A reusable QCR aptasensor for the detection of Brevetoxin-2 in shellfish.

Brevetoxins (BTX) are pharmacologically active, lipid soluble cyclic polyether neurotoxins that are known to cause a wide range of neurological symptoms in humans.Harvesting and consumption of infected molluscs provide an entry point for BTXs into, the food chain, causing long-term health effects on accumulation for individuals, commonly in people with a compromised immune system and existing allergies. This study is an acoustic assay that has been constructed using a 9 MHz AT-cut quartz crystal resonator modified by attaching a specific single-stranded DNA aptamer. The DNA oligo modifies its conformation to attach itself to the binding site of the incoming BTX molecule resulting in a change in frequency on the QCR. A small Δf value was observed for lower concentrations of BTX indicating a small change in mass deposited on the crystal surface, while the opposite was true for higher concentrations. Cross-species behavior was evaluated using samples of similar origin, molecular weight and a combination of two toxins. The LOD of the fabricated QCR is 220 nM which is lower than the maximum recommended residue limit in food samples. Fresh mussel samples were spiked with known concentrations of BTX to evaluate its sensitivity in a food matrix. No interaction with other compounds was observed. Overall, this sensor finds potential application in the food sector (fishing units) where mussels are tested and graded for allergens and toxins before reaching the customer.

Antiproliferative activity of Ontario grown onions against colorectal adenocarcinoma cells.

Cancer is the leading cause of mortality in Canada and other industrialized nations; the development of new/improved cancer therapies is desperately needed and continues to be a major focus of cancer research. Flavonoids, which are found in high levels in onions, have been shown to exert antiproliferative and potentially anti-cancer activities. To test their therapeutic potential, we assessed the antiproliferative, cytotoxic, apoptosis-inducing, and anti-migratory activities of five onion varieties grown in Ontario against human adenocarcinoma (Caco-2) cells. The properties of onion extracts were compared to pure extracts of flavonoids known to exhibit antiproliferative effects (quercetin, myricetin, and kaempferol). We compared more than one variety of onion, as agronomic and genetic factors influence the composition, as well as the quality of phytochemicals (e.g. flavonoids) in plant cultivars. We found that all onion varieties exhibited antiproliferative activity similar to purified flavonoids. The cytotoxic effects of the Stanley and Fortress onion varieties were strongest among the selected cultivars, as determined via lactate dehydrogenase (LDH) assays, while Safrane extracts showed the weakest activity. The Stanley and Lasalle cultivar extracts also had strong anti-migratory effects. Altogether these onion extracts may contain one or more compounds that may be effective anti-cancer therapies, while the Stanley extract showed the most comprehensive biological activities against Caco-2 cells.

Enhancing the bandwidth of piezoelectric composite transducers for air-coupled non-destructive evaluation.

This paper details the development of a novel method for increasing the operational bandwidth of piezocomposites without the need for lossy backing material, the aim being to increase fractional bandwith by geometrical design. Removing the need for lossy backing materials, should in turn increase the transmit efficiency in the desired direction of propagation. Finite element analysis has been employed to determine the mode of operation of the new piezocomposite devices and shows good correlation with that derived experimentally. Through a series of practical and analytical methods it has been shown that additional thickness mode resonances can be introduced into the structure by a simple machining process. The shaped composites described in this paper offer increased operational bandwidth. A simple example of a two step thickness design is described to validate and illustrate the principle. A more complex conical design is presented that illustrates a possible tenfold increase in bandwidth from 30kHz to 300kHz, operating in air without backing. An illustration of the applicability of this type of transducer technology for frequency agile guided mode non-destructive evaluation is then presented.

A simultaneous assessment metric for MAb quantity and glycan quality.

As a critical quality attribute, glycosylation represents an important consideration when analyzing the success of a glycoprotein production process. Though critical, glycosylation is not the only measure of culture success; other factors, including culture size, maintenance, and productivity, are also critical. A new metric was developed to address both product quality, as measured through glycosylation, and product quantity, as measured through product concentration. A monoclonal antibody Chinese hamster ovary cell culture model system was used to assess this metric across various media formulations. In a model test system, the metric discriminated that some media supplements had a net positive impact on productivity and glycosylation, while others had a net negative impact on productivity and glycosylation.

Time-of-flight measurement techniques for airborne ultrasonic ranging.

Airborne ultrasonic ranging is used in a variety of different engineering applications for which other positional metrology techniques cannot be used, for example in closed-cell locations, when optical line of sight is limited, and when multipath effects preclude electromagnetic-based wireless systems. Although subject to fundamental physical limitations, e.g., because of the temperature dependence of acoustic velocity in air, these acoustic techniques often provide a cost-effective solution for applications in mobile robotics, structural inspection, and biomedical imaging. In this article, the different techniques and limitations of a range of airborne ultrasonic ranging approaches are reviewed, with an emphasis on the accuracy and repeatability of the measurements. Simple time-domain approaches are compared with their frequency-domain equivalents, and the use of hybrid models and biologically inspired approaches are discussed.

Ultrasound nondestructive evaluation (NDE) imaging with transducer arrays and adaptive processing.

This paper addresses the challenging problem of ultrasonic non-destructive evaluation (NDE) imaging with adaptive transducer arrays. In NDE applications, most materials like concrete, stainless steel and carbon-reinforced composites used extensively in industries and civil engineering exhibit heterogeneous internal structure. When inspected using ultrasound, the signals from defects are significantly corrupted by the echoes form randomly distributed scatterers, even defects that are much larger than these random reflectors are difficult to detect with the conventional delay-and-sum operation. We propose to apply adaptive beamforming to the received data samples to reduce the interference and clutter noise. Beamforming is to manipulate the array beam pattern by appropriately weighting the per-element delayed data samples prior to summing them. The adaptive weights are computed from the statistical analysis of the data samples. This delay-weight-and-sum process can be explained as applying a lateral spatial filter to the signals across the probe aperture. Simulations show that the clutter noise is reduced by more than 30 dB and the lateral resolution is enhanced simultaneously when adaptive beamforming is applied. In experiments inspecting a steel block with side-drilled holes, good quantitative agreement with simulation results is demonstrated.

Simulation of ultrasonic lamb wave generation, propagation and detection for a reconfigurable air coupled scanner.

A computer simulator, to facilitate the design and assessment of a reconfigurable, air-coupled ultrasonic scanner is described and evaluated. The specific scanning system comprises a team of remote sensing agents, in the form of miniature robotic platforms that can reposition non-contact Lamb wave transducers over a plate type of structure, for the purpose of non-destructive evaluation (NDE). The overall objective is to implement reconfigurable array scanning, where transmission and reception are facilitated by different sensing agents which can be organised in a variety of pulse-echo and pitch-catch configurations, with guided waves used to generate data in the form of 2-D and 3-D images. The ability to reconfigure the scanner adaptively requires an understanding of the ultrasonic wave generation, its propagation and interaction with potential defects and boundaries. Transducer behaviour has been simulated using a linear systems approximation, with wave propagation in the structure modelled using the local interaction simulation approach (LISA). Integration of the linear systems and LISA approaches are validated for use in Lamb wave scanning by comparison with both analytic techniques and more computationally intensive commercial finite element/difference codes. Starting with fundamental dispersion data, the paper goes on to describe the simulation of wave propagation and the subsequent interaction with artificial defects and plate boundaries, before presenting a theoretical image obtained from a team of sensing agents based on the current generation of sensors and instrumentation.

Properties of photocured epoxy resin materials for application in piezoelectric ultrasonic transducer matching layers.

This paper describes the acoustic properties of a range of epoxy resins prepared by photocuring that are suitable for application in piezoelectric ultrasonic transducer matching layers. Materials, based on blends of diglycidyl ether of Bisphenol A and 1,4-cyclohexanedimethanol diglycidyl ether, are described. Furthermore, in order to vary the elastic character of the base resin, samples containing polymer microspheres or barium sulfate particles are also described. The acoustic properties of the materials are determined by a liquid coupled through transmission methodology, capable of determining the velocity and attenuation of longitudinal and shear waves propagating in an isotropic layer. Measured acoustic properties are reported which demonstrate materials with specific acoustic impedance varying in the range 0.88-6.25 MRayls. In the samples comprising blends of resin types, a linear variation in the acoustic velocities and density was observed. In the barium sulfate filled samples, acoustic impedance showed an approximately linear variation with composition, reflecting the dominance of the density variation. While such variations can be predicted by simple mixing laws, relaxation and scattering effects influence the attenuation in both the blended and filled resins. These phenomena are discussed with reference to dynamic mechanical thermal analysis and differential scanning calorimetry of the samples.

Theoretical analysis of ultrasonic vibration spectra from multiple particle-plate impacts.

Many industrial processes involve particles in a carrier fluid, and it is often of interest to monitor the size of these particles noninvasively. The aim of this paper is to develop a theoretical model of multiple particle-wall impact vibrations that can be used to recover the particle size from experimental data. These vibrations have been measured by an ultrasonic transducer attached to the exterior of a vessel containing a stirred-particle-laden fluid. A linear systems model is derived for the response of the piezoelectric ultrasonic transducer, which has a single matching layer. The acceleration power spectrum of these vibrations has been shown experimentally to contain information on the size of the impacting particle. In particular, the frequency of the main spectral lobe is inversely proportional to the particle size. We present a theoretical model that agrees with this empirically observed phenomenon. The theoretical model is then used to simulate multiple particle-wall impacts, with each particle impacting at a randomly chosen location. A set of theoretical vibration spectra arising from multiple particle-wall impacts are integrated and compared with the experimental data. The ability of this approach to distinguish between different particle sizes is clearly shown.

Estimating particle concentration using passive ultrasonic measurement of impact vibrations.

A method of obtaining particle size and concentration information, from ultrasonic transducer measurements of particle-wall interactions in a particle laden fluid, is presented. A mathematical model of the flexural vibrations of the vessel wall due to the particle impact is constructed. The key component of this model is the derivation of an analytic expression for the impact force amplitude. An analytic expression for the power spectrum is then obtained that shows its explicit dependency on the system parameters. This spectrum is then integrated over a specific frequency range and a comparison with experimental results is reported.

Simplex optimization of acoustic assay for plasminogen activators.

This article discusses the optimization of a newly developed method for measuring the activity of plasminogen activators using a thickness-shear-mode acoustic sensor. A variable-size simplex algorithm was used for optimization. Preliminary tests were performed to design the first simplex. A desirability function was defined to translate each performance value to a membership value of 0 to 1. If there was more than one performance variable, their membership values were translated to an aggregated membership value using another function that considers their individual influence on sensor performance. Two rounds of optimization were carried out for streptokinase followed by a single optimization for tissue-type plasminogen activator. In the last optimization, ratios of control variables were used in order to reduce the number of parameters and to formulate easily adjustable assay conditions. The results showed the usefulness of the simplex method for optimizing this type of assay, and the importance of preliminary tests and prior knowledge in providing rapid convergence using fewer experiments. The optimized plasminogen activator assay can be considered a reference method for measurement of all members of this drug class.

Acoustic determination of performance and equivalence of plasminogen activators.

A reliable method for the measurement of different plasminogen activators is of great interest for both manufacturing and clinical medicine. A one-step assay based on a thickness shear mode acoustic sensor has been developed for this purpose. Two separate mixtures of substrates (fibrinogen and plasminogen) and enzymes (thrombin and the plasminogen activator) were mixed, and placed on the acoustic sensor surface. During the assay, the resonant frequency of a quartz crystal oscillating in the thickness shear mode was measured and used to find a characteristic clot dissolution time, from the sample addition to the time at the maximum dissolution rate. Calibrations of the acoustic assay were done for tissue-type plasminogen activator (t-PA) as well as for the other plasminogen activators: urokinase (u-PA); streptokinase (SK) and staphylokinase (SAK). All gave relative standard deviations of about 12%. Since the same method was used for all of the activators, their activities were compared, resolving the differences between their unit definitions. Linear relationships were found between urokinase and streptokinase which activate plasminogen directly and between t-PA and staphylokinase which require fibrin as a cofactor. The relationship between the groups was found to curve, indicating the difference between the two mechanisms. The acoustic method, therefore, may be used as a rapid and cost-effective reference method for the standardization and comparison of different plasminogen activators.

One-step thickness shear mode acoustic assay for plasminogen activators.

A new procedure is presented for the measurement of plasminogen activators using a thickness shear mode sensor and a modified version of the fibrin plate assay at the micro-scale. Separate, well-mixed solutions of the substrates fibrinogen and plasminogen, and enzymes thrombin and the plasminogen activator sample were mixed together and placed on the sensor surface. The temperature and evaporation were controlled during the assay. The clot dissolution time correlated well with the quantity of the plasminogen activator in the sample. The average relative standard deviation was 12.5%.

Harmonic analysis of lossy, piezoelectric composite transducers using the plane wave expansion method.

Periodic composite ultrasonic transducers offer many advantages but the periodic pillar architecture can give rise to unwanted modes of vibration which interfere with the piston like motion of the fundamental thickness mode. In this paper, viscoelastic loss is incorporated into a three-dimensional plane wave expansion model (PWE) of these transducers. A comparison with experimental and finite element data is conducted and a design to damp out these lateral modes is investigated. Scaling and regularisation techniques are introduced to the PWE method to reduce ill-conditioning in the large matrices which can arise. The identification of the modes of vibration is aided by examining profiles of the displacements, electrical potential and Poynting vector. The dispersive behaviour of a 2-2 composite transducer with high shear attenuation in the passive phase is examined. The model shows that the use of a high shear attenuation filler material improves the frequency band gap surrounding the fundamental thickness mode.

Finite element technique for the assessment of 3-1 and "super 1-3" connectivity piezoelectric composite transducers.

This paper describes the use of finite element (FE) technique in the assessment of new types of multilayered piezoelectric composite structure using the PZFlex code. The background information leading to model configuration, including materials properties and boundary conditions, is discussed. This is coupled with an experimental program of model corroboration via a number of key stage prototypes to achieve a model-build-test methodology. Initially the 3-1 connectivity plate and multilayered piezoelectric composites are analyzed. Results from FE and experimental assessment indicate that the 3-1 plate devices offer no benefit over the conventional 1-3 connectivity arrangement. A simple, minimally diced, 3-1 connectivity multilayer device is analyzed and is shown to be suitable for the manufacture of wideband and efficient transducers operating in the 10-100 kHz frequency range for high power applications.

Theoretical modelling of frequency dependent elastic loss in composite piezoelectric transducers.

The large number of degrees of freedom in the design of piezoelectric transducers requires a theoretical model that is computationally efficient so that a large number of iterations can be performed in the design optimisation. The materials used are often lossy, and indeed loss can be used to enhance the operational characteristics of these designs. Motivated by these needs, this paper extends the one-dimensional linear systems model to incorporate frequency dependent elastic loss. The reception sensitivity, electrical impedance and electromechanical coupling coefficient of a 1-3 composite transducer, with frequency dependent loss in the polymer filler, are investigated. By plotting these operating characteristics as a function of the volume fraction of piezoelectric ceramic an optimum design is obtained. A device with a non-standard, high shear attenuation polymer is also simulated and this leads to an increase in the electromechanical coupling coefficient. A comparison with finite element simulations is then performed. This shows that the two methods are in reasonable agreement in their electrical impedance profiles in all the cases considered. The plots are almost identical away from the main resonant peak where the frequency location of the peaks are comparable but there is in some cases a 20% discrepancy in the magnitude of the peak value and in its bandwidth. The finite element model also shows that the use of a high shear attenuation polymer filler damps out the unwanted, low frequency modes whilst maintaining a reasonable impedance magnitude.

A theoretical analysis of a piezoelectric ultrasound device with an active matching layer.

This paper investigates the use of magnetically active materials in the matching layer of a piezoelectric transducer. This then allows the performance of the device to be dynamically altered by applying an external field. The effect that this new matching layer has on the performance of a typical device is theoretically investigated here. It transpires that the additional flexibility of an active matching layer can be used to maintain the efficiency of the device as the external load is varied.

Comparison of the frequency and physical nature of the lowest order parasitic mode in single crystal and ceramic 2-2 and 1-3 piezoelectric composite transducers.

This work describes an investigation into the first order parasitic mode (i.e., that closest to the fundamental thickness mode) that can occur in 2-2 and 1-3 thickness drive piezoelectric composite transducers. Specifically, the authors compare the performance of piezoceramic and piezocrystal composites with a common passive phase. A local Lamb wave approach is used to describe the generation of such modes, and the validity of this theory is investigated over the entire volume fraction range. It is shown that, when the parasitic mode is primarily generated by Lamb wave activity in the passive phase, both active materials demonstrate similar behavior. However, at higher volume fractions, the first order mode is related to the lateral resonance of the active material, and quite different behavior may be observed between the two sets of devices. The phase velocity of the parasitic modes in each device configuration was investigated by a combination of experimental measurement on a number of transducers along with simulations using the finite-element code PZFlex. Both 2-2 and 1-3 composites made from the single crystal materials pzn-4.5%pt, pzn-8%pt, and pmn-30%pt were investigated along with composites made from pzt5h ceramic. The PZFlex results are compared with experimental impedance analysis and laser scanning of surface displacement, with good agreement demonstrated. By comparing two very different active materials, additional insight into parasitic resonant activity within composite devices is demonstrated.

A block diagram model of the thickness mode piezoelectric transducer containing dual oppositely polarized piezoelectric zones.

A unidimensional, linear systems, block diagram model of a two-layer thickness mode piezoelectric transducer is presented. The layers are subject to opposing piezoelectric polarization and the device is assumed to be loaded by semi-infinite isotropic media at the two principal faces. Block diagram representations of the transducer acting as both a generator and a receiver of ultrasound are developed in conjunction with the equivalent model of the electrical admittance. When expressed in this manner, the underlying cause and effect relationships are identified, with the important contribution of the piezoelectric boundary highlighted. Comparisons with the conventional single-layer transducer are made throughout and the major physical differences in terms of transduction performance are discussed. The new model is compared with finite element analysis and good agreement is also demonstrated with experimental data. A key aspect of the methodology is the provision of a more intuitive understanding of such device behavior. Accordingly, emphasis has been placed on the physical relationships and this is considered a major contribution of the work.

Determination of lamb wave dispersion data in lossy anisotropic plates using time domain finite element analysis. Part I: theory and experimental verification.

A theoretical and experimental approach for extraction of guided wave dispersion data in plate structures is described. Finite element modeling is used to calculate the surface displacement data (in-plane and out-of-plane) when the plate is subject to either symmetrical or antisymmetrical impulsive force stimulation at one or both of the parallel faces. Fourier transformation of the resultant space-time displacement histories is then employed to obtain phase velocity as a function of frequency. Experimental verification in the case of antisymmetrical stimulation is provided by means of a high-power Q-switched laser source that is used to excite guided waves in the plate. The subsequent out-of-plane displacement data were then obtained by means of a scanning laser vibrometer, and good agreement between theory and experiment is demonstrated. Examples of dispersion data are provided for aluminum, and excellent correlation between the data sets and conventional Rayleigh-Lamb theory for plate structures was obtained. This was then extended to lossy polymeric plates, in addition to both unpolarized and polarized piezoelectric ceramic plates, again with good agreement between the finite element modeling and optical experiments. The last set of results prepares the way for a detailed investigation of the nonhomogeneous piezoelectric composite waveguides described in a companion paper (Part II).