A simple route to incorporate redox mediator into carbon nanotubes/Nafion composite film and its application to determine NADH at low potential

Through a new and simple ion-exchange route, two-electron redox mediator thionine has been deliberately incorporated into the carbon nanotubes (CNTs)/Nafion composite film due to the fact that there is strong interaction between any of two among the three materials (ion-exchange process between thionine and Nafion, strong adsorption of thionine by CNTs, and wrapping and solubilizing of CNTs with Nation). The good homogenization of electron conductor CNTs in the integrated films provides the possibility of three-dimensional electron conductive network. The resulting integrated films exhibited high and stable electrocatalytic activity toward NADH oxidation with the significant decrease of high overpotential, which responds more sensitively more than those modified by thioine or CNTs alone. Such high electrocatalytic activity facilitated the low potential determination of NADH (as low as -0.1 V), which eliminated the interferences from other easily oxidizable species. In a word, the immobilization approach is very simple, timesaving and effective, which could be extended to the immobilization of other cationic redox mediators into the CNTs/Nafion composite film. And these features may offer potential promise for the design of amperometric biosensors.

AMPEROMETRIC GLUCOSE SENSOR WITH FERROCENE AS AN ELECTRON-TRANSFER MEDIATOR

A glucose oxidase (GOD) electrode with ferrocene (Fc) used as an electron transfer mediator has been described. Using Nafion, Fc was modified on a glassy carbon (GC) electrode surface, and glucose oxidase was then immobilized on the Fc-Nafion film, forming a GOD-Fc-Nafion enzyme electrode. The preparation method was quite simple and rapid. The enzyme electrode showed a reversible reaction of the redox couple (Fc+/Fc), used in a biosensor system, displayed a sensitive catalytic current response (response time was less than 20 s) on variation of the glucose concentration, with a wide linear range up to 16 mM and with good repeatability. The enzyme electrode showed almost no deterioration over the course of three weeks. There was little or no interference from electro-active anions, such as ascorbic acid, to the determination of glucose based on Nafion film and lower oxidizing potentials of the enzyme electrode.

A Trabecular Bone Explant Model of Osteocyte–Osteoblast Co-Culture for Bone Mechanobiology

The osteocyte network is recognized as the major mechanical sensor in the bone remodeling process, and osteocyte-osteoblast communication acts as an important mediator in the coordination of bone formation and turnover. In this study, we developed a novel 3D trabecular bone explant co-culture model that allows live osteocytes situated in their native extracellular matrix environment to be interconnected with seeded osteoblasts on the bone surface. Using a low-level medium perfusion system, the viability of in situ osteocytes in bone explants was maintained for up to 4 weeks, and functional gap junction intercellular communication (GJIC) was successfully established between osteocytes and seeded primary osteoblasts. Using this novel co-culture model, the effects of dynamic deformational loading, GJIC, and prostaglandin E-2 (PGE(2)) release on functional bone adaptation were further investigated. The results showed that dynamical deformational loading can significantly increase the PGE(2) release by bone cells, bone formation, and the apparent elastic modulus of bone explants. However, the inhibition of gap junctions or the PGE(2) pathway dramatically attenuated the effects of mechanical loading. This 3D trabecular bone explant co-culture model has great potential to fill in the critical gap in knowledge regarding the role of osteocytes as a mechano-sensor and how osteocytes transmit signals to regulate osteoblasts function and skeletal integrity as reflected in its mechanical properties.

Low potential detection of glutamate based on the electrocatalytic oxidation of NADH at thionine/single-walled carbon nanotubes composite modified electrode

A glutamate biosensor based on the electrocatalytic oxidation of reduced nicotinamide adenine dinucleotide (NADH), which was generated by the enzymatic reaction, was developed via employing a single-walled carbon nanotubes/thionine (Th-SWNTs) nanocomposite as a mediator and an enzyme immobilization matrix. The biosensor, which was fabricated by immobilizing glutamate dehydrogenase (GIDH) on the surface of Th-SWNTs, exhibited a rapid response (ca. 5 s), a low detection limit (0.1 mu M), a wide and useful linear range (0.5-400 mu M), high sensitivity (137.3 +/- 15.7) mu A mM(-1) cm(-2), higher biological affinity, as well as good stability and repeatability. In addition, the common interfering species, such as ascorbic acid, uric acid, and 4-acetamidophenol, did not cause any interference due to the use of a low operating potential (190 mV vs. NHE). The biosensor can be used to quantify the concentration of glutamate in the physiological level. The Th-SWNTs system represents a simple and effective approach to the integration of dehydrogenase and electrodes, which can provide analytical access to a large group of enzymes for wide range of bioelectrochemical applications including biosensors and biofuel cells.

遮荫与外源脱落酸喷施对粗枝云杉(Picea asperata Mast.)不同种群抗旱性的影响

本研究通过粗枝云杉不同种群进行的温室半控制试验，采用植物生态学、生理学和生物化学的研究方法，系统地研究了粗枝云杉不同种群抗旱性的生长、形态、生理和生化机理，并结合有关研究进行综合分析，得出主要研究结论如下： 1.粗枝云杉对干旱胁迫的综合反应 粗枝云杉在干旱胁迫下的适应机制为：（1）相对生长速率及植株结构的调整：干旱胁迫下虽然植株相对生长速率显著降低，且有相对较多的生物量向根部分配，但并未发现细根/总根比增加。（2）粗枝云杉对干旱胁迫的光合作用表现为：干旱胁迫显著地降低了控制的理想条件下的气体交换，但干旱胁迫对PSII最大光化学效率（Fv/Fm）没有影响，表明干旱并未影响到光合机构。（3）干旱还影响了很多生理生化过程，包括渗透调解物质（游离脯氨酸）、膜脂过氧化产物、脱落酸（ABA）含量的增加，以及保护酶活性的升高。这些结果证明植物遭受干旱胁迫后发生了一系列的形态、生理和生化响应，这些变化能提高干旱时期植物的存活和生长能力。 2.粗枝云杉不同种群对干旱胁迫反应的种群差异 粗枝云杉三个种群－干旱种群（四川丹巴和甘肃迭部）和湿润种群（四川黑水）对干旱适应不同，这种不同应归因于它们采用的用水策略不同：在水分良好和干旱胁迫条件下，受试种群在相对生长速率和水分利用效率（WUE）方面都表现出显著的种群间差异。与湿润种群相比，干旱种群在两种水分条件下有更高的WUE。粗枝云杉不同种群的碳同位素组分（δ13C）只在干旱胁迫下有显著差异，并且这种差异在水分良好时比干旱胁迫条件下小，说明生理响应和干旱适应性之间的关系受植物内部抗旱机制和外部环境条件（如水分可利用性）或两者互作效应的影响。这些结果说明干旱种群和湿润种群所采用的用水策略不同。干旱种群有更强的抗旱能力，采用的是节水型的用水策略，而湿润种群抗旱能力较弱，采用的是耗水型的用水策略。 3. 遮荫对粗枝云杉不同种群抗旱性影响 干旱胁迫显著降低了全光条件下叶相对含水量（RWC）、相对生长速率、气体交换参数、PSII的有效量子产量（Y），提高了非光化学猝灭效率（qN）、水分利用效率、脯氨酸（PRO）积累、脱落酸（ABA）含量及保护酶活性。然而这种变化在遮荫条件下不明显。我们得出结论适度遮荫降低了干旱对植物的胁迫作用。另一方面，在干旱条件下，与湿润种群相比，干旱种群抗旱性更强，表现在干旱种群净光合速率与单位重量上叶氮含量（Nmass）降低较少。另外，干旱种群表现出更为敏感的气孔导度，更高的热耗散能力（qN）能力、用水效率、ABA积累、保护酶活性，以及更低的总用水量、相对生长速率。这一结果表明这两种群采用不同的生理策略对干旱和遮荫做出反应。许多生长和生理反应差异与这两个种群原产地气候条件相适应。 4. 外源脱落酸(ABA)喷施对粗枝云杉不同种群抗旱性影响 外源ABA喷施在干旱和水分良好条件下均不同程度地提高了根/茎比，表明根和茎对ABA敏感程度不同。实验结果还表明，外源ABA喷施对这两个种群在干旱胁迫期间影响不同。干旱胁迫期间，伴随着ABA喷施，湿润种群净光合速率(A)显著降低，而干旱种群净光合速率变化不明显。另一方面，外源ABA喷施显著提高了干旱条件下干旱种群的单位叶面积重（LMA）、根/茎比、细根/总根（Ft）比、水分利用效率（WUE）、ABA含量, 以及保护酶活性。然而，外源ABA喷施对湿润种群的上述测定指标没有显著影响。这一结果表明干旱种群对外源ABA喷施更为敏感, 反应在更大的气孔导度降低，更高的生物量可塑性，及更高的水分利用效率、ABA含量和保护酶活性。综上所述，我们得出结论，粗枝云杉对外源ABA敏感性因种群的不同而不同。该研究结果可为两个明显不同种群在适应分化方面提供强有力的证据。 Arid or semi-arid land covers more than half of China's land territory. In arid systems, severe shortages of soil water often coincide with periods of high temperatures and high solar radiation, producing multiple stresses on plant performance. Protection from high radiation loads in shaded microenvironments during drought may compensate for a loss of productivity due to reduced irradiance when water is available. Additionally, ABA, a well-known stress-inducible plant hormone, has long been studied as a potential mediator for induction of drought tolerance in plants. Picea asperata Mast., which is one of the most important tree species used for the production of pulp wood and timber, is a prime reforestation species in western China. In this experiment, different population of P. asperata were used as experiment material to study the adaptability to drought stress and population differences in adaptabiliy, and the effects of shade and exogenous abscisic acid (ABA) application on the drought tolerance. Our results cold provide a strong theoretical evidence and scientific direction for the afforestation, and rehabilitation of ecosystem in the arid and semi-arid area, and provide a strong evidence for adaptive differentiation of different populations, and so may be used as criteria for species selection and tree improvement. The results are as follows: 1. A large set of parallel response to drought stress Drought stress caused pronounced inhibition of the growth and increased relatively dry matter allocation into the root; drought stress also caused pronounced inhibition of photosynthesis, while drought showed no effects on the maximal quantum yield of PSII photochemistry (Fv/Fm) in dark-adapted leaves, indicating that drought had no effects on the primary photochemistry of PSII. However, in light-adapted leaves, drought reduced the quantum yield of PSII electron transport (Y) and increased the non-photochemical quenching (qN). Drought also affected many physiological and biochemical processes, including increases in superoxide dismutase (SOD), ascorbate peroxidase (APX) activities, malondialdehyde and ABA content. These results demonstrate that there are a large set of parallel changes in the morphological, physiological and biochemical responses when plants are exposed to drought stress; these changes may enhance the capability of plants to survive and grow during drought periods. 2. Difference in adaptation to drought stress between contrasting populations of Picea asperata There were significant population differences in growth, dry matter allocation and water use efficiency. Compared with the wet climate population (Heishui), the dry climate population (Dan ba and Jiebu) showed higher LMA, fine root/total root ratio and water use efficiency under drought-stressed treatments. The results suggested that there were different water-use strategies between the dry population and the wet population. The dry climate population with higher drought tolerance may employ a conservative water-use strategy, whereas the wet climate population with lower drought tolerance may employ a prodigal water-use strategy. These variations in drought responses may be used as criteria for species selection and tree improvement. 3. The effects of shade on the drought tolerance For both populations tested, drought resulted in lower needle relative water content (RWC), relative growth rate (RGR), gas exchange parameters and effective PSII quantum yield (Y), and higher non-photochemical quenching (qN), water use efficiency (WUE), proline (PRO) and abscisic acid (ABA) accumulation, superoxide dismutase (SOD), ascorbate peroxidase (APX) activities as well as malondialdehyde (MDA) levels and electrolyte leakage in sun plants, whereas these changes were not significant in shade plants. Our study results implied that shade, applied together with drought, ameliorated the detrimental effects of drought. On the other hand, compared with the wet climate population, the dry climate population was more tolerant to drought in the sun treatment, as indicated by less decreases in A and mass-based leaf nitrogen content (Nmass), more responsive stomata, greater capacity for non-radiative dissipation of excitation energy as heat (analysed by qN), and higher WUE，higher level of antioxidant enzyme activities，higher ABA accumulation as well as lower MDA content and electrolyte leakage. Many of the differences in growth and physiological responses reported here are consistent with the climatic differences between the locations of the populations of P. asperata. 4. The effects of exogenous abscisic acid (ABA) application on the drought tolerance For both populations tested, exogenous ABA application increased root/shoot ratio (Rs) under well-watered and drought-stressed conditions, indicating that there was differential sensitivity to ABA in the roots and shoots. However, it appeared that ABA application affected the two P. asperata populations very differently during drought. CO2 assimilation rate (A) was significantly decreased in the wet climate population, but only to a minor extent in the dry climate population following ABA application during soil drying. On the other hand, ABA application significantly decreased stomatal conductance (gs), transpiration rate (E) and malondialdehyde (MDA) content, and significantly increased leaf mass per area (LMA), Rs, fine root/total root ratio (Ft), water use efficiency (WUE), ABA contents, superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) activities under drought condition in the dry climate population, whereas ABA application did not significantly affect these parameters in the wet population plants. The results clearly demonstrated that the dry climate population was more responsive to ABA application than the wet climate population, as indicated by the strong stomata closure and by greater plasticity of LMA and biomass allocation, as well as by higher WUE, ABA content and anti-oxidative capacity to defense against oxidative stress, possibly predominantly by APX. We concluded that sensitivity to exogenous ABA application is population dependent in P. asperata. Our results provide strong evidence for adaptive differentiation between populations of P. asperata.

无介体双室微生物燃料电池产电性能初步研究

本文对无介体双室微生物燃料电池的产电性能进行了初步研究，并根据不同运行阶段产电性能的优劣，对其中微生物的差异性进行了比较分析。全文分为两个部分： 第一部分：以乙酸钠为阳极原料构建双室微生物燃料电池（MFC），研究不同阴极受体、外接电阻、乙酸钠浓度和pH等因素对电池产电性能的影响，研究结果表明：在500mL的阴阳极反应体系中，选用乙酸钠作为阳极底物，质量浓度为6.46 g/L， pH 7.0，接入500Ω外电阻，阴极电子受体选择高锰酸钾的情况下，微生物燃料电池产电性能最好，最大电功率密度达到294.72 mW/m2，库伦效率能达到25.87%。在确定最适外接电阻阻值的同时对MFC内阻进行测定，阻值为871.87Ω。 第二部分：微生物燃料电池运行中，比较以厌氧污泥作为接种源的第一阶段和只接入附着有大量微生物电极的第二阶段的产电性能，得出第二阶段产电性能优于第一阶段，最大电功率密度达到353.57mW/m2，比第一阶段提高58.85 mW/m2；库伦效率为39.35%，增幅达52％左右；针对微生物燃料电池运行过程中，底物CH3COONa可能存在其它的代谢途径，本实验进行了第二阶段产电性能与CH3COONa消耗率关系以及阳极液面上方气体成分和含量的研究，发现第二阶段50h前CH3COONa的大量消耗主要用于微生物的生长，在整个运行过程中，阳极液面上方含有CH4和CO2；对气体测定的同时还发现，振荡能增强电功率密度的输出；通过对电极上和污泥中微生物差异性分析得出，δ-变形菌纲、β-变形菌纲和拟杆菌门的菌种更适应微生物燃料电池的运行环境，能在电极上大量富集，提高电池的产电性能，只接入附着有大量微生物的电极能有效降低热袍菌纲的菌种数量，降低了CH3COONa的无为消耗，有效提高了电池的库伦效率。 Electricity production in the mediator-less two-chambered microbial fuel cell(MFC) was researched. Based on the result in the different operation phase in the MFC, the microbial diversity was analysed. The paper involved two parts: Part 1: A two-chambered microbial fuel cell (MFC) was constructed with high-concentration sodium acetate as fuel in the anode. The influence of different electron acceptors in the cathode, external resistance value, pH value and concentration of sodium acetate on electricity generation in MFC was investigated. The result showed that the maximum power density of 294.72 mW/m2 and the coulombic efficiency of 25.87% was achieved at sodium acetate concentration of 6.46 g/L, pH 7.0, external resistance 500Ωin the anode and when using potassium permanganate as electron acceptor in the cathode. While decided the value of resistor, we found that shaking has effect on electricity production in the MFC. Part 2: Comparing the electricity production in different operation phases when using anaerobic sludge as inoculum in the first phase and microbes in the anodic electrode as inoculum in the second phase, the result showed that electricity production in the second phase was more than that in the first phase, the maximum power density of 353.57 mW/m2 and the coulombic efficiency of 39.35% was achieved, 58.85 mW/m2 and 52％ more than that in the first phase, respectively. According to the fact that CH3COONa might be metabolized in other pathway in the running process in the MFC, we determining the relationship between electricity production and CH3COONa consumption, and the gas content in the anode, we found that CH3COONa was mainly used for microbe growth before 50h, and the anode contained CH4 and CO2. At the same time, we found that shaking could improve power density. The analysis on diversity of microbe in the anodic electrode and anaerobic sludge showed that δ-proteobacterium, β-proteobacterium and Bacteroidetes adapted themselves to the running environment of MFC. The anode could enrich them to improve the electricity production while reduced the quantity of Thermotogales, which were obligately anaerobic organotrophs with a fermentative metabolism, to increase the coulombic efficiency effectively.

Double proton transfer and one-electron oxidation behaviors in double H-bonded glycinamide-formamidine complex and comparison with biological base pair

The behaviors of double proton transfer (DPT) occurring in a representative glycinamide-formamidine complex have been investigated employing the B3LYP/6-311++G** level of theory. Computational results suggest that the participation of a formamidine molecule favors the proceeding of the proton transfer (PT) for glycinamide compared with that without mediator-assisted case. The DPT process proceeds with a concerted mechanism rather than a stepwise one since no zwitterionic complexes have been located during the DPT process. The barrier heights are 14.4 and 3.9 kcal/mol for the forward and reverse directions, respectively. However, both of them have been reduced by 3.1 and 2.9 kcal/mol to 11.3 and 1.0 kcal/mol with further inclusion of zero-point vibrational energy (ZPVE) corrections, where the lower reverse barrier height implies that the reverse reaction should proceed easily at any temperature of biological importance. Additionally, the one-electron oxidation process for the double H-bonded glycinamide-formamidine complex has also been investigated. The oxidated product is characterized by a distonic radical cation due to the fact that one-electron oxidation takes place on glycinamide fragment and a proton has been transferred from glycinamide to formamidine fragment spontaneously. As a result, the vertical and adiabatic ionization potentials for the neutral double H-bonded complex have been determined to be about 8.46 and 7.73 eV, respectively, where both of them have been reduced by about 0.79 and 0.87 eV relative to those of isolated glycinamide due to the formation of the intermolecular H-bond with formamidine. Finally, the differences between model system and adenine-thymine base pair have been discussed briefly.

Multilayer structured carbon nanotubes/poly-L-lysine/laccase composite cathode for glucose/O-2 biofuel cell

Multilayer film of laccase, poly-L-lysine (PLL) and multi-walled carbon nanotubes (MWNTs) were prepared by a layer-by-layer self-assembly technique. The results of the UV-vis spectroscopy and scanning electron microscopy studies demonstrated a uniform growth of the multilayer. The catalytic behavior of the modified electrode was investigated. The (MWNTs/PLL/laccase)(n) multilayer modified electrode catalyzed four-electron reduction of O-2 to water, without any mediator.

Magnetic control of bioelectrocatalytic processes based on assembled iron oxide particles

A facile magnetic control system was designed in bioelectrocatalytic process based on functionalized iron oxide particles. The iron oxide particles were modified with glucose oxidase, and ferrocene dicarboxylic acid was used as electron transfer mediator. Functionalized iron oxide particles can assemble along the direction of applied magnetic field, and the directional dependence of the assembled iron oxide particles can be utilized for device purposes. We report here how such functionalized magnetic particles are used to modulate the bioelectrocatalytic signal by changing the orientation of the applied magnetic field. (C) 2008 Elsevier B.V. All rights reserved.

Electrochemical characteristics of mediated laccase-catalysis and electrochemical detection of environmental pollutants

Laccase has been immobilized on the carbon nanotubes modified glassy carbon electrode surface by adsorption. As-prepared laccase retains good electrocatalytic activity to oxygen reduction by using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) as the mediator. It can be used as a biosensor for the determination of catechol with broad linear range.

Hydrogen peroxide biosensor based on horseradish peroxidase-Au nanoparticles at viologen grafted glassy carbon electrode

A novel hydrogen peroxide biosensor was fabricated that is based on horseradish peroxidase-Au nanoparticles immobilized on a viologen-modified glassy carbon electrode (GCE) by amino cation radical oxidation in basic solution. The immobilized BAPV acts as a mediator and a covalent linker between GCE and the Au nanoparticles. The biosensor exhibited fast response, good reproducibility, and long-term stability.

Synthesis of carbon nanofibers for mediatorless sensitive detection of NADH

Highly sensitive amperometric detection of dihydronicotinamide adenine dinucleotide (NADH) by using novel synthesized carbon nanofibers (CNFs) without addition of any mediator has been proposed. The CNFs were prepared by combination of electrospinning technique with thermal treatment method and were applied without any oxidation pretreatment to construct the electrochemical sensor. In amperometric detection of NADH, a linear range up to 11.45 mu M with a low detection limit of 20 nM was obtained with the CNF-modified carbon paste electrode (CNF-CPE).

Direct electron transfer of horseradish peroxidase and its electrocatalysis based on carbon nanotube/thionine/gold composites

Horseradish peroxidase (HRP) was incorporated into multiwalled carbon nanotube/thionine/Au (MTAu) composite film by electrostatic interactions between positively charged HRP and negatively charged MTAu composite. The results of electrochemical impedance spectroscopy (EIS) confirmed adsorption of HRP on the surface of MTAu modified GC electrode.