Microbial fuel cell (MFC)-based biosensor for combined heavy metals monitoring and associated bioelectrochemical process

To explore the feasibility and related mechanism of MFC biosensor for wastewater detection under the action of combined heavy metals. Cyclic voltammetry (CV) and scanning electrochemical impedance spectroscopy (EIS) were used to explore the related bioelectrochemical process. The response of the reactor to single/combined heavy metals, low/high heavy metal concentrations, and the differences in ohmic resistance (Rs) and charge transfer resistance (Rct) were investigated using Ni as the core heavy metal and the combined action of Cd, Cu and Zn. The results indicated that there was a linear relationship between the concentration and output voltage of the MFC biosensor under the action of combined heavy metals (R² = 0.8803–0.973). However, the internal resistance (Rint) of the MFC biosensor under the action of single heavy metal was far less than that of the combined heavy metal group, and the power density (19.849 W m^?3) was 4 times that of the combined heavy metal group (3.109–4.589 W m^?3). The Rs of the biosensors in the combined heavy metal group were 0.868Ω and 0.860, which were higher than 0.768Ω of the single heavy metal sensor. With the increase of the concentration of heavy metals in the influent, the increase of Rct was more obvious in the combined group, while the Rs in the single group significantly increased (P < 0.05). The results imply that it is possible for MFC biosensors to be used in the detection of actual water polluted by various heavy metals, but the biosensor performance is mainly limited by Rct, which needs to be further improved.     

An Atomistic Understanding of Allosteric Inhibition of Glutamate Racemase: a Dampening of Native Activation Dynamics

Glutamate racemases (GR) are members of the family of bacterial enzymes known as cofactor-independent racemases and epimerases and catalyze the stereoinversion of glutamate. D-amino acids are universally important for the proper construction of viable bacterial cell walls, and thus have been repeatedly validated as attractive targets for novel antimicrobial drug design. Significant aspects of the mechanism of this challenging stereoinversion remain unknown. The current study employs a combination of MD and QM/MM computational approaches to show that the GR from H. pylori must proceed via a pre-activation step, which is dependent on the enzyme's flexibility. This mechanism is starkly different from previously proposed mechanisms. These findings have immediate pharmaceutical relevance, as the H. pylori GR enzyme is a very attractive allosteric drug target. The results presented in this study offer a distinctly novel understanding of how AstraZeneca's lead series of inhibitors cripple the H. pylori GR's native motions, via prevention of this critical chemical pre-activation step. Our experimental studies, using SPR, fluorescence and NMR WaterLOGSY, show that H. pylori GR is not inhibited by the uncompetitive mechanism originally put forward by Lundqvist et al.. The current study supports a deep connection between native enzyme motions and chemical reactivity, which has strong relevance to the field of allosteric drug discovery.     

Fatigue strength evaluation of self-piercing riveted Al-5052 joints under different specimen configurations

In this study, static and fatigue tests were conducted using coach-peel, cross-tension and tensile–shear specimens with Al-5052 plates for evaluation of the fatigue strength of the SPR joints. For the coach-peel, cross-tension and tensile–shear geometries, the ratios of the fatigue endurance limit to static strength were 11%, 14% and 34%, respectively, assuming fatigue cycles of 10⁶ for an infinite lifetime. The equivalent stress intensity factor range can properly predict the current experimental fatigue lifetime. Fatigue crack initiation occurred due to fretting damage between the upper and lower sheets and between the rivet and these sheets.     

静电纺丝法制备ZnO构建酪氨酸酶生物传感器检测邻苯二酚

电化学生物传感器在环境监测、生物与食品分析等领域应用广泛。本文采用静电纺丝法制备了氧化锌微纳米纤维材料，并负载酪氨酸酶（Tyr）构建了检测邻苯二酚的生物传感电极。利用X射线粉末衍射（XRD）、扫描电子显微镜（SEM）表征了氧化锌微纳米纤维的形貌结构，采用循环伏安法（CV）和计时电流法（IT）优化了检测邻苯二酚的工作条件。结果表明，制备的Tyr/ZnO/CS/GCE生物传感电极具有良好的电化学性能，在反应过程中为扩散控制，对邻苯二酚的检测在5～50μmol/L的浓度范围内具有较好的线性关系，最低检测限为1.9041μmol/L，灵敏度为376.31μA/(mmol·L·cm²)，氧化锌的加入可增强酪氨酸酶的稳定性，在尿素、多巴胺和葡萄糖3种电化学活性相近物质存在的情况下仍对邻苯二酚的检测有较好的选择性，且具有良好的循环稳定性。     

A novel electrochemical aptasensor based on layer-by-layer assembly of DNA-Au@Ag conjugates for rapid detection of aflatoxin M1 in milk samples

Aflatoxin M₁ (AFM₁) is a common toxin in dairy products that causes acute and chronic human health disorders. Thus, the development of a rapid and accurate AFM₁ detection method is of vital importance for food safety monitoring. This work was to develop a novel electrochemical aptasensor for sensitive and specific determination of AFM_1. The dendritic-like nanostructure was formed on the gold electrode surface by layer-by-layer assembly of gold-silver core-shell nanoparticles modified with DNA conjugates. In the presence of AFM₁, the specific recognition between AFM₁ and Apt caused the disassociation of the DNA controlled dual Au@Ag conjugates from the surface of the electrode, causing less methylene blue to bind to the surface and weakening the electrochemical signal. The more AFM₁ there is, the weaker the electrochemical signal. Transmission electron microscope results showed that the successfully synthesized Au@Ag nanoparticles exhibited a core-shell structure with Au as core and Ag as shell, and their average diameter was about 30 nm. Under optimal conditions, the electrochemical aptasensor showed a wide detection ranging from 0.05 ng mL^?1 to 200 ng mL^?1, and a low detection limit of 0.02 ng mL^?1. Moreover, the proposed strategy has been successfully applied to the detection of AFM₁ in cow, goat, and sheep milk samples with satisfactory recoveries ranging from 91.10% to 104.05%. This work can provide a novel rapid detection method for AFM₁, and also provide a new sensing platform for the detection of other toxins.     

Measurement of hydrogen peroxide in an advanced oxidation process using an automated biosensor

A hydrogen peroxide biosensor was used to monitor hydrogen peroxide concentrations in a UV/hydrogen peroxide immobilised Fenton advanced oxidation process (AOP). The biosensor is based on gas phase monitoring and thus is more resistant to fouling from the liquid phase constituents of industrial processes. The biosensor is supplied with catalase continually, therefore overcoming any problems with enzyme degradation, which would occur in an immobilised enzyme biosensor. The biosensors response was linear within the experimental range 30-400mg H(2)O(2)l(-1) with a R(2) correlation of 0.99. The hydrogen peroxide monitor was used to monitor residual peroxide in an AOP, operated with a step overload of hydrogen peroxide, with correlation factors of 0.96-0.99 compared to offline hydrogen peroxide determinations by UV spectroscopy. Sparging the sample with nitrogen was found to be effective in reducing the interference from dissolved gases produced with the AOP itself. It is proposed that this biosensor could be used to improve the effectiveness of AOPs via hydrogen peroxide control.     

A microbial sensor for BOD

A microbial amperometric sensor for the determination of the biochemical oxygen demand (BOD) determination using Trichosporon cutaneum cells immobilized in polyvinylalcohol has been developed. This sensor allowed BOD measurement with very short response times (< 30 s), an operation stability of 48 days and a serial coefficient of variation of ±3.3% when a sample solution containing 22 mg l⁻¹ BOD was employed. A linear range was obtained up to 100 mg l⁻¹ BOD using a glucose/glutamic acid standard. The sensitivity and specificity was increased by incubation of the BOD sensor with the respective wastewater. Comparable results were obtained for BOD values estimated by the biosensor and those determined by the 5-day method.     

Bacterial communities in commercial aircraft high‐efficiency particulate air (HEPA) filters assessed by PhyloChip analysis

Abstract  Air travel can rapidly transport infectious diseases globally. To facilitate the design of biosensors for infectious organisms in commercial aircraft, we characterized bacterial diversity in aircraft air. Samples from 61 aircraft high‐efficiency particulate air (HEPA) filters were analyzed with a custom microarray of 16S rRNA gene sequences (PhyloChip), representing bacterial lineages. A total of 606 subfamilies from 41 phyla were detected. The most abundant bacterial subfamilies included bacteria associated with humans, especially skin, gastrointestinal and respiratory tracts, and with water and soil habitats. Operational taxonomic units that contain important human pathogens as well as their close, more benign relatives were detected. When compared to 43 samples of urban outdoor air, aircraft samples differed in composition, with higher relative abundance of Firmicutes and Gammaproteobacteria lineages in aircraft samples, and higher relative abundance of Actinobacteria and Betaproteobacteria lineages in outdoor air samples. In addition, aircraft and outdoor air samples differed in the incidence of taxa containing human pathogens. Overall, these results demonstrate that HEPA filter samples can be used to deeply characterize bacterial diversity in aircraft air and suggest that the presence of close relatives of certain pathogens must be taken into account in probe design for aircraft biosensors.

Practical Implications

A biosensor that could be deployed in commercial aircraft would be required to function at an extremely low false alarm rate, making an understanding of microbial background important. This study reveals a diverse bacterial background present on aircraft, including bacteria closely related to pathogens of public health concern. Furthermore, this aircraft background is different from outdoor air, suggesting different probes may be needed to detect airborne contaminants to achieve minimal false alarm rates. This study also indicates that aircraft HEPA filters could be used with other molecular techniques to further characterize background bacteria and in investigations in the wake of a disease outbreak.

     

Determination of pesticides using a low-temperature co-fired ceramic microfluidic platform

The aim of this work is the integration of an improved graphite–epoxy composite electrode modified with the acetylcholinesterase enzyme as a detector into a green tape ceramic microfluidic device for the online amperometric determination of pesticides. First, the operational conditions of the microfluidic system were optimized for the enzymatic substrate determination of acetylthiocholine. Good results were achieved for acetylthiocholine determination, obtaining a low detection limit and optimal sensitivity. The analytical performance of the microanalyzer was evaluated with organophosphorus and carbamate pesticides across a wide concentration range. Pesticides were determined indirectly measuring the enzymatic inhibition effect that they caused. The analysis was based on stopped-flow methodology with a three-step strategy: enzymatic substrate measurement, enzymatic inhibition by pesticide contact with the biosensor, and enzymatic substrate measurement after the inhibition process. The pesticide concentrations were determined by the percentage of inhibition produced. The microanalyzer was used for the analysis of spiked irrigation water samples using this indirect method. The low-temperature co-fired ceramic/epoxy–graphite–acetylcholinesterase microfluidic system showed low limits of detection and adequate sensitivity. The recoveries of the spiked water samples were approximately 100% with relative standard deviation values lower than 5% for three replicate measurements. All results obtained are indicative of a convenient, rapid, sensitive, and economic method for the determination of pesticides in environmental samples.