U-BTO, P-BTO). In vitro tests revealed that the internal ES from BTNPs was efficient at reducing the proliferative potential of cancer cells, particularly BC cells. In vitro experiments on MDA-MB-231, a typical TNBC cell line, further revealed that the internal wireless ES from BTNPs significantly inhibited cell growth and migration up to about 82% and 60%, respectively. In vivo evaluation of MDA-MB-231 tumor-bearing mice indicated that internal ES not only resisted almost 70% tumor growth but also significantly inhibited lung metastasis. More importantly, in vitro and in vivo studies demonstrated a favorable correlation between the anticancer impact and the intensities of ES. The underlying mechanism of MDA-MB-231 cell proliferation and metastasis inhibition caused by internal ES was also investigated. In summary, our results revealed the effect and mechanism of internal ES from piezoelectric nanoparticles on TNBC cell proliferation and migration regulation and proposed a promising noninvasive therapeutic strategy for TNBC with minimal side effects while exhibiting good therapeutic efficiency.

Purpose: Biopiezoelectric materials have good biocompatibility and excellent piezoelectric properties, and they can generate local currents in vivo to restore the physiological electrical microenvironment of the defect and promote bone regeneration. Previous studies of guided bone regeneration membranes have rarely addressed the point of restoring it, so this study prepared a Barium titanate/Polylactic acid (BT/PLA) piezoelectric composite membrane and investigated its bone-formation, with a view to providing an experimental basis for clinical studies of guided bone tissue regeneration membranes., Methods: BT/PLA composite membranes with different BT ratio were prepared by solution casting method, and piezoelectric properties were performed after corona polarization treatment. The optimal BT ratio was selected and then subjected to in vitro cytological experiments and in vivo osteogenic studies in rats. The effects on adhesion, proliferation and osteogenic differentiation of the pre-osteoblastic cell line (MC3T3-E1) were investigated. The effect of composite membranes on bone repair of cranial defects in rats was investigated after 4 and 12 weeks., Results: The highest piezoelectric coefficient d33 were obtained when the BT content was 20%, reaching (7.03 ± 0.26) pC/N. The value could still be maintained at (4.47±0.17) pC/N after 12 weeks, meeting the piezoelectric constant range of bone. In vitro, the MC3T3-E1 cells showed better adhesion and proliferative activity in the group of polarized 20%BT. The highest alkaline phosphatase (ALP) content was observed in cells of this group. In vivo, it promoted rapid bone regeneration. At 4 weeks postoperatively, new bone formation was evident at the edges of the defect, with extensive marrow cavity formation; after 12 weeks, the defect was essentially completely closed, with density approximating normal bone tissue and significant mineralization., Conclusion: The BT/PLA piezoelectric composite membrane has good osteogenic properties and provides a new idea for guiding the research of membrane materials for bone tissue regeneration., Competing Interests: The authors report no conflicts of interest for this work., (© 2022 Dai et al.)

Trivalent europium-based monochromatic red light-emitting phosphors are an essential component to realize high-performance smart lighting devices; however, the concentration and thermal quenching restrict their usage. Here, we report a series of efficient Eu 3+ -substituted Li 3 Y 3 BaSr(MoO 4 ) 8 red-emitting phosphors based on a stratified scheelite structure with negligible concentration and thermal quenching. All of the host and phosphor compositions crystallize in monoclinic crystal structure (space group C 2/ c ). All of the phosphor compositions produce narrow-band red emission (FWHM ∼6 nm), which is highly apparent to the human eyes, and lead to exceptional chromatic saturation of the red spectral window. Concurrently, detailed investigations were carried out to comprehend the concentration and thermal quenching mechanism. Absolute quantum yields as high as 88.5% were obtained for Li 3 Y 0.3 Eu 2.7 BaSr(MoO 4 ) 8 phosphor with virtuous thermal stability (at 400 K, retaining 87% of its emission intensity). The light-emitting diodes were constructed by coupling Li 3 BaSrY 0.3 Eu 2.7 (MoO 4 ) 8 red phosphor with a near-UV LED chip (395 nm) operated at 20 mA forward bias, and the hybrid white LED (an organic yellow dye + red Li 3 Y 3 BaSr(MoO 4 ) 8 :Eu 3+ phosphor integrated with an NUV LED chip) showed a low CCT (6645 K), high CRI (83) values, and CIE values of x = 0.303; y = 0.368, which indicated that the synthesized phosphors can be a suitable red component for white LEDs. In addition, we have systematically investigated the Sm 3+ and Sm 3+ , Eu 3+ activation in Li 3 Y 3 BaSr(MoO 4 ) 8 to display the latent use of the system in plant growth applications and establish that the phosphor exhibits orange red emission with an intense deep-red emission (645 nm ( 4 G 5/2 → 6 H 9/2 )). The phytochrome (Pr) absorption spectrum well matched the fabricated deep-red LED (by integrating a NUV LED + Li 3 Y 3 BaSr(MoO 4 ) 8 :Sm 3+ and Eu 3+ phosphor) spectral lines.

Hydroxyapatite (HA) is the most commonly used orthopedic implant material. In recent years, the emergence of cationic doped hydroxyapatite has revealed more possibilities for the biological application of HA. Conventional HA does not promote new bone formation because of its poor osteoinductive activity, and has a similar density to that of bone, leading to difficulty in distinguishing both via imaging. Magnesium ions are useful for regulating the cellular behavior and promoting bone regeneration. Ba ion related compounds, such as BaSO 4 , have a strong X-ray shielding effect. In this study, Ba/Mg@HA was synthesized to prepare Ba/Mg@HA/PLGA composites, and we aimed to investigate if Ba/Mg@HA/PLGA composites enhanced bone repair on osteoblasts and tibial defects, as well as the X-ray and CT imaging ability of bone implants in rats. The in vitro experimental results showed that the Ba/Mg@HA/PLGA composites significantly improved the attachment and osteogenic differentiation of MC3T3-E1 cells. These include the promotion of mineral deposition, enhancement of alkaline phosphatase activity, upregulation of OCN and COL-1 gene expression, and increase in COL-1 and OCN protein expression in a time- and concentration-dependent manner. The in vivo experimental results showed that the Ba/Mg@HA/PLGA composites significantly increased the rate of bone defect healing and the expression of BMP-2 and COL-1 in the bones of rats. X-ray and CT imaging results showed that the Ba/Mg@HA/PLGA composites enhanced the X-ray imaging ability. These findings indicate that the Ba/Mg@HA/PLGA composites can effectively promote bone formation and improve the X-ray and CT imaging abilities to a certain extent.

The aim of this study was cost-effective and greener synthesis of barium carbonate (BaCO 3 or witherite) nanoparticles with economic importance, and to evaluate their therapeutic potentials and biocompatibility with immune cells. Barium carbonate nanoparticles were biosynthesized using black elderberry extract in one step with non-toxic precursors and simple laboratory conditions; their morphologies and specific structures were analyzed using field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX). The therapeutic capabilities of these nanoparticles on the immune cells of murine macrophages J774 and promastigotes Leishmania tropica were evaluated. BaCO 3 nanoparticles with IC 50  = 46.6 µg/mL were more effective than negative control and glucantium (positive control) in reducing promastigotes (P < 0.01). Additionally, these nanoparticles with a high value of cytotoxicity concentration 50% (CC 50 ) were less toxic to macrophage cells than glucantime; however, they were significantly different at high concentrations compared to the negative control., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Degeneracy, the ability of multiple structural components to elicit the same characteristic functional properties, constitutes an elegant mechanism for achieving biological robustness. In this study, we sought electrophysiological signatures for the expression of ion-channel degeneracy in the emergence of intrinsic properties of rat hippocampal granule cells. We measured the impact of four different ion-channel subtypes-hyperpolarization-activated cyclic-nucleotide-gated (HCN), barium-sensitive inward rectifier potassium (K ir ), tertiapin-Q-sensitive inward rectifier potassium, and persistent sodium (NaP) channels-on 21 functional measurements employing pharmacological agents, and report electrophysiological data on two characteristic signatures for the expression of ion-channel degeneracy in granule cells. First, the blockade of a specific ion-channel subtype altered several, but not all, functional measurements. Furthermore, any given functional measurement was altered by the blockade of many, but not all, ion-channel subtypes. Second, the impact of blocking each ion-channel subtype manifested neuron-to-neuron variability in the quantum of changes in the electrophysiological measurements. Specifically, we found that blocking HCN or Ba-sensitive K ir channels enhanced action potential firing rate, but blockade of NaP channels reduced firing rate of granule cells. Subthreshold measures of granule cell intrinsic excitability (input resistance, temporal summation, and impedance amplitude) were enhanced by blockade of HCN or Ba-sensitive K ir channels, but were not significantly altered by NaP channel blockade. We confirmed that the HCN and Ba-sensitive K ir channels independently altered sub- and suprathreshold properties of granule cells through sequential application of pharmacological agents that blocked these channels. Finally, we found that none of the sub- or suprathreshold measurements of granule cells were significantly altered upon treatment with tertiapin-Q. Together, the heterogeneous many-to-many mapping between ion channels and single-neuron intrinsic properties emphasizes the need to account for ion-channel degeneracy in cellular- and network-scale physiology., (© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Hypoxia in a solid tumor microenvironment (TME) can lead to the overexpression of hypoxia-inducible factor-1α (HIF-1α), which correlates to tumor metastasis. Reactive oxygen species (ROS) induced tumor cell apoptosis is becoming a promising method in tumor treatment. Currently, the ROS generating systems, e.g., photodynamic treatment and sonodynamic treatment, highly depend on oxygen (O 2 ) in the tumor microenvironment (TME). However, the level of O 2 in TME is too low to produce enough ROS. Herein, we developed an ultrasmall DSPE-PEG 2000 coated barium titanate nanoparticle (P-BTO) for tumor treatment based on ultrasound triggered piezocatalysis and water splitting. Interestingly, irradiated by ultrasound, the surface of ultasmall P-BTO nanoparticles produced imbalance charges, which induced a cascade of redox reaction processes to simultaneously generate ROS and O 2 , the latter one was hardly generated in large-sized barium titanate nanoparticles. The as-synthesized P-BTO reached the highest accumulation in the tumor site at 4 h after intravenous injection. The results showed that the produced O 2 significantly alleviated the hypoxia of TME to down-regulate the expression of HIF-1α, and the produced ROS can efficiently kill tumor cells. Moreover, the tumor metastasis was also inhibited, providing a different way to treat triple-negative breast cancer, which was easily metastatic and lacked effective treatments in the clinic.

Cell death is a vital event in life. Infections and injuries cause lytic cell death, which gives rise to danger signals that can further induce cell death, inflammation, and tissue damage. The mevalonate (MVA) pathway is an essential, highly conserved and dynamic metabolic pathway. Here, we discover that farnesyl pyrophosphate (FPP), a metabolic intermediate of the MVA pathway, functions as a newly identified danger signal to trigger acute cell death leading to neuron loss in stroke. Harboring both a hydrophobic 15-carbon isoprenyl chain and a heavily charged pyrophosphate head, FPP leads to acute cell death independent of its downstream metabolic pathways. Mechanistically, extracellular calcium influx and the cation channel transient receptor potential melastatin 2 (TRPM2) exhibit essential roles in FPP-induced cell death. FPP activates TRPM2 opening for ion influx. Furthermore, in terms of a mouse model constructing by middle cerebral artery occlusion (MCAO), FPP accumulates in the brain, which indicates the function of the FPP and TRPM2 danger signal axis in ischemic injury. Overall, our data have revealed a novel function of the MVA pathway intermediate metabolite FPP as a danger signal via transient receptor potential cation channels., Competing Interests: The authors have declared that no competing interests exist.

Spiral ganglion neurons (SGNs) are the primary afferent neurons of the auditory system, and together with their attendant glia, form the auditory nerve. Within the cochlea, satellite glial cells (SGCs) encapsulate the cell body of SGNs, whereas Schwann cells (SCs) wrap their peripherally- and centrally-directed neurites. Despite their likely importance in auditory nerve function and homeostasis, the physiological properties of auditory glial cells have evaded description. Here, we characterized the voltage-activated membrane currents of glial cells from the mouse cochlea. We identified a prominent weak inwardly rectifying current in SGCs within cochlear slice preparations (postnatal day P5-P6), which was also present in presumptive SGCs within dissociated cultures prepared from the cochleae of hearing mice (P14-P15). Pharmacological block by Ba 2+ and desipramine suggested that channels belonging to the Kir4 family mediated the weak inwardly rectifying current, and post hoc immunofluorescence implicated the involvement of Kir4.1 subunits. Additional electrophysiological profiles were identified for glial cells within dissociated cultures, suggesting that glial subtypes may have specific membrane properties to support distinct physiological roles. Immunofluorescence using fixed cochlear sections revealed that although Kir4.1 is restricted to SGCs after the onset of hearing, these channels are more widely distributed within the glial population earlier in postnatal development (i.e., within both SGCs and SCs). The decrease in Kir4.1 immunofluorescence during SC maturation was coincident with a reduction of Sox2 expression and advancing neurite myelination. The data suggest a diversification of glial properties occurs in preparation for sound-driven activity in the auditory nerve., (© 2020 The Authors. Journal of Neuroscience Research published by Wiley Periodicals LLC.)

Barium (Ba 2+ ) is a classic permeant blocker of potassium (K + ) channels. The "external lock-in effect" in barium block experiments, whereby the binding of external K + impedes the forward translocation of the blocker, provides a powerful avenue to investigate the selectivity of the binding sites along the pore of potassium channels. Barium block experiments show that the external lock-in site is highly selective for K + over Na + . Wild-type KcsA was crystallized in low K + conditions, and the crystals were soaked in solutions containing various concentrations of barium. Structural analysis reveals open and closed gate conformations of the KcsA channel. Anomalous diffraction experiments show that Ba 2+ primarily binds to the innermost site S4 of the selectivity filter of the open-gate conformation and also the site S2, but no binding is detected with the closed-gate conformation. Alchemical free-energy perturbation calculations indicate that the presence of a Ba 2+ ion in the selectivity filter boosts the specificity of K + binding relative to Na + in the external sites S0-S2., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Total joint replacement implants are generally designed to physically mimic the biological environment to ensure compatibility with the host tissue. However, implant instability exposes patients to long recovery periods, high risk for revision surgeries, and high expenses. Introducing electrical stimulation to the implant site to accelerate healing is promising, but the cumbersome nature of wired devices is detrimental to the implant design. We propose a novel strategy to stimulate cells at the implant site by utilizing piezoelectric ceramics as electrical stimulation sources. The inherent ability of these materials to form electric surface potentials under mechanical load allows them to act as internal power sources. This characteristic is commonly exploited in non-biomedical applications such as transducers or sensors. We investigate calcium/zirconium-doped barium titanate (BCZT) ceramics in an in vitro environment to determine their potential as implant materials. BCZT exhibits low cytotoxicity with human osteoblast and endothelial cells as well as high piezoelectric responses. Microstructural adaptation was identified as a route for optimizing piezoelectric behavior. Our results show that BCZT is a promising system for biomedical applications. Its characteristic ability to autonomously generate electric surface potentials opens the possibility to functionalize existing bone replacement implant designs to improve implant ingrowth and long-term stability., (© 2019 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals, Inc.)

DNA can form many structures beyond the canonical Watson-Crick double helix. It is now clear that noncanonical structures are present in genomic DNA and have biological functions. G-rich G-quadruplexes and C-rich i-motifs are the most well-characterized noncanonical DNA motifs that have been detected in vivo with either proscribed or postulated biological roles. Because of their independent sequence requirements, these structures have largely been considered distinct types of quadruplexes. Here, we describe the crystal structure of the DNA oligonucleotide, d(CCAGGCTGCAA), that self-associates to form a quadruplex structure containing two central antiparallel G-tetrads and six i-motif C-C+ base pairs. Solution studies suggest a robust structural motif capable of assembling as a tetramer of individual strands or as a dimer when composed of tandem repeats. This hybrid structure highlights the growing structural diversity of DNA and suggests that biological systems may harbor many functionally important non-duplex structures., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)

We examined mechanisms of ammonia handling in the anterior, mid, and posterior intestine of unfed and fed freshwater rainbow trout (Oncorhynchus mykiss), with a focus on the Na + :K + :2Cl - co-transporter (NKCC), Na + :K + -ATPase (NKA), and K + channels. NKCC was localized by immunohistochemistry to the mucosal (apical) surface of enterocytes, and NKCC mRNA was upregulated after feeding in the anterior and posterior segments. NH 4 + was equally potent to K + in supporting NKA activity in all intestinal sections. In vitro gut sac preparations were employed to examine mucosal ammonia flux rates (Jm amm , disappearance from the mucosal saline), serosal ammonia flux rates (Js amm , appearance in the serosal saline), and total tissue ammonia production rates (Jt amm  = Js amm - Jm amm ). Bumetanide (10 -4 mol L -1 ), a blocker of NKCC, inhibited Js amm in most preparations, but this was largely due to reduction of Jt amm ; Jm amm was significantly inhibited only in the anterior intestine of fed animals. Ouabain (10 -4 mol L -1 ), a blocker of NKA, generally reduced both Jm amm and Js amm without effects on Jt amm in most preparations, though the anterior intestine was resistant after feeding. Barium (10 -2 mol L -1 ), a blocker of K + channels, inhibited Jm amm in most preparations, and Js amm in some, without effects on Jt amm . These pharmacological results, together with responses to manipulations of serosal and mucosal Na + and K + concentrations, suggest that NKCC is not as important in ammonia absorption as previously believed. NH 4 + appears to be taken up through barium-sensitive K + channels on the mucosal surface. Mucosal NH 4 + uptake via both NKCC and K + channels is energized by basolateral NKA, which plays an additional role in scavenging NH 4 + on the serosal surface to possibly minimize blood toxicity or enhance ion uptake and amino acid synthesis following feeding. Together with recent findings from other studies, we have provided an updated model to describe the current understanding of intestinal ammonia transport in teleost fish.

CaO-P 2 O 5 -SiO 2 -BaO-Fe 2 O 3 magnetic bioactive glasses were prepared via an optimized sol-gel method. This study is focused on investigating effects of magnetic content addition on the bioactive glass properties. To this aim, we evaluate the physical, rheological, and biocompatibility properties of synthesized magnetic bioactive glass. The morphology and composition of these glasses were studied using X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). The particle size was also determined using Laser Particle Size Analyzer (LPSA). The thermal measurements were carried out using Differential Thermal Analysis (DTA). For assessing the in-vitro bioactive character of synthesized glasses, the ability for apatite formation on their surface upon immersion in simulated body fluid (SBF) was checked using SEM, EDX and pH measurements. Furthermore, the Ca, Si, Ba and Fe ions in SBF were monitored using Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). The results showed that the addition of Ba and Fe in the glass composition affect formation of apatite layer onto the glass surfaces. Morphologies of the apatite layers were also different in which the bioactivity decreased with increasing Fe concentration, but the increase of Ba concentration led to an increase in bioactivity. However all of the synthesized glasses are still highly bioactive. Finally, this research demonstrates that the synthesized magnetic bioactive glasses are nontoxic and biocompatible and they can be used as thermoseeds for cancer hyperthermia studies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In the rodent cerebral circulation, inward rectifying K + (K IR ) channels set resting tone and the distance over which electrical phenomena spread along the arterial wall. The present study sought to translate these observations into human cerebral arteries obtained from resected brain tissue. Computational modeling and a conduction assay first defined the impact of K IR channels on electrical communication; patch-clamp electrophysiology, quantitative PCR, and immunohistochemistry then characterized K IR 2.x channel expression/activity. In keeping with rodent observations, computer modeling highlighted that K IR blockade should constrict cerebral arteries and attenuate electrical communication if functionally expressed. Surprisingly, Ba 2+ (a K IR channel inhibitor) had no effect on human cerebral arterial tone or intercellular conduction. In alignment with these observations, immunohistochemistry and patch-clamp electrophysiology revealed minimal K IR channel expression/activity in both smooth muscle and endothelial cells. This absence may be reflective of chronic stress as dysphormic neurons, leukocyte infiltrate, and glial fibrillary acidic protein expression was notable in the epileptic cortex. In closing, K IR 2.x channel expression is limited in human cerebral arteries from patients with epilepsy and thus has little impact on resting tone or the spread of vasomotor responses. NEW & NOTEWORTHY K IR 2.x channels are expressed in rodent cerebral arterial smooth muscle and endothelial cells. As they are critical to setting membrane potential and the distance signals conduct, we sought to translate this work into humans. Surprisingly, K IR 2.x channel activity/expression was limited in human cerebral arteries, a paucity tied to chronic brain stress in the epileptic cortex. Without substantive expression, K IR 2.x channels were unable to govern arterial tone or conduction.

A subset of neurons in the retrotrapezoid nucleus (RTN) function as respiratory chemoreceptors by regulating depth and frequency of breathing in response to changes in tissue CO 2 /H + . The activity of chemosensitive RTN neurons is also subject to modulation by CO 2 /H + -dependent purinergic signaling. However, mechanisms contributing to purinergic regulation of RTN chemoreceptors are not entirely clear. Recent evidence suggests adenosine inhibits RTN chemoreception in vivo by activation of A1 receptors. The goal of this study was to characterize effects of adenosine on chemosensitive RTN neurons and identify intrinsic and synaptic mechanisms underlying this response. Cell-attached recordings from RTN chemoreceptors in slices from rat or wild-type mouse pups (mixed sex) show that exposure to adenosine (1 µM) inhibits chemoreceptor activity by an A1 receptor-dependent mechanism. However, exposure to a selective A1 receptor antagonist (8-cyclopentyl-1,3-dipropylxanthine, DPCPX; 30 nM) alone did not potentiate CO 2 /H + -stimulated activity, suggesting activation of A1 receptors does not limit chemoreceptor activity under these reduced conditions. Whole-cell voltage-clamp from chemosensitive RTN neurons shows that exposure to adenosine activated an inward rectifying K + conductance, and at the network level, adenosine preferentially decreased frequency of EPSCs but not IPSCs. These results show that adenosine activation of A1 receptors inhibits chemosensitive RTN neurons by direct activation of a G-protein-regulated inward-rectifier K + (GIRK)-like conductance, and presynaptically, by suppression of excitatory synaptic input to chemoreceptors.

Peptic ulcer is prevalent in about 4% of the world population and nearly 10% of people have been affected by peptic ulcer at some point in their life. Therefore, there is a need for newer efficient and safe anti-ulcer agents. In the present strategy, we have prepared a novel bioactive glass containing 1.3 mol% of barium oxide (BaBG) and evaluated its antiulcer potential in gastroduodenal ulcer models. Prophylactic effect of BaBG pretreatment was evaluated for 5 days in ethanol, aspirin and pyloric ligation-induced gastric ulcer and cysteamine-induced duodenal ulcer models. Repeated treatment of 10 days of BaBG was evaluated in the healing ulcer model of acetic acid. BaBG significantly reduced the ulcerative damage against all the five tested ulcer models. Scanning electron microscope (SEM) images have shown that BaBG forms a physical protective barrier over the gastro-duodenal epithelium cell. In the pyloric-ligation, ethanol and aspirin models, BaBG showed significantly increased in gastric pH, indicating antacid like activity. BaBG treatment significantly increased cell proliferation in the pyloric model. Thus, BaBG mediates antiulcer action by forming a protective physical barrier against harsh luminal factors, acid neutralization and cell proliferation., (Copyright © 2018 Elsevier B.V. All rights reserved.)

At present, beneficial effects of melanin and harmful effects of barium have been reported. However, little is known about the adsorption of barium, and even less is known about the biological significance of adsorption of barium by melanin. In this study, we showed that there was a strong correlation between the digitalized level of skin pigmentation and barium level in murine skin compared to the correlations between skin pigmentation level and levels of homologous elements of barium (magnesium, calcium and strontium). The concentration of subcutaneously injected barium in skin with a high level of pigmentation was higher than that in skin with a low level of pigmentation. Our cell-free experiment using the Langmuir isotherm for adsorption of barium in synthetic melanin also provided direct evidence of adsorption of barium by melanin. We then investigated the biological significance of melanin-mediated barium adsorption. We found barium-mediated increase in transforming activity in pigmented melanocytes (melan-a) but not in unpigmented melanocytes (melan-c) after confirming that the barium level in melan-a melanocytes was 3.4-fold higher than that in melan-c melanocytes after culture of 5 μM barium for 24 h. Taken together, our results not only indicate adsorption of barium by melanin in mice, cells and cell-free systems but also suggest a disadvantageous effect of adsorption of barium by melanin on transforming activity in cultured cells., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Inward rectifier K + (Kir) channels contribute to a variety of physiological processes in insects and are emerging targets for insecticide development. Previous studies on insect Kir channels have primarily focused on dipteran species (e.g., mosquitoes, fruit flies). Here we identify and functionally characterize Kir channel subunits in a hemipteran insect, the soybean aphid Aphis glycines, which is an economically important insect pest and vector of soybeans. From the transcriptome and genome of Ap. glycines we identified two cDNAs, ApKir1 and ApKir2, encoding Kir subunits that were orthologs of insect Kir1 and Kir2, respectively. Notably, a gene encoding a Kir3 subunit was absent from the transcriptome and genome of Ap. glycines, similar to the pea aphid Acyrthosiphon pisum. Heterologous expression of ApKir1 and ApKir2 in Xenopus laevis oocytes enhanced K + -currents in the plasma membrane; these currents were inhibited by barium and the small molecule VU041. Compared to ApKir2, ApKir1 mediated currents that were larger in magnitude, more sensitive to barium, and less inhibited by small molecule VU041. Moreover, ApKir1 exhibited stronger inward rectification compared to ApKir2. Topical application of VU041 in adult aphids resulted in dose-dependent mortality within 24 h that was more efficacious than flonicamid, an established insecticide also known to inhibit Kir channels. We conclude that despite the apparent loss of Kir3 genes in aphid evolution, Kir channels are important to aphid survival and represent a promising target for the development of new insecticides., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Three divalent cations can elicit secretory responses in most neuroendocrine cells, including chromaffin cells. The extent to which secretion is elicited by the cations in intact depolarized cells was Ba 2+  > Sr 2+  ≥ Ca 2+ , contrasting with that elicited by these cations in permeabilized cells (Ca 2+  > Sr 2+  > Ba 2+ ). Current-clamp recordings show that extracellular Sr 2+ and Ba 2+ cause membrane depolarization and action potentials, which are not blocked by Cd 2+ but that can be mimicked by tetra-ethyl-ammonium. When applied intracellularly, only Ba 2+ provokes action potentials. Voltage-clamp monitoring of Ca 2+ -activated K + channels (K Ca ) shows that Ba 2+ reduces outward currents, which were enhanced by Sr 2+ . Extracellular Ba 2+ increases cytosolic Ca 2+ concentrations in Fura-2-loaded intact cells, and it induces long-lasting catecholamine release. Conversely, amperometric recordings of permeabilized cells show that Ca 2+ promotes the longest lasting secretion, as Ba 2+ only provokes secretion while it is present and Sr 2+ induces intermediate-lasting secretion. Intracellular Ba 2+ dialysis provokes exocytosis at concentrations 100-fold higher than those of Ca 2+ , whereas Sr 2+ exhibits an intermediate sensitivity. These results are compatible with the following sequence of events: Ba 2+ blocks K Ca channels from both the outside and inside of the cell, causing membrane depolarization that, in turn, opens voltage-sensitive Ca 2+ channels and favors the entry of Ca 2+ and Ba 2+ . Although Ca 2+ is less permeable through its own channels, it is more efficient in triggering exocytosis. Strontium possesses both an intermediate permeability and an intermediate ability to induce secretion.

Perceptual decisions require the accumulation of sensory information to a response criterion. Most accounts of how the brain performs this process of temporal integration have focused on evolving patterns of spiking activity. We report that subthreshold changes in membrane voltage can represent accumulating evidence before a choice. αβ core Kenyon cells (αβ c KCs) in the mushroom bodies of fruit flies integrate odor-evoked synaptic inputs to action potential threshold at timescales matching the speed of olfactory discrimination. The forkhead box P transcription factor (FoxP) sets neuronal integration and behavioral decision times by controlling the abundance of the voltage-gated potassium channel Shal (K V 4) in αβ c KC dendrites. αβ c KCs thus tailor, through a particular constellation of biophysical properties, the generic process of synaptic integration to the demands of sequential sampling., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Self-referencing H + -selective electrodes were used to measure extracellular H + fluxes from Müller (glial) cells isolated from the tiger salamander retina. A novel chamber enabled stable recordings using H + -selective microelectrodes in a self-referencing format using bicarbonate-based buffer solutions. A small basal H + flux was observed from the end foot region of quiescent cells bathed in 24 mM bicarbonate-based solutions, and increasing extracellular potassium induced a dose-dependent increase in H + flux. Barium at 6 mM also increased H + flux. Potassium-induced extracellular acidifications were abolished when bicarbonate was replaced by 1 mM HEPES. The carbonic anhydrase antagonist benzolamide potentiated the potassium-induced extracellular acidification, while 300 μM DIDS, 300 μM SITS, and 30 μM S0859 significantly reduced the response. Potassium-induced extracellular acidifications persisted in solutions lacking extracellular calcium, although potassium-induced changes in intracellular calcium monitored with Oregon Green were abolished. Exchange of external sodium with choline also eliminated the potassium-induced extracellular acidification. Removal of extracellular sodium by itself induced a transient alkalinization, and replacement of sodium induced a transient acidification, both of which were blocked by 300 μM DIDS. Recordings at the apical portion of the cell showed smaller potassium-induced extracellular H + fluxes, and removal of the end foot region further decreased the H + flux, suggesting that the end foot was the major source of acidifications. These studies demonstrate that self-referencing H + -selective electrodes can be used to monitor H + fluxes from retinal Müller cells in bicarbonate-based solutions and confirm the presence of a sodium-coupled bicarbonate transporter, the activity of which is largely restricted to the end foot of the cell. NEW & NOTEWORTHY The present study uses self-referencing H + -selective electrodes for the first time to measure H + fluxes from Müller (glial) cells isolated from tiger salamander retina. These studies demonstrate bicarbonate transport as a potent regulator of extracellular levels of acidity around Müller cells and point toward a need for further studies aimed at addressing how such glial cell pH regulatory mechanisms may shape neuronal signaling., (Copyright © 2017 the American Physiological Society.)

Aims: The vasorelaxant effects of the anti-diabetic drug, mitiglinide in phenylephrine (Phe)-pre-contracted aortic rings were examined., Materials and Methods: Arterial tone measurement was performed in aortic smooth muscle cells., Key Findings: Mitiglinide dose-dependently induced vasorelaxation. Application of the large-conductance Ca 2+ -activated K + (BK Ca ) channel blocker paxilline, inwardly rectifying K + (Kir) channel blocker Ba 2+ , and ATP-sensitive K + (K ATP ) channel blocker glibenclamide did not affect the vasorelaxant effect of mitiglinide. However, application of the voltage-dependent K + (Kv) channel blocker 4-AP, effectively inhibited mitiglinide-induced vasorelaxation. Although pretreatment with the Ca 2+ channel blocker nifedipine did not alter the mitiglinide-induced vasorelaxation, pretreatment with the sarcoplasmic/endoplasmic reticulum Ca 2+ -ATPase (SERCA) pump inhibitor thapsigargin and cyclopiazonic acid reduced the vasorelaxant effect of mitiglinide. In addition, the vasorelaxant effect of mitiglinide was not affected by the inhibitors of adenylyl cyclase, protein kinase A, guanylyl cyclase, or protein kinase G. Elimination of the endothelium and inhibition of endothelium-dependent vasorelaxant mechanisms also did not change the vasorelaxant effect of mitiglinide., Significance: We proposed that mitiglinide induces vasorelaxation via activation of Kv channels and SERCA pump. However, the vasorelaxant effects of mitiglinide did not involve other K + channels, Ca 2+ channels, PKA/PKG signaling pathways, or the endothelium., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In this study, we examined an ammonium conductance in the mouse thick ascending limb cell line ST-1. Whole cell patch clamp was performed to measure currents evoked by NH 4 Cl in the presence of BaCl 2 , tetraethylammonium, and BAPTA Application of 20 mmol/L NH 4 Cl induced an inward current (-272 ± 79 pA, n  = 9). In current-voltage ( I - V ) relationships, NH 4 Cl application caused the I - V curve to shift down in an inward direction. The difference in current before and after NH 4 Cl application, which corresponds to the current evoked by NH 4 Cl, was progressively larger at more negative potentials. The reversal potential for NH 4 Cl was +15 mV, higher than the equilibrium potential for chloride, indicating that the current should be due to NH 4 + We then injected ST-1 poly(A) RNA into Xenopus oocytes and performed two-electrode voltage clamp. NH 4 Cl application in the presence of BaCl 2 caused the I - V curve to be steeper. The NH 4 + current was retained at pH 6.4, where endogenous oocyte current was abolished. The NH 4 + current was unaffected by 10  μ mol/L amiloride but abolished after incubation in Na + -free media. These results demonstrate that the renal cell line ST-1 produces an NH 4 + conductance., (© 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

Objective The videofluoroscopic swallow study (VFSS) is the gold standard diagnostic tool to evaluate oropharyngeal dysphagia. Although objective measurements on VFSS have been described, there is no universal method of analysis, and the majority of clinicians use subjective interpretation alone. The purpose of this investigation was to evaluate the accuracy of subjective VFSS analysis. Study Design Double-blinded experiment. Setting Tertiary care laryngology center. Subjects and Methods Seventy-six de-identified videos from VFSS evaluations of patients with dysphagia were presented to blinded, experienced speech-language pathologists and laryngologists individually. Evaluators rated each video as normal or abnormal for hyoid elevation (HE), pharyngeal area (PA), pharyngeal constriction ratio (PCR), and pharyngoesophageal segment opening (PESo). A blinded investigator assessed evaluators' inter- and intrarater agreement and compared their responses to objectively measured results for these parameters to examine accuracy. Results Evaluators correctly classified only 61.5% of VFSS videos as normal or abnormal, with moderate interrater agreement (κ = 0.48, P < .0001). Intrarater agreement was highly variable (κ = 0.43-0.83). Accuracy was greatest for PCR (71.6%), with poorer performance for HE (61.3%), PESo (59.2%), and PA (45.3%). Interrater agreement was moderate for all parameters, with greater concordance for PCR (κ = 0.59) and PESo (κ = 0.54) and less for HE (κ = 0.40) and PA (κ = 0.44). Evaluators unanimously agreed on a correct interpretation of a VFSS only 28% of the time. Conclusion Subjective assessment of VFSS parameters is inconsistently accurate when compared with objective measurements, with accuracy ratings ranging from 45.3% to 71.6% for specific parameters. Inter- and intrarater reliability for subjective assessment was moderate and highly variable.

ATP-sensitive inwardly rectifying potassium (K ATP ) channels couple cellular metabolism to the membrane potential of the cell and play an important role in a variety of tissue types, including the insect dorsal vessel, making them a subject of interest not only for understanding invertebrate physiology, but also as a potential target for novel insecticides. Most of what is known about these ion channels is the result of work performed in mammalian systems, with insect studies being limited to only a few species and physiological systems. The goal of this study was to investigate the role that K ATP channels play in regulating cardiac function in a model social insect, the honey bee (Apis mellifera), by examining the effects that modulators of these ion channels have on heart rate. Heart rate decreased in a concentration-dependent manner, relative to controls, with the application of the K ATP channel antagonist tolbutamide and K ATP channel blockers barium and magnesium, whereas heart rate increased with the application of a low concentration of the K ATP channel agonist pinacidil, but decreased at higher concentrations. Furthermore, pretreatment with barium magnified the effects of tolbutamide treatment and eliminated the effects of pinacidil treatment at select concentrations. The data presented here confirm a role for K ATP channels in the regulation of honey bee dorsal vessel contractions and provide insight into the underlying physiology that governs the regulation of bee cardiac function., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The endolymphatic sac (ES) is a cystic structure of the inner ear connected to the cochlea and vestibule, which plays a role in regulating ion homeostasis in inner ear fluid. Disruption of ion homeostasis can cause inner ear disorders with hearing loss and dizziness, such as Meniere's disease. Herein, we found, for the first time, functional evidence for the involvement of β 1 - and β 2 -adrenergic receptors in apical electrogenic ion transport by human ES epithelium by using electrophysiological/pharmacological and molecular biological methods, which were dependent on K + and Cl - ion transport. The apical electrogenic transport was absent or very weak in ES epithelia of patients with Meniere's disease. These results suggested that adrenergic stimulation via β 1 - and β 2 -adrenergic receptors in the human ES was involved in regulation of inner ear fluid ion homeostasis and impairment of this response could be a pathological mechanism of Meniere's disease., Competing Interests: The authors declare no competing financial interests.

Mammalian colonic epithelia consist of cells that are capable of both absorbing and secreting Cl-. The present studies employing Ussing chamber technique identified two opposing short-circuit current (Isc) responses to basolateral bumetanide in rat distal colon. Apart from the transepithelial Cl--secretory Isc in early distal colon that was inhibited by bumetanide, bumetanide also stimulated Isc in late distal colon that had not previously been identified. Since bumetanide inhibits basolateral Na+-K+-2Cl- cotransporter (NKCC) in crypt cells and basolateral K+-Cl- cotransporter (KCC) in surface epithelium, we proposed this stimulatory Isc could represent a KCC-mediated Cl- absorptive current. In support of this hypothesis, ion substitution experiments established Cl- dependency of this absorptive Isc and transport inhibitor studies demonstrated the involvement of an apical Cl- conductance. Current distribution and RNA sequencing analyses revealed that this Cl- absorptive Isc is closely associated with epithelial Na+ channel (ENaC) but is not dependent on ENaC activity. Thus, inhibition of ENaC by 10 μM amiloride or benzamil neither altered the direction nor its activity. Physiological studies suggested that this Cl- absorptive Isc senses dietary Cl- content; thus when dietary Cl- was low, Cl- absorptive Isc was up-regulated. In contrast, when dietary Cl- was increased, Cl- absorptive Isc was down-regulated. We conclude that an active Cl- extrusion mechanism exists in ENaC-expressing late distal colon and likely operates in parallel with ENaC to facilitate NaCl absorption., Competing Interests: The authors have declared that no competing interests exist.

Several pharmacophore models have been proposed for 5-HT2A serotonin receptor antagonists. These typically consist of two aromatic/hydrophobic moieties separated by a given distance from each other, and from a basic amine. Although specified distances might vary, the models are relatively similar in their general construction. Because our preliminary data indicated that two aromatic (hydrophobic) moieties might not be required for such action, we deconstructed the serotonin-dopamine antipsychotic agent risperidone (1) into four smaller structural fragments that were thoroughly examined in 5-HT2A receptor binding and functional (i.e., two-electrode voltage clamp (TEVC) and intracellular calcium release) assays. It was apparent that truncated risperidone analogues behaved as antagonists. In particular, 6-fluoro-3-(1-methylpiperidin-4-yl)benzisoxazole (4) displayed high affinity for 5-HT2A receptors (Ki of ca. 12 nM) relative to risperidone (Ki of ca. 5 nM) and behaved as a potent 5-HT2A serotonin receptor antagonist. These results suggest that multiple aromatic (hydrophobic) moieties are not essential for high-affinity 5-HT2A receptor binding and antagonist activity and that current pharmacophore models for such agents are very much in need of revision.

It has been reported that blockade of the inward rectifier K(+) current (IK1) facilitates termination of ventricular fibrillation. We hypothesized that partial IK1 blockade destabilizes spiral wave (SW) re-entry, leading to its termination. Optical action potential (AP) signals were recorded from left ventricles of Langendorff-perfused rabbit hearts with endocardial cryoablation. The dynamics of SW re-entry were analyzed during ventricular tachycardia (VT), induced by cross-field stimulation. Intercellular electrical coupling in the myocardial tissue was evaluated by the space constant. In separate experiments, AP recordings were made using the microelectrode technique from right ventricular papillary muscles of rabbit hearts. Ba(2+) (10-50 μM) caused a dose-dependent prolongation of VT cycle length and facilitated termination of VT in perfused hearts. Baseline VT was maintained by a stable rotor, where an SW rotated around an I-shaped functional block line (FBL). Ba(2+) at 10 μM prolonged I-shaped FBL and phase-singularity trajectory, whereas Ba(2+) at 50 μM transformed the SW rotation dynamics from a stable linear pattern to unstable circular/cycloidal meandering. The SW destabilization was not accompanied by SW breakup. Under constant pacing, Ba(2+) caused a dose-dependent prolongation of APs, and Ba(2+) at 50 μM decreased conduction velocity. In papillary muscles, Ba(2+) at 50 μM depolarized the resting membrane potential. The space constant was increased by 50 μM Ba(2+) Partial IK1 blockade destabilizes SW rotation dynamics through a combination of prolongation of the wave length, reduction of excitability, and enhancement of electrotonic interactions, which facilitates termination of ventricular tachyarrhythmias., (Copyright © 2016 the American Physiological Society.)

Unlabelled: Cortical spreading depression (CSD) is a propagating event of neuronal depolarization, which is considered as the cellular correlate of the migraine aura. It is characterized by a change in the intrinsic optical signal and by a negative DC potential shift. Microglia are the resident macrophages of the CNS and act as sensors for pathological changes. In the present study, we analyzed whether microglial cells might sense CSD by recording membrane currents from microglia in acutely isolated cortical mouse brain slices during an experimentally induced CSD. Coincident with the change in the intrinsic optical signal and the negative DC potential shift we recorded an increase in potassium conductance predominantly mediated by K(+) inward rectifier (Kir)2.1, which was blocked by the NMDA receptor antagonist D-AP5. Application of NMDA and an increase in extracellular K(+) mimics the CSD-induced Kir activation. Application of D-AP5, but not the purinergic receptor antagonist RB2, blocks the NMDA-induced Kir activation. The K(+) channel blocker Ba(2+) blocks both the CSD- and the NMDA-triggered increase in Kir channel activity. In addition, we could confirm previous findings that microglia in the adult brain do not express functional NMDA receptors by recording from microglia cultured from adult brain. From these observations we conclude that CSD activates neuronal NMDA receptors, which lead to an increase in extracellular [K(+)] resulting in the activation of Kir channel activity in microglia., Significance Statement: Cortical spreading depression (CSD) is a wave of neuronal depolarization spreading through the cortex and is associated with the aura of migraine. Here we show that microglial cells, which are viewed as pathologic sensors of the brain, can sense this wave. The increase in the extracellular potassium concentration associated with that wave leads to the activation of an inward rectifying potassium conductance in microglia. The involvement of neuronal NMDA receptors is crucial because NMDA mimics that response and microglia do not express functional NMDA receptors. Although it is now evident that CSD leads to a signal in microglia, the consequences of this microglial activation during CSD needs to be explored., (Copyright © 2016 the authors 0270-6474/16/366165-10$15.00/0.)

Key Message: A GORK homologue K(+) channel from the ancient desert shrub Ammopiptanthus mongolicus (Maxim.) Cheng f. shows the functional conservation of the GORK channels among plant species. Guard cell K(+) release through the outward potassium channels eventually enables the closure of stomata which consequently prevents plant water loss from severe transpiration. Early patch-clamp studies with the guard cells have revealed many details of such outward potassium currents. However, genes coding for these potassium-release channels have not been sufficiently characterized from species other than the model plant Arabidopsis thaliana. We report here the functional identification of a GORK (for Gated or Guard cell Outward Rectifying K(+) channels) homologue from the ancient desert shrub Ammopiptanthus mongolicus (Maxim.) Cheng f. AmGORK was primary expressed in shoots, where the transcripts were regulated by stress factors simulated by PEG, NaCl or ABA treatments. Patch-clamp measurements on isolated guard cell protoplasts revealed typical depolarization voltage gated outward K(+) currents sensitive to the extracelluar K(+) concentration and pH, resembling the fundamental properties previously described in other species. Two-electrode voltage-clamp analysis in Xenopus lavies oocytes with AmGORK reconstituted highly similar characteristics as assessed in the guard cells, supporting that the function of AmGORK is consistent with a crucial role in mediating stomatal closure in Ammopiptanthus mongolicus. Furthermore, a single amino acid mutation D297N of AmGORK eventually abolishes both the voltage-gating and its outward rectification and converts the channel into a leak-like channel, indicating strong involvement of this residue in the gating and voltage dependence of AmGORK. Our results obtained from this anciently originated plant support a strong functional conservation of the GORK channels among plant species and maybe also along the progress of revolution.

Green synthesis of nanoparticles is a growing research area because of their potential applications in nanomedicine. Barium carbonate nanoparticles (BaCO3 NPs) were synthesized using an aqueous extract of Mangifera indica seed as a reducing agent. These particles were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Transmission electron microscopy (TEM), selected area electron diffraction (SAED), Energy-dispersive-X-ray (EDX) and X-ray photoelectron spectroscopy (XPS) analysis. HR-TEM images are confirmed that green synthesized BaCO3 NPs have spherical, triangular and uneven shapes. EDX analysis confirmed the presence of Ba, C and O. The peaks at 2θ of 19.45, 23.90, 24.29, 27.72, 33.71, 34.08, 34.60, 41.98, 42.95, 44.18, 44.85, and 46.78 corresponding to (110), (111), (021), (002), (200), (112), (130), (221), (041), (202), (132) and (113) showed that BaCO3 NPs average size was ~18.3 nm. SAED pattern confirmed that BaCO3 NPs are crystalline nature. BaCO3 NPs significantly inhibited cervical carcinoma cells, as evidenced by cytotoxicity assay. Immunofluorescence and fluorescence assays showed that BaCO3 NPs increased the expression and activity of caspase-3, an autocatalytic enzyme that promotes apoptosis. According to the results, green synthesis route has great potential for easy, rapid, inexpensive, eco-friendly and efficient development of novel multifunctional nanoparticles for the treatment of cancer., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The activation of γ-aminobutyric acid receptor subtype B (GABAB) receptors in the midbrain ventrolateral periaqueductal gray (vlPAG) induces both postsynaptic and presynaptic inhibition. Whereas the postsynaptic inhibition is mediated by G protein-coupled inwardly rectifying K channels, the presynaptic inhibition of neurotransmitter release is primarily mediated by voltage-gated calcium channels. Using whole-cell recordings from acute rat PAG slices, we report here that the bath application of 4-aminopyridine, a voltage-gated K channel blocker, increases the initial GABA and glutamate release probability (P) and reinstates P depressed by presynaptic GABAB receptor activation at inhibitory and excitatory synapses, respectively. However, Ba, which blocks G protein-coupled inwardly rectifying K channels, does not produce similar effects. Our data suggest that the blockade of presynaptic 4-aminopyridine-sensitive K channels in vlPAG facilitates neurotransmitter release and reinstates synaptic transmission that has been altered by presynaptic GABAB receptor activation. Because vlPAG is involved in the descending pain control system, the present results may have potential therapeutic applications.

This study aimed to develop and evaluate barium and calcium microcapsules as candidates for scaffolding in artificial dermal papilla. Dermal papilla cells (DPCs) were isolated and cultured by one-step collagenase treatment. The DPC-Ba and DPC-Ca microcapsules were prepared by using a specially designed, high-voltage, electric-field droplet generator. Selected microcapsules were assessed for long-term inductive properties with xenotransplantation into Sprague-Dawley rat ears. Both barium and calcium microcapsules maintained xenogenic dermal papilla cells in an immunoisolated environment and induced the formation of hair follicle structures. Calcium microcapsules showed better biocompatibility, permeability, and cell viability in comparison with barium microcapsules. Before 18 weeks, calcium microcapsules gathered together, with no substantial immune response. After 32 weeks, some microcapsules were near inflammatory cells and wrapped with fiber. A few large hair follicles were found. Control samples showed no marked changes at the implantation site. Barium microcapsules were superior to calcium microcapsules in structural and mechanical stability. The cells encapsulated in hydrogel barium microcapsules exhibited higher short-term viability. This study established a model to culture DPCs in 3D culture conditions. Barium microcapsules may be useful in short-term transplantation study. Calcium microcapsules may provide an effective scaffold for the development of artificial dermal papilla.

Hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) of the plasma membrane by phospholipase C (PLC) generates two critical second messengers, inositol-1,4,5-trisphosphate and diacylglycerol. For the enzymatic reaction, PIP2 binds to positively charged amino acids in the pleckstrin homology domain of PLC. Here we tested the hypothesis that positively charged divalent and multivalent cations accumulate around the negatively charged PIP2, a process called electrostatic charge shielding, and therefore inhibit electrostatic PIP2-PLC interaction. This charge shielding of PIP2 was measured quantitatively with an in vitro enzyme assay using WH-15, a PIP2 analog, and various recombinant PLC proteins (β1, γ1, and δ1). Reduction of PLC activity by divalent cations, polyamines, and neomycin was well described by a theoretical model considering accumulation of cations around PIP2 via their electrostatic interaction and chemical binding. Finally, the charge shielding of PIP2 was also observed in live cells. Perfusion of the cations into cells via patch clamp pipette reduced PIP2 hydrolysis by PLC as triggered by M1 muscarinic receptors with a potency order of Mg2+ < spermine4+ < neomycin6+. Accumulation of divalent cations into cells through divalent-permeable TRPM7 channel had the same effect. Altogether our results suggest that Mg2+ and polyamines modulate the activity of PLCs by controlling the amount of free PIP2 available for the enzymes and that highly charged biomolecules can be inactivated by counterions electrostatically.

Electrophysiological properties and molecular background of the zebrafish (Danio rerio) cardiac inward rectifier current (IK1) were examined. Ventricular myocytes of zebrafish have a robust (-6.7 ± 1.2 pA pF(-1) at -120 mV) strongly rectifying and Ba(2+)-sensitive (IC50 = 3.8 μM) IK1. Transcripts of six Kir2 channels (drKir2.1a, drKir2.1b, drKir2.2a, drKir2.2b, drKir2.3, and drKir2.4) were expressed in the zebrafish heart. drKir2.4 and drKir2.2a were the dominant isoforms in both the ventricle (92.9 ± 1.5 and 6.3 ± 1.5%) and the atrium (28.9 ± 2.9 and 64.7 ± 3.0%). The remaining four channels comprised together less than 1 and 7 % of the total transcripts in ventricle and atrium, respectively. The four main gene products (drKir2.1a, drKir2.2a, drKir2.2b, drKir2.4) were cloned, sequenced, and expressed in HEK cells for electrophysiological characterization. drKir2.1a was the most weakly rectifying (passed more outward current) and drKir2.2b the most strongly rectifying (passed less outward current) channel, whilst drKir2.2a and drKir2.4 were intermediate between the two. In regard to sensitivity to Ba(2+) block, drKir2.4 was the most sensitive (IC50 = 1.8 μM) and drKir2.1a the least sensitive channel (IC50 = 132 μM). These findings indicate that the Kir2 isoform composition of the zebrafish heart markedly differs from that of mammalian hearts. Furthermore orthologous Kir2 channels (Kir2.1 and Kir2.4) of zebrafish and mammals show striking differences in Ba(2+)-sensitivity. Structural and functional differences needs to be taken into account when zebrafish is used as a model for human cardiac electrophysiology, cardiac diseases, and in screening cardioactive substances.

G protein-activated inwardly rectifying potassium (GIRK) channels in 5-HT neurons are assumed to be principal effectors of 5-hydroxytryptamine 1A (5-HT1A) autoreceptors, but their pharmacology, subunit composition and the role in regulation of 5-HT neuron activity have not been fully elucidated. We sought for a pharmacological tool for assessing the functional role of GIRK channels in 5-HT neurons by characterizing the effects of drugs known to block GIRK channels in the submicromolar range of concentrations. Whole-cell voltage-clamp recording in brainstem slices were used to determine concentration-response relationships for the selected GIRK channel blockers on 5-HT1A autoreceptor-activated inwardly rectifying K+ conductance in rat dorsal raphe 5-HT neurons. 5-HT1A autoreceptor-activated GIRK conductance was completely blocked by the nonselective inwardly rectifying potassium channels blocker Ba2+ (EC50 = 9.4 μM, full block with 100 μM) and by SCH23390 (EC50 = 1.95 μM, full block with 30 μM). GIRK-specific blocker tertiapin-Q blocked 5-HT1A autoreceptor-activated GIRK conductance with high potency (EC50 = 33.6 nM), but incompletely, i.e. ~16% of total conductance resulted to be tertiapin-Q-resistant. U73343 and SCH28080, reported to block GIRK channels with submicromolar EC50s, were essentially ineffective in 5-HT neurons. Our data show that inwardly rectifying K+ channels coupled to 5-HT1A autoreceptors display pharmacological properties generally expected for neuronal GIRK channels, but different from GIRK1-GIRK2 heteromers, the predominant form of brain GIRK channels. Distinct pharmacological properties of GIRK channels in 5-HT neurons should be explored for the development of new therapeutic agents for mood disorders.

Inwardly rectifying potassium channels play essential roles in renal physiology across phyla. Barium-sensitive K(+) conductances are found on the basolateral membrane of a variety of insect Malpighian (renal) tubules, including Drosophila melanogaster. We found that barium decreases the lumen-positive transepithelial potential difference in isolated perfused Drosophila tubules and decreases fluid secretion and transepithelial K(+) flux. In those insect species in which it has been studied, transcripts from multiple genes encoding inwardly rectifying K(+) channels are expressed in the renal (Malpighian) tubule. In Drosophila melanogaster, this includes transcripts of the Irk1, Irk2, and Irk3 genes. The role of each of these gene products in renal tubule function is unknown. We found that simultaneous knockdown of Irk1 and Irk2 in the principal cell of the fly tubule decreases transepithelial K(+) flux, with no additive effect of Irk3 knockdown, and decreases barium sensitivity of transepithelial K(+) flux by ∼50%. Knockdown of any of the three inwardly rectifying K(+) channels individually has no effect, nor does knocking down Irk3 simultaneously with Irk1 or Irk2. Irk1/Irk2 principal cell double-knockdown tubules remain sensitive to the kaliuretic effect of cAMP. Inhibition of the Na(+)/K(+)-ATPase with ouabain and Irk1/Irk2 double knockdown have additive effects on K(+) flux, and 75% of transepithelial K(+) transport is due to Irk1/Irk2 or ouabain-sensitive pathways. In conclusion, Irk1 and Irk2 play redundant roles in transepithelial ion transport in the Drosophila melanogaster renal tubule and are additive to Na(+)/K(+)-ATPase-dependent pathways., (Copyright © 2015 the American Physiological Society.)

Two neuropeptides, orexin-A and orexin-B (also called hypocretin-1 and -2), have been implicated in sleep/wake regulation, feeding behaviors via the activation of two subtypes of G-protein-coupled receptors: orexin 1 and orexin 2 receptors (OX1R and OX2R). While the expression of orexins and orexin receptors is immunohistochemically revealed in retinal neurons, the function of these peptides in the retina is largely unknown. Using whole-cell patch-clamp recordings in rat retinal slices, we demonstrated that orexin-A increased L-type-like barium currents (IBa,L) in ganglion cells (GCs), and the effect was blocked by the selective OX1R antagonist SB334867, but not by the OX2R antagonist TCS OX2 29. The orexin-A effect was abolished by intracellular dialysis of GDP-β-S/GPAnt-2A, a Gq protein inhibitor, suggesting the mediation of Gq. Additionally, during internal dialysis of the phosphatidylinositol (PI)-phospholipase C (PLC) inhibitor U73122, orexin-A did not change the IBa,L of GCs, whereas the orexin-A effect persisted in the presence of the phosphatidylcholine (PC)-PLC inhibitor D609. The orexin-A-induced potentiation was not seen with internal infusion of Ca(2+)-free solution or when inositol 1,4,5-trisphosphate (IP3)-sensitive Ca(2+) release from intracellular stores was blocked by heparin/xestospongins-C. Moreover, the orexin-A effect was mimicked by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate, but was eliminated when PKC was inhibited by bisindolylmaleimide IV (Bis-IV)/Gö6976. Neither adenosine 3',5'-cyclic monophosphate (cAMP)-protein kinase A (PKA) nor guanosine 3',5'-cyclic monophosphate (cGMP)-protein kinase G (PKG) signaling pathway was likely involved, as orexin-A persisted to potentiate the IBa,L of GCs no matter these two pathways were activated or inhibited. These results suggest that, by activating OX1R, orexin-A potentiates the IBa,L of rat GCs through a distinct Gq/PI-PLC/IP3/Ca(2+)/PKC signaling pathway., (Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.)

Our previous study demonstrated that a large-conductance Ca2+-activated K+ current (BKCa), a voltage-gated TTX-sensitive sodium current (INa.TTX), and an inward rectifier K+ current (IKir) were heterogeneously present in most of human cardiac c-kit+ progenitor cells. The present study was designed to investigate the effects of these ion channels on cell cycling progression and migration of human cardiac c-kit+ progenitor cells with approaches of cell proliferation and mobility assays, siRNA, RT-PCR, Western blots, flow cytometry analysis, etc. It was found that inhibition of BKCa with paxilline, but not INa.TTX with tetrodotoxin, decreased both cell proliferation and migration. Inhibition of IKir with Ba2+ had no effect on cell proliferation, while enhanced cell mobility. Silencing KCa.1.1 reduced cell proliferation by accumulating the cells at G0/G1 phase and decreased cell mobility. Interestingly, silencing Kir2.1 increased the cell migration without affecting cell cycling progression. These results demonstrate the novel information that blockade or silence of BKCa channels, but not INa.TTX channels, decreases cell cycling progression and mobility, whereas inhibition of Kir2.1 channels increases cell mobility without affecting cell cycling progression in human cardiac c-kit+ progenitor cells.