Citations

ABSTRACTClinical mutations in patients diagnosed with Type 2A von Willebrand disease (VWD) have been identified that break the single disulfide bond linking N‐ and C‐termini in the vWF A1 domain. We have modeled the effect of these mutations on the disulfide‐bonded structure of A1 by reducing and carboxy‐amidating these cysteines. Solution biophysical studies show that loss of this disulfide bond induces a molten globule conformational state lacking global tertiary structure but retaining residual secondary structure. The conformational dependence of platelet adhesion to these native and molten globule states of A1 is quantitatively compared using real‐time high‐speed video microscopy analysis of platelet translocation dynamics under shear flow in a parallel plate microfluidic flow chamber. While normal platelets translocating on surface‐captured native A1 domain retain the catch‐bond character of pause times that increase as a function of shear rate at low shear and decrease as a function of shear rate at high shear, platelets that interact with A1 lacking the disulfide bond remain stably attached and do not translocate. Based on these findings, we propose that the shear stress‐sensitive regulation of the A1‐GPIb interaction is due to folding the tertiary structure of this domain. Removal of the tertiary structure by disrupting the disulfide bond destroys this regulatory mechanism resulting in high‐strength interactions between platelets and vWF A1 that are dependent only on residual secondary structure elements present in the molten globule conformation. Proteins 2014; 82:867–878. © 2013 Wiley Periodicals, Inc.

Bladder compliance is defined as bladder’s ability to expand in volume without a significant change in pressure. The bladder wall is a multi-layered structure including a muscular layer and fibrous connective tissue. As the percent of connective tissue in the bladder interstitium increases compared to smooth muscle, the bladder becomes more rigid and less capable of expanding during filling. Currently, urodynamic studies (UDS) are considered as the clinical gold standard for bladder compliance assessment. This procedure is invasive and is associated with patient discomfort and is expensive. Ultrasound bladder vibrometry (UBV) is a novel acoustic-radiation-forced-based method for noninvasive assessment of bladder compliance. In UBV, an impulsive acoustic radiation force is focused on the bladder wall (under B-mode ultrasound image guidance) to induce vibrations which may be modeled by as a Lamb wave. High frame rate ultrasound is then utilized to detect the induced waves. This wave motion is then used to estimate the viscoelastic properties and the compliance of the bladder wall. Our results reveal a remarkable agreement between UBV and UDS cystometry measures, suggesting the potential of UBV as a viable clinical tool for the assessment of bladder compliance.

Improved approximate frequency domain expressions are derived for cylindrical shear wave propagation in viscoelastic media. These expressions extend prior results that describe cylindrical wave propagation in lossless media. Previously, an analytical expression for a cylindrical wave was obtained in terms of a Hankel function and a large argument approximation was applied to the result. A leading frequency-dependent term was then treated as a constant with respect to frequency. In the improved expression, the frequency-dependence of the leading term is retained. For lossless media, the leading term is a fractional integrator, and for viscoelastic media, the leading term is either a fractional integrator or an integer-order integrator, depending on the frequency range. The lossless and the viscoelastic models are evaluated in the frequency domain for simplified source geometries and compared to numerical results. The comparison shows that the agreement between the analytical and the numerical models is excellent. Implications for time-domain calculations in viscous media will also be discussed. [This work was supported in part by NIH Grant Nos. R01 EB012079 and R01 DK092255.]

We validate a technique for model-free measurement of shear wave velocity and attenuation. A mechanical shaker was used to excite harmonic plane and cylindrical waves in phantoms and excised tissue. Radiation force was used to excite impulsive cylindrical waves. 2D FFT of the displacement yielded the k-space whose coordinates are frequency and the wave number. The shear wave velocity at each frequency was obtained by finding the maximum at the given frequency in k-space and dividing the frequency coordinate by the wave number coordinate. The attenuation (α) at a given frequency was calculated using α = FWHM × π/√3, where FWHM is the full width at half maximum of the k-space peak along the given frequency. This method was applied to measure shear wave velocity and attenuation of transplanted kidneys and livers, and in the thyroid tumor, and compare it to the healthy tissues. The velocities and attenuations at each frequency for various excitation methods agree within one standard deviation. The k-space estimates of velocity and attenuation agreed with those obtained using the phase gradient (velocity) and amplitude decay (attenuation). The transplanted organs and the thyroid tumor had higher velocity and lower attenuation than healthy tissues.

Measurements of viscoelasticity with shear wave velocity dispersion requires measurements over a large bandwidth. In this study, we explored the parameters that modulate the frequency characteristics of shear waves induced using radiation force push beams. We used a Verasonics ultrasound scanner equipped with a linear array transducer. We performed measurements of shear wave motion induced using both focused and unfocused ultrasound beams. Measurements were made in elastic phantoms with shear moduli of 1, 4, and 16 kPa. The number of elements used for the unfocused beams were varied from 8 to 24, and for the focused beams from 16 to 128. The shear wave motion was tracked using plane wave imaging, and a one-dimensional autocorrelation algorithm applied to the acquired in-phase/quadrature data. At each pixel we calculated the fast Fourier transform of the data and found the center frequency, center-of-gravity, and −3 dB bandwidth. We compared the frequency characteristics from the different push beams. The frequency characteristics were found to be spatially variant and dependent on the number of elements used as well as the shear modulus of the medium. The center frequency, center-of-gravity, and the bandwidth were found to be correlated to one another, and strongly associated with the stiffness of the medium.

Robust shear-wave motion detection is essential for producing reliable shear elasticity measurements for ultrasound shear-wave elastography (SWE). However, because shear-wave motion is extracted from radiofrequency signals which can be heavily contaminated by noise, shear-wave motion detection can be very challenging, especially for in vivo applications. This study investigated the implementation of harmonic imaging (HI) to facilitate more robust shear-wave tracking based on HI’s effectiveness in suppressing imaging noise associated with ultrasound reverberation, phase aberration, and clutter noise. A HI shear-wave tracking sequence was developed combining the pulse-inversion HI method with the plane wave imaging technique to transmit phase-inverted pulses at a high frame rate of several kilohertz. The backscattered ultrasound signals from phase-inverted pulses were added to suppress the fundamental and enhance the second harmonic component, from which the shear-wave motion was extracted. A pork belly phantom experiment showed that HI could significantly improve shear-wave motion detection by producing almost three-fold less underestimation of shear-wave motion and over ten-fold more precision for shear-wave speed measurements than fundamental imaging. An in vivo transthoracic case study of a human heart showed that HI substantially improved the success rate of shear-wave motion detection and could provide consistent estimates of the left ventricle myocardium stiffness.

Introduction: Ultrasound radiation force-based methods can quantitatively evaluate tissue viscoelastic material properties. A limitation of current methods includes neglecting the inherent anisotropy nature of tissue. To explore this phenomenon, we created a phantom incorporating fibrous material that has preferential orientations. Methods: Two phantoms were made in a cube-shaped mold using a fibrous material arranged in multiple layers and embedded in porcine gelatin using two different concentrations of the gelatin (8%, 14%). Shear wave measurements were made in the phantoms at different angles by rotating the phantom, where 0° and 180° were defined along the fibers, and 90° and 270° across the fibers. Measurements were performed using a Verasonics ultrasound system equipped with a linear array transducer. Results/Discussion: The mean shear wave speeds and mean standard deviations for 8% and 14% gelatin along the fibers (0°) were (3.60 ± 0.03 and 4.10 ± 0.11 m/s) and across the fibers (90°) were (3.18 ± 0.12 and 3.90 ± 0.02 m/s), respectively. Conclusion: The fibrous gelatin-based phantoms exhibited anisotropy that could be measured using quantitative shear waves speed measurements. Increasing the gelatin percentage increases the shear wave speed and anisotropic moduli. [This study was supported by NIH grant DK092255.]

Thyroid cancer is the fastest growing age and gender adjusted cancer in 2011 according the American Cancer Society. The majority of the clinically diagnosed thyroid nodules are benign while less than 5% represent intrathyroidal cancers. Currently, the clinical gold-standard procedure for assessing the thyroid nodules is needle biopsy, a procedure that is associated with significant financial burden as well as pain and risk for patients. Therefore, a noninvasive, affordable, and potentially widely available method to differentiate between benign and malignant thyroid nodules can play an important role in reducing the number of unnecessary biopsies. In this study, we investigate the feasibility of two acoustic radiation force elastography techniques, shear wave dispersion ultrasound vibrometry (SDUV) and comb-push ultrasound shear wave elastography (CUSE imaging), in identifying thyroid nodules (imaging) and differentiating between benign and malignant pathologies based on their elasticity and viscosity (SDUV measurements). Our preliminary results show that the measured shear elasticity and shear viscosity parameters depend on tissue type; hence, these measurements may be utilized to differentiate between healthy normal thyroid tissue, benign nodules, and malignant nodules. Further studies on a large population of patients is required to better evaluate the role of the combination of elasticity and viscosity properties of tissue in differentiating various thyroid nodules.

Diagnostic ultrasound images suffer from degradation due to tissues with sound speed inhomogeneities causing phase shifts of propagating waves. These phase shifts defocus the ultrasound beam, reducing spatial resolution and image contrast in the resulting image. We describe a phase aberration correction method that uses dynamic ultrasound radiation force to harmonically excite a medium using amplitude-modulated continuous wave ultrasound created by summing two ultrasound frequencies at f0=3.0 MHz and f0+Δf=3.0005 MHz. The phase of each element of a linear array transducer is sequentially adjusted to maximize the radiation force and obtain optimal focus of the ultrasound beam. The optimization is performed by monitoring the harmonic amplitude of the scatterer velocity in the desired focal region using Doppler techniques. Simulation results show the ability to regain a 3.0-MHz focused field after applying a phase screen with an rms time delay of 95.4 ns. The radiation force magnitude increased by 22 dB and the resolution of the field was regained. Simulation results show that the focus of the beam can be qualitatively and quantitatively improved with this method. [This study was supported in part by Grants EB002640 and EB002167 from the NIH.]