Adenosine-dependent assembly of aptazyme-functionalized gold nanoparticles and its application as a colorimetric biosensor

Previous work has shown that DNAzyme-directed assembly of gold nanoparticles can be utilized to make effective colorimetric biosensors. However, the method is restricted to analytes that are directly involved in phosphodiester cleavage. To expand the methodology to a broader range of analytes, a colorimetric adenosine biosensor based on the aptazyme-directed assembly of gold nanoparticles is reported here. The aptazyme is based on the 8-17 DNAzyme with an adenosine aptamer motif that can modulate the DNAzyme activity through allosteric interactions depending on the presence of adenosine. In the absence of adenosine, the aptazyme is inactive and the substrate strands can serve as linkers to assemble DNA-functionalized 13-nm-diameter gold nanoparticles, resulting in a blue color. However, the presence of adenosine activates the aptazyme, which cleaves the substrate strand, disrupting the formation of nanoparticle aggregates. A red color of separated gold nanoparticles is observed. Concentrations of adenosine of up to 1 mM can be measured semiquantitatively by the degree of blue to red color changes or quantitatively by the extinction ratio at 520 and 700 nm. Under the same conditions, 5 mM guanosine, cytidine, or uridine resulted in a blue color only, indicating good selectivity of the sensor. The color difference can be clearly observed by the naked eye by spotting the resulting sensor solution onto an alumina TLC plate. Since aptamers that can target many classes of important analytes have already been selected, they can be adapted into aptazyme systems through rational design or further selection. Thus, colorimetric biosensors for many analytes of interest can be designed using the method presented here, regardless of whether the analytes are directly involved in the cleavage reaction or not.     

Enzyme-catalyzed O2 removal system for electrochemical analysis under ambient air: application in an amperometric nitrate biosensor

Electroanalytical procedures are often subjected to oxygen interferences. However, achieving anaerobic conditions in field analytical chemistry is difficult. In this work, novel enzymatic systems were designed to maintain oxygen-free solutions in open, small volume electrochemical cells and implemented under field conditions. The oxygen removal system consists of an oxidase enzyme, an oxidase-specific substrate, and catalase for dismutation of hydrogen peroxide generated in the enzyme catalyzed oxygen removal reaction. Using cyclic voltammetry, three oxidase enzyme/substrate combinations with catalase were analyzed: glucose oxidase with glucose, galactose oxidase with galactose, and pyranose 2-oxidase with glucose. Each system completely removed oxygen for 1 h or more in unstirred open vessels. Reagents, catalysts, reaction intermediates, and products involved in the oxygen reduction reaction were not detected electrochemically. To evaluate the oxygen removal systems in a field sensing device, a model nitrate biosensor based on recombinant eukaryotic nitrate reductase was implemented in commercial screen-printed electrochemical cells with 200 μL volumes. The products of the aldohexose oxidation catalyzed by glucose oxidase and galactose oxidase deactivate nitrate reductase and must be quenched for biosensor applications. For general application, the optimum catalyst is pyranose 2-oxidase since the oxidation product does not interfere with the biorecognition element.     

Potential application of hemoglobin as an alternative to peroxidase in a phenol biosensor

This work describes a new amperometric biosensor for detecting phenolic compounds. The sensor was designed by immobilizing Hemoglobin (Hb) in a sol-gel matrix onto a carbon electrode. Using the peroxidase activity of Hb, the phenolic compound can be reduced in the presence of H₂O₂. The biosensor's performance in phenolic compound detection was based on mediated electron transfer by Hb. The direct electron transfer of Hb can be avoided by use of the sol-gel matrix. The proposed biosensor presents a very sensitive response for phenolic compounds at an applied potential of 0.0 mV vs. Ag/AgCl. The parameters of the fabrication process for the electrode were optimized. Experimental conditions influencing the biosensor performance, such as pH and potential, were investigated and assessed. Various types of phenolic compounds were detected. Among them, using the optimized conditions, a linearity for the detection of the phenol was observed from 5 μM to 50 μM. Biosensor response levels after 30 days were at more than 80% of their initial response readings level. The response time of the biosensor was about 10 s.     

Fabrication and electrochemical characterization of HRP-lipid Langmuir-Blodgett film and its application as an H2O2 biosensor

The formation of horseradish peroxidase-lipid Langmuir-Blodgett film and its applicability as a biosensor have been studied. HRP was spread directly onto the subphase covered with a layer of lipid in LB trough. Our experimental results showed that surface pressure of this film from liquid to solid state ranged between 15 to 25 mN/m. At surface pressure of 25 mN/m, the monolayer was successfully transferred onto the gold surface. In addition, electrochemical properties of this film showed that protein molecules still kept their natural structure and can give a well electrocatalytic activity to H₂O₂. As an H₂O₂ biosensor, this LB film was able to detect concentration of H₂O₂ which were 3 × 10^− 7 M.     

Facile synthesis of tremella-like MnO2 and its application as supercapacitor electrodes

In this work, three kinds of ultrathin tremella-like MnO₂ have been simply synthesized by decomposing KMnO₄ under mild hydrothermal conditions. When applied as electrode materials, they all exhibited excellent electrochemical performance. The as-prepared MnO₂ samples were characterized by means of XRD, SEM, TEM and XPS. Additionally, the relationship of the crystalline nature with the electrochemical performance was investigated. Among the three samples, the product with the poorest crystallinity had the highest capacitance of 220 F/g at a current density of 0.1 A/g. It is thought that the ultrathin MnO₂ nanostructures can serve as promising electrode materials for supercapacitors.     

Synthesis of the oxide NiSb2O6 and its electrical characterization in toxic atmospheres for its application as a gas sensor

In this work, nanoparticles of the trirutile-type oxide NiSb₂O₆ were synthesized for its application as a gas sensor using the colloidal method assisted by microwave radiation. The crystalline evolution of the powders was analyzed by X-ray diffraction, finding the phase NiSb₂O₆ at 600 °C. SEM micrographs revealed the growth of microspheres, microrods, and irregularly shaped particles. Using TEM, the average size of the nanoparticles was calculated at?~?17.1 nm. For dynamic tests, pellets and thick films were made from the powders calcined at 600 °C. For the thick films, alternating current was used at frequencies of 0.1 and 1 kHz in C₃H₈ and CO₂ atmospheres at 360 °C, where the material’s sensitivity magnitude in CO₂ was?~?2.61% (0.1 kHz) and?~?2.97% (1 kHz). In contrast, for C₃H₈, the sensitivity was?~?6.69% (0.1 kHz) and?~?5.12% (1 kHz) on average. For the pellets, direct current signals and volumetric flow rates of 100, 150, and 200 cm³/min of CO at 200 °C were applied, where the sensitivities were?~?24.37,?~?35.33, and?~?40.77%, respectively. In each test, the sensitivity visibly increased when the gases were injected. Likewise, the response and recovery times decreased when the frequency and gas concentration increased. The results obtained for the trirutile-type oxide NiSb₂O₆, which showed good stability, efficiency, and high sensitivity in CO₂, C₃H₈, and CO atmospheres, make it ideal as a toxic gas sensor.

     

Hydrogen substitutes for the in situ generation of H2O2: an application in the Fenton reaction

This study investigates the ability of formic acid, hydrazine and hydroxylamine to act as H₂ substitutes in conducting phenol degradation by Fenton reaction using in situ generated hydrogen peroxide. The processes were performed with semi-heterogeneous (Pd/Al₂O₃ + soluble Fe²⁺) and fully heterogeneous (FePd/Al₂O₃) catalytic systems under ambient conditions. In contrast to bulk H₂O₂ production conditions, hydrazine is able to produce H₂O₂in situ followed by phenol degradation using Pd/Al₂O₃ + Fe²⁺ at pH 3 without the need for halide ions. However, a degree of mineralization exceeding 37% could not be achieved. The significant production of in situ H₂O₂ at the inherent acidic pH of hydroxylammonium sulfate in the presence of Pd/Al₂O₃ + Fe²⁺ was also found to differ from the bulk production of H₂O₂, in which no H₂O₂ was detected. A remarkable degree of mineralization (ca. 65%) as well as fast phenol degradation during the reaction started at pH 7 over FePd/Al₂O₃ may be an advantage of using hydroxylamine. On the other hand, using formic acid, H₂O₂ was produced at a moderate rate, thereby achieving higher efficiency in the mineralization of phenol. Most importantly, the catalysts were more stable in the presence of formic acid than hydrazine or hydroxylamine.     

Application of H2O2 lifetime as an indicator of TCE Fenton-like oxidation in soils

Hydrogen peroxide decomposition and trichloroethylene (TCE) oxidation kinetics were studied through batch slurry experiments, performed on two TCE contaminated soils (a sandy soil and a clay soil), characterized by different texture and organic fraction; besides, experiments were also performed on sandy soil columns, in order to more closely reproduce the typical conditions of an in situ treatment. The results of the batch tests indicated that hydrogen peroxide lifetime was correlated to the oxidation efficiency; namely, complete TCE oxidation was achieved only for the conditions characterized by longer hydrogen peroxide lifetime, that was obtained by addition of a proper stabilizer (KH(2)PO(4)). The soil properties were also observed to influence both hydrogen peroxide decomposition and TCE oxidation kinetics, probably as a consequence of the different TOC content. The soil column experiments, performed on 10, 20, and 30 cm long columns, indicated that hydrogen peroxide decomposition, which was almost complete at 30 cm depth, was on the contrary negligible when the stabilizer was added. In agreement with this observation, the performance of TCE oxidation were greatly improved in the latter case. Based upon the collected results, it can be concluded that hydrogen peroxide experiments may be useful, at least in the first screening phase of the design activity, for selecting, among the different operating conditions, those that may be potentially more effective for the oxidation treatment.     

Freestanding nanosheets of 1T-2H hybrid MoS_2 as electrodes for efficient sodium storage

Molybdenum disulfide(MoS₂) is a promising electrode material for sodium-ion batteries as it offers a large capacity through a distinct conversion reaction.However,the electrochemical potential of MoS₂ is often restrained by the poor conductivity as the dominant 2 H phase is a semiconductor while the metallic1 T phase is thermodynamically unstable.In this work,we report a hybrid design and material preparation of freestanding nanosheets of MoS₂ composed of both 1 T and 2 H phases based on mild hydrothermal reaction.The introduction of the metallic 1 T-MoS₂ phase into 2 H-MoS₂ and their intimate hybridization enable a significant improvement in electronic conductivity,while the freestanding architecture avoids possible electrochemical aggregation.When used as electrodes for sodium storage,such a hybrid MoS₂ affords a high capacity of~500 mA h g^-1 at 0.5 A g^-1 after 300 cycles,and retains capacity of~200 mA h g^-1 at a high current rate of 4 A g^-1,thus demonstrating their potential in high-performance battery applications.     

Oxidized graphene in ionic liquids for assembling chemically modified electrodes: a structural and electrochemical characterization study

Dispersions of graphene oxide (GO) nanoribbons in ionic liquids, ILs (either 1-butyl-3-methylimidazolium chloride (BMIM-Cl-) or 1-butylpyridinium chloride (-Bupy-Cl-)) have been used to assemble modified screen printed electrodes (SPEs). The graphene oxide/ionic liquid dispersions have been morphologically and structurally characterized by the use of several techniques: X-ray photoelectron spectroscopy (XPS), Fourier transform-infrared (FT-IR) spectroscopy, high-resolution-transmission electron microscopy (HR-TEM). The assembled modified SPEs have then been challenged with various compounds and compared to several electro-active targets. In all cases high peak currents were detected, as well as significant potential shifts, especially in the detection of catecholamines and NADH, compared with the bare SPE and the conventional electrodes, such as glassy carbon (GC) and highly oriented pyrolitic graphite (HOPG). This opens the way to the assembly of new types of sensors and biosensors. The enhanced performances observed are attributed to electrocatalytic effects related to the high electrode surface area, to oxygen-assisted electron transfer, as well as to the disordering effect of the ILs, this latter related to the favorable π-π interactions with the ILs and the GO plane.     

聚吡咯膜电化学包埋固定酶及其复合酶生物传感器

采用电化学包埋法成功地将乳酸脱氢酶固定化在聚吡咯膜上,获得了具有生物活性和电活性的聚吡咯乳酸脱氢酶导电膜,并进一步制备了用以测定丙酮酸的聚吡咯乳酸脱氢酶金电极生物传感器。同时尝试用电化学包埋法将乳酸脱氢酶和辅酶NADH同时固定化在聚吡咯膜上,获得了具有电化学活性的聚吡咯乳酸脱氢酶NADH 金复合酶电极。深入的实验证实了复合电极生物传感器具有线性的工作曲线。     

Silsesquioxane as a new building block material for modified electrodes fabrication and application as neurotransmitters sensors

Nanostructured films comprising a 3-n-propylpyridinium silsesquioxane polymer (designated as SiPy+Cl-) and copper (II) tetrasulfophthalocyanine (CuTsPc) were produced using the Layer-by-Layer technique (LbL). To our knowledge this is the first report on the use of silsesquioxane derivative polymers as building blocks for nanostructured thin films fabrication. Deposition of the multilayers were monitored by UV-Vis spectroscopy revealing the linear increment in the absorbance of the Q-band from CuTsPc at 617 nm with the number of SiPy+Cl-/CuTsPc or CuTsPc/SiPy+Cl-bilayers. FTIR analyses showed that specific interactions between SiPy+Cl- and CuTsPc occurred between SO3- groups of tetrasulfophthalocyanine and the pyridinium groups of the polycation. Morphological studies were carried out using the AFM technique, which showed that the roughness and thickness of the films increase with the number of bilayers. The films displayed electroactivity and were employed to detection of dopamine (DA) and ascorbic acid (AA) using cyclic voltammetry, at concentrations ranging from 1.96 x 10(-4) to 1.31 x 10(-3) molL(-1). The number and the sequence of bilayers deposition influenced the electrochemical response in presence of DA and AA. Using differential pulse technique, films comprising SiPy+/-/CuTsPc were able to distinguish between DA and ascorbic acid (AA), with a potential difference of approximately with 500 mV, in the concentration range of 9.0 x 10(-5) to 2.0 x 10(-4) molL(-1), in pH 3.0.     

H2O2 production by cells on titanium and polystyrene surfaces using an in vivo model of exudate and surface related cell function

The determination of secreted levels of reactive oxygen species by implant-adherent cells in vivo is required for understanding of the role(s) of such reactive oxygen species for the tissue response around medical devices. A model with subcutaneous implants of c.p. titanium (Ti) or polystyrene (PS) (cell culture grade) inserted on the back of rats were used. Implants and associated cells were retrieved and assayed after 1, 3, 5, 7, 14, 21 and 28 days. Morphological analysis of exudate cells showed that polymorphonuclear leukocytes (PMN) predominated after one day whereas macrophages were predominant after three days. The number of implant-adherent cells, as reflected by measurement of DNA, decreased with time. Ultrastructural observations showed that macrophages were predominant cells in contact with the implant surface. Measurement of hydrogen peroxide (H₂O₂) secretion by implant-adherent cells during 40 min incubation ex vivo revealed a constitutive generation of 40–400 pmol H₂O₂/g DNA, depending on implantation time. Stimulation with protein kinase C agonist phorbol myristate acetate (PMA) caused an increased H₂O₂ generation by adherent cells at early (up to five days) but not later (7–28 days) time periods. No major differences between Ti and PS were observed. Taken together, these findings show that Ti and PS implant-adherent cells secrete H₂O₂ under in vivo conditions. Further, a reduced capacity to mount an enhanced H₂O₂ secretion upon stimulation was demonstrated at late time periods. The role of this mediator for biocompatibility remains to be established.     

Procedures for preparing Escherichia coli O157:H7 immunoliposome and its application in liposome immunoassay

Although Escherichia coli serotype O157:H7 was identified as a human pathogen in the ninth decade of the twentieth century, it has become recognized as a major foodborne pathogen. In the United States, the severity of E. coli O157:H7 infection in the young and the elderly has had a tremendous impact on human health, the food industry, and federal regulations regarding food safety. In laboratory diagnosis, most microbiologic assays rely on a single phenotype to selectively isolate this pathogen. However, the process is labor- and time-consuming. It is important eventually to develop new assay procedures to detect them. Immunoliposomes, anti-E. coli O157:H7 antibody-tagged liposomes, encapsulating a visible dye, sulforhodamine B, were used in the present study for the development of a field-portable colorimetric immunoassay to detect E. coli O157:H7. The N-succinimidyl-S-acetylthioacetate derivative of the antibodies (anti-E. coli O157: H7) was first conjugated through the reactive N-(kappa-maleimidoundecanoyloxy) sulfosuccinimide ester derivative of dipalmitoylphosphatidylethanolamine and subsequently incorported into liposomes to form the immunoliposomes. A plastic-backed nitrocellulose strip with two immobilized zones is the basis for a sandwich assay to detect E. coli O157:H7. The first zone is the antigen capture zone (AC zone), which is used in a sandwich (noncompetitive) assay format; the other is the biotin capture zone (BC zone), which is used as a positive control for the strip. During the capillary migration of the wicking reagent containing 50 microL of immunoliposomes and 90 microL of the test sample, E. coli O157:H7 with surface-bound immunoliposomes is captured at the AC zone, while the unbound immunoliposomes migrate and bind to the antibiotin antibodies coated on BC zone. The color density of the AC zone were directly proportional to the amount of E. coli O157:H7 in the test sample. The detection limit of the current assay with heat-killed E. coli O157:H7 was approximately 2500 cells. The selectivity of the newly developed biosensor system was investigated, and pathogens, including Salmonella typhimurium and Listeria genus specific, were proven to have no interference with the detection of E. coli O157:H7.     

PEDOT-PSSA as an alternative support for Pt electrodes in PEFCs

Poly (3,4-ethylenedioxythiophene) (PEDOT) and poly (styrene sulphonic acid) (PSSA) supported platinum (Pt) electrodes for application in polymer electrolyte fuel cells (PEFCs) are reported. PEDOT-PSSA support helps Pt particles to be uniformly distributed on to the electrodes, and facilitates mixed electronic and ionic (H⁺-ion) conduction within the catalyst, ameliorating Pt utilization. The inherent proton conductivity of PEDOT-PSSA composite also helps reducing Nafion content in PEFC electrodes. During prolonged operation of PEFCs, Pt electrodes supported onto PEDOT-PSSA composite exhibit lower corrosion in relation to Pt electrodes supported onto commercially available Vulcan XC-72R carbon. Physical properties of PEDOT- PSSA composite have been characterized by X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy and transmission electron microscopy. PEFCs with PEDOT-PSSA-supported Pt catalyst electrodes offer a peak power-density of 810 mW cm⁻² at a load current-density of 1800 mA cm⁻² with Nafion content as low as 5 wt.% in the catalyst layer. Accordingly, the present study provides a novel alternative support for platinized PEFC electrodes.     

Diatomite as high performance and environmental friendly catalysts for phenol hydroxylation with H2O2

A series of diatomite catalysts were treated and characterized. For the first time, the resulting materials were used in catalysis for the hydroxylation of phenol with H₂O₂ and showed very high hydroxylation activity due to the Fe species in the diatomite. The effect of HCl treatment, contents of catalysts and H₂O₂ were investigated and the active components of diatomite were discussed. The results show that diatomite is the promising candidate for industrial output due to their high catalytic activity, easy physical separation and very low costs.     

Terpyridine-based organic-inorganic hybrid nanofiber and its application as an adsorbent for protein

A terpyridine-based organic-inorganic hybrid amphiphile possessing a triethoxylsilane moiety was synthesized and following its self-assembly in the presence of Cu2+ ion, its binding ability of histidine and His-tag proteins was observed by UV-VIS spectroscopy, Circular dichroism spectroscopy and confocal fluorescence microscopy. The self-assembled 1 in the absence and the presence Cu2+ ion revealed a helical fiber structure, and the self-assembled 1 formed a 1:1 complex with Cu2+ ion. Histidine bound to the self-assembled 1-Cu2+ ion, but more interestingly, His-tag protein was selectively bound to the surface of the self-assembled 1-Cu2+ nanofiber. The finding indicates that the self-assembled 1-Cu2+ nanofiber selectively recognized the His-tag protein.     

Immobilization of the urease on eggshell membrane and its application in biosensor

Eggshell membrane is a natural material, essentially made up of protein fibers having flexibility in the aqueous solution and possessing gas and water permeability. It is used as a biomembrane for immobilization of urease for the development of a potentiometric urea biosensor. Eggshell membrane was treated with polyethyleneimine (PEI) to impart polycation characteristics. Urease was immobilized on the PEI treated eggshell membrane through adsorption. SEM study was carried out to observe the changes in surface morphology after immobilization. FTIR study of membrane was carried out to observe the changes in IR spectra after immobilization of enzyme. Immobilized membrane was associated with ammonium ion selective electrode. Biosensor exhibited sigmoidal responses for the urea concentration range from 0.5 to 10 mM. The response time of the biosensor was 120 s. A single membrane was reused for 270 reactions without loss of activity. The urease–eggshell membranes were stable for 2 months when stored in buffer even at room temperature.     

Hierarchical TiO2-B nanowire@α-Fe2O3 nanothorn core-branch arrays as superior electrodes for lithium-ion microbatteries

This paper reports a simple yet efficient method for the synthesis of hierarchical TiO2-B nanowire@α-Fe2O3 nanothorn core-branch arrays based on a stepwise hydrothermal approach. The as-fabricated hybrid arrays show impressive performance as a high-capacity anode for lithium-ion batteries. The key design in this study is a core-branch hybrid architecture, which not only provides large surface active sites for lithium ion insertion/extraction, but also enables fast charge transport owing to the reduced diffusion paths for both electrons and lithium ions. The peculiar combination of attributes of TiO2 （good structural stability） and Fe2O3 （large specific capacity） provides the hybrid array electrodes with several desirable electrochemical features： large reversible capacity （-800 mA.h.g^-1 for specific mass capacity and -750 μA.h-cm^-2 for specific areal＂ capacity）, good cycling stability, and high rate capability. The impressive electrochemical performance, together with the facile synthesis procedure, may provide an efficient platform to integrate the TiO2 nanowire@Fe2O3 nanothorn core-branch arrays as a three-dimensional thin film electrode for lithium-ion microbatteries.