H(2)O(2)-induced left ventricular dysfunction in isolated working rat hearts is independent of calcium accumulation

Reactive oxygen species (ROS) and intracellular Ca²⁺ overload play key roles in myocardial ischemia-reperfusion (IR) injury but the relationships among ROS, Ca²⁺ overload and LV mechanical dysfunction remain unclear. We tested the hypothesis that H₂O₂ impairs LV function by causing Ca²⁺ overload by increasing late sodium current (I_Na), similar to Sea Anemone Toxin II (ATX-II). Diastolic and systolic Ca²⁺ concentrations (d[Ca²⁺]_i and s[Ca²⁺]_i) were measured by indo-1 fluorescence simultaneously with LV work in isolated working rat hearts. H₂O₂ (100 μM, 30 min) increased d[Ca²⁺]_i and s[Ca²⁺]_i. LV work increased transiently then declined to 32% of baseline before recovering to 70%. ATX-II (12 nM, 30 min) caused greater increases in d[Ca²⁺]_i and s[Ca²⁺]_i. LV work increased transiently before declining gradually to 17%. Ouabain (80 μM) exerted similar effects to ATX-II. Late I_Na inhibitors, lidocaine (10 μM) or R56865 (2 μM), reduced effects of ATX-II on [Ca²⁺]_i and LV function, but did not alter effects of H₂O₂. The antioxidant, N-(2-mercaptopropionyl)glycine (MPG, 1 mM) prevented H₂O₂-induced LV dysfunction, but did not alter [Ca²⁺]_i. Paradoxically, further increases in [Ca²⁺]_i by ATX-II or ouabain, given 10 min after H₂O₂, improved function. The failure of late I_Na inhibitors to prevent H₂O₂-induced LV dysfunction, and the ability of MPG to prevent H₂O₂-induced LV dysfunction independent of changes in [Ca²⁺]_i indicate that impaired contractility is not due to Ca²⁺ overload. The ability of further increases in [Ca²⁺]_i to reverse H₂O₂-induced LV dysfunction suggests that Ca²⁺ desensitization is the predominant mechanism of ROS-induced contractile dysfunction.     

Machine learning–based inverse design for electrochemically controlled microscopic gradients of O2 and H2O2

A fundamental understanding of extracellular microenvironments of O₂ and reactive oxygen species (ROS) such as H₂O₂, ubiquitous in microbiology, demands high-throughput methods of mimicking, controlling, and perturbing gradients of O₂ and H₂O₂ at microscopic scale with high spatiotemporal precision. However, there is a paucity of high-throughput strategies of microenvironment design, and it remains challenging to achieve O₂ and H₂O₂ heterogeneities with microbiologically desirable spatiotemporal resolutions. Here, we report the inverse design, based on machine learning (ML), of electrochemically generated microscopic O₂ and H₂O₂ profiles relevant for microbiology. Microwire arrays with suitably designed electrochemical catalysts enable the independent control of O₂ and H₂O₂ profiles with spatial resolution of ∼10¹ μm and temporal resolution of ∼10° s. Neural networks aided by data augmentation inversely design the experimental conditions needed for targeted O₂ and H₂O₂ microenvironments while being two orders of magnitude faster than experimental explorations. Interfacing ML-based inverse design with electrochemically controlled concentration heterogeneity creates a viable fast-response platform toward better understanding the extracellular space with desirable spatiotemporal control.

Ubiquitous spatiotemporal heterogeneity of natural environments fosters the diverse and fascinating biology that our world embraces, and motivates researchers to mimic natural environments with high spatiotemporal resolution (1–5). Given their close relevance in biochemical metabolisms, dioxygen (O₂) and hydrogen peroxide (H₂O₂) as a surrogate of reactive oxygen species (ROS) are two ubiquitous biologically relevant species in extracellular medium (1, 6). Their extracellular spatial and temporal distributions, particularly at the microscopic scale ranging from 1 μm to 100 μm (7–11), are critical for signal transduction, protein expression, biochemical redox balance, and regulation for cellular metabolism with extensive ecological, environmental, and biomedical implications (Fig. 1A) (1, 3, 8–13). A programmable creation of the spatiotemporal concentration profiles of O₂ and H₂O₂ offers the freedom to mimic, control, and perturb the microenvironments of O₂ and H₂O₂ and hence advance our understanding in microbiology.Open in a separate windowFig. 1.AI-based inverse design of electrochemically generated O₂ and H₂O₂ heterogeneities. (A) The ubiquitous spatiotemporal heterogeneities of O₂ and H₂O₂ in microbiology and the challenges posed in this research topic. (B) The combination of electrochemistry and ML-based inverse design offers a viable approach to mimicking and controlling the heterogeneities of O₂ and H₂O₂ in microbiology. O, oxidant; R, reductant; E_appl (t), the time-dependent electrochemical voltages applied on electrodes. (C) The design of the electrochemically active microwire array electrodes for the generation of O₂ and H₂O₂ gradients; 4e⁻ ORR & 2e⁻ ORR, four-electron and two-electron oxygen reduction reaction into H₂O and H₂O₂, respectively. (D and E) The 45°-tilting images of SEM for the representative microwire arrays used for the training of the ML model (D) and the ones inversely designed for targeted O₂ and H₂O₂ gradients (E); k = (P, D, L), the morphological vector that includes the P, D, and L of the synthesized wire arrays in units of micrometers. (Scale bars, 20 μm.)Despite recent progress (14–18), there remain major technical challenges, particularly in the achievable spatiotemporal resolution and high-throughput design of concentration profiles to suit a plethora of scenarios in microbiology. Approaches based on microfluidics and hydrogels have been able to achieve concentration gradients of O₂ and H₂O₂ through the provision of either O₂/H₂O₂ source (14, 19–21), O₂/H₂O₂ scavenging agents (15, 22, 23), or a combination of both (24) across liquid-impermeable barriers such as agar layers or polymeric thin films (25, 26). Yet such approaches, dependent on passive mass transport and diffusion across more than 10² μm, are inherently incapable of achieving spatial features of less than 100 μm and temporal resolution smaller than ∼10¹ s, the prerequisites to investigate microbiology at cluster or single-cell levels (10–12). Moreover, the large variations of extracellular O₂ and H₂O₂ gradients in different microbial systems demand an inverse design strategy, which, with minimal expenditure, quickly programs a desired concentration profile catering to a specific biological scenario (2–5). The current lack of inverse design protocol impedes the adoption of controllable extracellular heterogeneity to mimic and investigate microbial systems that are of environmental, biomedical, and sustainability-related significance.We envision that the integration of electrochemically generated concentration gradients with inverse design based on machine learning (ML) will address the aforementioned challenges (Fig. 1B). Electrochemistry offers a venue for transducing electric signals into microscopic concentration profiles within ∼10⁰ μm to ∼10² μm away from electrodes’ surface, following the specific electrode reaction kinetics and the mass transport governing equations in the liquid phase (27). Proper designs of electrodes’ microscopic spatial arrangement and electrochemical kinetics lead to concentration gradients that are spatiotemporally programmable by time-dependent electric signals of varying voltages (28). Such benefits of electrochemically generated concentration gradients lead us to employ electrochemistry as a tool to spatiotemporally control the concentration profiles in the extracellular medium. In one example, we found that wire arrays electrochemically active toward O₂ reduction create anoxic microenvironment about 20 μm away from the aerobic external bulk environments, modulate the size and extent of O₂ depletion in the anoxic microenvironment by the wire array’s morphology and applied electrochemical potential (E_appl), and hence enable O₂-sensitive rhizobial N₂ fixation in ambient air powered by renewable electricity (29). Moreover, while not reported before as far as we know, electrochemically generated concentration heterogeneity is commensurate with ML-based inverse design (30, 31), thanks to the mathematically well-defined electrochemical processes that can be numerically simulated (32, 33). We recently reported neural networks, trained by numerically simulated data, that explore the influence of electrode geometry on electrochemical N₂ fixation and achieve optimized morphologies of wire array electrodes untenable without such an ML-based strategy (34). An inverse design for the electrochemically generated gradients will quickly program desirable microenvironments of O₂ and ROS with high spatiotemporal resolutions, thanks to the well-reported electrochemical transformation related to O₂ and H₂O₂ with high electrochemical selectivity (35, 36).In this work, we report an inverse design based on neural networks for independent electrochemical creation of O₂ and ROS microscopic gradients that are relevant, and mimic their extracellular heterogeneities in microbial systems. We hypothesize that careful design of electrocatalysis of O₂ reduction reaction (ORR) can either facilitate four-electron ORR on Pt electrocatalyst for a controllable O₂ spatiotemporal profile or promote two-electron ORR on Au electrocatalyst for a programmable generation of H₂O₂ gradient without significantly perturbing the O₂ one, thanks to their concentration differences in biological mediums (∼10⁻¹ μM to ∼10¹ μM for H₂O₂ and ∼10¹ μM to ∼10² μM for O₂) (2, 7–11). Electrochemically active microwire array electrodes as exemplary model systems (Fig. 1C) are experimentally shown to achieve tunable heterogeneities of O₂ and H₂O₂ independently, with spatial resolution of ∼10¹ μm and temporal resolution of ∼10° s, and are suitable as a platform for independently perturbing biologically relevant O₂ and H₂O₂ profiles in microbial systems. We further established and experimentally validated two neural networks that inversely design the wire array electrodes’ morphologies toward targeted microenvironments of O₂ and H₂O₂, respectively, which is at least one order of magnitude faster than trial-and-error numerical simulation and two orders of magnitude faster than experimental explorations. The demonstrated inverse design of electrochemically generated controlled gradients not only demonstrates a full electrochemical control of concentration profiles in an electrode’s proximity but also establishes an approach of spatiotemporally mimicking and perturbing extracellular space guided by artificial intelligence.     

Role of Nanocrystallites of Al-Based Glasses and H2O2 in Degradation Azo Dyes

Al-based metallic glasses have a special atomic structure and should have a unique degradation ability in azo dye solutions. The Al₈₈Ni₉Y₃ (Y3), Al₈₅Ni₉Y₆ (Y6) and Al₈₂Ni₉Y₉ (Y9) glassy ribbons are melt spun and used in degrading methyl orange (MO) azo dye solution with adding H₂O₂. With increasing c_Y, the as-spun ribbons have an increasing GFA (glass formability) and gradually decreased the degradation rate of MO solution. TEM (transmission electron microscopy) results show that the Y3 ribbon has nano-scale crystallites, which may form the channels to transport elements to the surface for degrading the MO solution. After adding H₂O₂, the degradation efficiency of Al-based glasses is improved and the Y6 ribbon has formed nano-scale crystallites embedded in the amorphous matrix and it has the largest improvement in MO solution degradation. These results indicate that forming nano-scale crystallites and adding H₂O₂ are effective methods to improve the degradation ability of Al-based glasses in azo dye solutions.     

Role of Eu2+ and Dy3+ Concentration in the Persistent Luminescence of Sr2MgSi2O7 Glass-Ceramics

In this study, glass-ceramics based on Sr₂MgSi₂O₇ phosphor co-doped with Eu/Dy were obtained from the sintering and crystallisation of glass powders. The glasses were melted in a gas furnace to simulate an industrial process, and the dopant concentration was varied to optimise the luminescence persistence times. The doped parent glasses showed red emission under UV light excitation due to the doping of Eu³⁺ ions, while the corresponding glass-ceramics showed persistent blue emission corresponding to the presence of Eu²⁺ in the crystalline environment. The dopant concentration had a strong impact on the sintering/crystallisation kinetics affecting the final glass-ceramic microstructure. The microstructures and morphology of the crystals responsible for the blue emission were observed by scanning electron microscopy–cathodoluminescence. The composition of the crystallised phases and the distribution of rare-earth (RE) ions in the crystals and in the residual glassy phase were determined by X-ray diffraction and energy dispersive X-ray analysis. The emission and persistence of phosphorescence were studied by photoluminescence.     

H(2)O(2) regulates cardiac myocyte phenotype via concentration-dependent activation of distinct kinase pathways

Reactive oxygen species (ROS) can act as signaling molecules to stimulate either hypertrophy or apoptosis in cardiac myocytes. We tested the hypothesis that the phenotypic effects of ROS are due to differential, concentration-dependent activation of specific kinase signaling pathways. Adult rat ventricular myocytes were exposed to H(2)O(2) over a broad concentration range (10-1000 microM). Low concentrations of H(2)O(2) (10-30 microM) increased protein synthesis without affecting survival. Higher concentrations of H(2)O(2) (100-200 microM) increased apoptosis (assessed by TUNEL). Still higher concentrations of H(2)O(2) (300-1000 microM) caused both apoptosis and necrosis. A hypertrophic concentration of H(2)O(2) (10 microM) increased the activity of ERK1/2, but not that of JNK, p38 kinase or Akt. An apoptotic concentration of H(2)O(2) (100 microM) activated JNK, p38 kinase and Akt, and further activated ERK1/2. The MEK1/2 inhibitor U0126 prevented the hypertrophic effect of 10 microM H(2)O(2). The apoptotic effect of 100 microM H(2)O(2) was inhibited bya dominant-negative JNK adenovirus, and was potentiated by U0126 or an Akt inhibitor. Thus, the concentration-dependent effects of ROS on myocyte hypertrophy and growth are due, at least in part, to the differential activation of specific kinase signaling pathways that regulate hypertrophy and apoptosis.     

Facile Fabrication of ZnO-ZnFe2O4 Hollow Nanostructure by a One-Needle Syringe Electrospinning Method for a High-Selective H2S Gas Sensor

Herein, a facile fabrication process of ZnO-ZnFe₂O₄ hollow nanofibers through one-needle syringe electrospinning and the following calcination process is presented. The various compositions of the ZnO-ZnFe₂O₄ nanofibers are simply created by controlling the metal precursor ratios of Zn and Fe. Moreover, the different diffusion rates of the metal oxides and metal precursors generate a hollow nanostructure during calcination. The hollow structure of the ZnO-ZnFe₂O₄ enables an enlarged surface area and increased gas sensing sites. In addition, the interface of ZnO and ZnFe₂O₄ forms a p-n junction to improve gas response and to lower operation temperature. The optimized ZnO-ZnFe₂O₄ has shown good H₂S gas sensing properties of 84.5 (S = R_a/R_g) at 10 ppm at 250 °C with excellent selectivity. This study shows the good potential of p-n junction ZnO-ZnFe₂O₄ on H₂S detection and affords a promising sensor design for a high-performance gas sensor.     

Atomic Layer Deposition of Ultrathin La2O3/Al2O3 Nanolaminates on MoS2 with Ultraviolet Ozone Treatment

Due to the chemically inert surface of MoS₂, uniform deposition of ultrathin high-κ dielectric using atomic layer deposition (ALD) is difficult. However, this is crucial for the fabrication of field-effect transistors (FETs). In this work, the atomic layer deposition growth of sub-5 nm La₂O₃/Al₂O₃ nanolaminates on MoS₂ using different oxidants (H₂O and O₃) was investigated. To improve the deposition, the effects of ultraviolet ozone treatment on MoS₂ surface are also evaluated. It is found that the physical properties and electrical characteristics of La₂O₃/Al₂O₃ nanolaminates change greatly for different oxidants and treatment processes. These changes are found to be associated with the residual of metal carbide caused by the insufficient interface reactions. Ultraviolet ozone pretreatment can substantially improve the initial growth of sub-5 nm H₂O-based or O₃-based La₂O₃/Al₂O₃ nanolaminates, resulting in a reduction of residual metal carbide. All results indicate that O₃-based La₂O₃/Al₂O₃ nanolaminates on MoS₂ with ultraviolet ozone treatment yielded good electrical performance with low leakage current and no leakage dot, revealing a straightforward approach for realizing sub-5 nm uniform La₂O₃/Al₂O₃ nanolaminates on MoS₂.     

Influence of interleukin-2 on Ca2+ handling in rat ventricular myocytes

In the present study, we examined the effect of interleukin-2 (IL-2) on cardiomyocyte Ca(2+) handling. The effects of steady-state and transient changes in stimulation frequency on the intracellular Ca(2+) transient were investigated in isolated ventricular myocytes by spectrofluorometry. In the steady state (0.2 Hz) IL-2 (200 U/ml) decreased the amplitude of Ca(2+) transients induced by electrical stimulation and caffeine. At 1.25 mM extracellular Ca(2+) concentration ([Ca(2+)](o)), when the stimulation frequency increased from 0.2 to 1.0 Hz, diastolic Ca(2+) level and peak intracellular Ca(2+) concentration ([Ca(2+)](i)), as well as the amplitude of the transient, increased. The positive frequency relationships of the peak and amplitude of [Ca(2+)](i) transients were blunted in the IL-2-treated myocytes. The effect of IL-2 on the electrically induced [Ca(2+)](i) transient was not normalized by increasing [Ca(2+)](o) to 2.5 mM. IL-2 inhibited the frequency relationship of caffeine-induced Ca(2+) release. Blockade of sarcoplasmic reticulum (SR) Ca(2+)-ATPase with thapsigargin resulted in a significant reduction of the amplitude-frequency relationship of the transient similar to that induced by IL-2. The restitutions were not different between control and IL-2 groups at 1.25 mM [Ca(2+)](o), which was slowed in IL-2-treated myocytes when [Ca(2+)](o) was increased to 2.5 mM. There was no difference in the recirculation fraction (RF) between control and IL-2-treated myocytes at both 1.25 and 2.5 mM [Ca(2+)](o). The effects of IL-2 on frequency relationship, restitution, and RF may be due to depressed SR functions and an increased Na(+)-Ca(2+) exchange activity, but not to any change in L-type Ca(2+) channels.     

Protection of peroxiredoxin II on oxidative stress-induced cardiomyocyte death and apoptosis

Peroxiredoxin II, a cytosolic isoform of the antioxidant enzyme family, has been implicated in cancer-associated cell death and apoptosis, but its functional role in the heart remains to be elucidated. Interestingly, the expression levels of peroxiredoxin II were decreased in mouse hearts upon ischemia-reperfusion, while they were elevated in two genetically modified hyperdynamic hearts with phospholamban ablation or protein phosphatase 1 inhibitor 1 overexpression. To delineate the functional significance of altered peroxiredoxin II expression, adenoviruses encoding sense or antisense peroxiredoxin II were generated; cardiomyocytes were infected, and then subjected to H₂O₂ treatment to mimic oxidative stress-induced cell death and apoptosis. H₂O₂ stimulation resulted in a significant decrease of endogenous peroxiredoxin II expression, along with reduced cell viability in control cells. However, overexpression of peroxiredoxin II significantly protected from H₂O₂-induced apoptosis and necrosis, while downregulation of this enzyme promoted the detrimental effects of oxidative stress in cardiomyocytes. The beneficial effects of peroxiredoxin II were associated with increased Bcl-2 expression, decreased expression of Bax and attenuated activity of caspases 3, 9 and 12. Furthermore, there were no significant alterations in the expression levels of the other five isoforms of peroxiredoxin, as well as active catalase or glutathione peroxidase-1 after ischemia-reperfusion or H₂O₂ treatment. These findings suggest that peroxiredoxin II may be a unique antioxidant in the cardiac system and may represent a potential target for cardiac protection from oxidative stress-induced injury. Returned for 1. Revision: 14 April 2008 1. Revision received: 25 September 2008     

Effect of Microstructure on Photo-Induced Current Characteristics of Eu2+-Doped SrAl2O4

Research interest is active in Eu²⁺/Dy³-doped SrAl₂O₄ phosphors, and the photocurrent characteristics of Eu²⁺-doped SrAl₂O₄ without Dy³⁺ was investigated extensively. However, it exhibits a low photocurrent of ~1 μA owing to high resistance. In this study, the changes in a photo-induced current with grain size were examined. The effect of the processing methods on the microstructure and photocurrent characteristics of Eu²⁺-doped SrAl₂O₄ phosphors was investigated. (Sr_0.99Eu_0.01)Al₂O₄ powders were synthesized using a conventional solid-state method and calcined at 1350 °C under a 3% H₂/(Ar + H₂) reducing gas atmosphere. Sample pellets were fabricated as conventional and hot-press-sintered bodies at 1400 °C. A thin film of the sample was deposited on an alumina substrate by radio-frequency magnetron sputtering. Scanning electron microscopy revealed that the hot-press-sintered body has larger grains than the conventionally sintered body. The photo-induced current of the hot-press-sintered body at 400 lx was ~100 and ~1000 times higher than those of the conventionally sintered body and thin film, respectively. Impedance analysis confirmed that this dramatic increase in the photo-induced current is closely related to the increase in the grain size and crystallinity of the sample. This study verifies the applicability of Eu²⁺-doped SrAl₂O₄ as both a phosphor and photosensor.     

The Influence of SiO2 + SiC + Al (H2PO4)3 Coating on Mechanical and Dielectric Properties for SiCf/MWCNTS/AlPO4 Composites

SiC fiber-reinforced AlPO₄ matrix (SiC_f/MWCNTs/AlPO₄) composites were fabricated using a hot laminating process with multi-walled carbon nanotubes (MWCNTs) as the absorber. A coating prepared from SiO₂ + SiC + Al (H₂PO₄)₃ was applied to the surface of the SiC_f/MWCNTs/AlPO₄ composites prior to an anti-oxidation test at 1273 K in air for 40 h. The anti-oxidation effect was verified by a three-point bending test, scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and a dielectric property test. Anti-oxidation mechanism investigations revealed that the coating effectiveness could be attributed to three substances, i.e., SiO₂, SiP₂O_7, and SiO₂ + AlPO₄ solid solution from the reactions of SiC + O₂→SiO₂ + CO, SiO₂ + P₂O₅→SiP₂O₇ and SiO₂ + AlPO₄→solid solution, respectively.     

Long-Lasting Binding of IT-066 to Human Histamine H2 Receptor

Based on animal models, IT-066, a histamine H₂-receptor antagonist, is reported to possess potent and long-lasting antagonisms on histamine H₂ receptor (H₂R) -mediated effects. However, no reports have been published concerning its interaction with the human H₂R. The aim of this study is to characterize its interaction with human H₂R. Chinese hamster ovary cell lines stably expressing human H₂Rs were obtained. The effects of IT-066, famotidine, and ranitidine on tiotidine binding and histamine-stimulated cAMP production were analyzed. IT-066 inhibited [³H]tiotidine binding and histamine-stimulated cAMP production more potently than famotidine or ranitidine. In addition, preincubation with 10^–5 M IT-066, but not with 10^–5 M famotidine or 10^–4 M ranitidine, had marked inhibitory effects long after extensive washing. Paraformaldehyde fixation of the cells blunted inhibition of [³H]tiotidine binding induced by preincubation with IT-066, but not that by preincubation with famotidine or ranitidine. IT-066 has potent and long-lasting antagonisms on human H₂R. At least one of the IT-066 binding sites is not shared by famotidine, ranitidine, or tiotidine and is affected by paraformaldehyde fixation.     

Severity of left ventricular dysfunction in heart failure patients affects the degree of serum-induced cardiomyocyte apoptosis. Importance of inflammatory response and metabolism

Background/Objectives

In heart failure pro-inflammatory cytokines contribute to cardiomyocytes loss by apoptosis and play a role in the remodelling of the extracellular matrix (ECM). Myocardial injury recruits endothelial progenitor cells (EPCs) to the site of damage and stimulates their differentiation, contributing to myocardial tissue repair. We investigated if the severity of left ventricular dysfunction in heart failure patients (HF) may influence the ability of serum to induce cardiomyocytes death and whether this effect is affected by inflammation and intracellular oxidative stress pathways.

Methods

Adult murine cardiomyocytes HL-5 were incubated with 2% human serum from patients with heart failure (NYHA classes I to IV). Apoptosis was analysed by two different methods. TNF-α, IL-1β, IL-6, matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinases 1 (TIMP-1) were measured in sera from patients.

Results

Cytokine levels were higher in sera from moderate-severe CHF compared to that of patients with mild CHF. Levels of CD117⁺ (c-Kit⁺) cells and EPCs were significantly lower in blood from moderate-severe HF patients. Serum from HF patients induced a significantly higher ROS production involving p38 MAPK signalling and apoptosis in cardiomyocytes. NAC treatment prevented serum-induced oxidative effects. The increase of AMPK phosphorylation showed an involvement of FFA β-oxidation during apoptotic stress.

Conclusions

All these alterations could be used as predictive factors of worsening in heart failure and culture of cardiomyocytes could be employed to test pharmacological effects.     

Crystal structure of Ca2+/H+ antiporter protein YfkE reveals the mechanisms of Ca2+ efflux and its pH regulation

Ca²⁺ efflux by Ca²⁺ cation antiporter (CaCA) proteins is important for maintenance of Ca²⁺ homeostasis across the cell membrane. Recently, the monomeric structure of the prokaryotic Na⁺/Ca²⁺ exchanger (NCX) antiporter NCX_Mj protein from Methanococcus jannaschii shows an outward-facing conformation suggesting a hypothesis of alternating substrate access for Ca²⁺ efflux. To demonstrate conformational changes essential for the CaCA mechanism, we present the crystal structure of the Ca²⁺/H⁺ antiporter protein YfkE from Bacillus subtilis at 3.1-Å resolution. YfkE forms a homotrimer, confirmed by disulfide crosslinking. The protonated state of YfkE exhibits an inward-facing conformation with a large hydrophilic cavity opening to the cytoplasm in each protomer and ending in the middle of the membrane at the Ca²⁺-binding site. A hydrophobic “seal” closes its periplasmic exit. Four conserved α-repeat helices assemble in an X-like conformation to form a Ca²⁺/H⁺ exchange pathway. In the Ca²⁺-binding site, two essential glutamate residues exhibit different conformations compared with their counterparts in NCX_Mj, whereas several amino acid substitutions occlude the Na⁺-binding sites. The structural differences between the inward-facing YfkE and the outward-facing NCX_Mj suggest that the conformational transition is triggered by the rotation of the kink angles of transmembrane helices 2 and 7 and is mediated by large conformational changes in their adjacent transmembrane helices 1 and 6. Our structural and mutational analyses not only establish structural bases for mechanisms of Ca²⁺/H⁺ exchange and its pH regulation but also shed light on the evolutionary adaptation to different energy modes in the CaCA protein family.     

Characterization of Al2O3 Matrix Composites Fabricated via the Slip Casting Method Using NiAl-Al2O3 Composite Powder

This work aimed to characterize Al₂O₃ matrix composites fabricated by the slip casting method using NiAl-Al₂O₃ composite powder as the initial powder. The composite powder, consisting of NiAl + 30 wt.% Al₂O₃, was obtained by mechanical alloying of Al₂O₃, Al, and Ni powders. The composite powder was added to the Al₂O₃ powder to prepare the final powder for the slip casting method. The stained composite samples presented high density. EDX and XRD analyses showed that the sintering process of the samples in an air atmosphere caused the formation of the NiAl₂O₄ spinel phase. Finally, the phase composition of the composites changed from the initial phases of Al₂O₃ and NiAl to Al₂O₃, Ni, and NiAl₂O₄. However, in the area of Ni, fine Al₂O₃ particles remaining from the initial composite powder were visible. It can be concluded that after slip casting, after starting with Al₂O₃ and the composite powder (NiAl-Al₂O₃) and upon sintering in air, ceramic matrix composites with Ni and NiAl₂O₄ phases, complex structures, high-quality sintered samples, and favorable mechanical properties were obtained.     

Layered Perovskites BaM2In2O7 (M = La,Nd): From the Structure to the Ionic (O2−, H+) Conductivity

The design of new oxide compounds that can be used as oxygen- or proton-conducting electrolytes for solid oxide fuel cells is actively in progress. Despite the intensive research activities regarding electrolytes with perovskite/fluorite structures, the search for other structural alternatives is of paramount importance. In this study we focus on a novel material with significantly improved properties for the electrochemical purposes. The two-layered BaNd₂In₂O₇ perovskite with a Ruddlesden–Popper structure was investigated as a protonic conductor for the first time. In detail, its local structure, water uptake, and the ionic (O²⁻, H⁺) conductivity were comprehensively studied. The nature of rare-earth elements (M = La, Nd) in the structure of BaM₂In₂O₇ on the structural and transport properties was revealed. The presented analysis showed that the composition of BaNd₂In₂O₇ is nearly pure proton conductor below 350 °C. This work opens up a new way in the design of protonic conductors with double-layered perovskite structure.     

The pH Value Control of Morphology and Luminescence Properties of Gd2O2S: Tb3+ Phosphors

Developing rare-earth doped oxysulfide phosphors with diverse morphologies has significant value in many research fields such as in displays, medical diagnosis, and information storage. All of the time, phosphors with spherical morphology have been developed in most of the related literatures. Herein, by simply adjusting the pH values of the reaction solution, Gd₂O₂S:Tb³⁺ phosphors with various morphologies (sphere-like, sheet-like, cuboid-like, flat square-like, rod-like) were synthesized. The XRD patterns showed that phosphors with all morphologies are pure hexagonal phase of Gd₂O₂S. The atomic resolution structural analysis by transmission electron microscopy revealed the crystal growth model of the phosphors with different morphology. With the morphological change, the band gap energy of Gd₂O₂S:Tb³⁺ crystal changed from 3.76 eV to 4.28 eV, followed by different luminescence performance. The samples with sphere-like and cuboid-like microstructures exhibit stronger cathodoluminescence intensity than commercial product by comparison. Moreover, luminescence of Gd₂O₂S:Tb³⁺ phosphors have different emission performance excited by UV light radiation and an electron beam, which when excited by UV light is biased towards yellow, and while excited by an electron beam is biased towards cyan. This finding provides a simple but effective method to achieve rare-earth doped oxysulfide phosphors with diversified and tunable luminescence properties through morphology control.     

Pulse Plasma Sintering of NiAl-Al2O3 Composite Powder Produced by Mechanical Alloying with Contribution of Nanometric Al2O3 Powder

NiAl-Al₂O₃ composites, fabricated from the prepared composite powders by mechanical alloying and then consolidated by pulse plasma sintering, were presented. The use of nanometric alumina powder for reinforcement of a synthetized intermetallic matrix was the innovative concept of this work. Moreover, this is the first reported attempt to use the Pulse Plasma Sintering (PPS) method to consolidate composite powder with the contribution of nanometric alumina powder. The composite powders consisting of the intermetallic phase NiAl and Al₂O₃ were prepared by mechanical alloying from powder mixtures containing Ni-50at.%Al with the contribution of 10 wt.% or 20 wt.% nanometric aluminum oxide. A nanocrystalline NiAl matrix was formed, with uniformly distributed Al₂O₃ inclusions as reinforcement. The PPS method successfully consolidated NiAl-Al₂O₃ composite powders with limited grain growth in the NiAl matrix. The appropriate sintering temperature for composite powder was selected based on analysis of the grain growth and hardness of Al₂O₃ subjected to PPS consolidation at various temperatures. As a result of these tests, sintering of the NiAl-Al₂O₃ powders was carried out at temperatures of 1200 °C, 1300 °C, and 1400 °C. The microstructure and properties of the initial powders, composite powders, and consolidated bulk composite materials were characterized by SEM, EDS, XRD, density, and hardness measurements. The hardness of the ultrafine-grained NiAl-Al₂O₃ composites obtained via PPS depends on the Al₂O₃ content in the composite, as well as the sintering temperature applied. The highest values of the hardness of the composites were obtained after sintering at the lowest temperature (1200 °C), reaching 7.2 ± 0.29 GPa and 8.4 ± 0.07 GPa for 10 wt.% Al₂O₃ and 20 wt.% Al₂O₃, respectively, and exceeding the hardness values reported in the literature. From a technological point of view, the possibility to use sintering temperatures as low as 1200 °C is crucial for the production of fully dense, ultrafine-grained composites with high hardness.     

The Effect of Bi2O3 and Fe2O3 Impurity Phases in BiFeO3 Perovskite Materials on Some Electrical Properties in the Low-Frequency Field

Pure bismuth ferrite (BFO) and BFO with impurity phases (Bi₂O₃ or Fe₂O₃) were synthesized by the hydrothermal method. Complex dielectric permittivity (ε) and electrical conductivity (σ) were determined by complex impedance measurements at different frequencies (200 Hz–2 MHz) and temperatures (25–290) °C. The conductivity spectrum of samples, σ(f), complies with Jonscher’s universal law and the presence of impurity phases leads to a decrease in the static conductivity (σ_DC); this result is correlated with the increased thermal activation energy of the conduction in impure samples compared to the pure BFO sample. The conduction mechanism in BFO and the effect of impurity phases on σ and ε were analyzed considering the variable range hopping model (VRH). Based on the VRH model, the hopping length (R_h), hopping energy (W_h) and the density of states at the Fermi level (N(E_F)) were determined for the first time, for these samples. In addition, from ε(T) dependence, a transition in the electronic structure of samples from a semiconductor-like to a conductor-like behavior was highlighted around 465–490 K for all samples. The results obtained are useful to explain the conduction mechanisms from samples of BFO type, offering the possibility to develop a great variety of electrical devices with novel functions.     

橙皮素对H2O2诱导H9c2心肌细胞凋亡的抑制效应研究

目的 探讨橙皮素(HES)对H₂O₂诱导的H9c2心肌细胞凋亡有何影响。方法 采用H₂O₂建立H9c2心肌细胞氧化应激损伤模型,实验分为四组：正常对照组(Control组),H₂O₂损伤组(H₂O₂组),单纯橙皮素处理组(HES组),橙皮素预处理 H₂O₂组(HES H₂O₂组)。H₂O₂(400 μM)处理2 h建立心肌细胞氧化应激损伤模型,HES H₂O₂组于建模前1 h加入40 μM橙皮素。采用CCK-8法确定H9c2细胞活性;DCFH-DA 探针检测细胞活性氧簇(ROS)水平,流式细胞术检测心肌细胞凋亡,分光光度计检测Caspase-3活性。结果 橙皮素预处理可明显改善H₂O₂诱导的H9c2心肌细胞活性降低,并且呈浓度依赖性;给予40 μM橙皮素预处理后ROS的产生明显减少,Caspase-3活性显著下降,心肌细胞凋亡率下降到30%左右。结论 橙皮素对氧化应激诱导的心肌细胞凋亡具有抑制效应。