Affordable automated phenotypic antibiotic susceptibility testing method based on a contactless conductometric sensor.

User-friendly phenotypic antibiotic susceptibility testing (AST) methods are urgently needed in many fields including clinical medicine, epidemiological studies and drug research. Herein, we report a convenient and cost-effective phenotypic AST method based on online monitoring bacterial growth with a developed 8-channel contactless conductometric sensor (CCS). Using E. coli and V. parahaemolyticus as microorganism models, as well as enoxacin, florfenicol, ampicillin, kanamycin and sulfadiazine as antibiotic probes. The minimum inhibitory concentration (MIC) determination was validated in comparison with standard broth microdilution (BMD) assay. The total essential agreements between the CCS AST assays and the reference BMD AST assays are 68.8-92.3%. The CCS has an approximate price of $9,000 (USD). Requiring neither chemical nor biotic auxiliary materials for the assay makes the cost of each sample < $1. The MICs obtained with the automated CCS AST assays are more precise than those obtained with the manual BMD. Moreover, in 72 percent of the counterpart, the MICs obtained with the CCS AST assays are higher than that obtained with the BMD AST assays. The proposed CCS AST method has advantages in affordability, accuracy, sensitivity and user-friendliness.

Simultaneous Determination of Inorganic Anions and Cations in Water and Biological Samples by Capillary Electrophoresis with a Capacitive Coupled Contactless Conductivity Detector Using Capillary Filling Method.

An analytical method for concurrent analysis of inorganic anions and cations has been developed using a capillary electrophoresis (CE)-capacitively coupled contactless conductivity detector (C4D) system. Although hydrodynamic and electrokinetic injection techniques have been widely used in CE, we employed a capillary filling method (CFM) for the analysis of inorganic ions. The procedure is relatively simple and has the advantage that CMF does not require pressure control and vial exchange. Three anions (chloride, sulfate, nitrate) and five cations (ammonium, potassium, sodium, magnesium, calcium) were successfully separated and detected at ppm levels within 80 s using a 9 mM histidine/15 mM malic acid (pH 3.6) containing 50 mM N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate as background electrolyte. Applying this analytical condition, the electroosmotic flow is negligible and anions and cations were migrated concurrently to different polarities according to their electrophoretic mobility. Obtained raw data showed stepwise increases in detected conductivity due to the migration of sample components, which expresses as peak profiles by differentiation of electropherograms. The RSD values of the peak area and migration times for the anions and cations were satisfactory and were less than 5.15 and 2.04%, respectively. The developed method was applied for the analysis of inorganic anions and cations in commercial mineral waters, tap water, urine, and exhaled breath condensate. These results indicate that the CE-C4D system with CFM is suitable for the rapid analysis of inorganic anions and cations in various samples.

Conductometric biosensor for arginine determination in pharmaceutics.

A new conductometric biosensor based on coimmobilized urease and arginase has been developed for arginine determination in pharmaceutics. First, the main parameters of the selected method of immobilization (concentrations of arginase, urease, and glutaraldehyde, time of incubation) were optimized. An influence of the solution parameters (buffer ionic strength, capacity, pH, Mn2+ concentration) on the biosensor operation was studied, working conditions were optimized. After biosensor optimization, the main analytical characteristics were as follows. The limit of detection - 2.5 µM, the linear range - 2.5-500 µM, the sensitivity to arginine 13.4 ±â€¯2.4 µS/mM, the response time - 20 s. The signals repeatability and operational stability in continuous exploitation were studied over one working day and during one week. Additionally, the selectivity of the developed biosensor towards arginine was essayed relative to other amino acids. The developed biosensor has been used to measure arginine concentrations in some drugs. The results obtained were in high correlation with the characteristics declared by producers.

A sandwich-type electrochemical immunoassay for ultrasensitive detection of non-small cell lung cancer biomarker CYFRA21-1.

Many studies confirm that the aberrant expression of Cytokeratin 19 fragment 21-1 (CYFRA21-1) is highly correlated with non-small cell lung cancer (NSCLC), especially for squamous cell carcinoma. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to test CYFRA21-1 selectively and sensitively. The proposed immunosensor fabricated by three-dimensional graphene (3D-G), chitosan (CS) and glutaraldehyde (GA) composite on the glass carbon electrode (GCE) with a large surface area is prepared to immobilize primary antibodies (Ab1) and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with amino-functionalized carbon nanotube (MWCNT-NH2) nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab2) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CYFRA21-1 with a wide linear range from 0.1 to 150ng·mL-1 and a low detection limit (LOD) of 43pg·mL-1. Furthermore, satisfactory results are obtained for the determination of CYFRA21-1 in real clinical serum samples, indicating the potential of the immunoassay to be applied in clinical analysis.

Sensitive detection of pyoverdine with an electrochemical sensor based on electrochemically generated graphene functionalized with gold nanoparticles.

The design and development of an electrochemical sensor for the sensitive and selective determination of pyoverdine, a virulence factor secreted by Pseudomonas aeruginosa, bacteria involved in nosocomial infections is presented in this work. The presence of pyoverdine in water and body fluids samples can be directly linked to the presence of the Pseudomonas bacteria, thus being a nontoxic and low cost marker for the detection of water pollution as well as for the biological contamination of other media. The sensor was elaborated using layer-by-layer technique for the deposition of a graphene­gold nanoparticles composite film on the graphite-based screen printed electrode, from aqueous suspension. Under optimal conditions, the electrochemical signal corresponding to the pyoverdine oxidation process was proportional to its concentration, showing a wide linear range from 1 to 100µmolL-1 and a detection limit of 0.33µmolL-1. This sensor discriminate with satisfactory recoveries the target analyte in different real matrices and also exhibited low response to other interfering species, proving that this technique is promising for medical and environmental applications. In addition, the proposed nanocomposite platform presented good reproducibility, high and long term stability, the sensitivity for pyoverdine remain unchanged after being stored at 4°C for four weeks.

Adsorption/desorption of biomacromolecules involved in catalytic hydrogen evolution.

Previously, it has been shown that proteins and some polysaccharides (PSs) catalyse hydrogen evolution, producing electrochemical signals on mercury electrodes. The catalytic hydrogen evolution reaction (CHER) of the above-mentioned biomacromolecules was studied by voltammetric and chronopotentiometric stripping (CPS) methods. To obtain more information about electrode processes involving CHER, here we used protein such as BSA, and chitosan as a PS; in addition, we investigated dextran as a control PS not involved in CHER. We studied biomacromolecules by phase-sensitive alternating current (AC) voltammetry. Using phase-in AC voltammetry, for CHER-involved biomacromolecules we observed a CHER peak at highly negative potentials, similar to that observed with other voltammetric and CPS methods. On the other hand, by means of the adsorption/desorption processes studied in phase-out AC voltammetry, we uncovered a sharp and narrow decrease of capacitive current in the potential range of the CHER peak, denominated as the tensammetric minimum. This minimum was closely related to the CHER peak, as demonstrated by similar dependences on specific conditions affecting the CHER peak such as buffer capacity and pH. A tensammetric minimum was not observed for dextran. Our results suggest specific organization of biopolymer layers at negative potentials observed only in biomacromolecules involved in CHER.

The strip method and the microelectrode technique in assessing dental plaque pH.

BACKGROUND: The pH strip method has been proposed for use in a clinical setting for the evaluation of dental plaque pH. The aim of this in-vivo study was to compare the reliability of the pH-indicator strip method to the microelectrode technique in plaque pH measurement. METHODS: 136 subjects (7-9 years) were enrolled and, for each subject, measurements of plaque pH were made at two interproximal sites; a total of 272 sites were analyzed. Plaque pH was assessed in triplicate using pH-indicator strips (pH range: 4.0-7.0) and an iridium microelectrode with a diameter of 0.1 mm. The caries status of 544 proximal surfaces between two primary molars, in proximity at the plaque pH sites, was assessed. All measurements were performed before and 2, 5, 10, 15, 20, and 30 minutes after a mouth rinse with water solution of sucrose (10%). One-way ANOVA was performed to analyze statistically significant differences between the two techniques. Instrument reliability was measured by assessing the intraclass correlation coefficient. RESULTS: Comparable, not statistically significant pH values (P values range from 0.98 to 1.00) were obtained by the two methods for AUC5.7, AUC6.2, maximum pH fall and minimum pH. High intraclass coefficients were recorded (Pearson's r=0.96, Yule's Q=0.99), implying an almost perfect association when the measurements were grouped as a carious lesion being present or not on the surface near the site of measurement. CONCLUSIONS: The use of the pH strip method has high validity and may replace the traditional microtouch electrode technique for clinical use.

Nano-Composite Foam Sensor System in Football Helmets.

American football has both the highest rate of concussion incidences as well as the highest number of concussions of all contact sports due to both the number of athletes and nature of the sport. Recent research has linked concussions with long term health complications such as chronic traumatic encephalopathy and early onset Alzheimer's. Understanding the mechanical characteristics of concussive impacts is critical to help protect athletes from these debilitating diseases and is now possible using helmet-based sensor systems. To date, real time on-field measurement of head impacts has been almost exclusively measured by devices that rely on accelerometers or gyroscopes attached to the player's helmet, or embedded in a mouth guard. These systems monitor motion of the head or helmet, but do not directly measure impact energy. This paper evaluates the accuracy of a novel, multifunctional foam-based sensor that replaces a portion of the helmet foam to measure impact. All modified helmets were tested using a National Operating Committee Standards for Athletic Equipment-style drop tower with a total of 24 drop tests (4 locations with 6 impact energies). The impacts were evaluated using a headform, instrumented with a tri-axial accelerometer, mounted to a Hybrid III neck assembly. The resultant accelerations were evaluated for both the peak acceleration and the severity indices. These data were then compared to the voltage response from multiple Nano Composite Foam sensors located throughout the helmet. The foam sensor system proved to be accurate in measuring both the HIC and Gadd severity index, as well as peak acceleration while also providing additional details that were previously difficult to obtain, such as impact energy.

Graphene Field-Effect Transistors for the Sensitive and Selective Detection of Escherichia coli Using Pyrene-Tagged DNA Aptamer.

This study reports biosensing using graphene field-effect transistors with the aid of pyrene-tagged DNA aptamers, which exhibit excellent selectivity, affinity, and stability for Escherichia coli (E. coli) detection. The aptamer is employed as the sensing probe due to its advantages such as high stability and high affinity toward small molecules and even whole cells. The change of the carrier density in the probe-modified graphene due to the attachment of E. coli is discussed theoretically for the first time and also verified experimentally. The conformational change of the aptamer due to the binding of E. coli brings the negatively charged E. coli close to the graphene surface, increasing the hole carrier density efficiently in graphene and achieving electrical detection. The binding of negatively charged E. coli induces holes in graphene, which are pumped into the graphene channel from the contact electrodes. The carrier mobility, which correlates the gate voltage to the electrical signal of the APG-FETs, is analyzed and optimized here. The excellent sensing performance such as low detection limit, high sensitivity, outstanding selectivity and stability of the graphene biosensor for E. coli detection paves the way to develop graphene biosensors for bacterial detection.

Novel synthesis and characterization of pristine Cu nanoparticles for the non-enzymatic glucose biosensor.

Non enzymatic electrochemical glucose sensing was developed based on pristine Cu Nanopartilces (NPs)/Glassy Carbon Electrode (GCE) which can be accomplished by simple green method via ocimum tenuiflorum leaf extract. Then, the affect of leaf extract addition on improving Structural, Optical and electrochemical properties of pristine cu NPs was investigated. The synthesized Cu NPs were characterized with X-ray diffraction (X-ray), Uv-Visible spectroscopy (Uv-Vis), Fourier transformation infrared spectroscopy (FTIR), Particle size distribution (PSA), Scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), Transmission electron microscopy (TEM) for structural optical and morphological studies respectively. The synthesized Cu NPs were coated over glassy carbon electrode (GCE) to study the electrochemical response of glucose by cyclic voltammetry and ampherometer. The results indicates that the modified biosensor shows a remarkable sensitivity (1065.21 µA mM-1 cm-2), rapid response time (<3s), wide linear range (1 to 7.2 mM), low detection limit (0.038 µM at S/N = 3). Therefore, the prepared Cu NPs by the Novel Bio-mediated route were exploited to construct a non-enzymatic glucose biosensor for sustainable clinical field applications.

Glucose monitoring system using nanopellets.

The combination of the fields of software engineering, gadgets, and science has stood out among the most revolutionary future innovations. Health issues have been the focus of various engaging and explanatory studies. One such health-related dilemma is diabetes. Diabetes at its serious stage results in impaired vision. Increase in the glucose level is a critical parameter that could result in hyperglycaemia, hypoglycaemia, massive heart attack, strokes, and aneurysms. Monitoring the glucose level in blood is one of the control measures for diabetes in the affected population. A glucose monitoring framework interminably measures and screens the glucose level in blood. A novel framework for measuring the glucose level is proposed in this study. This study employs nanopellets that evaluate the glucose level. When the glucose level increases or decreases, it is continuously recorded and displayed using a microcontroller (mixed signal processor (MSP) 430). The data are then sent to the physician through global system for mobile communication. The typical blood glucose level of human being ranges from 70 to 110 mg/dl. When the insulin level builds up to certain point, hyperglycaemia occurs. When decreases, hypoglycaemia occurs. Hyperglycaemia leads to cataracts, oedema, hypertension, polyuria, and polydipsia. Hypoglycaemia causes perplexity, energy, insensateness, coma, and death.

Novel photoelectrochemical immunosensor for disease-related protein assisted by hemin/G-quadruplex-based DNAzyme on gold nanoparticles to enhance cathodic photocurrent on p-CuBi2O4 semiconductor.

A novel p-type semiconductor material (p-CuBi2O4) is designed for the construction of split-type photoelectrochemical (PEC) immunosensor for alpha-fetoprotein (AFP) with the hemin assistant to enhance the cathodic photocurrent. Initially, the photocathode of PEC immunosensor is fabricated by p-CuBi2O4 on a layer of gold nanoparticles (AuNPs, as a front contact of p-CuBi2O4) to enhance the efficiency of charge separation. In the presence of target AFP, a sandwich-type immunoreaction was carried out in capture antibody-coated microplate by using detection antibody and hemin-based G-quadruplex (labeled on the AuNP) as the signal probe. Upon exonuclease I (Exo I) introduction, the enzyme digested the hemin/G-quadruplex-based DNAzyme to release the hemin[Fe(III)], which captured the generated electrons of p-CuBi2O4-based photocathode to enhance photocurrent via the reduction of hemin[Fe(III)] to hemin[Fe(II)] in PEC detection system. Under the optimal conditions, the split-type photocathodic immunosensor showed a wide linear dynamic range from 50pgmL-1 to 20ngmL-1 at a limit of detection (LOD) of 14.7pgmL-1 toward target AFP. Moreover, the PEC immunosensor also displayed high specificity and good reproducibility. Favorably, method accuracy was evaluated to analyze human serum specimens, and gave matched-well results in comparison with commercially available enzyme-linked immunosorbent assay (ELISA) method.

Gait Phase Estimation Based on Noncontact Capacitive Sensing and Adaptive Oscillators.

This paper presents a novel strategy aiming to acquire an accurate and walking-speed-adaptive estimation of the gait phase through noncontact capacitive sensing and adaptive oscillators (AOs). The capacitive sensing system is designed with two sensing cuffs that can measure the leg muscle shape changes during walking. The system can be dressed above the clothes and free human skin from contacting to electrodes. In order to track the capacitance signals, the gait phase estimator is designed based on the AO dynamic system due to its ability of synchronizing with quasi-periodic signals. After the implementation of the whole system, we first evaluated the offline estimation performance by experiments with 12 healthy subjects walking on a treadmill with changing speeds. The strategy achieved an accurate and consistent gait phase estimation with only one channel of capacitance signal. The average root-mean-square errors in one stride were 0.19 rad (3.0% of one gait cycle) for constant walking speeds and 0.31 rad (4.9% of one gait cycle) for speed transitions even after the subjects rewore the sensing cuffs. We then validated our strategy in a real-time gait phase estimation task with three subjects walking with changing speeds. Our study indicates that the strategy based on capacitive sensing and AOs is a promising alternative for the control of exoskeleton/orthosis.

Amperometric L-lysine enzyme electrodes based on carbon nanotube/redox polymer and graphene/carbon nanotube/redox polymer composites.

Highly sensitive L-lysine enzyme electrodes were constructed by using poly(vinylferrocene)-multiwalled carbon nanotubes-gelatine (PVF/MWCNTs-GEL) and poly(vinylferrocene)-multiwalled carbon nanotubes-gelatine-graphene (PVF/MWCNTs-GEL/GR) composites as sensing interfaces and their performances were evaluated. Lysine oxidase (LO) was immobilized onto the composite modified glassy carbon electrodes (GCE) by crosslinking using glutaraldehyde and bovine serum albumin. Effects of pH value, enzyme loading, applied potential, electrode composition, and interfering substances on the amperometric response of the enzyme electrodes were discussed. The analytical characteristics of the enzyme electrodes were also investigated. The linear range, detection limit, and sensitivity of the LO/PVF/MWCNTs-GEL/GCE were 9.9 × 10-7-7.0 × 10-4 M, 1.8 × 10-7 M (S/N = 3), and 13.51 µA mM-1 cm-2, respectively. PVF/MWCNTs-GEL/GR-based L-lysine enzyme electrode showed a short response time (<5 s) and a linear detection range from 9.9 × 10-7 to 7.0 × 10-4 M with good sensitivity of 17.8 µA mM-1 cm-2 and a low detection limit of 9.2 × 10-8 M. The PVF/MWCNTs-GEL/GR composite-based L-lysine enzyme electrode exhibited about 1.3-fold higher sensitivity than its MWCNTs-based counterpart and its detection limit was superior to the MWCNTs-based one. In addition, enzyme electrodes were successfully applied to determine L-lysine in pharmaceutical sample and cheese.

Electrochemical techniques reveal that total ammonium stress increases electron flow to anode respiration in mixed-species bacterial anode biofilms.

When anode-respiring bacteria (ARB) respire electrons to an anode in microbial electrochemical cells (MXCs), they harvest only a small amount of free energy. This means that ARB must have a high substrate-oxidation rate coupled with a high ratio of electrons used for respiration compared to total electrons removed by substrate utilization. It also means that they are especially susceptible to inhibition that slows anode respiration or lowers their biomass yield. Using several electrochemical techniques, we show that a relatively high total ammonium-nitrogen (TAN) concentration (2.2 g TAN/L) induced significant stress on the ARB biofilms, lowering their true yield and forcing the ARB to boost the ratio of electrons respired per electrons consumed from the substrate. In particular, a higher respiration rate, measured as current density (j), was associated with slower growth and a lower net yield, compared to an ARB biofilm grown with a lower ammonium concentration (0.2 g TAN/L). Further increases in influent TAN (to 3 and then to 4.4 g TAN/L) caused nearly complete inhibition of anode respiration. However, the ARB could recover from high-TAN inhibition after a shift of the MXC's feed to 0.2 g TAN/L. In summary, ARB biofilms were inhibited by a high TAN concentration, but could divert more electron flow toward anode respiration with modest inhibition and recover when severe inhibition was relieved. Biotechnol. Bioeng. 2017;114: 1151-1159. © 2017 Wiley Periodicals, Inc.

Noncontact Capacitive Sensing-Based Locomotion Transition Recognition for Amputees With Robotic Transtibial Prostheses.

Recent advancement of robotic transtibial prostheses can restore human ankle dynamics in different terrains. Automatic locomotion transitions of the prosthesis guarantee the amputee's safety and smooth motion. In this paper, we present a noncontact capacitive sensing-based approach for recognizing locomotion transitions of amputees with robotic transtibial prostheses. The proposed sensing system is designed with flexible printed circuit boards which solves the walking instability brought by our previous system when using robotic prosthesis and improves the recognition performance. Six transtibial amputees were recruited and performed tasks of ten locomotion transitions with the robotic prosthesis that we recently constructed. The capacitive sensing system was integrated on the prosthesis and worked in combination with on-prosthesis mechanical sensors. With the cascaded classification method, the proposed system achieved 95.8% average recognition accuracy by support vector machine (SVM) classifier and 94.9% accuracy by quadratic discriminant analysis (QDA) classifier. It could accurately recognize the upcoming locomotion modes from the stance phase of the transition steps. In addition, we proved that adding capacitance signals could significantly reduce recognition errors of the robotic prosthesis in locomotion transition tasks. Our study suggests that the fusion of capacitive sensing system and mechanical sensors is a promising alternative for controlling the robotic transtibial prosthesis.

Estimation of Finger Joint Angles Based on Electromechanical Sensing of Wrist Shape.

An approach to finger motion capture that places fewer restrictions on the usage environment and actions of the user is an important research topic in biomechanics and human-computer interaction. We proposed a system that electrically detects finger motion from the associated deformation of the wrist and estimates the finger joint angles using multiple regression models. A wrist-mounted sensing device with 16 electrodes detects deformation of the wrist from changes in electrical contact resistance at the skin. In this study, we experimentally investigated the accuracy of finger joint angle estimation, the adequacy of two multiple regression models, and the resolution of the estimation of total finger joint angles. In experiments, both the finger joint angles and the system output voltage were recorded as subjects performed flexion/extension of the fingers. These data were used for calibration using the least-squares method. The system was found to be capable of estimating the total finger joint angle with a root-mean-square error of 29-34 degrees. A multiple regression model with a second-order polynomial basis function was shown to be suitable for the estimation of all total finger joint angles, but not those of the thumb.

Glucose Monitoring in Individuals With Diabetes Using a Long-Term Implanted Sensor/Telemetry System and Model.

OBJECTIVE: The use of a fully implanted first-generation prototype sensor/telemetry system is described for long-term monitoring of subcutaneous tissue glucose in a small cohort of people with diabetes. METHODS: Sensors are based on a membrane containing immobilized glucose oxidase and catalase coupled to oxygen electrodes and a telemetry system, integrated as an implant. The devices remained implanted for up to 180 days, with signals transmitted every 2 min to external receivers. RESULTS: The data include signal recordings from glucose clamps and spontaneous glucose excursions, matched, respectively, to reference blood glucose and finger-stick values. The sensor signals indicate dynamic tissue glucose, for which there is no independent standard, and a model describing the relationship between blood glucose and the signal is, therefore, included. The values of all model parameters have been estimated, including the permeability of adjacent tissues to glucose, and equated to conventional mass transfer parameters. As a group, the sensor calibration varied randomly at an average rate of -2.6%/week. Statistical correlation indicated strong association between the sensor signals and reference glucose values. CONCLUSION: Continuous long-term glucose monitoring in individuals with diabetes is feasible with this system. SIGNIFICANCE: All therapies for diabetes are based on glucose control, and therefore, require glucose monitoring. This fully implanted long-term sensor/telemetry system may facilitate a new era of management of the disease.

Thermoelectric detection and imaging of propagating graphene plasmons.

Controlling, detecting and generating propagating plasmons by all-electrical means is at the heart of on-chip nano-optical processing. Graphene carries long-lived plasmons that are extremely confined and controllable by electrostatic fields; however, electrical detection of propagating plasmons in graphene has not yet been realized. Here, we present an all-graphene mid-infrared plasmon detector operating at room temperature, where a single graphene sheet serves simultaneously as the plasmonic medium and detector. Rather than achieving detection via added optoelectronic materials, as is typically done in other plasmonic systems, our device converts the natural decay product of the plasmon-electronic heat-directly into a voltage through the thermoelectric effect. We employ two local gates to fully tune the thermoelectric and plasmonic behaviour of the graphene. High-resolution real-space photocurrent maps are used to investigate the plasmon propagation and interference, decay, thermal diffusion, and thermoelectric generation.

Artificial Nose Technology: Status and Prospects in Diagnostics.

Biomimetic crossreactive sensor arrays have been used to detect and analyze a wide variety of vapor and liquid components in applications such as food science, public health and safety, and diagnostics. As technology has advanced over the past three decades, these systems have become selective, sensitive, and affordable. Currently, the need for noninvasive and accurate devices for early disease diagnosis remains a challenge. This Opinion article provides an overview of the various types of biomimetic crossreactive sensor arrays (also referred to as electronic noses or tongues in the literature), their current use and future directions, and an outlook for future technological development.