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Measuring the activity of low-abundance enzymes, down to a few molecules in one living cell, is important but challenging to elucidate their biological function. Here, an electrochemical molecule trap is established at the tip of a nanopipette with an electrochemical detector, in which the diffusion of the molecules away from the electrochemical detector is prevented by electro-osmotic flow (EOF). Accordingly, a limited amount of enzymes is trapped to continuously catalyze the conversion of the substrate to generate a sufficient amount of the byproduct hydrogen peroxide for electrochemical measurements. The resistive pulse sensing of the enzymes in single liposomes validates the detection sensitivity down to 15 molecules. Using this ultrasensitive electrochemical strategy, the activity of 60 sphingomyelinase molecules inside single unstimulated living J774 cells is measured, which was hardly detected by previous methods. The established electrochemical molecule trap-based sensing approach opens the door toward single-molecule electrochemical detection in one living cell. This success will solve the long-standing problem regarding the study of the activity of low-abundance proteins in cells in their native physiological state and greatly enhance the understanding of the roles of proteins in cellular behavior.

Elemental doping confined in atomically-thin 2D semiconductors offers a compelling strategy for constructing high performance photocatalysts. Although impressive progress has been achieved based on co-thermolysis method, the choices of dopants as well as semiconductor hosts are still quite limited to yield the elaborate photocatalyst with atomic-layer-confined doping defects, owing to the difficulty in balancing the reaction kinetics of different precursors. This study shows that the cation exchange reaction, which is dictated by the Pearson's hard and soft acids and bases (HSAB) theory and allowed to proceed at mild temperatures, can be developed into a conceptually new protocol for engineering elemental doping confined in semiconductor atomic layers. To this aim, the two atomic layers of a new type of 2D photocatalyst PdSeO3 (PdSeO3 2Als, 1.1 nm) are created by liquid exfoliation and exploited as a proof-of-concept prototype. It is demonstrated that the Mn(II) dopants with controlled concentrations can be incorporated into PdSeO3 2ALs via topological Mn2+-for-Pd2+ cation exchange performed in water/isopropanol solution at 30 °C. The resulting Mn-doped PdSeO3 2ALs present enhanced capacity for driving photocatalytic oxidation reactions in comparison with their undoped counterparts. The findings here suggest that the new route mediated by post synthetic cation exchange promises to give access to manifold 2D confined-doping photocatalysts, with little perturbations on the thickness, morphology, and crystal structure of the atomically-thin semiconductor hosts.

This study aims to resolve the difficult problem of manufacturers’ service outsourcing strategy decision-making, with services divided into two types: hygienic and motivational. We build a value creation model for the manufacturer, service contractor, and customer under different outsourcing strategies, and we examine optimal outsourcing decision-making in various scenarios and its effect on the value creation of related entities. The results indicate that motivational services have a lower bound characterized by service utility. When the manufacturer derives services higher than this lower bound, it should operate its own motivational service and outsource the hygienic service to create greater value. When the manufacturer is limited by resources or capabilities and is required to outsource the motivational service, it can maintain value creation by setting an appropriate contract coefficient. The service contractor should undertake motivational services within the contract boundary negotiated by the manufacturer and conduct hygienic services outside the boundary. Theoretically, outsourcing both service types can maximize customer value, but this may not achieve the manufacturers’ optimal value creation.

Water electrolysis is one of the most promising approaches for producing hydrogen. However, it has been hindered by the sluggishness of the anodic oxygen evolution reaction. In this work, we fabricated Ru-Co-Mn trimetallic alloy nanoparticles on N-doped carbon support (RuCoMn@NC) via the pyrolysis-adsorption-pyrolysis process using ZIF-67 as a precursor. The RuCoMn@NC catalyst exhibited excellent electrocatalytic performance for the hydrogen evolution reaction (HER) over a wide range of pH and glucose oxidation reaction in alkaline media. It showed exceptional HER activity in alkaline medium, superior to that of the commercial Pt/C catalyst (20 wt%), and good electrochemical stability. Further, a two-electrode alkaline electrolyzer pairing RuCoMn@NC as both cathode and anode was employed, and only a cell voltage of 1.63 V was required to attain a current density of 10 mA cm−2 in glucose electrolysis, which is about 270 mV lower than that in the overall water-splitting electrolyzer. This paper provides a promising method for developing efficiently bifunctional electrocatalysts driving redox electro-catalysis, and it would be beneficial to energy-saving electrolytic H2 production.

Combination of the strong light-absorbing power of plasmonic metals with the superior charge carrier dynamics of halide perovskites is appealing for bio-inspired solar-energy conversion due to the potential to acquire long-lived plasmon-induced hot electrons. However, the direct coupling of these two materials, with Au/CsPbBr

The analysis of biomolecules in a 3D cell model is crucial for the collection of spatial information close to the actual organ. In this work, a highly sensitive platinized open carbon nanocavity electrode is fabricated to investigate reactive oxygen species (ROS) in three regions (proliferating zone, quiescent zone, and necrotic core) of a 3D CT26 cell model. The presence of a nanocavity permits more frequent collisions of ROS on the Pt surface, accelerating electron transfer, and thus pushes the detection limit down to 1 nM. This improved detection sensitivity guarantees the spatial investigation of the ROS distribution in a 3D cell sphere, including a high concentration in the outer proliferating layer even without any external stimulus, a low concentration in the quiescent layer, and almost no ROS at the center. The observation of ROS in the cell sphere without the stimulus reveals the presence of oxygen stress in the 3D cancer cell model, which is obviously different from the previous observation in living cultured 2D cells. This discovery provides direct evidence about the discrepancy about the metabolism in 2D and 3D cells, which could also direct a new study in cell electroanalysis to achieve more actual molecular information in life study.

Conventional photoelectrochemical (PEC) analysis mostly utilizes photoactive material modified planar indium tin oxides (ITOs) to obtain photocurrent responses for the measurement of analytes in solution. In this work, a CdS quantum dot (QD) modified nanopipette was prepared for the PEC analysis of the alkaline phosphatase (ALP) activity in single MCF-7 cells. The nanopipette was filled with ascorbic acid 2-phosphate (AAP) that was egressed outside the nanopipette by electrochemical pumping. Next, AAP was catalyzed by ALP to generate ascorbic acid (AA), which is an efficient electron donor for CdS QDs under illumination. Based on the result that the nanopipette showed a linear photocurrent response to AA, a nearly linear correlation between the photocurrent and the activity of ALP was established. Accordingly, using these CdS QD modified nanopipettes, the ALP activity in single MCF-7 cells was determined to be 0.12 U mL−1 by PEC analysis. This work does not expand the application of PEC bioanalysis, but offers a new strategy for single cell analysis.

Self-assembly of colloidal nanocrystals (NCs) into large-scale superlattices with complex and controllable structures has attracted extensive attention due to their collective properties and promising device applications. Plasmonic NCs are very popular for long-range ordered superstructures by virtue of their collective nanogaps for electromagnetic field enhancement, in particular bulk-scale single-layer assembly. Large-area two-dimensional (2D) quasinanosheets (QNSs) composed of mono-component Au NCs or multi-component Au@ZnS core-shell hetero-nanocrystals (HNCs) were successfully prepared, via careful solvent evaporation-induced interfacial self-assembly. The entire self-assembly process was carried out on the liquid-air surface and mediated simply by tuning the operating temperatures and concentrations of the NCs. Specifically, monolayer and double-layer 2D QNSs in tens of micrometers scale with different stacking models were fabricated by precisely controlling the solvent evaporation rate and colloidal concentration.

Here we report a synthetic strategy for controllable construction of yolk-shell and core-shell plasmonic metal@semiconductor hybrid nanocrystals through modulating the kinetics of sulfurization reaction followed by cation exchange. The yielded yolk-shell structured products feature exceptional crystallinity and more importantly, the intimately adjoined and sharp interface between plasmonic metal and semiconductor which facilitates efficient charge carrier communications between them. By exploiting the system composed of Au nanorods and p-type PbS as a demonstration, we show that the Au@PbS yolk-shell nanorods manifest notable improvement in visible and near infrared light absorption compared to the Au@PbS core-shell nanorods as well as hollow PbS nanorods. Moreover, the photocathode constituted by Au@PbS yolk-shell nanorods affords the highest photoelectrochemical activities both under simulated sunlight and λ < 700 nm light irradiation. The superior performance of Au@PbS yolk-shell nanorods is considered arising from the combination of the favorable structural advantages of yolk-shell configuration and the surface plasmon resonance enhancement effect. We envision that the reported synthetic strategy can offer a valuable means to create hybrid nanocrystals with desirable structures and functions that enable to harness the photogenerated charge carriers, including the plasmonic hot holes, in wide-range solar-to-fuel conversion.

Cation exchange has evolved into a powerful tactic for synthesis of semiconductor nanocrystals (NCs) that are not readily accessible otherwise. Here we have investigated the In3+-for-Cu+ cation exchange in the dodecahedral-shaped Cu7S4 NCs and found that surface passivation, either caused by excess guest cations or ligand molecules, can be exploited to engineer the structural properties of the NCs. By monitoring the parallel reactions carried out under systematically varied conditions, together with the positron annihilation spectroscopy investigation, we have demonstrated that the key element accounting for the observed surface passivation is associated with the copper vacant sites on the surface of Cu7S4 NCs. The reduction in In3+/Cu+ ratio and the presence of strong-binding ligands could enhance the density of surface copper vacancies and boost cation exchange reaction, which therefore alters the competition between the in-diffusion of In3+ and out-diffusion of Cu+ ions. Such capability to control the inter-diffusion balance in cation exchange (and the accompanying Kirkendall effect) enables the creation of a series of CuInS2 (and Cu7S4@CuInS2) NCs with various exotic structures, which show different photocatalytic abilities in singlet oxygen generation. This study can not only add more structural complexity and diversity to the semiconductor NCs achievable by cation exchange, but also presents an important guideline for establishing a unifying mechanistic understanding of the reaction kinetics in cation exchange process.

In 2018, the two-dimensional (2D) PdSeO3 monolayer was predicted to be a highly promising photocatalyst for direct overall water splitting in the light of density functional theory (DFT) computations. Herein, we present the first report on the synthesis of 2D PdSeO3 nanosheets by using the quaternary ammonium intercalation-assisted electrochemical exfoliation method. The resulting atomically thin PdSeO3 nanosheets possess a moderate band gap with suitable band edge alignments for water splitting, and display excellent activity and stability in photocatalytic H2 evolution.

Maneuvering the architecture and composition of semiconductors is essential to optimizing their performance in photocatalytic solar-to-fuel conversion. Here, we show that ion exchange, having a disparate mechanism with direct nucleation and growth of semiconductor crystals, can provide a new platform for rational control over the geometry and electronic structures of chalcogenide semiconductor photocatalysts. As a demonstration, the ZnSe nanocubes possessing a hollowed architecture and doped with a controllable amount of Ag

High glucose(HG)-induced excessive proliferation and migration of the media vascular smooth muscle cell(VSMC) are the main pathological characteristics in diabetes related vascular injuries. Previous studies have shown that microRNA-34a (miR-34a) is involved in cancer metastasis, proliferation and invasion and plays an essential role in cardiovascular disease. However, little is known about the regulating role miR-34a in HG-induced proliferation and migration of VSMC. Here we demonstrated that miR-34a was downregulated at different timepoints under HG stimulation. Then, HG induced proliferation and migration was found to be impaired by miR-34a overexpression using transwell, CCK8 and RT-qPCR assays. Furthermore, the HG-induced depression of “contractile” VSMC-specific markers were reversed by the overexpression of miR-34a. Moreover, we confirmed that miR-34a regulated HG-induced VSMC proliferation and migration through its target gene, Notch1, which has been shown to be associated with cell proliferation and migration in previous studies. Taken together, we propose that the miR34a-Notch1 axis plays an important role in regulating HG-induced VSMC proliferation and migration.

The chemical reaction in a confined space is known to be accelerated due to a high collision probability; however, the study of this confinement effect in a supersmall space down to femtoliter (fL) is seldom reported. Here, an adjustable volume [from picoliter (pL) to fL] of the aqueous phase is retrained at the tip of a nanopipette by an organic solvent so that the confinement effect on the specific activity of glucose oxidase is investigated. The activity is determined by the amount of hydrogen peroxide generated from the reaction between the oxidase and glucose using a nanoelectrode inside the nanopipette. As compared with the activity in bulk solution (82 U/mg), the activity increases up to 7500 U/mg in a 105 fL space. The 2 orders of magnitude increase in the enzymatic activity is the highest amplification in the volume-confined enzyme reaction as reported. A near-exponential drop in the activity is observed with the increase in the space volume, revealing the dominant enhancement in the confined space at the fL level for the first time. The established electrochemical nanopipettes should not only provide a strategy for the study of the enzymatic activity in supersmall confined space but also help understand the confinement effect of enzyme-catalyzed reactions.

Ononin (ON) is an isoflavone with numerous reported bioactivities, including anti-oxidative, anti-inflammatory and neuroprotective effects. Autophagy is a critical homeostatic process in the body that has been reported to closely associate with the apoptotic processes of cardiomyocytes. Using flow cytometry, western blotting, echocardiography and Masson's staining, the present study investigated the effects of ON on H2O2-induced cardiomyocyte apoptosis and myocardial infarction, in addition to any potential underlying molecular mechanisms. H2O2 treatment reliably induced apoptosis in H9C2 cells. The anti-apoptotic effects of ON were revealed by flow cytometry results and by the downregulation of cleaved-caspase 3. Further investigations indicated that ON may alleviate apoptosis by enhancing autophagy, as evidenced by increased microtubule-associated proteins 1A/1B light chain 3B expression and p62 degradation. Activation of the 5' AMP-activated protein kinase (AMPK)/mTOR pathway was observed after ON administration following H2O2-induced cardiomyocyte injury. However, these anti-apoptotic effects mediated by ON were lost after autophagy inhibition by chloroquine or AMPK inhibition by Compound C. Finally, the protective effects of ON on cardiomyocytes in vitro could also be observed in vivo. A myocardial infarction model was established by ligating the left anterior descending branch of the rat heart. Using echocardiography and Masson's staining, ON was shown to increase the ejection fraction and decrease cardiac fibrosis in rats with myocardial infarction. These results suggest that ON exerts cardioprotective effects by improving autophagy via the AMPK/mTOR signaling pathway.

Background Left-right (LR) asymmetry is an essential feature of bilateral animals. Studies in vertebrates show that LR asymmetry formation comprises three major steps: symmetry breaking, asymmetric gene expression, and LR morphogenesis. Although much progress has been made in the first two events, mechanisms underlying asymmetric morphogenesis remain largely unknown due to the complex developmental processes deployed by vertebrate organs. Results We here addressed this question by studying Pitx gene function in the basal chordate amphioxus whose asymmetric organogenesis, unlike that in vertebrates, occurs essentially in situ and does not rely on cell migration. Pitx null mutation in amphioxus causes loss of all left-sided organs and incomplete ectopic formation of all right-sided organs on the left side, whereas Pitx partial loss-of-function leads to milder phenotypes with only some LR organs lost or ectopically formed. At the N1 to N3 stages, Pitx expression is gradually expanded from the dorsal anterior domain to surrounding regions. This leads to activation of genes like Lhx3 and/or Prop1 and Pit, which are essential for left-side organs, and downregulation of genes like Hex and/or Nkx2.1 and FoxE4, which are required for right-side organs to form ectopically on the left side. In Pitx mutants, the left-side expressed genes are not activated, while the right-side genes fail to decrease expression on the left side. In contrast, in embryos overexpressing Pitx genes, the left-side genes are induced ectopically on the right side, and the right-side genes are inhibited. Several Pitx binding sites are identified in the upstream sequences of the left-side and right-side genes which are essential for activation of the former and repression of the latter by Pitx. Conclusions Our results demonstrate that (1) Pitx is a major (although not the only) determinant of asymmetric morphogenesis in amphioxus, (2) the development of different LR organs have distinct requirements for Pitx activity, and (3) Pitx controls amphioxus LR morphogenesis probably through inducing left-side organs and inhibiting right-side organs directly. These findings show much more dependence of LR organogenesis on Pitx in amphioxus than in vertebrates. They also provide insight into the molecular developmental mechanism of some vertebrate LR organs like the lungs and atria, since they show a right-isomerism phenotype in Pitx2 knockout mice like right-sided organs in Pitx mutant amphioxus. Our results also explain why some organs like the adenohypophysis are asymmetrically located in amphioxus but symmetrically positioned in vertebrates.

In a typical intracellular electroanalytical measurement, a nanoelectrode is located inside a living cell and a reference electrode outside the cell. This setup faces a problem to drop a certain potential across the cellular plasma membrane that might interrupt the cellular activity. To solve this problem, a self-referenced nanopipette is assembled by incorporating a reference electrode inside the nanocapillary, with a Pt ring at the tip as the electrochemical surface. The potential applied between the Pt ring and the reference electrode is restricted inside the capillary and thus has a negligible effect on the surrounding cellular environment. Using this new setup, the nanopipette pierces into the nucleus of a single living cell for the measurement of hydrogen peroxide under oxidative stress. It is found that a lesser amount of hydrogen peroxide is measured in the nucleus compared with the cytoplasm, revealing uneven oxidative stress inside the cell. The result will not only greatly improve the current setup for intracellular electrochemical analysis but also provide biological information of the compartment inside the living cell.

For realizing scalable solar hydrogen synthesis, the development of visible-light-absorbing photocatalysts capable of overall water splitting is essential. Metal sulfides can capture visible light efficiently; however, their utilization in water splitting has long been plagued by the poor resilience against hole oxidation. Herein, we report that the ZnIn2S4 monolayers with dual defects (Ag dopants and nanoholes) accessed via cation exchange display stoichiometric H2 and O2 evolution in pure water under visible light irradiation. In-depth characterization and modeling disclose that the dual-defect structure endows the ZnIn2S4 monolayers with optimized light absorption and carrier dynamics. More significantly, the dual defects cooperatively function as active sites for water oxidation (Ag dopants) and reduction (nanoholes), thus leading to steady performance in photocatalytic overall water splitting without the assistance of cocatalysts. This work demonstrates a feasible way for fulfilling "all-in-one" photocatalyst design and manifests its great potential in addressing the stability issues associated with sulfide-based photocatalysts.

Here we show a novel strategy for tailoring the synergistic electrical properties of metal@semiconductor hybrid nanocrystals (HNCs) based on cation exchange-enabled electronic doping. As a demonstration, the conductive nature of Au@CdS core–shell HNCs was changed from n- to p-type by introducing Cu dopants into the CdS shell. The dependence of the conductivity type on the dopant concentration in the HNCs is disclosed by combined photoelectrochemical (PEC) studies. Moreover, a tandem PEC cell consisting of an undoped Au@CdS photoanode and a Cu-doped Au@CdS photocathode respectively modified with cocatalysts is fabricated, which displayed stable H2 and O2 evolution in unassisted PEC overall water splitting.

In this study, a simple and sensitive LC-tandem mass spectrometric method was developed and validated for the determination of LNP023 in rat plasma. The plasma sample was precipitated with acetonitrile and then separated on an ACQUITY HSS T3 column (50 mm × 2.1 mm, 1.8 μm) using 0.1% formic acid in water and acetonitrile as the mobile phase. The MS detection was performed in positive multiple-reaction monitoring mode with precursor-to-product ion transitions of m/z 423.3 → 174.1 and m/z 435.3 → 367.1 for LNP023 and olaparib (internal standard), respectively. The developed assay was validated in the linear range of 0.1-1000 ng/mL with correlation coefficient (r) greater than 0.9992. The validation parameters were all within the acceptable limits. The validated method has been successfully used to investigate the pharmacokinetics of LNP023 in rat plasma, and our results indicated that LNP023 showed low clearance and high bioavailability (62.2%). Furthermore, four minor metabolites from rat plasma were detected and identified by LC combined with high-resolution mass spectrometry. The metabolic pathways were O-deethylation (M1), hydroxylation (M4), oxidation (M3), and acyl-glucuronidation (M2).

In 2018, the two-dimensional (2D) PdSeO

Resistive-pulse sensing is a technique widely used to detect single nanoscopic entities such as nanoparticles and large molecules that can block the ion current flow through a nanopore or a nanopipette. Although the species of interest, e.g., antibodies, DNA, and biological vesicles, are typically produced by living cells, so far, they have only been detected in the bulk solution since no localized resistive-pulse sensing in biological systems has yet been reported. In this report, we used a nanopipette as a scanning ion conductance microscopy (SICM) tip to carry out resistive-pulse experiments both inside immobilized living cells and near their surfaces. The characteristic changes in the ion current that occur when the pipet punctures the cell membrane are used to monitor its insertion into the cell cytoplasm. Following the penetration, cellular vesicles (phagosomes, lysosomes, and/or phagolysosomes) were detected inside a RAW 264.7 macrophage. Much smaller pipettes were used to selectively detect 10 nm Au nanoparticles in the macrophage cytoplasm. The in situ resistive-pulse detection of extracellular vesicles released by metastatic human breast cells (MDA-MB-231) is also demonstrated. Electrochemical resistive-pulse experiments were carried out by inserting a conductive carbon nanopipette into a macrophage cell to sample single vesicles and measure reactive oxygen and nitrogen species (ROS/RNS) contained inside them.

Electrochemical analysis of intracellular hydrogen peroxide in submicrometer scaled cellular protrusion is challenging and requires a highly sensitive nanoelectrode and advanced electrochemical detection system. In this technical note, a resistive analysis based on acrylic acid polymerization in the nanopipets is established to measure hydrogen peroxide in one axon of single neuron. Upon the position of nanopipet tip inside the axon, hydrogen peroxide is electrokinetically loaded into the pipet to react with ferrous ions generating hydroperoxyl radicals. These radicals initial the polymerization of acrylic acid for the elevation of capillary resistance, as reflected by the drop of the ion current. A 0.3-0.5 nA of current drop is observed supporting the successful analysis of intracellular hydrogen peroxide in the axon. Similar current drops are observed at the body and the axon after the physical loading or physiological stimulation, which suggests even distribution of hydrogen peroxide in the neuron. As compared with the classical amperometric analysis using nanoelectrodes, this new strategy avoids the complexity in the electrode fabrication and the measurement that should facilitate electrochemical analysis of the intracellular molecule at the subcellular level.

Resistive-pulse sensing with biological or solid-state nanopores and nanopipettes has been widely employed in detecting single molecules and nanoparticles. The analytical signal in such experiments is the change in ionic current caused by the molecule/particle translocation through the pipet orifice. This paper describes a new version of the resistive-pulse technique based on the use of carbon nanopipettes (CNP). The measured current is produced by electrochemical oxidation/reduction of redox molecules at the carbon surface and responds to the particle translocation. In addition to counting single entities, this technique enables qualitative and quantitative analysis of the electroactive material they contain. Using liposomes as a model system, we demonstrate the capacity of CNPs for (1) conventional resistive-pulse sensing of single liposomes, (2) electrochemical resistive-pulse sensing, and (3) electrochemical identification and quantitation of redox species (e.g., ferrocyanide, dopamine, and nitrite) contained in a single liposome. The small physical size of a CNP suggests the possibility of single-entity measurements in biological systems.

The development of more intricate devices for the analysis of enzyme activity in single cells, and even in individual intracellular compartments, would advance the knowledge of cellular heterogeneity and protein function at subcellular locations. This chapter describes construction and implementation of nano-capillary electrodes kits, named as "Nanokits," for unprecedented cost-effective analysis of enzyme activity within single living cells and even at single lysosomes. The nanokit that is specific for the target enzyme is assembled in a nanometer-sized capillary with a working electrode, and electrochemically loaded into the cell allowing the electrochemical quantification of the enzyme activity. Furthermore, the enzyme activity in individual intracellular compartments can be characterized by reversed electrochemical pumping to confine the targeted organelle in the nano-capillary tip with the nanokit. The use of commercially available reagent kits marketed for cell population studies permits direct application of the nano-capillary electrode for targeting a variety of enzymes in single cells. This protocol will likely spark expansion in the number of groups embarking on enzyme activity investigations by clearly providing a cookbook description of new cost-effective technology for single cell analysis.

Significance The quantification of protein activity in individual lysosomes in living cells is realized using a nanocapillary designed to electrochemically analyze internal solution, in which a single lysosome is sorted from the cell and the target protein is reacted with the corresponding kit components to generate hydrogen peroxide for measurement. The ability to sort and assay multiple lysosomes from the same cell allows direct study of protein function at subcellular resolution and provides unprecedented information about the homogeneity within the lysosomal population of a single cell., The protein activity in individual intracellular compartments in single living cells must be analyzed to obtain an understanding of protein function at subcellular locations. The current methodology for probing activity is often not resolved to the level of an individual compartment, and the results provide an extent of reaction that is averaged from a group of compartments. To address this technological limitation, a single lysosome is sorted from a living cell via electrophoresis into a nanocapillary designed to electrochemically analyze internal solution. The activity of a protein specific to lysosomes, β-glucosidase, is determined by the electrochemical quantification of hydrogen peroxide generated from the reaction with its substrate and the associated enzymes preloaded in the nanocapillary. Sorting and assaying multiple lysosomes from the same cell shows the relative homogeneity of protein activity between different lysosomes, whereas the protein activity in single lysosomes from different cells of the same type is heterogeneous. Thus, this study for the analysis of protein activity within targeted cellular compartments allows direct study of protein function at subcellular resolution and provides unprecedented information about the homogeneity within the lysosomal population of a single cell.

The objective of this work is to prepare a hyperbranched polyethylenimine (HBPEI) grafted activated carbon obtained from waste fiberboard for effectively removing Cr(VI) from aqueous solution. The ...