Acetylcholinesterase-based biosensor electrodes for organophosphate pesticide detection. I. Modification of carbon surface for immobilization of acetylcholinesterase

Screen-printed carbon electrodes modified with the dialdehydes, glutaraldehyde and terephthaldicarboxaldehyde, and then polyethyleneimine have been utilized for production of pesticide biosensors based on acetylcholinesterase. To improve the extent of dialdehyde modification, the electrodes were NH2-derivatized, initially by electrochemical reduction of 4-nitrobenzenediazonium to a nitroaryl radical permitting attachment to the carbon surface. Subsequent reduction of the 4-nitrobenzene yields a 4-aminobenzene modified carbon surface. Drosophila melanogaster acetylcholinesterase was immobilized either covalently onto dialdehyde modified electrodes or non-covalently onto polyethyleneimine modified electrodes. Internal diffusion limitations due to the dialdehyde and polyethyleneimine modifications increased the apparent Km of the immobilized enzyme. The thiocholine sensitivity was about 90% for dialdehyde modified electrodes and about 10% for polyethyleneimine modified electrodes as compared with non-modified carbon electrodes. The detection limit of the biosensors produced by non-covalent immobilization of acetylcholinesterase onto polyethyleneimine modified carbon electrodes was found to be about 10(-10) M for the organophosphate pesticide dichlorvos.     

Mechanical Property Evolution of Silicon Composite Electrodes Studied by Environmental Nanoindentation

Mechanical degradation is largely responsible for the short cycle life of silicon (Si)‐based electrodes for future lithium‐ion batteries. An improved fundamental understanding of the mechanical behavior of Si electrodes, which evolves, as demonstrated in this paper, with the state of charge (SOC) and the cycle number, is a prerequisite for overcoming mechanical degradation and designing high capacity and durable Si‐based electrodes. In this study, Young's modulus (E) and hardness (H) of Si composite electrodes at different SOCs and after different cycle numbers are measured by nanoindentation under both dry and wet (liquid electrolyte) conditions. Unlike electrodes made of Si alone, E and H values of Si composite electrodes increase with increasing Li concentration. The composite electrodes under wet conditions are softer than that under dry conditions. Both E and H decrease with the cycle number. These findings highlight the effects of porosity, liquid environment, and degradation on the mechanical behavior of composite electrodes. The methods and results of this study on the mechanical property evolution of Si/polyvinylidene fluoride electrodes form a basis for exploring more effective binders for Si‐based electrodes. Furthermore, the evolving nature of the mechanical behavior of composite electrodes should be taken into consideration in future modeling efforts of porous composite electrodes.     

Factors influencing the properties of voltammetric carbon fibre electrodes: the importance of the pH of the medium used for the electrical treatment and of the resin coating of the fibres

Electrical treatment of resin-coated voltammetric carbon fibre electrodes with triangular voltage at low (1.1) pH resulted in electrodes almost insensitive to ascorbic acid (AA) and dihydroxyphenylacetic acid (DOPAC) while their response to 5-hydroxyindoleacetic acid (5-HIAA) was practically the same as after treatment in the usually employed pH 7.4 medium. Electrodes treated at high pH (12.0), on the other hand, were more sensitive to AA and DOPAC than those treated at pH 7.4 and less sensitive to 5-HIAA. Exposing resin-coated electrodes to the treatment media without electrical treatment was not sufficient to obtain the same results as with the application of the current. Electrodes without resin coating were sensitive to AA without electrical treatment while coated electrodes were not. Electrical treatment increased the sensitivity of non-coated electrodes and rendered coated electrodes even more sensitive than non-coated ones. Treatment of coated electrodes for a maximum sensitivity to 5-HIAA was found to require less time than to obtain maximum sensitivity to the other compounds. Present results suggest that it is possible to prepare selective voltammetric electrodes by choosing the right parameters for their electrical pretreatment.     

Branched EMG electrodes for stable and selective recording of single motor unit potentials in humans.

Branched surface EMG electrodes are bipolar electrodes with the hot signal pole referenced to two or more short-circuited leading-off surfaces. This technique provides stable recording of single motor unit potentials during real movements, up to maximal muscle contractions. The selective characteristic of branched electrodes is based on the same principles as the double differential detection system and spatial filtering technique proposed later. Equi-weight calculations to assess the selectivity of different electrode types and their position are used. The main advantage of branched electrodes, especially high stability, is achieved by the wire electrode version. The design, manufacture, implementation, and application of wire electrodes are discussed in detail. During recording of motor unit potentials, electrodes are positioned subcutaneously over the muscle fascia. This positioning maximizes electrode stability. Appropriate orientation of the electrode relative to the muscle architecture ensures adequate selectivity for single motor unit recordings. Branched electrodes require ordinary EMG equipment (two or even one amplifier).     

Enhancing the yield of high-density electrode arrays through automated electrode selection

Recently developed CMOS-based microprobes contain hundreds of electrodes on a single shaft with inter-electrode distances as small as 30 μm. So far, neuroscientists needed to select electrodes manually from hundreds of electrodes. Here we present an electronic depth control algorithm that allows to select electrodes automatically, hereby allowing to reduce the amount of data and locating those electrodes that are close to neurons. The electrodes are selected according to a new penalized signal-to-noise ratio (PSNR) criterion that demotes electrodes from becoming selected if their signals are redundant with previously selected electrodes. It is shown that, using the PSNR, interneurons generating smaller spikes are also selected. We developed a model that aims to evaluate algorithms for electronic depth control, but also generates benchmark data for testing spike sorting and spike detection algorithms. The model comprises a realistic tufted pyramidal cell, non-tufted pyramidal cells and inhibitory interneurons. All neurons are synaptically activated by hundreds of fibers. This arrangement allows the algorithms to be tested in more realistic conditions, including backgrounds of synaptic potentials, varying spike rates with bursting and spike amplitude attenuation.     

EMG signals detection and processing for on-line control of functional electrical stimulation.

The surface EMG signal detected from voluntarily activated muscles can be used as a control signal for functional neuromuscular electrical stimulation. A proper positioning of the recording electrodes in relation to the stimulation electrodes, and a proper processing of the recorded signals is required to reduce the stimulus artefact and the non-voluntary contribution (M-wave). Six orientations and six locations of the recording electrodes were investigated in the present work. A comb filter (with and without a blanking windowing) was applied to remove the signal components synchronously correlated to the stimulus. An operative definition of the signal to noise ratio and an efficiency index were implemented. It resulted that when the recording electrodes were located within the two stimulation electrodes the best orientation was perpendicular to the longitudinal line. However the best absolute indexes were obtained when the recording electrodes were located externally of the stimulation electrodes, and in that case the best orientation was longitudinal. Concerning the filtering procedure, the use of a blanking window before the application of the comb filter, gave the best performance.     

Which Disposable Chest Electrode?

Chest electrodes are preferred to limb electrodes for cardiac monitoring, as limb movements are not restricted and produce less interference of the E.C.G. trace. Eight types of disposable chest electrodes were investigated to compare their performance, skin reactions, cost, ease of application, size, and skin–electrode impedance.Elema-Schonander electrodes were found to be the most efficient and the most expensive. In their application care was required to avoid severe skin reactions.Dracard electrodes were simple to attach, worked well without severe skin reactions, and were cheap. They are recommended for routine use. Smith and Nephew electrodes, a type of “multipoint electrodes” which do not require electrode jelly, frequently produced severe skin reactions, making them unsuitable for monitoring for periods exceeding 12 hours.     

A motorized microdrive for recording of neural ensembles in awake behaving rats

The recording of neural ensembles in awake, behaving rats has been an extremely successful experimental paradigm, providing demonstrable scientific advances. Dynamic control of the position of the implanted electrodes is of key importance as mobile electrodes provide a better signal-to-noise ratio and a better cell/ electrode yield than nonmobile electrodes. Here we describe the use of low cost, soon to be commercially available dc motors to successfully control the depth of electrodes. The prototype designed is approximately 30 mm in diameter and 50 mm in length and weighed about 30 gms. This paper presents the results of linear displacements of electrodes achievable with this motorized microdrive.     

Assessing the validity of surface electromyography for recording muscle activation patterns from serratus anterior

PurposeNo direct evidence exists to support the validity of using surface electrodes to record muscle activity from serratus anterior, an important and commonly investigated shoulder muscle. The aims of this study were to determine the validity of examining muscle activation patterns in serratus anterior using surface electromyography and to determine whether intramuscular electromyography is representative of serratus anterior muscle activity.MethodsSeven asymptomatic subjects performed dynamic and isometric shoulder flexion, extension, abduction, adduction and dynamic bench press plus tests. Surface electrodes were placed over serratus anterior and around intramuscular electrodes in serratus anterior. Load was ramped during isometric tests from 0% to 100% maximum load and dynamic tests were performed at 70% maximum load. EMG signals were normalised using five standard maximum voluntary contraction tests.ResultsSurface electrodes significantly underestimated serratus anterior muscle activity compared with the intramuscular electrodes during dynamic flexion, dynamic abduction, isometric flexion, isometric abduction and bench press plus tests. All other test conditions showed no significant differences including the flexion normalisation test where maximum activation was recorded from both electrode types. Low correlation between signals was recorded using surface and intramuscular electrodes during concentric phases of dynamic abduction and flexion.ConclusionsIt is not valid to use surface electromyography to assess muscle activation levels in serratus anterior during isometric exercises where the electrodes are not placed at the angle of testing and dynamic exercises. Intramuscular electrodes are as representative of the serratus anterior muscle activity as surface electrodes.     

Comparison of colloidal gold electrode fabrication methods: the preparation of a horseradish peroxidase enzyme electrode.

In order to prepare biosensing electrodes which respond to hydrogen peroxide, horseradish peroxidase has been adsorbed to colloidal gold sols and electrodes prepared by deposition of these enzyme-gold sols onto glassy carbon using three methods: evaporation, electrodeposition and electrolyte deposition. In the latter method the enzyme-gold sol is applied to the surface of a glassy carbon disk electrode followed by an equal volume of 2 mM CaCl2. The electrolyte causes the sol to precipitate on the electrode surface, producing an immobilized enzyme electrode. Satisfactory electrodes which gave an electrochemical response to hydrogen peroxide in the presence of the electron transfer mediator ferrocenecarboxylic acid were produced by all three methods. Evaporation of horseradish peroxidase-gold sols produced electrodes with the best reproducibility and the widest linear amperometric response range. These electrodes can also easily be stored in a dry state. Although not as good as evaporation, electrodeposition also produced satisfactory electrodes. Electro-deposition provides the added advantage that it lends itself to the preparation of multi-enzyme/multi-analyte electrodes by the adsorption of different enzymes to separate gold sols, followed by sequential electrodeposition onto discrete areas of a multichannel electrode.     

Preparation of nanoparticles by electrocoagulation from soluble exopolysaccharide produced by Claviceps viridis

Electrocoagulation is an evolving technology that has been effectively applied for wastewater treatment but its applications in biotechnology and nanotechnology are very limited. This method was applied for the preparation of nanoparticles from soluble exopolysaccharide (EPS) produced by Claviceps viridis in a submerged batch culture. A cathode/anode pair electrode (Al or Fe) system was used for determination of the separation rates of electrocoagulation and the yields of EPS nanoparticles production. The separation rates of 0.170 +/- 0.003 mg EPS/sec (Fe electrodes) and 0.250 +/- 0.003 mg EPS/sec (Al electrodes) were calculated for voltage gradient 1 V/1 cm of electrodes distance and were constant during experiments. The specific yield of EPS nanoparticles production based on the consumed electric power was dependent on the material of the electrodes and its value was determined as 0.71 +/- 0.01 mg EPS/W for Fe electrodes and 0.91 +/- 0.01 mg EPS/W for Al electrodes, respectively.     

Electrical contacting of redox proteins by nanotechnological means

Redox enzymes in bioelectronic devices usually lack direct electrical contact with electrodes, owing to the spatial separation of their redox centers from the conductive surfaces by the protein shells. The reconstitution of apo-enzymes on cofactor-functionalized nanostructures associated with electrodes provides a means to align the biocatalysts on the conductive surface and to electrically contact redox enzymes with electrodes. The reconstitution of apo-enzymes on cofactor-functionalized gold nanoparticles or carbon nanotubes has led to effective electrical communication between the redox proteins and the electrodes. Alternatively, the reconstitution of redox enzymes on molecular wires that enable electron tunneling or dynamic charge shuttling represent supramolecular biocatalytic nanostructures exhibiting electrical contact. The bioelectrocatalytic activities of the electrically wired reconstituted enzymes on electrodes have allowed the development of amperometric biosensors and biofuel cell elements.     

Do surface electrodes validly represent lower trapezius activation patterns during shoulder tasks?

Because of its superficial location, surface electrodes are commonly used to record lower trapezius activity. Recent evidence, however, would suggest that surface electromyography is not a valid to record activity from other superficially placed shoulder muscles. Therefore, the aim of this study was to determine the validity of using surface electrodes to record lower trapezius activity. Ten asymptomatic subjects performed ramped isometric (0–100% maximum load) and dynamic (70% maximum load) shoulder tasks. Intramuscular electrodes were inserted into lower trapezius and rhomboid major. Surface electrodes were placed over lower trapezius around the intramuscular electrodes. Differences in the recorded activity of lower trapezius between surface and intramuscular electrodes were tested using a 2 factor repeated measures analysis of variance with factors: test and electrode type. Similarity in the recorded activity patterns between the two electrodes was tested using Pearson’s correlation coefficient (r). Results indicated that there was no difference in lower trapezius activity levels (p = 0.98) or activation patterns (r ≥ 0.74) recorded by the intramuscular and surface electrodes. The results of this study indicate that any potential crosstalk contamination in the surface electrode signal is having little influence on the recorded activity from lower trapezius and therefore, support the common practice of surface electromyography to investigate lower trapezius function.     

Characteristics of spinal cord-evoked responses in man

The averaged electrical potentials evoked by the stimulation of the peripheral nerves were recorded with surface electrodes over the lumbosacral, lower thoracic and cervical spine and with epidurally placed electrodes in the cervical area. The waveforms of the lumbosacral and cervical spinal cord potentials show similar complexity reflecting peripheral and central generators. The larger negative wave with at least two components is followed by a slower positive deflection. Evoked potentials recorded over the cervical segments of the spinal cord with epidural electrodes are of much higher amplitude and more complex waveform than those recorded with surface electrodes.     

Electrochemical reduction of cytochromes P450 using electrodes containing immobilized substrates

A new approach for the electrochemical reduction of cytochromes P450 (P450, CYPs) with electrodes chemically modified with CYP appropriate substrates (“reverse” electrodes) has been proposed. The method is based on the analysis of cyclic voltammograms, square wave voltammograms, amperograms and determination of such electrochemical characteristics as catalytic current and redox potential. The differences of maximal current and potentials in square wave voltammograms and catalytic current in amperometric measurements are more sensitive and reliable. The planar mode of screen-printed electrodes permits to use 20–60 μl of electrolyte volume. We have investigated P450 2B4-benzphetamine or P450scc-cholesterol enzyme-substrate pairs. Electrochemical parameters of electrodes with nonspecific P450 substrate were differed from the electrodes with appropriate substrates.     

Site-specific covalent attachment of heme proteins on self-assembled monolayers

Naturally occurring hemin cofactor has been functionalized to introduce two terminal alkyne groups. This modified hemin has been successfully covalently attached to mixed self-assembled monolayers of alkanethiols and azide-terminated alkanethiols on gold electrodes using a Cu(I)-catalyzed 1,3-cycloaddition reaction. However these hemin-modified electrodes could not be used to reconstitute apomyoglobin on gold electrodes owing to the hydrophobicity of the alkane thiol self-assembled monolayer. Modification of existing techniques allowed covalent attachment of alkyne-terminated electroactive species onto mixed monolayers of azidothiols and carboxylatoalkanethiols on electrodes using the same Cu(I)-catalyzed 1,3-cycloaddition reaction. Apomyoglobin could be reconstituted using the hemin covalently attached to these hydrophilic electrodes. The electrochemical data, UV-vis absorption data, surface-enhanced resonance Raman spectroscopy data, and atomic force microscopy data indicate the presence of these modified myoglobin proteins on these electrodes. The direct attachment of the heme cofactor of these modified myoglobin proteins to the electrode allows fast electron transfer to the heme center from the electrode and affords efficient O(2)-reducing bioelectrodes under physiological conditions.     

Development of a biochip using antibody-coated gold nanoparticles to detect specific bioparticles

This study developed a method of detecting bioparticles such as Salmonella that exist in the biological samples. The method employed a substrate with interlaced comb-like electrodes into which the mixtures of biological samples and antibody-coated gold nanoparticles were added. The alternative signals with appropriate frequency bands were then conducted into the comb-like electrodes to change the dielectrophoresis force. The gold-modified Salmonella can be adsorbed on the edges of the electrodes and isolated from various biological samples. The impedance of the adsorbed Salmonella on the edges of the electrodes was measured and comparison of the impedance between the electrodes with and without Salmonella can quantify the amount of the adsorbed Salmonella.     

Electrocatalytic oxidation of guanine, guanosine, and guanosine monophosphate

The electrochemical behavior of guanine, guanosine, and guanosine monophosphate (GMP) at redox polymer film modified indium tin oxide electrodes is examined by voltammetry and redox titration. Utilizing the redox polymer-coated electrodes as indicator electrodes, a new method for measuring the oxidation potentials, based on monitoring their catalytic oxidation by different redox polymer coated electrodes at different pH, was proposed in this work. The oxidation potentials of 0.81 V and 1.02 V versus normal hydrogen electrode were determined for guanine and guanosine/GMP under physiological conditions, the lowest oxidation potentials ever reported, to our knowledge.     

An extracellular microelectrode array for monitoring electrogenic cells in culture

This paper describes a planar array of microelectrodes developed for monitoring the electrical activity of cells in culture. The device allows the incorporation of surface topographical features in an insulating layer above the electrodes. Semiconductor technology is employed for the fabrication of the gold electrodes and for the deposition and patterning of an insulating layer of silicon nitride. The electrodes have been tested using a cardiac cell culture of chick embryo myocytes, and the physical beating of the cultured cells correlated with the simultaneous extracellular voltage measurements obtained. It was found that extracellular stimulation of the cells was possible via the same electrodes used for recording.     

Anterior zygomatic electrodes: a special electrode for the study of temporal lobe epilepsy

The position of a surface electrode over the anterior zygomatic arch is described. Epileptiform activity recorded at this position is compared with that recorded by sphenoidal electrodes in 21 cases of temporal lobe epilepsy. In 100% of the cases abnormalities were detected with both electrodes, although in 11% of the cases the findings could not be definitely described as epileptiform in the anterior zygomatic electrodes.