Inhibition of B16F10 Cancer Cell Growth by Exposure to the Square Wave with 7.83+/-0.3Hz Involves L- and T-Type Calcium Channels.

Extremely low-frequency electromagnetic field (ELF-EMF) exposure influences many biological systems; these effects are mainly related to the intensity, duration, frequency, and pattern of the ELF-EMF. In this study, exposure to square wave with 7.83±0.3 Hz (sweep step 0.1 Hz) was shown to inhibit the growth of B16F10 melanoma tumor cells. In addition, the distribution of the magnetic field was calculated by Biot-Savart Law and plotted using MATLAB. In vitro studies demonstrated a decrease in B16F10 cell proliferation and an increase of Ca2+ influx after 48 h of exposure to the square wave. Ca2+ influx was also partially blocked by inhibition of voltage-gated L- and T-type Ca2+ channels. The data confirmed that the specific time-varying ELF-EMF had an anti-proliferation effect on B16F10 cells and that the inhibition is related to Ca2+ and voltage-gated L- and T-type Ca2+ channels.

Effect of extremely low frequency electromagnetic field parameters on the proliferation of human breast cancer.

Extremely low-frequency electromagnetic field (ELF-EMF) exposures influence many biological systems. These effects are mainly related to the intensity, duration, frequency, and pattern of the ELF-EMF. Our intent was to characterize the effect of specific pulsed electromagnetic fields on the in vitro proliferation of MCF-7 adenocarcinoma and MDA-MB-231 breast cancer cell lines and one non-cancerous M10 breast epithelial cell line. The following four important parameters of ELF-EMF were examined: frequencies (7.83 ± 0.3, 23.49 ± 0.3, and 39.15 ± 0.3 Hz), flux density (0.5 and 1 mT), exposure duration (12, 24, and 48 h), and the exposure methodology (continuous exposure versus switching exposure). The viability of MDA-MB-231 cells exposed to the optimized ELF-EMF pattern (7.83 ± 0.3 Hz, 1 mT, and 6 h switching exposure) was 40.1%. By contrast, the optimized ELF-EMF parameters that were most cytotoxic to breast cancer MDA-MB-231 cells were not damaging to normal M10 cells. In vitro studies also showed that exposure of MDA-MB-231 cells to the optimized ELF-EMF pattern promoted Ca2+ influx and resulted in apoptosis. These data confirm that exposure to this specific ELF-EMF pattern can influence cellular processes and inhibit cancer cell growth. The specific ELF-EMF pattern determined in this study may provide a potential anti-cancer treatment in the future.

Effects of extremely low-frequency electromagnetic fields on B16F10 cancer cells.

This paper presents a method to inhibit B16F10 cancer cells using extremely low-frequency electromagnetic fields (ELF-EMFs) and to evaluate cell viability using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The study examined the effect of a natural EMF resonance frequency (7.83 Hz) and a power line frequency (60 Hz) on B16F10 cancer cells for 24 and 48 h. The B16F10 cancer cells were also exposed to sweep frequencies in several sweep intervals to quantitatively analyze the viability of cancer cells. The results yielded a 17% inhibition rate under 7.83 Hz compared with that of the control group. Moreover, sweep frequencies in narrow intervals (7.83 ± 0.1 Hz for the step 0.05 Hz) caused an inhibition rate of 26.4%, and inhibitory effects decreased as frequency sweep intervals increased. These results indicate that a Schumann resonance frequency of 7.83 Hz can inhibit the growth of cancer cells and that using a specific frequency type can lead to more effective growth inhibition.

A complete high-to-low spin state transition of trivalent cobalt ion in octahedral symmetry in SrCo0.5Ru0.5O(3-δ).

The complex metal oxide SrCo0.5Ru0.5O(3-δ) possesses a slightly distorted perovskite crystal structure. Its insulating nature infers a well-defined charge distribution, and the six-fold coordinated transition metals have the oxidation states +5 for ruthenium and +3 for cobalt as observed by X-ray spectroscopy. We have discovered that Co(3+) ion is purely high-spin at room temperature, which is unique for a Co(3+) in an octahedral oxygen surrounding. We attribute this to the crystal field interaction being weaker than the Hund's-rule exchange due to a relatively large mean Co-O distances of 1.98(2) Å, as obtained by EXAFS and X-ray diffraction experiments. A gradual high-to-low spin state transition is completed by applying high hydrostatic pressure of up to 40 GPa. Across this spin state transition, the Co Kß emission spectra can be fully explained by a weighted sum of the high-spin and low-spin spectra. Thereby is the much debated intermediate spin state of Co(3+) absent in this material. These results allow us to draw an energy diagram depicting relative stabilities of the high-, intermediate-, and low-spin states as functions of the metal-oxygen bond length for a Co(3+) ion in an octahedral coordination.

Battery-powered portable instrument system for single-cell trapping, impedance measurements, and modeling analyses.

A battery-powered portable instrument system for the single-HeLa-cell trapping and analyses is developed. A method of alternating current electrothermal (ACET) and DEP are employed for the cell trapping and the method of impedance spectroscopy is employed for cell characterizations. The proposed instrument (160 mm × 170 mm × 110 mm, 1269 g) equips with a highly efficient energy-saving design that promises approximately 120 h of use. It includes an impedance analyzer performing an excitation voltage of 0.2-2 Vpp and a frequency sweep of 11-101 kHz, function generator with the sine wave output at an operating voltage of 1-50 Vpp with a frequency of 4-12 MHz, cell-trapping biochip, microscope, and input/output interface. The biochip for the single cell trapping is designed and simulated based on a combination of ACET and DEP forces. In order to improve measurement accuracy, the curve fitting method is adopted to calibrate the proposed impedance spectroscopy. Measurement results from the proposed system are compared with results from a precision impedance analyzer. The trapped cell can be modeled for numerical analyses. Many advantages are offered in the proposed instrument such as the small volume, real-time monitoring, rapid analysis, low cost, low-power consumption, and portable application.

Key role of bismuth in the magnetoelastic transitions of Ba3BiIr2O9 and Ba3BiRu2O9 as revealed by chemical doping.

The key role played by bismuth in an average intermediate oxidation state in the magnetoelastic spin-gap compounds Ba3BiRu2O9 and Ba3BiIr2O9 has been confirmed by systematically replacing bismuth with La(3+) and Ce(4+). Through a combination of powder diffraction (neutron and synchrotron), X-ray absorption spectroscopy, and magnetic properties measurements, we show that Ru/Ir cations in Ba3BiRu2O9 and Ba3BiIr2O9 have oxidation states between +4 and +4.5, suggesting that Bi cations exist in an unusual average oxidation state intermediate between the conventional +3 and +5 states (which is confirmed by the Bi L3-edge spectrum of Ba3BiRu2O9). Precise measurements of lattice parameters from synchrotron diffraction are consistent with the presence of intermediate oxidation state bismuth cations throughout the doping ranges. We find that relatively small amounts of doping (∼10 at%) on the bismuth site suppress and then completely eliminate the sharp structural and magnetic transitions observed in pure Ba3BiRu2O9 and Ba3BiIr2O9, strongly suggesting that the unstable electronic state of bismuth plays a critical role in the behavior of these materials.

Domination of second-sphere shrinkage effect to improve photoluminescence of red nitride phosphors.

Red Ca0.99Al(1-4δ/3-x)Si(1+δ+x)N(3-x)C(x):Eu(2+)0.01 (δ = 0.345; x = 0-0.2) nitride phosphors exhibit a blue-shifted emission with increased eye sensitivity function and excellent thermal stability. The variations in the photoluminescence in the Ca0.99Al(1-4δ/3-x)Si(1+δ+x)N(3-x)C(x):Eu(2+)0.01 (δ = 0.345; x = 0-0.2) system are thoroughly investigated. The enhanced emission energy and the improved thermal stability with increasing x are dominated by the second-sphere shrinkage effect via the substitution of small Si(4+) for large Al(3+) with simultaneous charge compensation. Related proofs of the second-sphere shrinkage effect control for photoluminescence are confirmed via high-resolution neutron powder diffraction, EXAFS, and (29)Si solid-state NMR techniques.

Anion disorder in lanthanoid zirconates Gd(2-x)Tb(x)Zr2O7.

The pyrochlore-defect fluorite order-disorder transition has been studied for a series of oxides of the type Gd(2-x)Tb(x)Zr2O7 by a combination of diffraction and spectroscopy techniques. Synchrotron X-ray diffraction data suggest an abrupt transition from the coexistence of pyrochlore and defect fluorite phases to a single defect fluorite phase with increasing Tb content. However neutron diffraction data, obtained at λ ≈ 0.497 Å for all Gd-containing samples to minimize absorption, not only provide evidence for independent ordering of the anion and cation sublattices but also suggest that the disorder transition across the pyrochlore-defect fluorite boundary of Ln2Zr2O7 is rather gradual. Such disorder was also evident in X-ray absorption measurements at the Zr L3-edge, which showed a gradual increase in the effective coordination number of the Zr from near 6-coordinate in the pyrochlore rich samples to near 7-coordinate in the Tb rich defect fluorites. These results indicate the presence of ordered domains throughout the defect fluorite region, and demonstrate the gradual nature of the order-disorder transition across the Gd(2-x)Tb(x)Zr2O7 series.

Biogeochemical reductive release of soil embedded arsenate around a crater area (Guandu) in northern Taiwan using X-ray absorption near-edge spectroscopy.

This study investigates biogeochemical reductive release of arsenate from beudantite into solution in a crater area in northern Taiwan, using a combination of X-ray absorption near-edge structure (XANES) and atomic absorption spectrometry. Total arsenic (As) concentrations in the soil were more than 200 mg/kg. Over four months of laboratory experiments, less than 0.8% As was released into solution after reduction experiments. The 71% to 83% As was chemically reduced into arsenite (As(III)) and partially weathering into the soluble phase. The kinetic dissolution and re-precipitation of As, Fe, Pb and sulfate in this area of paddy soils merits further study.

Does local disorder occur in the pyrochlore zirconates?

The zirconates Ln(2)Zr(2)O(7) (Ln = lanthanoid) have been studied using a combination of Zr L-edge X-ray absorption near edge structure (XANES) and synchrotron X-ray and neutron powder diffraction methods. These studies demonstrate that as the size of the lanthanoid cation decreases, the local structure evolves smoothly from the ideal pyrochlore toward the defect fluorite rather than undergoing an abrupt transformation. The Zr L-edge spectrum is found to be extremely sensitive to changes in the local coordination environment and demonstrates an increase in local disorder across the pyrochlore oxides. The sensitivity of the XANES measurements enables us to identify the progressive nature of the transition that could not be detected using bulk diffraction techniques.

Insight into the dinuclear {Fe(NO)2}10{Fe(NO)2}10 and mononuclear {Fe(NO)2}10 dinitrosyliron complexes.

The reversible redox transformations [(NO)(2)Fe(S(t)Bu)(2)](-) ⇌ [Fe(µ-S(t)Bu)(NO)(2)](2)(2-) ⇌ [Fe(µ-S(t)Bu)(NO)(2)](2)(-) ⇌ [Fe(µ-S(t)Bu)(NO)(2)](2) and [cation][(NO)(2)Fe(SEt)(2)] ⇌ [cation](2)[(NO)(2)Fe(SEt)(2)] (cation = K(+)-18-crown-6 ether) are demonstrated. The countercation of the {Fe(NO)(2)}(9) dinitrosyliron complexes (DNICs) functions to control the formation of the {Fe(NO)(2)}(10){Fe(NO)(2)}(10) dianionic reduced Roussin's red ester (RRE) [PPN](2)[Fe(µ-SR)(NO)(2)](2) or the {Fe(NO)(2)}(10) dianionic reduced monomeric DNIC [K(+)-18-crown-6 ether](2)[(NO)(2)Fe(SR)(2)] upon reduction of the {Fe(NO)(2)}(9) DNICs [cation][(NO)(2)Fe(SR)(2)] (cation = PPN(+), K(+)-18-crown-6 ether; R = alkyl). The binding preference of ligands [OPh](-)/[SR](-) toward the {Fe(NO)(2)}(10){Fe(NO)(2)}(10) motif of dianionic reduced RRE follows the ligand-displacement series [SR](-) > [OPh](-). Compared to the Fe K-edge preedge energy falling within the range of 7113.6-7113.8 eV for the dinuclear {Fe(NO)(2)}(9){Fe(NO)(2)}(9) DNICs and 7113.4-7113.8 eV for the mononuclear {Fe(NO)(2)}(9) DNICs, the {Fe(NO)(2)}(10) dianionic reduced monomeric DNICs and the {Fe(NO)(2)}(10){Fe(NO)(2)}(10) dianionic reduced RREs containing S/O/N-ligation modes display the characteristic preedge energy 7113.1-7113.3 eV, which may be adopted to probe the formation of the EPR-silent {Fe(NO)(2)}(10)-{Fe(NO)(2)}(10) dianionic reduced RREs and {Fe(NO)(2)}(10) dianionic reduced monomeric DNICs in biology. In addition to the characteristic Fe/S K-edge preedge energy, the IR ν(NO) spectra may also be adopted to characterize and discriminate [(NO)(2)Fe(µ-S(t)Bu)](2) [IR ν(NO) 1809 vw, 1778 s, 1753 s cm(-1) (KBr)], [Fe(µ-S(t)Bu)(NO)(2)](2)(-) [IR ν(NO) 1674 s, 1651 s cm(-1) (KBr)], [Fe(µ-S(t)Bu)(NO)(2)](2)(2-) [IR ν(NO) 1637 m, 1613 s, 1578 s, 1567 s cm(-1) (KBr)], and [K-18-crown-6 ether](2)[(NO)(2)Fe(SEt)(2)] [IR ν(NO) 1604 s, 1560 s cm(-1) (KBr)].

The Subjective and Objective Improvement of Non-Invasive Treatment of Schumann Resonance in Insomnia-A Randomized and Double-Blinded Study.

Purpose: Accumulated studies revealed that electromagnetic field can affect human brain and sleep, and the extremely low-frequency electromagnetic field, Schumann resonance, may have the potential to reduce insomnia symptoms. The purpose of this study was to investigate the responses of patients with insomnia to a non-invasive treatment, Schumann resonance (SR), and to evaluate its effectiveness by subjective and objective sleep assessments. Patients and Methods: We adopted a double-blinded and randomized design and 40 participants (70% female; 50.00 ± 13.38 year) with insomnia completed the entire study. These participants were divided into the SR-sleep-device group and the placebo-device group and were followed up for four weeks. The study used polysomnography (PSG) to measure objective sleep and used sleep diaries, Pittsburgh Sleep Quality Inventory (PSQI), Epworth Sleepiness Scale (ESS), and visual analogy of sleep satisfaction to measure subjective sleep. The 36-Item Short-Form Health Survey (SF-36) was used to evaluate quality of life. Chi-square test, Mann-Whitney U-test, and Wilcoxon test were used to analyze the data. Results: About 70% of the subjects were women, with an average age of 50±13.38 years and an average history of insomnia of 9.68±8.86 years. We found that in the SR-sleep-device group, objective sleep measurements (sleep-onset-latency, SOL, and total-sleep-time, TST) and subjective sleep questionnaires (SOL, TST, sleep-efficiency, sleep-quality, daytime-sleepiness, and sleep-satisfaction) were significantly improved after using the SR-sleep-device; in the placebo-device group, only such subjective sleep improvements as PSQI and sleep-satisfaction were observed. Conclusion: This study demonstrates that the SR-sleep-device can reduce the insomnia symptoms through both objective and subjective tests, with minimal adverse effects. Future studies can explore the possible mechanism of SR and health effects and, with a longer tracking time, verify the effectiveness and side effects.

Experimental study of dielectrophoresis and liquid dielectrophoresis mechanisms for particle capture in a droplet.

This work presents a microfluidic system that can transport, concentrate, and capture particles in a controllable droplet. Dielectrophoresis (DEP), a phenomenon in which a force is exerted on a dielectric particle when it is subjected to a non-uniform electric field, is used to manipulate particles. Liquid dielectrophoresis (LDEP), a phenomenon in which a liquid moves toward regions of high electric field strength under a non-uniform electric field, is used to manipulate the fluid. In this study, a mechanism of droplet creation presented in a previous work that uses DEP and LDEP is improved. A driving electrode with a DEP gap is used to prevent beads from getting stuck at the interface between air and liquid, which is actuated with an AC signal of 200 V(pp) at a frequency of 100 kHz. DEP theory is used to calculate the DEP force in the liquid, and LDEP theory is used to analyze the influence of the DEP gap. The increment of the actuation voltage due to the electrode with a DEP gap is calculated. A set of microwell electrodes is used to capture a bead using DEP force, which is actuated with an AC signal of 20 V(pp) at a frequency of 5 MHz. A simulation is carried out to investigate the dimensions of the DEP gap and microwell electrodes. Experiments are performed to demonstrate the creation of a 100-nL droplet and the capture of individual 10-µm polystyrene latex beads in the droplet.

Discrimination of mononuclear and dinuclear dinitrosyl iron complexes (DNICs) by S K-edge X-ray absorption spectroscopy: insight into the electronic structure and reactivity of DNICs.

In addition to probing the formation of dinitrosyl iron complexes (DNICs) by the characteristic Fe K-edge pre-edge absorption energy ranging from 7113.4 to 7113.8 eV, the distinct S K-edge pre-edge absorption energy and pattern can serve as an efficient tool to unambiguously characterize and discriminate mononuclear DNICs and dinuclear DNICs containing bridged-thiolate and bridged-sulfide ligands. The higher Fe-S bond covalency modulated by the stronger electron-donating thiolates promotes the Fe → NO π-electron back-donation to strengthen the Fe-NO bond and weaken the NO-release ability of the mononuclear DNICs, which is supported by the Raman ν(Fe-NO) stretching frequency. The Fe-S bond covalency of DNICs further rationalizes the binding preference of the {Fe(NO)(2)} motif toward thiolates following the trend of [SEt](-) > [SPh](-) > [SC(7)H(4)SN](-). The relative d-manifold energy derived from S K-edge XAS as well as the Fe K-edge pre-edge energy reveals that the electronic structure of the {Fe(NO)(2)}(9) core of the mononuclear DNICs [(NO)(2)Fe(SR)(2)](-) is best described as {Fe(III)(NO(-))(2)}(9) compared to [{Fe(III)(NO(-))(2)}(9)-{Fe(III)(NO(-))(2)}(9)] for the dinuclear DNICs [Fe(2)(µ-SEt)(µ-S)(NO)(4)](-) and [Fe(2)(µ-S)(2)(NO)(4)](2-).

OsMFS1/OsHOP2 Complex Participates in Rice Male and Female Development.

Meiosis plays an essential role in the production of gametes and genetic diversity of posterities. The normal double-strand break (DSB) repair is vital to homologous recombination (HR) and occurrence of DNA fragment exchange, but the underlying molecular mechanism remain elusive. Here, we characterized a completely sterile Osmfs1 (male and female sterility 1) mutant which has its pollen and embryo sacs both aborted at the reproductive stage due to severe chromosome defection. Map-based cloning revealed that the OsMFS1 encodes a meiotic coiled-coil protein, and it is responsible for DSB repairing that acts as an important cofactor to stimulate the single strand invasion. Expression pattern analyses showed the OsMFS1 was preferentially expressed in meiosis stage. Subcellular localization analysis of OsMFS1 revealed its association with the nucleus exclusively. In addition, a yeast two-hybrid (Y2H) and pull-down assay showed that OsMFS1 could physically interact with OsHOP2 protein to form a stable complex to ensure faithful homologous recombination. Taken together, our results indicated that OsMFS1 is indispensable to the normal development of anther and embryo sacs in rice.

Ferromagnetic CoPt3 nanowires: structural evolution from fcc to ordered L1(2).

This investigation demonstrates the magnetic properties and nanostructures of CoPt(3) wire arrays that were fabricated by electrodeposition using a porous alumina template. The X-ray absorption analysis clearly verified the occurrence of a phase transition in CoPt(3) nanowires. This phase transition significantly influences the magnetic properties and enhances coercivity and squareness. The phase transition of CoPt(3) nanowires was from a random alloy distribution to anisotropically ordered CoPt(3) (L1(2)). The thermally induced phase transition of CoPt(3) nanowires to ordered L1(2) CoPt(3) through a "cluster-in-cluster" intermediate state via interdiffusion processes is revealed. The mechanism exhibited by these CoPt(3) nanowires is proposed to explain the strong correlation between their magnetic character and their atomic distribution.

Fabrication of protein chips based on 3-aminopropyltriethoxysilane as a monolayer.

Although 3-aminopropyltriethoxysilane (APTES) is widely adopted as a monolayer in biosensors, experimental silanization takes at least 1 h at high temperature. Therefore, the feasibility of the silanization with APTES in a short reaction time and at room temperature was investigated. The surface modification of glass slides using a self-assembled monolayer of APTES with a concentration of 10% was studied by immobilizing FITC. APTES was successfully immobilized on the glass slide. The effect of reaction temperature and time of silanization were investigated. Various silanization conditions of APTES were examined by contact angle measurement and fluorescence microscopy. The surface of glass patterns with a gold thin film as background was characterized by determining the fluorescent intensities following the immobilization of fluorescein isothiocyanate (FITC), protein A-FITC, antimouse IgG-FITC and sheep anti-bovine albumin-FITC. The normalized fluorescent intensity indicated that a short period (4 min) of silanization at 25 degrees C suffices to form an APTES thin film by the immobilization of protein A on a glass surface. Such a condition does not require microheaters and temperature sensors in a microfluidic system, which will significantly reduce the manufacturing process, cost, and reaction time in the future.

Effect of electrode geometry on performance of EWOD device driven by battery-based system.

This study develops a driving system for an electrowetting-on-dielectric (EWOD) device comprising a 9 V battery, an ATmega8535 microprocessor, a DC/DC converter, two regulator ICs and a switch circuit. The driving system greatly improves the portability of the EWOD device and is capable of generating a square wave with voltages ranging from 50~100 V(pp) and frequencies in the range 1~5 kHz. A series of experimental and numerical investigations are performed to investigate the effect of the conducting electrode geometry on the droplet velocity in the EWOD device. Three different electrode configurations are considered, namely a linear array of square electrodes, a series of interdigitated electrodes having either two or three fingers, and a series of interdigitated electrodes having five or six fingers. The experimental results show that the corresponding droplet velocities are 7.25 mm/s, 8.17 mm/s and 7.82 mm/s, respectively. The simulation results indicate that the pressure difference induced within the droplets actuated by the square, interdigitated (2323) and interdigitated (5656) electrodes has a value of 15.5 N/m², 262 N/m² and 141.1 N/m², respectively. The corresponding droplet velocities are 33.8 mm/s, 72.7 mm/s and 64.5 mm/s, respectively. Overall, the experimental and numerical results indicate that the interdigitated (2323) electrode optimizes the transportation of the droplets in the EWOD device. The improved droplet velocity obtained using this particular electrode configuration is attributed to an increased length of the contact line between the droplet and the actuating electrode, which in turn increases the driving force.

Effect of actuation sequence on flow rates of peristaltic micropumps with PZT actuators.

Many biomedical applications require the administration of drugs at a precise and preferably programmable rate. The flow rate generated by the peristaltic micropumps used in such applications depends on the actuation sequence. Accordingly, the current study performs an analytical and experimental investigation to determine the correlation between the dynamic response of the diaphragms in the micropump and the actuation sequence. A simple analytical model of a peristaltic micropump is established to analyze the shift in the resonant frequency of the diaphragms caused by the viscous damping effect. The analytical results show that this damping effect increases as the oscillation frequency of the diaphragm increases. A peristaltic micropump with three piezoelectric actuators is fabricated on a silicon substrate and is actuated using 2-, 3-, 4- and 6-phase actuation sequences via a driving system comprising a microprocessor and a phase controller. A series of experiments is conducted using de-ionized water as the working fluid to determine the diaphragm displacement and the flow rates induced by each of the different actuation sequences under phase frequencies ranging from 50 Hz to 1 MHz. The results show that the damping effect of actuation sequences influences diaphragm resonant frequency, which in turn affects the profiles of flow rates.

Oxygen Electrocatalysis at MnIII-O x-C Hybrid Heterojunction: An Electronic Synergy or Cooperative Catalysis?

The interface at the metal oxide-carbon hybrid heterojunction is the source to the well-known "synergistic effect" in catalysis. Understanding the structure-function properties is key for designing more advanced catalyst-support systems. Using a model MnIII-O x single-layer catalyst on carbon, we herein report a full elucidation to the catalytic synergism at the hybrid heterojunction in the oxygen reduction reaction (ORR). The successful fabrication of the single-layer catalyst from bottom-up is fully characterized by the X-ray absorption fine structure and high-resolution transmission electron microscopy. For oxygen electrocatalysis over this model hybrid heterostructure, our results, from both theory and experiment, show that the synergistic ORR truly undergoes a cooperated two-step electrocatalysis with catalytic promotion (Δ Eonset = 60 mV) near the heterojunction and over the single-layer catalyst through an interfacial electronic interplay, rather than an abstruse transition towards a one-step dissociative pathway. Finally, we report a superior peroxide-reducing activity of 432.5 mA cm-2 mg(M)-1 over the MnIII-O x single-layer.