Label-Free, Highly Sensitive Electrochemical Aptasensors Using Polymer-Modified Reduced Graphene Oxide for Cardiac Biomarker Detection

Acute myocardial infarction (AMI), also recognized as a ???heart attack,??? is one leading cause of death globally, and cardiac myoglobin (cMb), an important cardiac biomarker, is used for the early assessment of AMI. This paper presents an ultrasensitive, label-free electrochemical aptamer-based sensor (aptasensor) for cMb detection using polyethylenimine (PEI)-functionalized reduced graphene oxide (PEI???rGO) thin films. PEI, a cationic polymer, was used as a reducing agent for graphene oxide (GO), providing highly positive charges on the rGO surface and allowing direct immobilization of negatively charged single-strand DNA aptamers against cMb via electrostatic interaction without any linker or coupling chemistry. The presence of cMb was detected on Mb aptamer-modified electrodes using differential pulse voltammetry via measuring the current change due to the direct electron transfer between the electrodes and cMb proteins (Fe3+/Fe2+). The limits of detection were 0.97 pg mL???1 (phosphate-buffered saline) and 2.1 pg mL???1 (10-fold-diluted human serum), with a linear behavior with logarithmic cMb concentration. The specificity and reproducibility of the aptasensors were also examined. This electrochemical aptasensor using polymer-modified rGO shows potential for the early assessment of cMb in point-of-care testing applications

Flexible electrical aptasensor using dielectrophoretic assembly of graphene oxide and its subsequent reduction for cardiac biomarker detection

Cardiac troponin T (cTnT) is considered a clinical standard for its high specificity and sensitivity when diagnosing acute myocardial infarction; however, most studies on the electrical sensors of cardiac troponin biomarkers have focused on cTnI rather than cTnT. This study presents label-free, low-cost, transparent, and flexible aptamer-based immunosensors for the electrical detection of cTnT using reduced graphene oxide (rGO) sheets. GO was first deposited by AC dielectrophoresis between two predefined source and drain electrodes on a 3-aminopropyltriethoxylsilane-modified polyethylene terephthalate substrate. The GO was then reduced using hydrazine vapour without damaging the substrate, resulting in uniform, controlled, and stable deposition of rGO sheets, and demonstrating more stability than those directly deposited by dielectrophoresis. Amine-modified single-strand DNA aptamers against cTnT were immobilized onto the rGO channels. The relative resistance change of this sensor owing to the attachment of cTnT was quantified as the cTnT concentration decreased from 10???ng/mL to 1???pg/mL in phosphate buffered saline (PBS) and 10-fold diluted human serum in PBS, with the limits of detection being 1.2???pg/mL and 1.7???pg/mL, respectively, which is sufficiently sensitive for clinical applications. High-yield and rapid fabrication of the present rGO sensors will have significant influences on scaled-up fabrication of graphene-based sensors

Mal-Netminer: Malware Classification Approach based on Social Network Analysis of System Call Graph

As the security landscape evolves over time, where thousands of species of malicious codes are seen every day, antivirus vendors strive to detect and classify malware families for efficient and effective responses against malware campaigns. To enrich this effort, and by capitalizing on ideas from the social network analysis domain, we build a tool that can help classify malware families using features driven from the graph structure of their system calls. To achieve that, we first construct a system call graph that consists of system calls found in the execution of the individual malware families. To explore distinguishing features of various malware species, we study social network properties as applied to the call graph, including the degree distribution, degree centrality, average distance, clustering coefficient, network density, and component ratio. We utilize features driven from those properties to build a classifier for malware families. Our experimental results show that influence-based graph metrics such as the degree centrality are effective for classifying malware, whereas the general structural metrics of malware are less effective for classifying malware. Our experiments demonstrate that the proposed system performs well in detecting and classifying malware families within each malware class with accuracy greater than 96%.Comment: Mathematical Problems in Engineering, Vol 201

Cost-Effective and Handmade Paper-Based Immunosensing Device for Electrochemical Detection of Influenza Virus

Although many studies concerning the detection of influenza virus have been published, a paper-based, label-free electrochemical immunosensor has never been reported. Here, we present a cost-effective, handmade paper-based immunosensor for label-free electrochemical detection of influenza virus H1N1. This immunosensor was prepared by modifying paper with a spray of hydrophobic silica nanoparticles, and using stencil-printed electrodes. We used a glass vaporizer to spray the hydrophobic silica nanoparticles onto the paper, rendering it super-hydrophobic. The super-hydrophobicity, which is essential for this paper-based biosensor, was achieved via 30-40 spray coatings, corresponding to a 0.39-0.41 mg cm(-2) coating of nanoparticles on the paper and yielding a water contact angle of 150 degrees +/- 1 degrees. Stencil-printed carbon electrodes modified with single-walled carbon nanotubes and chitosan were employed to increase the sensitivity of the sensor, and the antibodies were immobilized via glutaraldehyde cross-linking. Differential pulse voltammetry was used to assess the sensitivity of the sensors at various virus concentrations, ranging from 10 to 10(4) PFU mL(-1), and the selectivity was assessed against MS2 bacteriophages and the influenza B viruses. These immunosensors showed good linear behaviors, improved detection times (30 min), and selectivity for the H1N1 virus with a limit of detection of 113 PFU mL(-1), which is sufficiently sensitive for rapid on-site diagnosis. The simple and inexpensive methodologies developed in this study have great potential to be used for the development of a low-cost and disposable immunosensor for detection of pathogenic microorganisms, especially in developing countries

Long-term measurement of PM2.5 mass concentration using an electrostatic particle concentrator-based quartz crystal microbalance integrated with carbon dioxide aerosol jets for PM sensing in remote areas

Fine particulate matter (PM2.5) is a major environmental health risk. Several instruments based on the quartz crystal microbalance (QCM) have been developed for PM2.5 measurement because of their accurate, sensitive, real-time, and low-cost mass measurements. However, prolonged or non-uniform deposition on the quartz crystal can cause nonlinear responses between frequency shifts and mass deposition, and its frequent manual cleaning with wet sponges is required. These disable long-term measurements of the instruments, thus limiting their applications in remote areas. Herein, we present a new PM2.5 instrument called qEPC-Snow. This instrument consists of a QCM crystal embedded in an electrostatic particle concentrator (EPC) for collection and sensing of PM2.5 and a carbon dioxide aerosol (snow) jet unit for residue-free, rapid, effective, and non-destructive cleaning of the crystal. Laboratory tests were conducted with aerosolized 100-nm and 2-mu m polystyrene latex microspheres as PM2.5 representatives to evaluate (i) frequency responses and (ii) mass sensitiveness of qEPC-Snow, (iii) particle removal efficiencies, and (iv) reuse of the used crystals. Experimental results demonstrated high removal efficiencies (approximately 99.9% for both particle sizes) and statistical similarity between the initial and cleaned QCM crystals in the frequency shift-mass deposition relationship, thereby enabling measurement for more than one month without demounting the crystals. The mass sensitivity was 57.34 (Hz/mu g) with R-2 = 0.9904, corresponding to 0.05667 [(Hz/min)/(mu g/m(3))] in mass concentration sensitivity for the PM2.5 representatives. The influence of particle sizes on qEPC-Snow's frequency behaviors will also be discussed in detail

Simultaneous determination of position and mass in the cantilever sensor using transfer function method

We present the simultaneous measurement of mass and position of micro-beads attached to the cantilever-based mass sensors using the transfer function method. 10 ??m diameter micro-beads were placed on micro-cantilevers and the cantilevers were excited by lead-zirconate-titanate through low-pass filtered random voltages. The cantilever vibration was measured via a laser Doppler vibrometer before and after applying the beads. From the measured transfer function, the bead position was identified using its influence on the cantilever kinetic energy. The bead mass was then obtained by analyzing the wave propagation near the beads. The predicted position and mass agreed well with actual values.open0

Measurement of PM2.5 Mass Concentration Using an Electrostatic Particle Concentrator-Based Quartz Crystal Microbalance

Particulate matter (PM) is one of the most critical air pollutants, and various instruments have been developed to measure PM mass concentration. Of these, quartz crystal microbalance (QCM) based instruments have received much attention. However, these instruments are subject to significant drawbacks: particle bounce due to poor adhesion, need for frequent cleanings of the crystal electrode, and non-uniform distribution of collected particles. In this study, we present an electrostatic particle concentrator (EPC)-based QCM (qEPC) instrument capable of measuring the mass concentration of PM 2.5 (PM smaller than 2.5 ??m), while avoiding the drawbacks. Experimental measurements showed high collection efficiencies (~99% at 1.2 liters/min), highly uniform particle distributions for long sampling periods (up to 120 min at 50 ??g/m 3 ), and high mass concentration sensitivity [0.068(Hz/min)/(??g/m 3 )]. The enhanced uniformity of particle deposition profiles and mass concentration sensitivity were made possible by the unique flow and electrical design of the qEPC instrument

Rapid and selective concentration of bacteria, viruses, and proteins using alternating current signal superimposition on two coplanar electrodes

Dielectrophoresis (DEP) is usually effective close to the electrode surface. Several techniques have been developed to overcome its drawbacks and to enhance dielectrophoretic particle capture. Here we present a simple technique of superimposing alternating current DEP (high-frequency signals) and electroosmosis (EO; low-frequency signals) between two coplanar electrodes (gap: 25 mu m) using a lab-made voltage adder for rapid and selective concentration of bacteria, viruses, and proteins, where we controlled the voltages and frequencies of DEP and EO separately. This signal superimposition technique enhanced bacterial capture (Escherichia coli K-12 against 1-mu m-diameter polystyrene beads) more selectively (>99%) and rapidly (similar to 30 s) at lower DEP (5 Vpp) and EO (1.2 Vpp) potentials than those used in the conventional DEP capture studies. Nanometer-sized MS2 viruses and troponin I antibody proteins were also concentrated using the superimposed signals, and significantly more MS2 and cTnI-Ab were captured using the superimposed signals than the DEP (10 Vpp) or EO (2 Vpp) signals alone (p < 0.035) between the two coplanar electrodes and at a short exposure time (1 min). This technique has several advantages, such as simplicity and low cost of electrode fabrication, rapid and large collection without electrolysis

Determination of Fluid Density and Viscosity by Analyzing Flexural Wave Propagations on the Vibrating Micro-cantilever

The determination of fluid density and viscosity using most cantilever-based sensors is based on changes in resonant frequency and peak width. Here, we present a wave propagation analysis using piezoelectrically excited micro-cantilevers under distributed fluid loading. The standing wave shapes of microscale-thickness cantilevers partially immersed in liquids (water, 25% glycerol, and acetone), and nanoscale-thickness microfabricated cantilevers fully immersed in gases (air at three different pressures, carbon dioxide, and nitrogen) were investigated to identify the effects of fluid-structure interactions to thus determine the fluid properties. This measurement method was validated by comparing with the known fluid properties, which agreed well with the measurements. The relative differences for the liquids were less than 4.8% for the densities and 3.1% for the viscosities, and those for the gases were less than 6.7% for the densities and 7.3% for the viscosities, showing better agreements in liquids than in gases