Brain-wide mapping of resting-state networks in mice using high-frame rate functional ultrasound.

Functional ultrasound (fUS) imaging is a method for visualizing deep brain activity based on cerebral blood volume changes coupled with neural activity, while functional MRI (fMRI) relies on the blood-oxygenation-level-dependent signal coupled with neural activity. Low-frequency fluctuations (LFF) of fMRI signals during resting-state can be measured by resting-state fMRI (rsfMRI), which allows functional imaging of the whole brain, and the distributions of resting-state network (RSN) can then be estimated from these fluctuations using independent component analysis (ICA). This procedure provides an important method for studying cognitive and psychophysiological diseases affecting specific brain networks. The distributions of RSNs in the brain-wide area has been reported primarily by rsfMRI. RSNs using rsfMRI are generally computed from the time-course of fMRI signals for more than 5 min. However, a recent dynamic functional connectivity study revealed that RSNs are still not perfectly stable even after 10 min. Importantly, fUS has a higher temporal resolution and stronger correlation with neural activity compared with fMRI. Therefore, we hypothesized that fUS applied during the resting-state for a shorter than 5 min would provide similar RSNs compared to fMRI. High temporal resolution rsfUS data were acquired at 10 Hz in awake mice. The quality of the default mode network (DMN), a well-known RSN, was evaluated using signal-noise separation (SNS) applied to different measurement durations of rsfUS. The results showed that the SNS did not change when the measurement duration was increased to more than 210 s. Next, we measured short-duration rsfUS multi-slice measurements in the brain-wide area. The results showed that rsfUS with the short duration succeeded in detecting RSNs distributed in the brain-wide area consistent with RSNs detected by 11.7-T MRI under awake conditions (medial prefrontal cortex and cingulate cortex in the anterior DMN, retrosplenial cortex and visual cortex in the posterior DMN, somatosensory and motor cortexes in the lateral cortical network, thalamus, dorsal hippocampus, and medial cerebellum), confirming the reliability of the RSNs detected by rsfUS. However, bilateral RSNs located in the secondary somatosensory cortex, ventral hippocampus, auditory cortex, and lateral cerebellum extracted from rsfUS were different from the unilateral RSNs extracted from rsfMRI. These findings indicate the potential of rsfUS as a method for analyzing functional brain networks and should encourage future research to elucidate functional brain networks and their relationships with disease model mice.

Functional ultrasound reveals effects of MRI acoustic noise on brain function.

Loud acoustic noise from the scanner during functional magnetic resonance imaging (fMRI) can affect functional connectivity (FC) observed in the resting state, but the exact effect of the MRI acoustic noise on resting state FC is not well understood. Functional ultrasound (fUS) is a neuroimaging method that visualizes brain activity based on relative cerebral blood volume (rCBV), a similar neurovascular coupling response to that measured by fMRI, but without the audible acoustic noise. In this study, we investigated the effects of different acoustic noise levels (silent, 80 dB, and 110 dB) on FC by measuring resting state fUS (rsfUS) in awake mice in an environment similar to fMRI measurement. Then, we compared the results to those of resting state fMRI (rsfMRI) conducted using an 11.7 Tesla scanner. RsfUS experiments revealed a significant reduction in FC between the retrosplenial dysgranular and auditory cortexes (0.56 ± 0.07 at silence vs 0.05 ± 0.05 at 110 dB, p=.01) and a significant increase in FC anticorrelation between the infralimbic and motor cortexes (-0.21 ± 0.08 at silence vs -0.47 ± 0.04 at 110 dB, p=.017) as acoustic noise increased from silence to 80 dB and 110 dB, with increased consistency of FC patterns between rsfUS and rsfMRI being found with the louder noise conditions. Event-related auditory stimulation experiments using fUS showed strong positive rCBV changes (16.5% ± 2.9% at 110 dB) in the auditory cortex, and negative rCBV changes (-6.7% ± 0.8% at 110 dB) in the motor cortex, both being constituents of the brain network that was altered by the presence of acoustic noise in the resting state experiments. Anticorrelation between constituent brain regions of the default mode network (such as the infralimbic cortex) and those of task-positive sensorimotor networks (such as the motor cortex) is known to be an important feature of brain network antagonism, and has been studied as a biological marker of brain disfunction and disease. This study suggests that attention should be paid to the acoustic noise level when using rsfMRI to evaluate the anticorrelation between the default mode network and task-positive sensorimotor network.

Characteristic X-ray absorptiometry applied to the assessment of tissue-engineered cartilage development.

BACKGROUND: Transmission and tomographic X-ray measurements are useful in assessing bone structures, but only a few studies have examined cartilage growth because of the poor contrast in conventional X-ray imaging. OBJECTIVE: In this study, we attempted to use the linear attenuation coefficient (LAC) as a metric of tissue-engineered cartilage development, which would be useful in high-throughput screening of cartilage products. METHODS: Assuming that the LAC is related to the amount of extracellular matrix (ECM) in terms of the density and its atomic components, we measured X-ray absorption through tissue-engineered cartilage constructs. Characteristic X-ray beams from a molybdenum microfocus X-ray tube were employed to avoid beam hardening. The correlation of the LAC with mechanical properties was analyzed for verification. RESULTS: The LAC was higher for chondrocyte constructs and lower for fibroblast-dominant constructs and was consistent with the quantification of toluidine blue staining, which is a proof of ECM production. The LAC was positively correlated with the bending modulus but negatively correlated with the dynamic elastic modulus and stiffness, possibly because of the remaining scaffold. CONCLUSIONS: The LAC has the potential to be used as a metric of development of tissue-engineered cartilage. However, the calcified regions should be excluded from analysis to avoid decreasing the correlation between the LAC and the amount of ECM.

Towards fully automated robotic platform for remote auscultation.

BACKGROUND: Since most developed countries are facing an increase in the number of patients per healthcare worker due to a declining birth rate and an ageing population, relatively simple and safe diagnosis tasks may need to be performed using robotics and automation technologies, without specialists and hospitals. This study presents an automated robotic platform for remote auscultation, which is a highly cost-effective screening tool for detecting abnormal clinical signs. METHOD: The developed robotic platform is composed of a 6-degree-of-freedom cooperative robotic arm, LiDAR camera, and a spring-based mechanism holding an electric stethoscope. The platform enables autonomous stethoscope positioning based on external body information acquired using the LiDAR camera-based multi-way registration; the platform also ensures safe and flexible contact, maintaining the contact force within a certain range through the passive-actuated mechanism. RESULTS: Our preliminary results confirm that the robotic platform enables estimation of the landing positions required for cardiac examinations based on the depth and landmark information of the body surface. It also handles the stethoscope while maintaining the contact force without relying on the push-in displacement by the robotic arm. CONCLUSION: The developed robotic platform enables the estimation of the landing positions and handling the stethoscope while maintaining the contact force, which promises the potential of automatic remote auscultation.

Acoustic radiation force impulse-induced lung hemorrhage: investigating the relationship with peak rarefactional pressure amplitude and mechanical index in rabbits.

PURPOSE: The safety of acoustic radiation force impulse (ARFI) elastography, which applies higher acoustic power with a longer pulse duration (PD) than conventional diagnostic ultrasound, is yet to be verified. We assessed the ARFI-induced lung injury risk and its relationship with peak rarefactional pressure amplitude (PRPA) and mechanical index (MI). METHODS: Eighteen and two rabbits were included in the ARFI (0.3-ms push pulses) and sham groups, respectively. A 5.2-MHz linear probe was applied to the subcostal area and aimed at both lungs through the liver for 30 ARFI emissions. The derated PRPA varied among the six ARFI groups-0.80 MPa, 1.13 MPa, 1.33 MPa, 1.70 MPa, 1.91 MPa, and 2.00 MPa, respectively. RESULTS: The occurrence of lung hemorrhage and the mean lesion area among all samples in the seven groups were 0/6, 0/6, 1/6 (1.7 mm2), 4/6 (8.0 mm2), 4/6 (11.2 mm2), 5/6 (23.8 mm2), and 0/4 (sham), respectively. Logistic regression analysis showed that derated PRPA was significantly associated with lung injury occurrence (odds ratio: 207, p < 0.01), with the threshold estimated to be 1.1 MPa (MI, 0.5). Spearman's rank correlation showed a positive correlation between derated PRPA and lesion area (r = 0.671, p < 0.01). CONCLUSION: This study demonstrated that the occurrence and severity of ARFI-induced lung hemorrhage increased with a rise in PRPA under clinical conditions in rabbits. This indicates a potential risk of lung injury due to ARFI elastography, especially when ARFI is unintentionally directed to the lungs during liver, heart, or breast examinations.

In Vivo Temperature Rise Measurements of Rabbit Liver and Femur Bone Surface Exposed to an Acoustic Radiation Force Impulse.

Acoustic radiation force impulse (ARFI) imaging and shear wave elastography use a "push pulse." The push pulse, which is referenced as an ARFI in this study, has a longer duration than that of conventional diagnostic pulses (several microseconds). Therefore, there are concerns regarding thermal safety in vivo. However, few in vivo studies have been conducted using living animals. In this study, to suggest a concept for deciding an ARFI output and cooling time while considering thermal safety, the liver (with and without an ultrasound contrast agent) and femur bone surface of living rabbits were exposed to an ARFI, and the maximum temperature rise, temperature rise for 5-min duration, and cooling time were measured via a thermocouple. While testing within the regulation limits of diagnostic ultrasound outputs, a maximum temperature rise on the femur bone surface exceeded the allowable temperature rise (1.5°C) in the British Medical Ultrasound Society (BMUS) statement. However, using the linear relationships between the pulse intensity integral (PII) of a single pulse and the above three temperature parameters, PII may be determined so that the maximum temperature rise is within the allowable temperature rise in the BMUS statement. The cooling time can be estimated from the PII.

A review of physical and engineering factors potentially affecting shear wave elastography.

It has been recognized that tissue stiffness provides useful diagnostic information, as with palpation as a screening for diseases such as cancer. In recent years, shear wave elastography (SWE), a technique for evaluating and imaging tissue elasticity quantitatively and objectively in diagnostic imaging, has been put into practical use, and the amount of clinical knowledge about SWE has increased. In addition, some guidelines and review papers regarding technology and clinical applications have been published, and the status as a diagnostic technology is in the process of being established. However, there are still unclear points about the interpretation of shear wave speed (SWS) and converted elastic modulus in SWE. To clarify these, it is important to investigate the factors that affect the SWS and elastic modulus. Therefore, physical and engineering factors that potentially affect the SWS and elastic modulus are discussed in this review paper, based on the principles of SWE and a literature review. The physical factors include the propagation properties of shear waves, mechanical properties (viscoelasticity, nonlinearity, and anisotropy), and size and shape of target tissues. The engineering factors include the region of interest depth and signal processing. The aim of this review paper is not to provide an answer to the interpretation of SWS. It is to provide information for readers to formulate and verify the hypothesis for the interpretation. Therefore, methods to verify the hypothesis for the interpretation are also reviewed. Finally, studies on the safety of SWE are discussed.

Acoustic radiation force impulse under clinical conditions with single infusion of ultrasound contrast agent evoking arrhythmias in rabbit heart.

PURPOSE: We previously reported that acoustic radiation force impulse (ARFI) with concomitant administration of perfluorobutane as an ultrasound contrast agent (UCA)-induced arrhythmias at a mechanical index (MI) of 1.8 or 4.0 in a rabbit model. The present study identified the location of arrhythmias with a MI < 1.8 using a new system that can transmit ARFI with B-mode imaging. METHODS: Under general anesthesia, six male Japanese white rabbits were placed in a supine position. Using this system, we targeted ARFI to the exact site of the heart. ARFI exposure with MI 0.9-1.2 was performed to the right or left ventricle of the heart 2 min after UCA injection. RESULTS: ARFI with a MI lower than previously reported to rabbit heart evoked extrasystolic waves with single UCA infusion. Arrhythmias were not observed using ARFI without UCA. Extrasystolic waves were observed significantly more frequently in the right ventricle group than in the left ventricle group, with arrhythmias showing reversed shapes. No fatal arrhythmias were observed. CONCLUSION: ARFI applied to simulate clinical conditions in rabbit heart evoked extrasystolic waves with single UCA infusion. The right ventricle group was significantly more sensitive to ARFI exposure, resulting in arrhythmias, than the left ventricle group. The shapes of PVCs that occurred in the right ventricle group and the left ventricle group were reversed. Ultrasound practitioners who use ARFI should be aware of this adverse reaction, even if the MI is below the previously determined value of 1.9.

Intervendor variability of carotid intima-media thickness measurement: validation study using newly developed ultrasound phantom.

PURPOSE: Ultrasonography-derived carotid artery intima-media thickness (IMT) has been established as an early atherosclerotic imaging biomarker. The IMT reference value of a healthy person is approximately 0.1 × (every 10 years of age) + 0.2 (mm); accordingly, it requires an accuracy of at least 0.1 mm. However, one concern of IMT measurement is intervendor variability. In this study, we aimed to verify the intervendor variability using an IMT phantom. METHODS: An improved IMT phantom was developed, and it was possible to analyze the IMT by software for all vendors. RESULTS: With the vendor-specific software, the maximum difference between the devices was 0.08 mm, and the difference in quartile range was 0.06 mm. On the other hand, with the vendor-independent offline software, the maximum difference between the devices was 0.16 mm, and the quartile range of variation was 0.06 mm. CONCLUSION: The intervendor variability assessed using our IMT phantom was less than 0.10 mm, and the on-board vendor-specific software was shown to reduce the difference between the devices significantly compared with the vendor-independent offline software. To further improve the vender difference, adjustment by means of vendor-specific software based on a standardized IMT phantom is warranted.

Correction to: Intervendor variability of carotid intima-media thickness measurement: validation study using newly developed ultrasound phantom.

In the Original publication of the article Table 3 was published incorrectly.

Application of the novel estimation method by shear wave elastography using vibrator to human skeletal muscle.

In recent years, non-invasive measurement of tissue stiffness (hardness) using ultrasound elastography has attracted considerable attention. It has been used to evaluate muscle stiffness in the fields of rehabilitation, sports, and orthopedics. However, ultrasonic diagnostic devices with elastography systems are expensive and clinical use of such devices has been limited. In this study, we proposed a novel estimation method for vibration-based shear wave elastography measurement of human skeletal muscle, then determined its reproducibility and reliability. The coefficient of variation and correlation coefficient were used to determine reproducibility and reliability of the method by measuring the shear wave velocities in konjac phantom gels and agar phantom gels, as well as skeletal muscle. The intra-day, day-to-day, and inter-operator reliabilities were good when measuring the shear wave velocities in phantom gels. The intra-day and day-to-day reliabilities were good when measuring the shear wave velocities in skeletal muscle. The findings confirmed adequate reproducibility and reliability of the novel estimation method for vibration-based shear wave elastography. Therefore, the proposed measurement method may be a useful tool for evaluation of muscle stiffness.

Ultrasonic measurement of sound velocity fluctuations in biological tissue due to ultrasonic heating and estimation of thermo-physical properties.

PURPOSE: Tissue characterization in terms of the differences in thermo-physical properties of biological tissues was investigated in this study. The objective was to measure the ratio of variation in sound velocity due to ultrasonic heating and to derive the relational expression between the ratio and thermo-physical properties. METHODS: The ratio of sound velocity variation before and after the temperature rise of tissue samples exposed to ultrasound was measured by ultrasonic pulse echo method. The thermo-physical properties were estimated for a tissue-mimicking material and porcine muscle and fat tissues due to theoretical expression. The transducer for heating had a resonance frequency of 3.2 MHz, and the transducer for measurement of sound velocity variation had a resonance frequency of 5.2 MHz. RESULTS: In the phantom study, the measured values of the temperature rise agreed with the values calculated by a finite element method (FEM). The estimated values of the temperature rise from the sound velocity variations of muscle and fat tissues were 0.36 °C and 1.1 °C, respectively. Also, the estimated values of thermo-physical properties agreed with the reference values within an error of 10%. CONCLUSIONS: The thermo-physical properties of the porcine tissues were measured by sound velocity variation due to ultrasonic heating within the safety regulations.

Volumetric q-space imaging by 3D diffusion-weighted MRI.

High b-value diffusion magnetic resonance imaging (MRI) enables us to detect far smaller architectures, by using q-space analysis, than the resolution in conventional MRI. Average displacement, one of the q-space parameters, quantitatively reflects architecture size and is very useful in observing small changes in microstructures in vivo (e.g., neurodegeneration, tumor heterogeneity, and others). Diffusion-weighted imaging (DWI) is performed by a two-dimensional (2D) multislice method; however, due to finite slice thickness and slice gap, there is a partial-volume effect that makes it difficult to detect the net q-space signal. On the other hand, three-dimensional (3D) MRI, having the advantages of very thin slice thickness and no slice gap (contiguous slices), allows volumetric evaluation acquired in a small isotropic voxel, as compared to 2D multislice imaging. Little is known about the isotropic high-resolution 3D DWI application to q-space analysis. In this study, we have developed and implemented a high b-value 3D DWI sequence, applied q-space analysis to study the reliability of high b-value 3D DWI and obtained a microscopic analytical map with isotropic high resolution and less contamination.

Development of a robust diffusion-MR elastography (dMRE) technique to mitigate intravoxel phase dispersion.

Diffusion-magnetic resonance elastography (dMRE) is an emerging practical technique that can acquire diffusion magnetic resonance imaging and MRE simultaneously. However, a signal loss attributable to intravoxel phase dispersion (IVPD) interferes with the calculation of the apparent diffusion coefficient (ADC). This study presents an approach to dMRE that reduces the influence of IVPD by introducing a new pulse sequence. The existing and proposed techniques were performed using a phantom comprising five rods with different elasticities at 60 Hz vibration to investigate the accuracy of previous and proposed dMRE techniques. The measures of ADC and stiffness, obtained by using both dMRE techniques, were compared with conventional spin-echo (SE) diffusion and SE-MRE. Then, we evaluated those differences by using the mean of absolute differences (MAD) in each rod within the phantom. The results of the MAD of the stiffness from both dMRE techniques showed almost no difference. In contrast, the value of the ADC MAD (MAD ≒ 0.16 × 10-3 mm2/s), obtained in the soft region within the phantom with the previous dMRE technique, was large. This value was about 2.7 times that of the value produced by the proposed dMRE technique. This difference must reflect the degree of influence of IVPD in both techniques. These results demonstrate that our dMRE technique is a robust method for addressing the signal loss attributable to IVPD.

In vivo isotropic 3D diffusion tensor mapping of the rat brain using diffusion-weighted 3D MP-RAGE MRI.

The purpose of this study was to examine the potential of diffusion-weighted (DW) three-dimensional (3D) MP-RAGE MRI for diffusion-tensor mapping of the rat brain in vivo. A DW-3D-MP-RAGE (3D-DWI) sequence was implemented at 2.0 T using six gradient orientations and a b value of 1000 s/mm2. In this sequence, the preparation sequence with a "90 degrees RF-motion proving gradient (MPG): MPG-180 degrees RF-MPG-90 degrees RF" pulse train (DW driven equilibrium Fourier transform) was used to sensitize the magnetization to diffusion. A centric k-space acquisition order was necessary to minimize saturation effects (T1 contamination) from tissues with short relaxation time. The image matrix was 128x128x128 (interpolated from 64x64x64 acquisitions), which resulted in small isotropic DW image data (voxel size: 0.273x0.273x0.273 mm3). Moreover, 3D-DWI-derived maps of the fractional anisotropy (FA), relative anisotropy (RA) and main-diffusion direction were completely free of susceptibility-induced signal losses and geometric distortions. Two well-known commissural fibers, the corpus callosum and anterior commissure, were indicated and shown to be in agreement with the locations of these known stereotaxic atlases. The experiment took 45 min, and shorter times should be possible in clinical application. The 3D-DWI sequence allows for in vivo 3D diffusion-tensor mapping of the rat brain without motion artifacts and susceptibility to distortion.

The effect of ultrasound with acoustic radiation force on rabbit lung tissue: a preliminary study.

PURPOSE: Acoustic radiation force (ARF) elastography has recently become available. The previous animal studies have revealed lung injuries induced by diagnostic ultrasound, but the effects on the lung resulting from exposure to ultrasound with ARF are unknown. This study aimed to assess the risk of lung injury associated with ultrasound with ARF. METHODS: A focused 2.5-MHz transducer that emits ultrasound with ARF was used. A rabbit was anesthetized, and the transducer was placed in the right subcostal region. Exposure settings of mechanical index (MI) 0.80, pulse duration 10 ms, pulse repetition time 5 s, and exposure time 150 s were applied. RESULTS: One red spot (7 × 6 mm) was observed on the surface of the right lung corresponding to the area of exposure. Alveolar hemorrhage was observed microscopically. This lesion was visible across a range of 20-170 µm in depth from the pleural surface. CONCLUSION: The first example of lung hemorrhage induced by ultrasound with ARF was observed in this study. This observation suggests the possibility of lung injury in humans when ARF elastography is applied with the transducer directed toward the lung. Further studies are needed to determine the safety of this modality.